[(boron or boric or boronic or borate) and inflammation]
________________________________________________________________

Boron-containing compounds on neurons: Actions and potential applications 
for treating neurodegenerative diseases.

Barrón-González M, Montes-Aparicio AV, Cuevas-Galindo ME, Orozco-Suárez S, 
Barrientos R, Alatorre A, Querejeta E, Trujillo-Ferrara JG, 
Farfán-García ED, Soriano-Ursúa MA

J Inorg Biochem. 2023 Jan;238:112027

Boron-containing compounds (BCC) exert effects on neurons. After the expanding 
of both the identification and synthesis of new BCC, novel effects in living 
systems have been reported, many of these involving neuronal action. In this 
review, the actions of BCC on neurons are described; the effects have been 
inferred by boron deprivation or addition. Also, the effects can be related to 
those mediated by interaction on ionic channels, G-protein coupled receptors, or 
other receptors exerting modification on neuronal behavior. Additionally, BCC 
have exhibited effects by the modulation of inflammation or oxidative processes. 
BCC are expanding as drugs. Deprivation of boron sources from the diet shows the 
role of some natural BCC. However, the observations of several new synthesized 
compounds suggest their ability to act with attractive potency, efficacy, and 
long-term action on neuronal receptors or processes related with the origin and 
evolution of neurodegenerative processes. The details of BCC-target interactions 
are currently being elucidated in progress, as those observed from BCC-protein 
crystal complexes. Taking all of the above into account, the expansion is 
presumably near to having studies on the application of BCC as drugs on specific 
targets for treating neurodegenerative diseases.
________________________________________________________________

The effects of boron-supplemented diets on adipogenesis-related gene 
expressions, anti-inflammatory, and antioxidative response in high-fat fed rats.

Kucukkurt I, Ince S, Eryavuz A, Demirel HH, Arslan-Acaroz D, 
Zemheri-Navruz F, Durmus I

J Biochem Mol Toxicol. 2022 Nov 23:e23257 

The fatty liver syndrome caused by nutritional factors is a common cause of 
hepatic dysfunction globally. This research was designed to study the shielding 
effect of boron in rats fed a diet having high fat. Overall, 40 Wistar albino 
male rats were placed into one control and four treatment groups, that is, each 
having eight rats. Group I was provided with a standard rat diet while group II 
was only provided a high-fat diet for 60 days. Groups III, IV, and V were 
provided with 5, 10, and 20 mg/kg/day boron, respectively, by gastric gavage 
besides a high-fat diet for 60 days. Malondialdehyde was increased significantly 
in rats' blood and tissue because of high-fat diets. Glutathione was decreased 
significantly in blood and tissues because of a high-fat diet. Moreover, the 
activities of superoxide dismutase (SOD) and catalase (CAT) were decreased in 
the blood and tissues of the high-fat-fed rats. The genes expression for 
C-reactive protein, interleukin-1β, leptin, and tumor necrosis factor-α were 
increased while gene expression for peroxisome proliferator-activated 
receptors was decreased in the liver of rats fed with a high-fat diet. 
Contrariwise, boron supplementation improves antioxidative response in terms of 
increased SOD and CAT activities, gene expression regulation, and improved 
anti-inflammatory activities. In a nutshell, boron has dose-dependent shielding 
antioxidative and tissue regenerative effects in rats.
________________________________________________________________

Therapeutic Effects of Boric Acid in a Septic Arthritis Model 
Induced by Escherichia coli in Rats.

Yıldız K, Makav M, Adalı Y, Bulut M

Biol Trace Elem Res. 2022 Nov;200(11):4762-4770 
Erratum in Biol Trace Elem Res. 2022 Feb 7 

The study aimed to evaluate the therapeutic effect of boric acid (BA) in 
experimentally induced septic arthritis. A total of 30 rats, 6 rats in each 
group (5 groups), were used in the study. No treatment was applied to the rats 
in the control group. Only BA was administered intraperitoneally (IP) to the 
rats in the bor group. Escherichia coli was administered at a single dose of 25 
μL, 1 × 1010 cfu/rat from the right foot pad of the rats, via intra-articular 
route, to the mice in the arthritis, arthritis-bor, and arthritis-antb groups. 
Then, BA at a dose of 50 mg/kg and cefazolin at a dose of 25 mg/kg were 
administered to the rats in the arthritis-bor and arthritis-antb groups, 
respectively, for 7 days via the IP route. At the end of the study, all animals 
were euthanized following the ethical rules. Blood and tissue samples were taken 
from the rats for biochemical and histopathological analyses. The levels of GSH, 
MDA, Endoglin, Endocan, and TNF-β markers were measured in the blood samples 
taken. A significant decrease was observed in MDA and Endoglin levels in the 
boric acid-administered group compared with the arthritis group, while a 
significant increase was observed at the GSH level. Histopathologically, it was 
determined that the reactive surrounding tissue response in the bor group was 
significantly reduced. As a result, a significant decrease in inflammation was 
found biochemically and histopathologically in the groups treated with BA.
________________________________________________________________

Boric Acid Reverses Nicotine-Induced Cytokine 
Expressions of Human Gingival Fibroblasts.

Bozkurt SB, Nielsen FH, Hakki SS

Biol Trace Elem Res. 2022 Apr 20 

Nicotine, the major bioactive ingredient in tobacco, is a major risk factor for 
periodontal disease and destruction. Nicotine has been shown to stimulate the 
production of cytokines that are priming agents for inflammation that induces 
tissue destruction, such as IL-1β, IL-6, and IL-8, by gingival keratinocytes and 
human gingival fibroblasts (HGF). Boron as boric acid has been found to decrease 
pro-inflammatory cytokines and increase anti-inflammatory cytokines in cells 
with inflammatory stress. Thus, a study was performed to determine whether boric 
acid reverses negative effects of nicotine on human gingival fibroblasts (HGFs). 
The viability and cytokine expressions of HGFs cultured for 24 and 72 h in 
control medium with no nicotine or boric acid added and in media containing only 
nicotine, only boric acid, or a combination of BA and nicotine were determined. 
Nicotine in concentrations of 10-1, 10-2, 10-3,10-4, 10-5, and 10-6 mM 
significantly reduced cell viability compared to the control. Boric acid at 10 
and 50 ng/mL in the media partially restored and 100 ng/mL in the media fully 
restored the nicotine-depressed HGF cell viability to the same level as the 
control group. Nicotine elevated the expression of pro-inflammatory cytokines 
TNF-α, IL-1β, IL-6, IL-8, and IL-17 and decreased the anti-inflammatory IL-10 in 
HGFs at 24 and 72 h. Boric acid at 100 ng/mL in the medium prevented the changes 
induced by nicotine alone. The findings indicate that boric acid can inhibit or 
reverse nicotine-induced pathology in periodontal tissue and thus may help 
maintain oral and periodontal health in tobacco users.
________________________________________________________________

Possible Curative Effects of Boric Acid and Bacillus clausii Treatments 
on TNBS-Induced Ulcerative Colitis in Rats.

Özkoç M, Can B, Şentürk H, Burukoğlu Dönmez D, Kanbak G

Biol Trace Elem Res. 2022 Mar 29 

Crohn's disease (CD) and ulcerative colitis (UC) are two chronic relapsing 
inflammatory bowel diseases (IBD). Although there are several treatment options 
available to improve the symptoms of IBD patients, there is no effective 
treatment that provides a definitive solution. In the present study, we aim to 
investigate the antioxidative/anti-inflammatory effects of oral administration 
of boric acid and Bacillus clausii in a rat trinitrobenzenesulfonic acid 
(TNBS)-induced colitis model. The effects of boric acid and B. clausii were 
examined in serum and colon tissues with the help of some biochemical and 
histological analyses. Elevated inflammation and oxidative damage were found in 
the blood and colon tissue samples in the TNBS-induced group according to the 
complete blood count (CBC), tumor necrosis factor (TNF) alpha, interleukin-35 
(IL-35), malondialdehyde (MDA), glutathione peroxidase (GPx), myeloperoxidase 
(MPO), nitric oxide (NO), and histological findings. Particularly, the highest 
IL-35 level (70.09 ± 12.62 ng/mL) in the combined treatment group, highest 
catalase activity (5322 ± 668.1 U/mg protein) in the TNBS-induced group, and 
lower relative expression of inducible nitric oxide synthase in the TNBS-induced 
group than the control group were striking findings. According to our results, 
it can be concluded that boric acid showed more curative effects, even if B. 
clausii probiotics was partially ameliorative.
________________________________________________________________

Boron Intake and decreased risk of mortality in kidney transplant recipients.

Kremer D, Post A, Seidel U, Huebbe P, van der Veen Y, Groothof D, Gomes-Neto AW, 
Knobbe TJ, Lüersen K, Eisenga MF, Navis GJ, Rimbach G, Bakker SJL; 
TransplantLines Investigators.

Collaborators: Kremer D, Knobbe TJ, Annema-de Jong JH, Berger SP, Blokzijl J, 
Bodewes FAJA, de Boer MT, Damman K, De Borst MH, Diepstra A, Dijkstra G, Douwes 
RM, Eisenga MF, Erasmus ME, Gan CT, Gomes Neto AW, Grootjans H, Hak E, 
Heiner-Fokkema MR, Hepkema BG, Klont F, Leuvenink HGD, Lexmond WS, de Meijer VE, 
Niesters HGM, van Pelt LJ, Pol RA, Porte RJ, Ranchor AV, Sanders JSF, Schutten 
JC, Siebelink MJ, Slart RHJA, Swarte JC, Timens W, Touw DJ, van den Heuvel MC, 
van Leer-Buter C, van Londen M, Verschuuren EAM, Vos MJ, Weersma RK, Bakker SJL.

Eur J Nutr. 2022 Mar;61(2):973-984 

PURPOSE: In a search for potentially modifiable factors to improve long-term 
outcome among kidney transplant recipients (KTR), we hypothesized that boron 
exposure is associated with improved long-term outcome in KTR.
METHODS: We determined 24 h urinary boron excretion using inductively coupled 
plasma mass spectrometry as a measure of boron exposure in 693 stable KTR (57% 
male, mean age 53y), enrolled in the TransplantLines Food and Nutrition Biobank 
and Cohort Study. Dietary intake was assessed using validated food-frequency 
questionnaires.
RESULTS: Linear regression analyses showed that dietary intake of fruit, wine 
and nuts were key determinants of boron excretion. In addition, boron excretion 
was negatively correlated with homocysteine and inflammatory parameters. In 
total, 73 (32%), 47 (20%) and 30 (13%) KTR died among the lowest, middle and 
highest tertiles of 24 h urinary boron excretion, respectively 
(Plog-rank < 0.001). Cox regression analyses showed that high boron excretion 
was strongly associated with lower risk of mortality, independent of age, sex, 
estimated glomerular filtration rate and history of cardiovascular disease (HR 
per doubling: 0.51, 95% CI: 0.40 to 0.66, P < 0.001).
CONCLUSION: Boron may be an overlooked target to improve long-term survival 
among KTR and potentially other patients, likely through pathways other than 
inflammation or the methionine-homocysteine cycle that were previously 
suggested. Interventional trials are warranted to confirm the potential of 
dietary boron supplementation in KTR and other patient populations.
________________________________________________________________

Anti- inflammatory effect of boric acid on cytokines 
in ovariectomy-induced rats.

Tekeli H, Ekren Asıcı GS, Bildik A

Cell Mol Biol (Noisy-le-grand). 2022 Jan 2;67(4):313-320 

The increase in the rate of inflammation in the post-menopause period also leads 
to a significant increase in the use of anti-inflammatory agents. This study 
aimed to investigate the effect of BA supplementation on pro-and 
anti-inflammatory cytokines in ovariectomy (OVX) induced rats. A total of 48 
nonpregnant female Wistar albino rats (80-100 g) were used in the experiment. 
Forty-eight rats were divided into six equal groups (n=8): Control, OVX, OVX+5 
mg/kg BA (OVX+BA5), OVX+10 mg/kg BA (OVX+BA10), 5 mg/kg BA (BA5), 10 mg/kg BA 
(BA10). Serum TNF-α cytokine levels of rats in the OVX group were higher than in 
control rats (P<0.05). TNF-α levels were significantly reduced in the 
OVX-induced rats with 5 mg/kg BA and 10 mg/kg BA supplementation (P<0.05). While 
serum IL-1α and IL-6 levels were not different between OVX and control rats, 
serum IL-3 levels were low (P<0.05) and not affected by 5 mg/kg and 10 mg/kg BA 
supplementation. Serum IL-11 levels increased significantly in the OVX rats with 
5 mg/kg and 10 mg/kg BA supplementation (P<0.05). As far as we know, certain 
doses (5 and 10 mg/kg) of BA are the first study on the prevention of increased 
inflammation in rats induced by OVX. Results suggest that the supplementation of 
BA regulates the inflammatory changes associated with OVX and thus has 
beneficial for menopause management.
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Borate Ameliorates Sodium Nitrite-Induced Oxidative Stress Through Regulation 
of Oxidant/Antioxidant Status: Involvement of the Nrf2/HO-1 and NF-κB Pathways.

Soliman MM, Aldhahrani A, Elshazly SA, Shukry M, Abouzed TK

Biol Trace Elem Res. 2022 Jan;200(1):197-205 

The widespread industrial use of nitrite in preservatives, colorants, and 
manufacturing rubber products and dyes increases the possibilities of organ 
toxicity. Lithium borate (LB) is known as an antioxidant and an oxidative stress 
reliever. Therefore, this study is aimed at examining the effect of LB on 
nitrite-induced hepatorenal dysfunction. Twenty-eight male Swiss mice were 
divided into four equal groups. Group 1, the control group, received saline. 
Group 2 received LB orally for 5 consecutive days at a dose of 15 mg/kg bw. 
Group 3, the nitrite group, received sodium nitrite (NaNO2) on Day 5 (60 mg/kg 
bw intraperitoneally). Group 4, the protective group (LB + NaNO2 group), 
received LB for 5 days and then a single dose of NaNO2 intraperitoneally on Day 
5, the same as in Groups 2 and 3, respectively. Samples of blood and kidney were 
taken for serum analysis of hepatorenal biomarkers, levels of antioxidants and 
cytokines, and the expression of genes associated with oxidative stress and 
inflammation. NaNO2 intoxication increased markers of liver and kidney functions 
yet decreased reduced glutathione (GSH), superoxide dismutase (SOD), and 
catalase activities in blood. NaNO2 also increased the expression of tumor 
necrosis factor (TNF-α), interleukin-1β and interleukin-6 (IL-1β and IL-6). 
Pre-administration of LB protected mice from oxidative stress, lipid 
peroxidation, and the decrease in antioxidant enzyme activity. Moreover, LB 
protected mice from cytokine changes, which remained within normal levels. LB 
ameliorated the changes induced by NaNO2 on the mRNA of nuclear factor erythroid 
2-related factor 2 (Nfr2), heme oxygenase-1 (HO-1), nuclear factor-kappa B 
(NF-κB), transforming growth factor-beta 2 (TGF-β2), and 
glutathione-S-transferase (GST) as determined using quantitative real-time PCR 
(qRT-PCR). These results collectively demonstrate that LB ameliorated 
NaNO2-induced oxidative stress by controlling the oxidative stress biomarkers 
and the oxidant/antioxidant state through the involvement of the Nrf2/HO-1 and 
NF-κB signaling pathways.
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The Impact of Trace Elements on Osteoarthritis.

Li G, Cheng T, Yu X

Front Med (Lausanne). 2021 Dec 23;8:771297 

Osteoarthritis (OA) is a progressive degenerative disease characterized by 
cartilage degradation, synovial inflammation, subchondral sclerosis and 
osteophyte formation. It has a multifactorial etiology with potential 
contributions from heredity, endocrine function, abnormal mechanical load and 
nutrition. Of particular considerations are trace element status. Several trace 
elements, such as boron and magnesium are essential for normal development of 
the bone and joint in human. While cadmium correlates with the severity of OA. 
The present review focuses on the roles of trace elements (boron, cadmium, 
copper, iron, magnesium, manganese, selenium, zinc) in OA and explores the 
mechanisms by which they act.
________________________________________________________________

Cytokines and Chemokines Involved in Osteoarthritis Pathogenesis.

Molnar V, Matišić V, Kodvanj I, Bjelica R, Jeleč Ž, Hudetz D, Rod E, 
Čukelj F, Vrdoljak T, Vidović D, Starešinić M, Sabalić S, Dobričić B, 
Petrović T, Antičević D, Borić I, Košir R, Zmrzljak UP, Primorac D 

Int J Mol Sci. 2021 Aug 26;22(17):9208 

Osteoarthritis is a common cause of disability worldwide. Although commonly 
referred to as a disease of the joint cartilage, osteoarthritis affects all 
joint tissues equally. The pathogenesis of this degenerative process is not 
completely understood; however, a low-grade inflammation leading to an imbalance 
between anabolic and katabolic processes is a well-established factor. The 
complex network of cytokines regulating these processes and cell communication 
has a central role in the development and progression of osteoarthritis. 
Concentrations of both proinflammatory and anti-inflammatory cytokines were 
found to be altered depending on the osteoarthritis stage and activity. In this 
review, we analyzed individual cytokines involved in the immune processes with 
an emphasis on their function in osteoarthritis.
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Calcium Fructoborate Prevents Skin Cancer Development in Balb-c Mice: 
Next Part, Reverse Inflammation, and Metabolic Alteration.

Kisacam MA, Kocamuftuoglu GO, Ozan IE, Yaman M, Ozan S

Biol Trace Elem Res. 2021 Jul;199(7):2627-2634 

Metabolic alterations and inflammation are regarded as hallmarks of cancer. 
Glycolytic flux and intermediate accumulation lead to the production of building 
blocks and NADPH which is important in protecting the cell from oxidative 
damage. Inflammation causes the release of mediators responsible for regulating 
molecular mechanism affecting metabolic pathways. CaFB due to its cis-diol-rich 
feature may have the potential to interact with molecules taking part in cancer 
development. This study was aimed to investigate the effects of CaFB on 
metabolic alterations and inflammation in 7,12-dimethylbenz(a)anthracene 
(DMBA)/12-O-tetradecanoylphorbol-13-acetate (TPA)-induced skin cancer. For this 
purpose, 92 Balb-c mice were distributed into 6 groups as control, CaFB, 
DMBA/TPA (D-T), treatment 1 (T1), 2 (T2), and 3(T3). Apart from control and CaFB 
in other groups, tumors initiated with 97.5-nmol DMBA and 6.5-nmol TPA. 
Treatment groups received 3 mg/kg/day CaFB with DMBA (T1), with TPA (T2), and 
after tumor formation (T3). In the D-T group, glyceraldehyde-3-phosphate 
dehydrogenase (GAPDH) activity, 6-phosphogluconate dehydrogenase (PGD), 
glutathione (GSH), interleukin 6 (IL-6), (IL-1β), tumor necrosis factor-α 
(TNF-α) levels increased (p < 0.001) while malondialdehyde (MDA) levels 
decreased (p < 0.001) compared with that in control. CaFB application 
ameliorated DMBA-TPA effect according to the distribution time. It is noteworthy 
to consider CaFB as a potential preventive agent in skin cancer development.
________________________________________________________________

Preclinical and histological study of boron-containing compounds 
hydrogels on experimental model of periodontal disease.

Mitruţ I, Cojocaru MO, Scorei IR, Biţă A, Mogoşanu GD, 
Popescu M, Olimid DA, Manolea HO.

Rom J Morphol Embryol. 2021 Jan-Mar;62(1):219-226 

Periodontitis is a disease that affects a wide group of people, and there has 
been an increased interest in the research of finding useful materials that help 
reduce inflammation and the further loss of tissue. In this study, we have 
tested a boron-containing compound (BCC) Calcium Fructoborate (CaFB) and Boric 
Acid (BA) hydrogels on the gingival level on Wistar rats. First, we have induced 
the periodontal disease at the lower incisors, we have applied the hydrogels and 
after a week, we have euthanized the rats. Next, the oral soft tissue reaction 
was clinically and then histologically investigated. Our study has shown good 
clinical response of the oral tissue, and we have noticed lower levels of 
inflammation on the experimental groups treated with the BCCs hydrogels. Despite 
the generally good response of the biological structures to the presence of BA 
and CaFB on periodontal level, more scientifically proved information is needed 
to obtain the desired biological responses in all clinical situations.
________________________________________________________________

Boric acid inhibits alveolar bone loss in rat experimental periodontitis 
through diminished bone resorption and enhanced osteoblast formation.

Shalehin N, Hosoya A, Takebe H, Hasan MR, Irie K

J Dent Sci. 2020 Dec;15(4):437-444 

BACKGROUND/PURPOSE: Inhibition of bone resorption is essential for periodontal 
treatment. Recently, it has been suggested that boric acid suppresses 
periodontitis, but the mechanism of this inhibition is still not well 
understood. Therefore, to analyze the cellular response to boric acid 
administration, we histologically evaluated alveolar bone in experimental 
periodontitis of rats administered boric acid.
MATERIALS AND METHODS: 5-0 silk ligatures were placed around the cervix of the 
second maxillary molars of 4 week-old rats treated with or without boric acid. 
Five and 14 days after ligature placement, the periodontal tissues between first 
and second molars were investigated histologically and immunohistochemically 
using antibodies to CD68, cathepsin K, and α-smooth muscle actin (SMA).
RESULTS: Five days after the beginning of the experiment, many CD68-positive 
cells appeared in the periodontal tissues with ligature placement without boric 
acid administration. Also, the number of cathepsin K-positive osteoclasts had 
increased on the surface of alveolar bone. However, boric acid administration 
prevented severe bone resorption and reduced the number of cells positive for 
CD68 and cathepsin K. At day 14 post treatment, cells positive for α-SMA were 
seen in the periodontal tissues after boric acid administration, whereas no such 
cells were found around the alveolar bone without the administration of boric 
acid.
CONCLUSION: Boric acid inhibited the inflammation of ligature-induced 
periodontitis. This agent might reduce bone resorption by inhibiting 
osteoclastogenesis and also could accelerate osteoblastogenesis.
________________________________________________________________

Evaluation of oxidative stress and immune parameters 
of boron exposed males and females.

Başaran N, Duydu Y, Bacanlı M, Gül Anlar H, Dİlsİz SA, 
Üstündağ A, Yalçın CÖ, Schwerdtle T, Bolt HM

Food Chem Toxicol. 2020 Aug;142:111488 

There is growing evidence that boron (B) and B compounds are essential nutrients 
for animals and humans. Besides, B compounds have been suggested to treat 
inflammation and oxidative stress. As a part of our "Boron Project II" on 
B-exposed persons in Bandırma and Bigadic (Turkey) between 2014 and 2017, 
anti-oxidant/pro-oxidant and inflammatory parameters were assessed. In this 
first large-scale human study biomarkers of oxidative stress such as the enzyme 
activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase 
(GPx), glutathione reductase (GR) and the levels of malondialdehyde (MDA), 
glutathione (GSH) and 8-hydroxy-2'-deoxy-guanosine (8-OH-dG) were investigated, 
in relation to B exposure. The immune biomarkers interleukin (IL)-1ra, IL-6, 
IL-8 and nuclear factor kappa B (NF-κB) levels were included. There was no 
influence of human exposure to B on the parameters of oxidative stress and 
inflammation.
________________________________________________________________

The Role of Oxidative Stress, Renal Inflammation, and Apoptosis 
in Post Ischemic Reperfusion Injury of Kidney Tissue: 
the Protective Effect of Dose-Dependent Boric Acid Administration.

Kar F, Hacioglu C, Senturk H, Donmez DB, Kanbak G

Biol Trace Elem Res. 2020 May;195(1):150-158 

Ischemia/reperfusion (I/R) injury is associated with a strong inflammatory and 
oxidative stress response to hypoxia and reperfusion that impair organ function. 
We aimed to investigate the role of oxidative stress, renal inflammation, and 
apoptosis in the injury of the kidney tissue after ischemic reperfusion, and the 
protective effect of dose-dependent boric acid administration. For this purpose, 
35 Sprague Dawley albino rats were divided into five groups of seven animals in 
each group: Sham, I/R and I/R + boric acid (BA) (i.p at doses of 50, 100, and 
200 mg/kg). All animals underwent nephrectomy (the right kidney was removed) and 
were expected to recover for 15 days. After recovery, each animal received 
45 min of ischemia. BA was injected intraperitoneally 10 min before reperfusion 
and a 24-h reperfusion procedure was performed. Sham group only underwent 
surgical stress procedure. In order to investigate the oxidative stress induced 
by I/R injury and antioxidant effects of different BA doses in the kidney 
tissue, TAS, TOS, MDA, SOD, CAT, and GSH levels were measured. DNA 
fragmentation, cytochrome C levels, caspase 3 activity were measured to 
determine apoptotic index in tissue. IL-6 and TNF-α levels were measured in the 
evaluation of inflammation. Hematoxylin-eosin and TUNEL staining was performed 
for histopathological examinations. As a result, increased oxidative stress, 
inflammation, and apoptosis after I/R were decreased with different doses of BA 
treatment. The application of high-dose BA was found to be lower in 
anti-apoptotic, anti-inflammatory, and antioxidant effects than in the low-dose 
groups.
________________________________________________________________

Propolis and Its Combination with Boric Acid Protect Against 
Ischemia/Reperfusion-Induced Acute Kidney Injury by Inhibiting Oxidative Stress, 
Inflammation, DNA Damage, and Apoptosis in Rats.

Geyikoglu F, Koc K, Colak S, Erol HS, Cerig S, Yardimci BK, 
Cakmak O, Dortbudak MB, Eser G, Aysin F, Ozek NS, Yildirim S

Biol Trace Elem Res. 2019 Dec;192(2):214-221 

Ischemia reperfusion (I/R) injury which causes kidney dysfunction is one of the 
most studied diseases directly linked to oxidative stress. In this regard, it is 
important to protect cells against damage by inducing antioxidant response. 
Herein, we aimed to evaluate the therapeutic roles and possible mechanisms of 
propolis and boric acid in kidney I/R injury based on relevant basic research 
and clinical studies. Sprague-Dawley rats were subjected to 50 min of ischemia 
followed by 3 h of reperfusion. Animals were randomly divided into a control 
group (the abdominal wall was just opened and closed), an I/R injury group, the 
propolis intervention group (200 mg/kg, intragastric administration, 1 h before 
ischemia), boric acid intervention group (14 mg/kg, intragastric administration 
1 h before ischemia), and the propolis + boric acid intervention group 
(intragastric administration 1 h before ischemia). Kidney function, the 
antioxidant defensive system, and renal damage were assessed. In addition, the 
oxidative stress and inflammatory status were estimated in renal tissue. 
Furthermore, DNA damageand apoptosis were detected by immunohistochemistry. When 
compared with I/R group, propolis alone and especially propolis + boric acid 
groups significantly improved functional parameters. While the antioxidant 
response was increased, renal injury size and apoptosis were significantly 
decreased in both groups. Also, the MDA and TNF-α levels besides the 8-OHdG 
formation were downregulated. According to these outcomes, it can be said that 
especially propolis together with boric acid ameliorates kidney injury caused by 
I/R through acting as an antioxidant, anti-inflammatory, and antiapoptotic 
agent. In conclusion, propolis alone and its combination with boric acid could 
be developed as therapeutic agents against serious renal I/R injuries.
________________________________________________________________

The propolis and boric acid can be highly suitable, alone/or 
as a combinatory approach on ovary ischemia-reperfusion injury.

Geyikoglu F, Koc K, Erol HS, Colak S, Ayer H, 
Jama S, Eser G, Dortbudak MB, Saglam YS 

Arch Gynecol Obstet. 2019 Nov;300(5):1405-1412 

PURPOSE: Ovarian ischemia-reperfusion (IR) damage continues to be a serious 
infertility problem. The oxidative stress plays central role in the development 
of IR injuries. Activation of antioxidants decreases IR injuries; however, the 
efficacy of antioxidant agents remains controversial. Unfortunately, there has 
been no evidence for medicinal use of boric acid (BA) and propolis (Prop) on 
ovarian IR injury on rats so far. This study will provide to reveal the 
potential applications of the Prop and BA in ovarian IR therapy.
METHODS: The Sprague-Dawley rats were randomized into five groups: I-control, 
II-IR, 3 h of ischemia and 3 h of reperfusion, III and IV-a signal dose of oral 
BA (7 mg/kg) and Prop (100 mg/kg) alone 1 h before induction of IR, V-Prop and 
BA together 1 h before induction of IR. SOD (superoxide dismutase), CAT 
(catalase), GSH (glutathione), MPO (myeloperoxidase), MDA (malondialdehyde), and 
IL-6 (interleukin-6) levels were quantified by ELISA and the TNF-α (tumor 
necrosis factor-α), 8-OHdG (8-hydroxylo-2'-deoxyguanosin) and Caspase-3 
expressions were performed by immunohistochemical analyses.
RESULTS: BA and Prop pretreatment significantly reduced MPO, MDA, and IL-6 
levels and pathologic score in IR rats, with no effects in control group. These 
agents used in therapy also decreased TNF-α, 8-OHdG and Caspase-3 protein 
expressions increased by IR. Furthermore, BA and Prop combination showed 
significant ameliorative effects on ovary injury caused by IR through acting as 
an antioxidant, anti-inflammatory and antiapoptotic agent.
CONCLUSION: BA and Prop alone and especially in combination could be developed 
as therapeutic agents against ovary IR injury.
________________________________________________________________

Effects of boron-containing compounds on immune responses: 
review and patenting trends.

Romero-Aguilar KS, Arciniega-Martínez IM, Farfán-García ED, 
Campos-Rodríguez R, Reséndiz-Albor AA, Soriano-Ursúa MA

Expert Opin Ther Pat. 2019 May;29(5):339-351 

INTRODUCTION: Boron-containing compounds induce effects on immune responses. 
Such effects are interesting to the biomedical field for the development of 
therapeutic tools to modulate the immune system.
AREAS COVERED: The scope of BCC use to modify immune responses is expanding, 
mainly with regard to inflammatory diseases. The information was organized to 
demonstrate the breadth of reported effects. BCCs act as modulators of innate 
and adaptive immunity, with the former including regulation of cluster 
differentiation and cytokine production. In addition, BCCs exert effects on 
inflammation induced by infectious and noninfectious agents, and there are also 
reports regarding their effects on mechanisms involving hypersensitivity and 
transplants. Finally, the authors discuss the beneficial effects of BCCs on 
pathologies involving various targets and mechanisms.
EXPERT OPINION: Some BCCs are currently used as drugs in humans. The mechanisms 
by which these BCCs modulate immune responses, as well as the required 
structure-activity relationship for each observed mechanism of action, should be 
clarified. The former will allow for the development of improved 
immunomodulatory drugs with extensive applications in medicine. Patenting trends 
involve claims concerning the synthesis and actions of identified molecules with 
a defined profile regarding cytokines, cell differentiation, proliferation, and 
antibody production.
________________________________________________________________

Boron ameliorates arsenic-induced DNA damage, proinflammatory cytokine gene 
expressions, oxidant/antioxidant status, and biochemical parameters in rats.

Ince S, Kucukkurt I, Acaroz U, Arslan-Acaroz D, Varol N

J Biochem Mol Toxicol. 2019 Feb;33(2):e22252 

Arsenic, an element found in nature, causes hazardous effects on living 
organisms. Meanwhile, natural compounds exhibit protective effects against 
hazardous substances. This study evaluated the effects of boron against 
arsenic-induced genotoxicity and altered biochemical parameters in rats. 
Thirty-five male Wistar albino rats were equally divided into five groups, and 
the experimental period lasted 30 days. One group was used as the control, and 
another group was treated with 100 mg/L arsenic in drinking water. The other 
groups were orally treated with 5, 10, and 20 mg/kg boron plus arsenic (100 mg/L 
via drinking water). Arsenic caused changes in biochemical parameters, total 
oxidant/antioxidant status, and DNA damage in mononuclear leukocytes. Moreover, 
it increased IFN-γ, IL-1β, TNF-α, and NFκB mRNA expression levels in rat tissue. 
However, boron treatment improved arsenic-induced alterations in biochemical 
parameters and increases in DNA damage and proinflammatory cytokine gene 
expressions.
________________________________________________________________

Effects of boron-containing compounds on cardiovascular 
disease risk factors - a review.

Donoiu I, Militaru C, Obleagă O, Hunter JM, 
Neamţu J, Biţă A, Scorei IR, Rogoveanu OC

J Trace Elem Med Biol. 2018 Dec;50:47-56 

Boron is considered to be a biological trace element but there is substantial 
and growing support for it to be classified as an essential nutrient for animals 
and humans, depending on its speciation. Boron-containing compounds have been 
reported to play an important role in biological systems. Although the exact 
biochemical functions of boron-containing compounds have not yet been fully 
elucidated, previous studies suggest an active involvement of these molecules in 
the mediation of inflammation and oxidative stress. Chronic inflammation and 
oxidative stress are known to amplify the effects of the main cardiovascular 
risk factors: smoking, diet, obesity, arterial hypertension, dyslipidemia, type 
2 diabetes (as modifiable risk factors), and hyperhomocysteinemia and age (as 
independent risk factors). However, the role of boron-containing compounds in 
cardiovascular systems and disease prevention has yet to be established. This 
paper is a review of boron-containing compounds' existence in nature and their 
possible functions in living organisms, with a special focus on certain 
cardiovascular risk factors that may be diminished by intake of these compounds, 
leading to a reduction of cardiovascular morbidity and/or mortality.
________________________________________________________________

Profile of three boron-containing compounds on the body weight, 
metabolism and inflammatory markers of diabetic rats.

López-Cabrera Y, Castillo-García EL, Altamirano-Espino JA, 
Pérez-Capistran T, Farfán-García ED, Trujillo-Ferrara JG, Soriano-Ursúa MA

J Trace Elem Med Biol. 2018 Dec;50:424-429 

It has been reported that boron induces changes in carbohydrate and lipid 
metabolism, body weight and inflammatory processes. This is relevant to the 
biomedical field due to the requirement for developing therapeutic tools with 
potential application in metabolic disorders affecting humankind. However, most 
of the reported data from both humans and animals were obtained after boron was 
administered as borax or boric acid. In this work, we determined the effects of 
boric, cyclohexylboronic (CHB) and phenylboronic (PBA) acids (10 mg/kg of body 
weight/daily for two weeks) on the body weight, metabolism and inflammatory 
markers in the blood of control, fat-feeding and experimental diabetic rats. In 
particular, we observed the effects of the administration of these compounds on 
glycaemia and cholesterol, triglyceride, insulin, IL-6 and C-reactive protein 
levels, as well as visceral fat and body weight. We found different profiles for 
each boron-containing compound: boric acid induced decreasing body weight, 
insulin and IL-6 levels; CHB administration induced an increase in body weight 
and cholesterol but decreased IL-6 levels; and PBA administration induced a 
decrease in visceral fat and glucose and insulin levels. These results can 
improve the understanding of boron as a metabolic regulator and help develop new 
potential strategies to use compounds with this trace element for therapeutic 
purposes.
________________________________________________________________

The ameliorative effects of boron against acrylamide-induced oxidative stress, 
inflammatory response, and metabolic changes in rats.

Acaroz U, Ince S, Arslan-Acaroz D, Gurler Z, Kucukkurt I, 
Demirel HH, Arslan HO, Varol N, Zhu K

Food Chem Toxicol. 2018 Aug;118:745-752 

Acrylamide (ACR) is a hazardous substance associated with the accumulation of 
excessive reactive oxygen species and causes oxidative stress. Presence of ACR 
in foods leads to public health concerns due to its known neurotoxic, genotoxic, 
and carcinogenic effects. The present study investigated the ameliorative 
effects of boron (B) against ACR exposed rats. Forty Wistar albino male rats, 
fed with low-boron diet, were randomly and equally allocated into 5 groups. The 
control group was orally treated with physiological saline as placebo, the 
second group was orally given 15 mg/kg ACR. The other groups were orally treated 
with 15 mg/kg ACR and B at the levels of 5, 10, and 20 mg/kg/day for 60 days, 
respectively. ACR-treatment significantly increased malondialdehyde levels 
whereas decreased glutathione levels in rat tissues. Also, ACR-treatment 
increased the activities of superoxide dismutase and catalase in erythrocytes 
and tissues. Meanwhile, mRNA expression levels of NFĸB, IFN-γ, IL-1β, and TNF-α 
in liver and brain of rats were increased under ACR treatment. Additionally, ACR 
caused a significant decrease in the level of high-density lipoprotein, with 
increase in the levels of low-density lipoprotein, triglyceride, cholesterol, 
glucose, urea nitrogen, and creatinine. Lastly, B alleviated histopathological 
alterations induced by ACR in rat tissues.
________________________________________________________________

Cytotoxic and inflammatory effects of contact lens solutions 
on human corneal epithelial cells in vitro.

Oh S, McCanna DJ, Subbaraman LN, Jones LW

Cont Lens Anterior Eye. 2018 Jun;41(3):282-289 

PURPOSE: To ascertain the effect that four contact lens (CL) multipurpose 
solutions (MPS) have on the viability and release of pro-inflammatory cytokines 
from human corneal epithelial cells (HCEC).
METHODS: HCEC were exposed to four different MPS at various concentrations for 
18 hours. The cells were also exposed to phosphate buffer, borate buffer, and 
PHMB. The cell viability was evaluated using the alamarBlue assay. The release 
of pro-inflammatory cytokines was measured using a Multiplex 
electrochemiluminescent assay.
RESULTS: MPS-A, MPS-B and MPS-C all reduced cell metabolic activity p < 0.05 
from control with MPS-A showing the greatest cytotoxic effect (maximum 
reduction, 90.6%). In contrast, MPS-D showed no significant reductions in 
cytotoxicity except at the highest concentration tested (19% reduction at 20% 
MPS concentration). Of the four cytokines evaluated MPS-C showed a substantial 
increase in the release of IL-1β, IL-6, IL-8, and TNF-α at higher concentrations 
when compared to control p < 0.05. At the 20% concentration of MPS-A and MPS-B 
the release of IL-1 β increased p < 0.05 but the release of IL-6, IL-8, and 
TNF-α decreased. MPS-D did not cause a change in the release of cytokines IL-1β, 
IL-6, IL-8 and TNF-α p > 0.05. Exposing the cells to borate buffer and PHMB 
caused an increase in the release of TNF-α p < 0.05.
CONCLUSIONS: This investigation demonstrates that at different concentration 
levels, several of the MPS tested showed a decrease in viability and an increase 
in the release of inflammatory cytokines from HCEC. The borate buffer component 
as well as PHMB appears to contribute to this pro-inflammatory reaction.
________________________________________________________________

Ototoxicity of boric acid powder in a rat animal model.

Salihoglu M, Dogru S, Cesmeci E, Caliskan H, 
Kurt O, Kuçukodaci Z, Gungor A 

Braz J Otorhinolaryngol. 2018 May-Jun;84(3):332-337 

INTRODUCTION: Boric acid, which has antiseptic and acidic properties, is used to 
treat external and middle ear infections. However, we have not found any 
literature about the effect of boric acid powder on middle ear mucosa and inner 
ear.
OBJECTIVE: The purpose of this study is to investigate possible ototoxic effects 
of boric acid powder on cochlear outer hair cell function and histological 
changes in middle ear mucosa in a rat animal model.
METHODS: Twenty healthy, mature Wistar albino rats were used in this study. The 
rats were divided into two groups, Group A and Group B, each of which consisted 
of 10 rats. Initially, the animals in each group underwent distortion product 
otoacoustic emissions testing of their right and left ears. After the first 
distortion product otoacoustic emissions test, a surgical microscope was used to 
make a small perforation in both ears of the rats in each group, and a second 
distortion product otoacoustic emissions test was used to measure both ears in 
all of the rats. Boric acid powder was applied to the right middle ear of the 
rats using tympanic membrane perforation, and the distortion product otoacoustic 
emissions were measured immediately after the boric acid powder application. The 
histological changes and distortion product otoacoustic emissions were evaluated 
three days later in Group A and 40 days later in Group B.
RESULTS: No significant differences were found at all of the distortion product 
otoacoustic emissions frequencies. In Group A, mild inflammation of the middle 
ear mucosa was found on the third day after boric acid powder application. In 
Group B, boric acid powder caused mild inflammatory changes on the 40th day, 
which declined over time. Those changes did not lead to significant fibrosis 
within the mucosa.
CONCLUSION: In rats, boric acid powder causes mild inflammation in middle ear 
mucosa and it has no ototoxic effects on cochlear outer hair cell function in 
the inner ear of rats.

________________________________________________________________

Boron-Based Inhibitors of the NLRP3 Inflammasome.

Baldwin AG, Rivers-Auty J, Daniels MJD, White CS, Schwalbe CH, Schilling T, 
Hammadi H, Jaiyong P, Spencer NG, England H, Luheshi NM, Kadirvel M, 
Lawrence CB, Rothwell NJ, Harte MK, Bryce RA, Allan SM, Eder C, Freeman S, Brough D

Cell Chem Biol. 2017 Nov 16;24(11):1321-1335.e5 

NLRP3 is a receptor important for host responses to infection, yet is also known 
to contribute to devastating diseases such as Alzheimer's disease, diabetes, 
atherosclerosis, and others, making inhibitors for NLRP3 sought after. One of 
the inhibitors currently in use is 2-aminoethoxy diphenylborinate (2APB). 
Unfortunately, in addition to inhibiting NLRP3, 2APB also displays non-selective 
effects on cellular Ca2+ homeostasis. Here, we use 2APB as a chemical scaffold 
to build a series of inhibitors, the NBC series, which inhibit the NLRP3 
inflammasome in vitro and in vivo without affecting Ca2+ homeostasis. The core 
chemical insight of this work is that the oxazaborine ring is a critical feature 
of the NBC series, and the main biological insight the use of NBC inhibitors led 
to was that NLRP3 inflammasome activation was independent of Ca2+. The NBC 
compounds represent useful tools to dissect NLRP3 function, and may lead to 
oxazaborine ring-containing therapeutics.
________________________________________________________________

Calcium Fructoborate for Bone and Cardiovascular Health.

Mogoşanu GD, Biţă A, Bejenaru LE, Bejenaru C, Croitoru O, Rău G, 
Rogoveanu OC, Florescu DN, Neamţu J, Scorei ID, Scorei RI 

Biol Trace Elem Res. 2016 Aug;172(2):277-281 

Calcium fructoborate (CF), a natural sugar-borate ester found in fresh fruits 
and vegetables, is a source of soluble boron. CF contains three forms of borate 
(diester, monoester, and boric acid) and all are biologically active, both at 
the intracellular (as free boric acid) and extracellular level (as 
fructose-borate diester and monoester). At the cellular and molecular level, CF 
is superior to the boric acid/borate, exhibiting a complex "protective" effect 
against inflammatory response. CF is commercially available in the USA as a 
"nature-identical" complex, an active compound for dietary supplements. It 
provides effective and safe support against the discomfort and lack of 
flexibility associated with osteoarticular conditions (arthritis and joint 
degeneration), and improves Western Ontario and McMaster Universities 
Osteoarthritis (WOMAC) and McGill indexes. In addition, orally administered CF 
is effective in ameliorating symptoms of physiological response to stress, 
including inflammation of the mucous membranes, discomfort associated with 
osteoarthritis disorders, and bone loss, and also for supporting cardiovascular 
health. Clinical studies have exhibited the ability of CF to significantly 
modulate molecular markers associated with inflammatory mechanisms, mainly on 
the elevated serum levels of C-reactive protein (CRP).
________________________________________________________________

Comparison of Boric Acid and Combination Drug of Polymyxin, Neomycin and 
Hydrocortisone (polymyxin NH) in the Treatment of Acute Otitis Externa.

Amani S, Moeini M 

J Clin Diagn Res. 2016 Jul;10(7):MC01-4 

INTRODUCTION: Acute otitis externa is an inflammation of the external auditory 
canal known as "swimmer's ear". Direct costs including medical treatment, 
painkillers, antibiotics, steroids or both and indirect costs are also 
remarkable.
AIM: The aim of this study was to compare the effect of boric acid and 
polymyxin, neomycin and hydrocortisone composition in the treatment of acute 
otitis externa.
MATERIALS AND METHODS: This randomized clinical trial was carried out on 80 
patients aged more than 17-year-old who were referred to Kashani hospital clinic 
with a diagnosis of acute otitis externa by otolaryngologist. The patients were 
randomly allocated to two groups (A: Boric acid and B: polymyxin NH ear drops) 
and Painkiller was prescribed and administered orally for all patients and in 
the presence of fever, cellulitis around the ears and neck adenopathy, 
broad-spectrum systemic antibiotics were used besides topical treatment. 
Symptoms of patients who were evaluated by a physician includes pain, discharge 
from the ear, swelling of the ear canal, auricle swelling, tenderness, and ear 
itching. In addition, pain was evaluated in patients and was recorded by Macgill 
Pain Questionnaire, in the first, third, seventh and tenth days.
RESULTS: Results showed that itching on third day (p=0.007) and swelling of the 
ear canal in the examination of the third day (p=0.006) and the seventh day 
(p=0.001) in the polymyxin NH group was more than those of boric acid group. 
Overall mean pain based on McGill questionnaire was 11.10±1.49 in boric acid 
group in the examination on the first day and was 4.05±0.22 in the examination 
on the tenth day and in the polymyxin NH group, it was 10.9±0.99 on the first 
day and 4.20±0.40 on the tenth day. In both groups, pain relief was the same and 
there was no significant difference between two groups (p=0.075).
CONCLUSION: The findings of this study showed slight differences in the 
effectiveness of the boric acid drug and combination of polymyxin, neomycin and 
hydrocortisone in the treatment of patients with acute otitis externa that is of 
clinical significance.
________________________________________________________________

Boron Induces Lymphocyte Proliferation and Modulates the Priming Effects of 
Lipopolysaccharide on Macrophages. [Comment: while boron compounds in many 
settings are anti-inflammatory, yet here they are immune stimulating.]

Routray I, Ali S 

PLoS One. 2016 Mar 2;11(3):e0150607 

Chemical mediators of inflammation (CMI) are important in host defense against 
infection. The reduced capacity of host to induce the secretion of these 
mediators following infection is one of the factors in host susceptibility to 
infection. Boron, which has been suggested for its role in infection, is 
reported in this study to increase lymphocyte proliferation and the secretion of 
CMI by the lipopolysaccharide (LPS)-stimulated peritoneal macrophages in BALB/c 
mice. Boron was administered to mice orally as borax at different doses for 10 
consecutive days, followed by the stimulation of animals with ovalbumin and 
isolation of splenocytes for proliferation assay. The lymphocyte subsets were 
determined by flow cytometry in spleen cell suspension. The mediators of 
inflammation, TNF-α, IL-6, IL-1β and nitric oxide (NO), were measured in culture 
supernatant of LPS-primed macrophages isolated from borax treated mice. TNF and 
ILs were measured by ELISA. NO was determined by Griess test. The expression of 
inducible nitric oxide synthase (iNOS) in macrophages was studied by confocal 
microscopy. Results showed a significant increase in T and B cell populations, 
as indicated by an increase in CD4 and CD19, but not CD8, cells. Boron further 
stimulated the secretion of TNF-α, IL-6, IL-1β, NO and the expression of iNOS by 
the LPS-primed macrophages. The effect was dose dependent and most significant 
at a dose level of 4.6 mg/kg b. wt. Taken together, the study concludes that 
boron at physiological concentration induces lymphocyte proliferation and 
increases the synthesis and secretion of pro-inflammatory mediators by the 
LPS-primed macrophages, more specifically the M1 macrophages, possibly acting 
through Toll-like receptor. The study implicates boron as a regulator of the 
immune and inflammatory reactions and macrophage polarization, thus playing an 
important role in augmenting host defense against infection, with possible role 
in cancer and other diseases.
________________________________________________________________

Effect of absorbable gelatin sponge in the middle ear: 
in vitro and in vivo animal model. 

Goncalves S, Chiossone-Kerdel JA, Bianco AS, Ercolino JM, Hernandez-Rojas J.

Acta Otolaryngol. 2015 Jan;135(1):14-25 

CONCLUSION: GelitaSpon gelatin sponge (GS) showed faster reabsorption than 
gelfoam (GF) in vitro, and GS-packed middle ears resulted in a faster hearing 
recovery and less inflammation than the ears packed with GF soaked in saline. 
Soaking GF sponges with boric acid (BA) appeared to offset the inflammatory 
response of saline-soaked GF, making this inflammatory response comparable to 
that of GS-packed ears.
OBJECTIVE: To describe the reabsorption and inflammatory effects of absorbable 
gelatin sponge in the middle ear.
METHODS: For in vitro evaluation, GF and GS were used as disks immersed in 
saline solution or 3% BA and placed in gel. Images were captured with a 
microscope and processed using image processing Toolbox. For in vivo tests, 16 
female albino Sprague Dawley rats were divided into four groups: bulla opening; 
GF + 0.9% saline; GF + BA; and GS + 0.9% saline. An anterior approach to the 
right bulla was used for surgery. Preoperative and postoperative auditory 
brainstem response thresholds were measured.
RESULTS: In vitro, there was marked degradation of GF by day 14, while GS showed 
complete degradation by the third day. In vivo, hearing recovery occurred by day 
21. GF produced a more severe inflammatory response, which could be reduced by 
treating the GF with BA.
________________________________________________________________

Borax partially prevents neurologic disability and oxidative stress 
in experimental spinal cord ischemia/reperfusion injury.

Koc ER, Gökce EC, Sönmez MA, Namuslu M, Gökce A, Bodur AS 

J Stroke Cerebrovasc Dis. 2015 Jan;24(1):83-90 

OBJECTIVES: The aim of this study is to investigate the potential effects of 
borax on ischemia/reperfusion injury of the rat spinal cord.
METHODS: Twenty-one Wistar albino rats were divided into 3 groups: sham (no 
ischemia/reperfusion), ischemia/reperfusion, and borax (ischemia/reperfusion + 
borax); each group was consist of 7 animals. Infrarenal aortic cross clamp was 
applied for 30 minutes to generate spinal cord ischemia. Animals were evaluated 
functionally with the Basso, Beattie, and Bresnahan scoring system and 
inclined-plane test. The spinal cord tissue samples were harvested to analyze 
tissue concentrations of nitric oxide, nitric oxide synthase activity, xanthine 
oxidase activity, total antioxidant capacity, and total oxidant status and to 
perform histopathological examination.
RESULTS: At the 72nd hour after ischemia, the borax group had significantly 
higher Basso, Beattie, and Bresnahan and inclined-plane scores than those of 
ischemia/reperfusion group. Histopathological examination of spinal cord tissues 
in borax group showed that treatment with borax significantly reduced the degree 
of spinal cord edema, inflammation, and tissue injury disclosed by light 
microscopy. Xanthine oxidase activity and total oxidant status levels of the 
ischemia/reperfusion group were significantly higher than those of the sham and 
borax groups (P < .05), and total antioxidant capacity levels of borax group 
were significantly higher than those of the ischemia/reperfusion group (P < 
.05). There was not a significantly difference between the sham and borax groups 
in terms of total antioxidant capacity levels (P > .05). The nitric oxide levels 
and nitric oxide synthase activity of all groups were similar (P > .05).
CONCLUSIONS: Borax treatment seems to protect the spinal cord against injury in 
a rat ischemia/reperfusion model and improve neurological outcome.
________________________________________________________________

Chromatin immunoprecipitation analysis of bortezomib-mediated inhibition 
of NFκB recruitment to IL-1β and TNFα gene promoters in human macrophages.

Sanacora S, Chang TP, Vancurova I.

Methods Mol Biol. 2014;1172:315-27 

Interleukin-1β (IL-1) and tumor necrosis factor-α (TNF) are important 
pro-inflammatory cytokines involved in the mediation of the immune response, 
inflammation, tissue repair, and tumor progression. Regulation of IL-1 and TNF 
expression is mediated at the level of transcription by the transcription factor 
NFκB. Inhibition of NFκB activity by the proteasome inhibitor bortezomib (BZ) 
has been used as a frontline therapy in multiple myeloma and other hematological 
malignancies. In this chapter, we describe a protocol that uses chromatin 
immunoprecipitation (ChIP) to analyze the NFκB recruitment to endogenous IL-1 
and TNF promoters in BZ-treated human macrophages. Corresponding to the 
BZ-suppressed mRNA levels of IL-1 and TNF, we show that BZ inhibits p65 NFκB 
recruitment to IL-1 and TNF promoters. This study specifically uses U937 
macrophages, but the protocol could be easily modified to analyze the regulation 
of NFκB recruitment in other cell types.
________________________________________________________________

Effects of boric acid and 2-aminoethoxydiphenyl borate 
on necrotizing enterocolitis.

Yazıcı S, Akşit H, Korkut O, Sunay B, Çelik T.

J Pediatr Gastroenterol Nutr. 2014 Jan;58(1):61-7 

OBJECTIVE: The aim was to study the effects of boric acid (BA) and 
2-aminoethoxydiphenyl borate (2-APB) on oxidative stress and inflammation in an 
experimental necrotizing enterocolitis (NEC) rat model.
METHODS: Experimental NEC was induced in 40 newborn Sprague-Dawley rats by 
asphyxia and hypothermia applied in 3 consecutive days. Rats were subdivided 
into 4 subgroups as NEC, NEC+BA, NEC+2-APB, and controls. BA and 2-APB were 
applied daily before the procedure. Serum total antioxidant status, superoxide 
dismutase (SOD), tumor necrosis factor (TNF)-α, interleukin (IL)-6, and 
erythrocyte glutathione (GSH) levels were measured. Pathological changes for NEC 
in intestinal architecture were evaluated by a grading system.
RESULTS: Pretreatment with BA and 2-APB resulted in a decrease in NEC incidence. 
In all of the NEC groups, decreased serum levels of GSH and SOD were measured. 
Boron limited GSH consumption but had no effect on SOD levels. Total antioxidant 
status levels were not statistically different among groups. In our experimental 
NEC model, BA, but not 2-APB, prevented the increase of TNF-α. Pretreatment with 
BA and 2-APB downregulated the activity levels of IL-6 in NEC.
CONCLUSIONS: In the experimental NEC model, BA and 2-APB partly prevent NEC 
formation, modulate the oxidative stress parameters, bring a significant 
decrease in GSH consumption, and enhance the antioxidant defense mechanism, but 
have no effect on total antioxidant status. BA inhibits the hypoxia and 
hypothermia-induced increase in both IL-6 and TNF-a, but 2-APB only in IL-6. 
Boron may be beneficial in preventing NEC.
________________________________________________________________

Calcium fructoborate helps control inflammation associated 
with diminished bone health.

Scorei ID, Scorei RI.

Biol Trace Elem Res. 2013 Dec;155(3):315-21 

Inflammation has been identified as a possible contributory factor to disruption 
of the normal bone remodeling process, a process essential to healthy bone 
mineral density. Several large population-based clinical studies have 
specifically shown that levels of C-reactive protein, an immune recognition 
protein that is a sensitive marker of inflammation, are inversely and 
independently associated with total bone mineral density. The evidence suggests 
that control of C-reactive protein levels may contribute to bone health by 
protecting against inflammation's disruption of the equilibrium between bone 
resorption and bone deposition. Calcium fructoborate, a patented complex of 
calcium, fructose, and boron found naturally in fresh and dried fruits, 
vegetables and herbs, and wine, is a sugar-borate ester. A growing body of 
peer-reviewed, published clinical research indicates that the calcium 
fructoborate significantly reduces serum levels of the C-reactive protein in 
humans, suggesting that this unique plant-mineral complex may contribute to bone 
health by controlling the inflammation associated with loss of bone mineral 
density.
________________________________________________________________

Attenuation of early atherogenesis in low-density lipoprotein 
receptor-deficient mice by proteasome inhibition.

Wilck N, Fechner M, Dreger H, Hewing B, Arias A, 
Meiners S, Baumann G, Stangl V, Stangl K, Ludwig A.

Arterioscler Thromb Vasc Biol. 2012 Jun;32(6):1418-26 

OBJECTIVE: Low and nontoxic proteasome inhibition has anti-inflammatory, 
antiproliferative, and antioxidative effects on vascular cells in vitro and in 
vivo. We hypothesized that low-dose inhibition of the proteasome could provide 
antiatherogenic protection. The present study investigated the effect of 
low-dose proteasome inhibition on early lesion formation in low-density 
lipoprotein receptor-deficient mice fed a Western-type diet.
METHODS AND RESULTS: Male low-density lipoprotein receptor-deficient mice, 10 
weeks old, were fed a Western-type diet for 6 weeks with intraperitoneal 
injections of bortezomib or solvent. Bortezomib was injected at a dose of 50 
μg/kg body weight. Cholesterol plasma levels were not affected by bortezomib 
treatment. En face Oil Red O staining of aortae and aortic root cryosections 
demonstrated significant reduction of atherosclerotic lesion coverage in 
bortezomib-treated animals. Bortezomib significantly reduced vascular cellular 
adhesion molecule-1 expression and macrophage infiltration as shown by 
histological analysis. Bortezomib treatment resulted in a significant reduction 
of superoxide content, lipid peroxidation and protein oxidation products, serum 
levels of monocyte chemoattractant protein-1, and interleukin-6. Gene expression 
microarray analysis showed that expressional changes induced by Western-type 
diet were attenuated by treatment with low-dose bortezomib.
CONCLUSIONS: Low-dose proteasome inhibition exerts antioxidative and 
anti-inflammatory effects and attenuates development of atherosclerotic lesions 
in low-density lipoprotein receptor-deficient mice.
________________________________________________________________

Comparative effects of daily and weekly boron supplementation 
on plasma steroid hormones and proinflammatory cytokines.

Naghii MR, Mofid M, Asgari AR, Hedayati M, Daneshpour MS.

J Trace Elem Med Biol. 2011 Jan;25(1):54-8 

Boron possesses widespread properties in biochemistry and nutrition. Acute 
supplementation with 11.6 mg of boron resulted in a significant increase in 
plasma boron concentration. Given such a fast bioavailability, the objective was 
to determine whether acute (hourly or daily), and weekly supplementation could 
have any significant biological effects on the steroid hormones and further on 
some inflammatory biomarkers. Eight healthy male volunteers attended the 
laboratory on three occasions (days 0, 1 and 7). On the first day (day 0), a 
blood sample collection at 8.00 A.M was followed by ingestion of placebo with 
the breakfast. On the next day (supplementation-day 1), similar procedure was 
followed by ingestion of a capsule containing 10mg of boron. On both occasions 
blood was collected every 2h for the next 6h. Subjects were requested to consume 
a capsule of 10mg boron every day with their breakfast, and on the day 7, the 
blood collection was carried out at 8.00 A.M, again. Boron in plasma increased 
significantly following hours and weekly consumption. Six hours supplementation 
showed a significant decrease on sex hormone binding globulin (SHBG), high 
sensitive CRP (hsCRP) and TNF-α level. After one week (in samples taken at 8.00 
A.M, only), the mean plasma free testosterone increased and the mean plasma 
estradiol decreased significantly. Dihydrotestosterone, cortisol and vitamin D 
was elevated. Also, concentrations of all three inflammatory biomarkers 
decreased after supplementation. Of note, despite decreased proinflammatory 
cytokines, based on recent clinical data, this must be the first human study 
report to show an increase level of free testosterone after boron consumption.
________________________________________________________________

Inhibition of NF-κB signaling retards eosinophilic dermatitis 
in SHARPIN-deficient mice.

Liang Y, Seymour RE, Sundberg JP.

J Invest Dermatol. 2011 Jan;131(1):141-9 

The NF-κB pathway performs pivotal roles in diverse physiological processes such 
as immunity, inflammation, proliferation, and apoptosis. NF-κB is kept inactive 
in the cytoplasm through association with inhibitors (IκB), and translocates to 
the nucleus to activate its target genes after the IκBs are phosphorylated and 
degraded. Here, we demonstrate that loss of function of SHANK-associated RH 
domain interacting protein (SHARPIN) leads to activation of NF-κB signaling in 
skin, resulting in the development of an idiopathic hypereosinophilic syndrome 
(IHES) with eosinophilic dermatitis in C57BL/KaLawRij-Sharpin(cpdm)/RijSunJ 
mice, and clonal expansion of B-1 B cells and CD3(+)CD4(-)CD8(-) T cells. 
Transcription profiling in skin revealed constitutive activation of classical 
NF-κB pathways, predominantly by overexpressed members of IL1 family. 
Compound-null mutants for both the IL1 receptor accessory protein 
(Il1rap(tm1Roml)) and SHARPIN (Sharpin(cpdm)) resulted in mice having decreased 
skin disease severity. Inhibition of IκBA degradation by the proteasome 
inhibitor bortezomib alleviated the dermatitis in Sharpin(cpdm) mice. These 
results indicate that absence of SHARPIN causes IHES with eosinophilic 
dermatitis by NF-κB activation, and bortezomib may be an effective treatment for 
skin problems of IHES.
________________________________________________________________

Discovery and optimization of boronic acid based inhibitors of autotaxin.

Albers HM, van Meeteren LA, Egan DA, van Tilburg EW, Moolenaar WH, Ovaa H.

J Med Chem. 2010 Jul 8;53(13):4958-67 

Autotaxin (ATX) is an extracellular enzyme that hydrolyzes 
lysophosphatidylcholine (LPC) to produce the lipid mediator lysophosphatidic 
acid (LPA). The ATX-LPA signaling axis has been implicated in diverse 
physiological and pathological processes, including vascular development, 
inflammation, fibrotic disease, and tumor progression. Therefore, targeting ATX 
with small molecule inhibitors is an attractive therapeutic strategy. We 
recently reported that 2,4-thiazolidinediones inhibit ATX activity in the 
micromolar range. Interestingly, inhibitory potency was dramatically increased 
by introduction of a boronic acid moiety, designed to target the active site 
threonine in ATX. Here we report on the discovery and further optimization of 
boronic acid based ATX inhibitors. The most potent of these compounds inhibits 
ATX-mediated LPC hydrolysis in the nanomolar range (IC(50) = 6 nM). The finding 
that ATX can be targeted by boronic acids may aid the development of ATX 
inhibitors for therapeutic use.
________________________________________________________________

Boronic acid-based inhibitor of autotaxin reveals rapid turnover 
of LPA in the circulation.

Albers HM, Dong A, van Meeteren LA, Egan DA, Sunkara M, van Tilburg EW, 
Schuurman K, van Tellingen O, Morris AJ, Smyth SS, Moolenaar WH, Ovaa H.

Proc Natl Acad Sci U S A. 2010 Apr 20;107(16):7257-62 

Autotaxin (ATX) is a secreted nucleotide pyrophosphatase/phosphodiesterase that 
functions as a lysophospholipase D to produce the lipid mediator 
lysophosphatidic acid (LPA), a mitogen, chemoattractant, and survival factor for 
many cell types. The ATX-LPA signaling axis has been implicated in angiogenesis, 
chronic inflammation, fibrotic diseases and tumor progression, making this 
system an attractive target for therapy. However, potent and selective nonlipid 
inhibitors of ATX are currently not available. By screening a chemical library, 
we have identified thiazolidinediones that selectively inhibit ATX-mediated LPA 
production both in vitro and in vivo. Inhibitor potency was approximately 
100-fold increased (IC(50) approximately 30 nM) after the incorporation of a 
boronic acid moiety, designed to target the active-site threonine (T210) in ATX. 
Intravenous injection of this inhibitor into mice resulted in a surprisingly 
rapid decrease in plasma LPA levels, indicating that turnover of LPA in the 
circulation is much more dynamic than previously appreciated. Thus, boronic 
acid-based small molecules hold promise as candidate drugs to target ATX.
________________________________________________________________

Potent anti-inflammatory effects of low-dose proteasome 
inhibition in the vascular system.

Ludwig A, Fechner M, Wilck N, Meiners S, 
Grimbo N, Baumann G, Stangl V, Stangl K 

J Mol Med (Berl). 2009 Aug;87(8):793-802 
Comment in J Mol Med (Berl). 2009 Aug;87(8):749-51.

Proteasome inhibitors are considered to have anti-inflammatory therapeutic 
potential. However, recent reports addressing proteasome inhibition in the 
vascular system are controversial, ranging from beneficial anti-inflammatory and 
anti-oxidative effects to potentiation of inflammation and oxidative stress. 
This study was based on the hypothesis that the divergent effects might be a 
result of a differential and dose-dependent responsiveness of vascular cells to 
proteasome inhibitors. We tested whether low doses of proteasome inhibitors 
would favor anti-inflammatory effects in vascular cells in vitro and in vivo. 
Human umbilical vein endothelial cells (HUVEC) were preincubated with proteasome 
inhibitors MG132 and MG262 at concentrations that did not affect cell viability 
during a 24-h treatment. Upon addition of tumor necrosis factor alpha 
(TNF-alpha) the induced expression of adhesion molecules and the adhesion of 
monocytic THP-1 cells to HUVECs was significantly lowered. However, nuclear 
translocation of NF-kappaB was only slightly diminished. Low-dose pretreatment 
with proteasome inhibitors decreased TNF-alpha-induced generation of reactive 
oxygen species in HUVEC. Bortezomib was administered at a dose of 50 microg/kg 
body weight to Dahl salt-sensitive rats (DSSR) on high-salt diet. This low-dose 
proteasome inhibition led to decreased hypertension-induced oxidative stress and 
reduced expression of vascular cell adhesion molecule 1 (VCAM-1) in the aortae.
________________________________________________________________

Effect of proteasome inhibitors on endotoxin-induced diaphragm dysfunction.

Supinski GS, Vanags J, Callahan LA.

Am J Physiol Lung Cell Mol Physiol. 2009 Jun;296(6):L994-L1001 

Infections produce severe respiratory muscle dysfunction. It is known that the 
proteasome proteolytic system is activated in skeletal muscle in sepsis, and it 
has been postulated that this degradative pathway is responsible for inducing 
skeletal muscle weakness and wasting. The objective of this study was to 
determine if administration of proteasomal inhibitors (MG132, epoxomicin, 
bortezomib) can prevent sepsis-induced diaphragm weakness. Rats were given 
either 1) saline (0.5 ml ip), 2) endotoxin (12 mg/kg ip), 3) endotoxin plus 
MG132 (2.5 mg/kg), 4) endotoxin plus epoxomicin (1 micromol/kg), or 5) endotoxin 
plus bortezomib (0.05 mg/kg). Animals were killed either 48 or 96 h after 
injections, and assessments were made of diaphragm proteolysis, force-frequency 
relationships, mass, protein content, and caspase activation. Endotoxin 
increased proteolysis (P <0.001). MG132, epoxomicin, and bortezomib each 
prevented the endotoxin-induced increase in proteolysis (P <0.01). Endotoxin 
induced severe reductions in diaphragm force generation by 48 h (P <0.01); none 
of the proteasomal inhibitors prevented loss of force. Endotoxin induced 
significant reductions in diaphragm mass and protein content by 96 h (P <0.01); 
neither MG132 nor epoxomicin prevented loss of mass or protein, but bortezomib 
attenuated the reduction in protein content (P <0.05). Endotoxin increased 
diaphragm caspase-3 activity (P <0.01); caspase-3 activity remained high when 
either MG132, epoxomicin, or bortezomib were given. These data suggest 
proteasomal inhibitors are not an adequate treatment to prevent 
endotoxin-induced diaphragmatic dysfunction.
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Inhibition of experimental periodontitis by a topical boron-based antimicrobial.

Luan Q, Desta T, Chehab L, Sanders VJ, Plattner J, Graves DT.

J Dent Res. 2008 Feb;87(2):148-52 

AN0128 is a boron-containing compound with antibacterial and anti-inflammatory 
properties. To test its potential effectiveness in treating periodontal disease, 
we induced experimental periodontitis in the rat by placing ligatures and 
assessed the impact of AN0128 and positive and negative controls by micro-CT and 
histologic measurements. The formation of an inflammatory infiltrate was 
measured in hematoxylin-and-eosin-stained sections. Daily application of AN0128 
(1%) compared with controls reduced bone loss by 38 to 44% (P < 0.05), while 
vehicle alone had no effect (P > 0.05). The reduction in bone loss with AN0128 
was similar to that achieved with a NSAID, ketorolac, and Total toothpaste 
containing triclosan. AN0128 also reduced the level of gingival inflammation 42% 
compared with the ligature only (P < 0.05), whereas vehicle alone had no effect 
(P > 0.05). The results indicate that AN0128 significantly reduces the formation 
of an inflammatory infiltrate and reduces bone loss, measured histologically and 
by micro-CT.
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Targeting NF-kappaB: a promising molecular therapy in inflammatory arthritis.

Roman-Blas JA, Jimenez SA.

Int Rev Immunol. 2008;27(5):351-74 

The nuclear factor-kappa B family of transcription factors is intimately 
involved in the regulation of the inflammatory responses that play a fundamental 
role in the damage of articular tissues. Thus, many studies have examined the 
important contributions of components of the NF-kappaB signaling pathways to the 
pathogenesis of various rheumatic diseases and their pharmacologic modulation. 
Currently available therapeutic agents including nonsteroidal anti-inflammatory 
drugs, corticosteroids, nutraceuticals, and disease-modifying antirheumatic 
drugs, as well as novel specific small-molecule inhibitors have been employed. 
In addition, promising nucleic acid-based strategies have shown encouraging 
results. However, further research will be needed before NF-kappaB-aimed 
strategies become an effective therapy for inflammatory arthritis.
________________________________________________________________

Boric acid inhibits LPS-induced TNF-alpha formation 
through a thiol-dependent mechanism in THP-1 cells.

Cao J, Jiang L, Zhang X, Yao X, Geng C, Xue X, Zhong L.

J Trace Elem Med Biol. 2008;22(3):189-95

Oxidative stress plays an important role during inflammatory diseases and 
antioxidant administration to diminish oxidative stress may arrest inflammatory 
processes. Boron has been implicated to modulate certain inflammatory mediators 
and regulate inflammatory processes. Here we investigated the role of the 
tripeptide glutathione (GSH) in modulating the effects of boric acid (BA) on 
lipopolysaccharide (LPS)-induced tumor necrosis factor alpha (TNF-alpha) 
formation in THP-1 monocytes. Interestingly, we found that BA had no significant 
effects on both TNF-alpha production and intracellular GSH contents, whereas it 
could inhibit LPS-induced TNF-alpha formation and ameliorated the 
d,l-buthionine-S,R-sulfoximine (BSO)-induced GSH depletion. Twenty-four hour 
incubation with BSO induced a decrease of the intracellular GSH and an increase 
of TNF-alpha. Treatment with N-acetyl-l-cysteine (NAC) did not significantly 
increase intracellular content of GSH but significantly reduced the secretion of 
TNF-alpha. BSO-pretreatment for 24h enhanced the LPS-induced secretion and mRNA 
expression of TNF-alpha further. BA inhibited LPS-stimulated TNF-alpha formation 
was also seen after GSH depletion by BSO. These results indicate that BA may 
have anti-inflammatory effect in the LPS-stimulated inflammation and the effect 
of BA on TNF-alpha secretion may be induced via a thiol-dependent mechanism.
________________________________________________________________

Proteasome inhibitor attenuates skeletal muscle reperfusion injury 
by blocking the pathway of nuclear factor-kappaB activation.

Park JW, Qi WN, Cai Y, Urbaniak JR, Chen LE.

Plast Reconstr Surg. 2007 Dec;120(7):1808-1818 

BACKGROUND: Nuclear factor-kappaB is a key transcriptional factor in the 
regulation of inflammatory factors that are involved in tissue reperfusion 
injury, but conflicting data have been presented in the literature. The 
proteasome regulates proteins that control cell-cycle progression and apoptosis, 
and inhibition of the proteasome has been shown to reduce nuclear factor-kappaB 
activation and reperfusion injury. Although bortezomib is a potent proteasome 
inhibitor, its role in skeletal muscle reperfusion injury has not been 
documented, and its effects on the regulation of inflammatory factors in 
reperfused tissue are unclear. In this study, the authors investigated the role 
of nuclear factor-kappaB in skeletal muscle reperfusion injury and the effect of 
bortezomib (a proteasome inhibitor) on reperfusion injury.
METHODS: Pedicled cremaster muscle flaps from bortezomib-treated and 
phosphate-buffered saline-treated control mice were subjected to 4.5 hours of 
ischemia and 90 minutes of reperfusion.
RESULTS: During reperfusion, arterial diameters and blood flow recovered earlier 
and more completely in bortezomib-treated muscle than in controls. Compared with 
controls, Western blot analysis demonstrated a significant reduction in 
degradation of nuclear factor-kappaB inhibitory protein and expression of 
inducible nitric oxide synthase protein in bortezomib-treated muscle at the end 
of reperfusion. Immunohistochemistry showed decreased nuclear factor-kappaB 
p65-binding activity and down-regulated protein expression of intercellular 
adhesion molecule-1 and nitrotyrosine, accompanied by less muscle edema and 
inflammation as proven by histologic examination.
CONCLUSIONS: Bortezomib effectively blocks nuclear factor-kappaB activation in 
attenuating muscle reperfusion injury through inhibiting nuclear factor-kappaB 
inhibitory protein degradation. Therefore, inhibition of proteasome activity may 
provide a novel therapeutic strategy for the treatment of skeletal muscle 
reperfusion injury.
________________________________________________________________

Differential involvement of NF-kappaB and MAP kinase pathways 
in the generation of inflammatory cytokines by human neutrophils.

Cloutier A, Ear T, Blais-Charron E, Dubois CM, McDonald PP.

J Leukoc Biol. 2007 Feb;81(2):567-77 

The ability of human neutrophils to express a variety of genes encoding 
inflammatory mediators is well documented, and mounting evidence suggests that 
neutrophil-derived cytokines and chemokines contribute to the recruitment of 
discrete leukocyte populations at inflammatory sites. Despite this, our 
understanding of the signaling intermediates governing the generation of 
inflammatory cytokines by neutrophils remains fragmentary. Here, we report that 
inhibitors of the p38 MAPK and MEK pathways substantially diminish the release 
of (and in the case of p38 inhibitors, the gene expression of) several 
inflammatory cytokines in neutrophils stimulated with LPS or TNF. In addition, 
various NF-kappaB inhibitors were found to profoundly impede the inducible gene 
expression and release of inflammatory cytokines in these cells. The MAPK 
inhibitors did not affect NF-kappaB activation; instead, the transcriptional 
effects of the p38 MAPK inhibitor appear to involve transcriptional factor IID. 
Conversely, the NF-kappaB inhibitors failed to affect the activation of MAPKs. 
Finally, the MAPK inhibitors were found to prevent the activation a key 
component of the translational machinery, S6 ribosomal protein, in keeping with 
their post-transcriptional impact on cytokine generation. To our knowledge, this 
constitutes the first demonstration that in neutrophils, the inducible 
expression of proinflammatory cytokines by physiological stimuli largely 
reflects the ability of the latter to activate NF-kappaB and selected MAPK 
pathways. Our data also raise the possibility that NF-kappaB or MAPK inhibitors 
could be useful in the treatment of inflammatory disorders in which neutrophils 
predominate.
________________________________________________________________

NF-kappaB and not the MAPK signaling pathway regulates 
GADD45beta expression during acute inflammation.

Zhang N, Ahsan MH, Zhu L, Sambucetti LC, Purchio AF, West DB.

J Biol Chem. 2005 Jun 3;280(22):21400-8 

The GADD45 (growth arrest and DNA damage-inducible) family of genes is involved 
in the regulation of cell cycle progression and apoptosis. To study signaling 
pathways affecting GADD45beta expression and to examine systematically in vivo 
the GADD45beta expression in tissues following various toxic stresses, we 
created a transgenic mouse by fusing the GADD45beta promoter to firefly 
luciferase (Gadd45beta-luc). In vivo GADD45beta expression was assessed by 
measuring the luciferase activity in the Gadd45beta-luc transgenic mouse using a 
non-invasive imaging system (IVIS Imaging System, Xenogen Corporation). We found 
that a number of agents that induce oxidative stress, such as sodium arsenite, 
CCl4, lipopolysaccharide (LPS), or tumor necrosis factor-alpha, are able to 
induce luciferase expression throughout the entire animal. In liver, spleen, 
lung, intestine, kidney, and heart, we observed an induction of luciferase 
activity after LPS treatment, which correlates with an increase of GADD45beta 
mRNA in these tissues. Processes that induce DNA damage activate the NF-kappaB 
signaling pathway. Several inhibitors of the NF-kappaB signaling pathway, 
including dexamethasone, thalidomide, and a proteasome inhibitor, bortezomib, 
showed inhibitory effects on LPS-induced GADD45beta expression as indicated by a 
decrease of the luciferase activity. Northern blot analysis confirmed a broad 
inhibitory effect of bortezomib on LPS-induced GADD45beta mRNA expression in 
spleen, lung, and intestine. In liver of bortezomib-treated mice, we observed a 
reverse correlation between the luciferase activity and the GADD45beta mRNA 
level. We speculate that such a discrepancy could be due to severe liver 
toxicity caused by bortezomib and LPS co-treatment. MAPK inhibitors had 
transient and inconsistent effects on LPS-induced luciferase expression. Our 
data are consistent with the notion that NF-kappaB, but not the MAPK signaling 
pathways, is involved in the in vivo regulation of GADD45beta expression. Thus, 
NF-kappaB signaling involves induction of GADD45beta expression, which supports 
the proposed role of GADD45beta in protecting cells against DNA damaged under 
various stress conditions.
________________________________________________________________

Small-molecule inhibitors of proteasome activity.

Gaczynska M, Osmulski PA.

Methods Mol Biol. 2005;301:3-22 

The fast-track approval of a proteasome inhibitor, PS-341, to treat multiple 
myeloma spurred a wave of interest in both the proteasome itself and 
small-molecule compounds blocking its activities. Besides being candidates for 
drugs against cancer, autoimmune diseases, inflammation, or stroke, specific 
proteasome inhibitors are indispensable tools for biochemical and cell biology 
investigations of the proteasome and proteasome-ubiquitin system. Numerous 
synthetic peptide derivatives, such as boronates, epoxides, aldehydes, vinyl 
sulfones, cyclic peptides, and lactones, block the N-terminal threonine-type 
active centers of the enzyme, halting the cleavage of proteasomal protein 
substrates both in vitro and in vivo. Because some of the proteasomal inhibitors 
exhibit a high specificity toward only one particular type of an active center 
of the proteasome, they constitute valuable probes for testing the mechanism of 
proteolysis catalyzed by the enzyme. In this chapter we discuss the most common 
applications of available proteasome inhibitors. In addition to the best-known 
competitive inhibitors, we also describe the benefits from the use of allosteric 
inhibitors, which induce distinct but less understood in vitro and in vivo 
effects on the proteasomal machinery. Finally, we present the application of the 
basic biochemical procedures to decipher the mechanism of interactions of a 
novel compound with the proteasome.
________________________________________________________________

Effect of boron supplementation of pig diets on the production 
of tumor necrosis factor-alpha and interferon-gamma.

Armstrong TA, Spears JW.

J Anim Sci. 2003 Oct;81(10):2552-61 

Two experiments were conducted to determine the effects of dietary B on the 
production of cytokines following an endotoxin challenge. In both experiments, 
pigs were obtained from litters generated from sows fed low-B (control) or 
B-supplemented (5 mg/ kg, as-fed basis) diets. In Exp. 1 and 2, 28 and 35 pigs, 
respectively (21 d old), remained with their littermates throughout a 49-d 
nursery phase and were fed either a control or B-supplemented diet. In Exp. 1, 
12 pigs per treatment were moved to individual pens at the completion of the 
nursery phase and fed their respective experimental diet. On d 99 of the study, 
pigs were injected with 150 microg of phytohemagglutinin (PHA) to evaluate a 
local inflammatory response. Pigs receiving the B-supplemented diet had a 
decreased (P < 0.01) inflammatory response following PHA injection. Peripheral 
blood monocytes were isolated from six pigs per treatment on d 103 and cultured 
in the presence of lipopolysaccharide (LPS) to determine the effect of dietary B 
on tumor necrosis factor-alpha (TNF-alpha) production from monocytes. Isolated 
monocytes from pigs that received the B-supplemented diet had a numerically 
greater (P = 0.23) production of TNF-alpha. In Exp. 2, pigs were group housed 
with their littermates following the nursery phase for 43 d, after which 10 pigs 
per treatment were moved to individual pens. In Exp. 1 and 2, pigs were assigned 
randomly within dietary treatment to receive either an i.m. injection of saline 
or LPS on d 117 and d 109, respectively. The dose of LPS in Exp. 1 and 2 was 100 
and 25 microg of LPS/kg of BW, respectively. In Exp. 1, serum TNF-alpha was 
increased (P < 0.01) at 2 h and tended to be increased (P < 0.11) at 6 and 24 h 
after injection by dietary B; however, only numerical trends existed for a 
B-induced increase in TNF-alpha in Exp. 2. Serum interferon-gamma (IFN-gamma) 
was increased (P < 0.01) at 6 h and tended to be increased (P < 0.08) at 24 h 
after injection in Exp. 1. In Exp. 2, dietary B also numerically increased 
IFN-alpha. These data indicate that dietary B supplementation increased the 
production of cytokines following a stress, which indicates a role of B in the 
immune system; however, these data do not explain the reduction in localized 
inflammation following an antigen challenge in pigs.
________________________________________________________________

Arginase and autoimmune inflammation in the central nervous system.

Xu L, Hilliard B, Carmody RJ, Tsabary G, Shin H, Christianson DW, Chen YH.

Immunology. 2003 Sep;110(1):141-8 

Using a high throughput gene microarray technology that detects approximately 22 
000 genes, we found that arginase I was the most significantly up-regulated gene 
in the murine spinal cord during experimental autoimmune encephalomyelitis 
(EAE). By Northern blot and arginase enzyme assay, we detected high levels of 
arginase I mRNA and protein, respectively, in the spinal cord of EAE mice, but 
not in the spinal cord of normal mice or mice that had recovered from EAE. In 
vitro, both microglia and astrocytes produced arginase and nitric oxide 
synthase, two enzymes that are involved in arginine metabolism. To explore the 
roles of arginase in EAE, we injected the arginase inhibitor 
amino-6-boronohexanoic acid (ABH) into mice during the inductive and effector 
phases of the disease. Compared with mice that received vehicle control, mice 
treated with ABH developed milder EAE with delayed onset, reduced disease score 
and expedited recovery. Spleen mononuclear cells from ABH-treated mice produced 
more nitric oxide and secreted less interferon-gamma and tumour necrosis 
factor-alpha as compared to control mice. These results indicate that arginase 
plays important roles in autoimmune inflammation in the central nervous system.
________________________________________________________________

Proteasome inhibition: a new anti-inflammatory strategy.

Elliott PJ, Zollner TM, Boehncke WH.

J Mol Med (Berl). 2003 Apr;81(4):235-45 

The ubiquitin-proteasome pathway has a central role in the selective degradation 
of intracellular proteins. Among the key proteins modulated by the proteasome 
are those involved in the control of inflammatory processes, cell cycle 
regulation, and gene expression. Consequently proteasome inhibition is a 
potential treatment option for cancer and inflammatory conditions. Thus far, 
proof of principle has been obtained from studies in numerous animal models for 
a variety of human diseases including cancer, reperfusion injury, and 
inflammatory conditions such as rheumatoid arthritis, asthma, multiple 
sclerosis, and psoriasis. Two proteasome inhibitors, each representing a unique 
chemical class, are currently under clinical evaluation. Velcade (PS-341) is 
currently being evaluated in multiple phase II clinical trials for several solid 
tumor indications and has just entered a phase III trial for multiple myeloma. 
PS-519, representing another class of inhibitors, focuses on the inflammatory 
events following ischemia and reperfusion injury. Since proteasome inhibitors 
exhibit anti-inflammatory and antiproliferative effects, diseases characterized 
by both of these processes simultaneously, as is the case in rheumatoid 
arthritis or psoriasis, might also represent clinical opportunities for such 
drugs.
________________________________________________________________

The proteasome: a new target for novel drug therapies.

Elliott PJ, Ross JS.

Am J Clin Pathol. 2001 Nov;116(5):637-46 

The proteasome is an enzyme present in all cells, from yeast to human, and has a 
central role in the proteolytic degradation of the vast majority of 
intracellular proteins. Among the key proteins modulated by the proteasome are 
those involved in controlling inflammatory processes, cell cycle regulation, and 
gene expression. As such, agents that inhibit the proteasome have been shown to 
be active in numerous animal models of inflammation and cancer Two proteasome 
inhibitors are under clinical evaluation. PS-519 is being studied for the 
treatment of reperfusion injury that occurs following cerebral ischemia and 
myocardial infarction. The other, PS-341, has recently entered multiple phase 2 
clinical trials for the treatment of multiple myeloma, chronic lymphocytic 
leukemia, and a variety of solid tumors. The proteasome may have an important 
role in the evolution of HIV-related disorders including AIDS and inflammatory 
disorders. Therapeutic strategies using proteasome inhibitors for the treatment 
of these conditions have now entered preclinical development.
________________________________________________________________

The anti-inflammatory activity of boron derivatives in rodents.

Hall IH, Burnham BS, Chen SY, Sood A, Spielvogel BF, Morse KW.

Met Based Drugs. 1995;2(1):1-12 

Acyclic amine-carboxyboranes were effective anti-inflammatory agents in mice at 
8 mg/kg x 2. These amine-carboxyboranes were more effective than the standard 
indomethacin at 8 mg/kg x 2, pentoxifylline at 50 mg/kg x 2, and phenylbutazone 
at 50 mg/kg x 2. The heterocyclic amine derivatives as well as 
amine-carbamoylboranes, carboalkoxyboranes, and cyanoboranes were generally less 
active. However, selected aminomethyl-phosphonate-N-cyanoboranes demonstrated 
greater than 60% reduction of induced inflammation. The boron compounds were 
also active in the rat induced edema, chronic arthritis, and pleurisy screens, 
demonstrating activity similar to the standard indomethacin. The compounds were 
effecive in reducing local pain and decreased the tail flick reflex to pain. The 
derivatives which demonstrated good anti-inflammatory activity were effective 
inhibitors of hydrolytic lysosomal, and proteolytic enzyme activities with 
IC(50) 50 values equal to (-6)M in mouse macrophages, human leukocytes, and Be 
Sal osteofibrolytic cells. In these same cell lines, the agents blocked 
prostaglandin cyclooxygenase activity with IC(50) values of (-6)M. In mouse 
macrophage and human leukocytes, 5' lipoxygenase activity was also inhibited by 
the boron derivatives with IC(50) values of 10(-6)M. These IC(50) values for 
inhibition of these enzyme activities are consistent with published values of 
known anti-inflammatory agents which target these enzymes.
________________________________________________________________

Anti-inflammatory and anti-osteoporotic activities of base-boronated 
nucleosides and phosphate-boronated nucleotides in rodents.

Rajendran KG, Burnham BS, Chen SY, Sood A, Spielvogel BF, Shaw BR, Hall IH.

J Pharm Sci. 1994 Oct;83(10):1391-5 

The 2'-deoxyribonucleoside cyanoboranes were effective anti-inflammatory agents 
in rodents at 2-8 mg/kg; they blocked induced edema, septic shock, and pleurisy. 
Overall compounds 3',5'-O-(bis- (triisopropylsilyl)-2'-deoxyinosine (1), 
3',5'-O-bis(triisopropylsilyl)-2'-deoxycytidine (10), 
N3-(cyanoboryl)-2'-deoxycytidine (11), N7-(cyanoboryl)-N2-isobutyryl- 
3',5'-O-bis(triisopropylsilyl)-2'-deoxyguanosine (20), and N7-(cyanoboryl)-N2- 
isobutyryl-5'-O-(4,4'-dimethoxytrityl)-3'-O-(triisopropylsilyl)-2' 
-deoxyguanosine (22) were the most active when all the anti-inflammatory screens 
are considered. The agents also blocked both local and central pain caused by 
inflammation. These nucleosides blocked calcium resorption but were less 
effective compared to other amine carboxyboranes. The inflammation process 
appeared blocked by these compounds because of their effectiveness in reducing 
both hydrolytic lysosomal enzyme and proteolytic enzyme activities. The agents 
were also dual inhibitors of prostaglandin cyclooxygenase and 5'-lipoxygenase 
activities in leukocytes and macrophages. These agents at 10(-4) M demonstrated 
no specific organ toxicity to ileum mucosa cells grown in tissue culture.
________________________________________________________________

Peptide boronic acids, substrate analogs, inhibit chymase, 
and histamine release from rat mast cells.

Kato Y, Kido H, Fukusen N, Katunuma N 

J Biochem. 1988 May;103(5):820-2 

Peptide boronic acids, such as methoxysuccinyl-Ala-Ala-Pro-(L)boro-Phe-OH, its 
pinacol ester, and t-butyloxycarbonyl-Phe-Pro-(L)boro-Phe-pinacol, inhibited the 
activity of chymase from connective tissue mast cells approximately 40- to 
80-fold more than atypical chymase from mucosal mast cells, and did not inhibit 
trypsin. Only peptide boronic acids containing "L" forms of boronic acids were 
inhibitory. The Ki values of these peptide boronic acids for chymase were in the 
60-170 nM concentration range, like those of the natural inhibitors tested, but 
all the natural inhibitors tested except Eglin C and chymostatin inhibited both 
chymase and trypsin. Thus these peptide boronic acids should be useful for 
selective inhibition of chymase with less inhibitory activity for atypical 
chymase and without inhibition of trypsin. These peptide boronic acids markedly 
inhibited histamine release induced by anti-rat immunoglobulin E, suggesting 
that chymase in connective tissue mast cells plays some role in the process of 
histamine release. These peptides are assumed to be therapeutically useful for 
treatment of allergic inflammations catalyzed by chymase.
________________________________________________________________