Antibacterial effects of microbial synthesized silver-copper nanoalloys on Escherichia coli, Burkholderia cepacia, Listeria monocytogenes and Brucella abortus.

BACKGROUND AND OBJECTIVES: Bacterial resistance is an emerging public health problem worldwide. Metallic nanoparticles and nanoalloys open a promising field due to their excellent antimicrobial effects. The aim of the present study was to investigate the antibacterial effects of Ag-Cu nanoalloys, which were biosynthesized by Lactobacillus casei ATCC 39392, on some of the important bacterial pathogens, including Escherichia coli, Burkholderia cepacia, Listeria monocytogenes and Brucella abortus. MATERIALS AND METHODS: Ag-Cu nanoalloys were synthesized through the microbial reduction of AgNO3 and CuSO4 by Lactobacillus casei ATCC39392. Furthermore, they were characterized by Fourier-Transform Infrared Spectrometer (FTIR) and Field Emission Scanning Electron Microscopy (FESEM) analysis in order to investigate their chemical composition and morphological features, respectively. The minimum inhibitory and minimum bactericidal concentrations of Ag-Cu nanoalloys were determined against each strain. The bactericidal test was conducted on the surface of MHA supplemented with 1, 0.1, and 0.01 µg/µL of the synthesized nanoalloy. The antimicrobial effects of synthesized nanoalloy were compared with ciprofloxacin, ampicillin and ceftazidime as positive controls. RESULTS: Presence of different chemical functional groups, including N-H, C-H, C-N and C-O on the surface of Ag-Cu nanoalloys was recorded by FTIR. FESEM micrographs revealed uniformly distributed nanoparticles with spherical shape and size ranging from 50 to 100 nm. The synthesized Ag-Cu nanoalloys showed antibacterial activity against L. monocytogenes PTCC 1298, E. coli ATCC 25922 and B. abortus vaccine strain. However, no antibacterial effects were observed against B. cepacia ATCC 25416. CONCLUSION: According to the findings of the present research, the microbially synthesized Ag-Cu nanoalloy demonstrated antibacterial effects on the majority of the bacteria studied even at 0.01 µg/µL. However, complementary investigations should be conducted into the safety of this nanoalloy for in vivo or systemic use.

The protective effect of Lactobacillus and Bifidobacterium as the gut microbiota members against chronic urticaria.

BACKGROUND: Chronic Urticaria is a common disorder which is defined by recurrent occurrence of wheals and sometimes angioedema. It has a notable influence on the patients' quality of life. Regulation of the immune system is one of the important roles of the gut microbiota. The effect of dysbiosis considering some members of gut microbiota in patients with chronic urticaria has been demonstrated in our previous study. OBJECTIVE: Comparing the frequency and bacterial load of Lactobacillus, Bifidobacterium, and Bacteroides between patients with chronic urticaria and healthy controls. METHODS: 20 patients with chronic urticaria and 20 age and sex matched healthy individuals were included in the present study. Stool samples were analyzed for determining the frequency and bacterial load of Lactobacillus, Bifidobacterium, and Bacteroides genera. RESULTS: There were no significant differences among the frequencies of detectable Lactobacillus, Bifidobacterium, or Bacteroides in stool samples of patients with chronic urticaria and healthy controls. The relative amounts of Lactobacillus and Bifidobacterium were significantly higher in fecal samples from controls compared to patients with chronic urticaria (P = 0.038 and 0.039, respectively). CONCLUSION: It is the first study on the implication of Lactobacillus, Bifidobacterium, and Bacteroides genera as gut microbiota members in patients with chronic urticaria.

Emergence of vancomycin-intermediate and -resistant Staphylococcus aureus among methicillin-resistant S. aureus isolated from clinical specimens in the northwest of Iran.

OBJECTIVES: The aim of this study was to evaluate the frequency as well as the phenotypic and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant S. aureus (VRSA) isolates from clinical specimens at three university teaching hospitals in Urmia, Northwest Iran, from 2012-2015. METHODS: Following identification of the isolates, antibiotic susceptibility testing was performed. The presence of the mecA, vanA and pvl genes was evaluated, and staphylococcal cassette chromosome mec (SCCmec) typing was performed. RESULTS: A total of 177 S. aureus isolates were collected from various clinical specimens. Antibiotic susceptibility testing revealed high resistance rates to penicillin (98.9%), followed by erythromycin (61.6%). A total of 95 isolates (53.7%) were confirmed as MRSA. Among the initially screened vancomycin-intermediate S. aureus (VISA) isolates, one isolate with a minimum inhibitory concentration (MIC) of 6µg/mL harboured the vanA gene. Eleven MRSA isolates (11.6%) were also VRSA. A majority (23/95; 24.2%) of MRSA were classified as SCCmec type III. Only 6 MRSA isolates (6.3%) harboured the pvl gene. CONCLUSIONS: This study highlights the presence of MRSA along with VISA and VRSA in our setting. To our knowledge, this is the first report showing that a strain can be defined as VISA phenotypically and as VRSA by molecular analysis. Such a finding raises major concerns with regard to control measures and reliable laboratory tests for screening of resistant strains.

Association of altered gut microbiota composition with chronic urticaria.

BACKGROUND: An altered gut microbiota composition has recently been linked to some types of allergies. OBJECTIVE: To compare the relative amounts of Akkermansia muciniphila, Clostridium leptum, Faecalibacterium prausnitzii, and Enterobacteriaceae as members of gut microbiota among patients with chronic urticaria (CU) and healthy controls. METHODS: A total of 20 patients with CU and 20 healthy individuals matched by age and sex participated in the study. Fresh fecal samples were collected, and DNA extracted from stool samples was analyzed by real-time polymerase chain reaction for the qualitative and quantitative assays of the so-called bacteria. RESULTS: The frequencies of A muciniphila, C leptum, and F prausnitzii in healthy controls' stool samples were significantly more than those of patients with CU (P < .001, P < .01, and P < .05, respectively), whereas the Enterobacteriaceae family was detected in all patients and healthy controls' stool samples. The relative amounts of A muciniphila in healthy control positive samples were significantly higher than those of samples from patients with CU (P < .001). Furthermore, there was a corresponding increase of relative amounts of C leptum and F prausnitzii in healthy control positive samples compared with those of patients with CU (P = .09 and P = .08, respectively). The mean of the relative amounts of Enterobacteriaceae family in the stool samples from patients with CU was more than that of healthy controls; however, the difference was nearly significant (P = .12). CONCLUSION: The results reveal a change of frequency and relative amounts of A muciniphila, C leptum, and F prausnitzii in patients with CU compared with healthy controls. This is the first study, to our knowledge, to show the change of microbiota composition in patients with CU.

Polymorphisms in Beta-2 Adrenergic Receptor Gene and Association with Tuberculosis.

PURPOSE: Genetic susceptibility for tuberculosis in human has been previously demonstrated. Polymorphisms in genes involved in immune responses may alter the susceptibility of individuals to tuberculosis. Polymorphisms of beta-2 adrenergic receptor (ADRB2) gene can be possibly an important risk factor in tuberculosis. In this study, the association between rs1042713 (Arg16Gly +46A>G) and rs1042714 (Gln27Glu +79C>G) polymorphisms in ADRB2 gene and tuberculosis was evaluated. METHODS: Genotype distributions of the rs1042713 (Arg16Gly +46A>G) and rs1042714 (Gln27Glu +79C>G) polymorphisms in ADRB2 gene in 106 patients with pulmonary tuberculosis and 88 healthy subjects were studied by PCR-RFLP method in an Iranian population. RESULTS: The frequency of rs1042713*G and rs1042714*G alleles in ADRB2 gene in tuberculosis patients was significantly different from healthy controls [odds ratio (OR) 0.176, 95% confidence interval (CI) 0.065-0.48, P value <0.001 and OR 0.45, 95% CI 0.247-0.825, P value = 0.009, respectively]. There were no significant differences in haplotype analysis between the patients and control subjects. CONCLUSION: The association was reported between rs1042713 and rs1042714 polymorphisms in ADRB2 gene and tuberculosis for the first time. rs1042713*G and rs1042714*G polymorphisms in ADRB2 gene makes people more susceptible to develop the disease.

Mutation analysis of the phenylalanine hydroxylase gene in Azerbaijani population, a report from West Azerbaijan province of Iran.

OBJECTIVES: Phenylketonuria (PKU) is a genetic inborn error of phenylalanine (Phe) metabolism resulting from insufficiency in the hepatic enzyme, phenylalanine hydroxylase (PAH), which leads to elevated levels of Phe in the blood. The present study was carried out for mutation analysis of the PAH gene in West Azerbaijan province of Iran. MATERIALS AND METHODS: A total of 218 alleles from 40 PKU families were studied using restriction fragment length polymorphism-polymerase chain reaction (RFLP-PCR) method. RESULTS: The frequencies of IVS10-11, S67P, R261Q, R252W, IVS11nt-1 g>c, R408Q, and Q232Q mutations were 28(35), 17(21.25), 15(18.75), 3(3.75), 3(3.75), 2(2.5), and 1(1.25), in cases group, and 51(23.4), 31(14.2), 27(12.4), 6(2.75), 6(2.75), 4(1.83), and 2(0.92) in total group, respectively. The mutations of R243Q, 364delG, L333F, 261X, I65T, and R408W were not detected in our samples. CONCLUSION: It can be concluded that the IVS10-11 mutation has the highest frequency in the tested population. To our knowledge, this report is the first in its own kind and provides better understanding of the genetic heterogeneity, the origin and distributions of PAH mutations in West Azerbaijan province of Iran.

Frequency of the VNTR-Polymorphisms at the PAH Gene in the Iranian Azeri Turkish Patients with Phenylketonuria.

INTRODUCTION: This study was carried out to determine the frequency of the VNTR-polymorphisms at the PAH gene in the Iranian Azeri Turkish patients with phenylketonuria (PKU) and normal controls. MATERIAL AND METHODS: The VNTR-polymorphisms were determined by PCR in 43 PKU patients as well as 43 controls. OUTCOMES: The frequencies of VNTR-alleles were 13(15.1%), 3(3.49%), 64(74.4%), 5(5.81%), and 1(1.16%) in the patients and 43(50%), 0(0%), 42(48.8%), 0(0%), and 1(1.16%) in the controls regarding 3, 7, 8, 9, and 11 repeat copies, respectively. The VNTR alleles with 12 and 13 repeats were not found in our samples. The frequencies of VNTR-genotypes were 25(58.1%), 1(2.33%), 1(2.33%), 10(23.3%), 2(4.65%), 2(4.65%), 1(2.33%), 1(2.33%), and 0(0%) in the patients and 13(30.2%), 13(30.2%), 0(0%), 16(37.2%), 0(0%), 0(0%), 0(0%), 0(0%) and 1(2.33%) in the controls regarding VNTR8/VNTR8, VNTR3/VNTR3, VNTR3/VNTR9, VNTR8/VNTR3, VNTR8/VNTR9, VNTR7/VNTR9, VNTR7/ VNTR8, VNTR8/VNTR11, and VNTR3/VNTR11 genotypes, respectively. The comparisons of VNTRpolymorphisms imply that there are statistically significant differences between the patients and controls regarding VNTR3, VNTR8, and VNTR9 alleles as well as VNTR8/VNTR8 and VNTR3/VNTR3 genotypes (all P-Value <0.05). The frequency of "risk-associated genotype of VNTR8/VNTR8" was significantly higher in the cases. CONCLUSION: It is concluded that this position is heterozygous and there were statistically significant differences between patients and controls concerning the VNTR8/VNTR8 genotype. We found higher frequencies of disease-associated genotype in our samples than controls. This report is the first in its own type in the west Azerbaijani population. Further studies require assessing how this genotype predicts adverse outcomes in tested population.

Association Between PAH Mutations and VNTR Alleles in the West Azerbaijani PKU Patients.

INTRODUCTION: We report the frequency of IVS10nt546, R261Q, S67P, R252W, and R408W mutations linked to PAH VNTR alleles in the west Azerbaijani PKU patients. MATERIAL AND METHODS: VNTR alleles and IVS10nt546, R261Q, S67P, R252W, R408W mutations were studied in a total of 20 PKU patients by PCR and RFLP-PCR. OUTCOMES: Our analysis showed that 95% of cases were homozygote for an allele containing eight-repeat VNTR (VNTR8); while 5% were homozygote for an allele containing three-repeat VNTR (VNTR3). The IVS10nt546, R252W, and R261Q mutations were associated with VNTR8 allele, and also, R252W and S67P mutations were associated with VNTR3 allele. VNTR8 was common among mutant alleles as were IVS10nt546-VNTR8 (50%), R252W-VNTR8 (2.5%), and R261Q-VNTR8 (22.5%). The association of VNTR3 was found as R252W-VNTR3 (2.5%) and S67P-VNTR3 (2.5%) among studied cases. The frequency of IVS10nt546-VNTR8/IVS10nt546-VNTR8, IVS10nt546-VNTR8/ND-VNTR8, IVS10nt546-VNTR8/R252W-VNTR8, R261Q-VNTR8/R261Q-VNTR8, R261Q-VNTR8/ND-VNTR8, and S67P-VNTR3/ R252W-VNTR3 were 30%, 35%, 5%, 20%, 5%, and 5%, respectively. R408W mutation was not found in this study. CONCLUSIONS: The present report is the first in its own kind in the west Azerbaijani population (Iran) and implies that the most common PKU mutation in this population, IVS10nt546, is exclusively associated with VNTR8 allele, and IVS10nt546-VNTR8 alleles testing should be considered for routine carrier screening and prenatal diagnostic setting.

Lack of Association of Vitamin D Receptor FokI (rs10735810) (C/T) and BsmI (rs1544410) (A/G) Genetic Variations with Polycystic Ovary Syndrome Risk: a Case-control Study from Iranian Azeri Turkish Women.

INTRODUCTION: In this study we evaluate the involvement of Vitamin D Receptor (VDR) FokI (rs10735810) Exon 2 (C/T) and BsmI (rs1544410) Intron 8 (A/G) gene variations in genetic susceptibility to polycystic ovary syndrome (PCOS) in Iranian Azeri Turkish women. MATERIALS AND METHODS: The RFLP-PCR method was performed on peripheral blood lymphocyte for a total of 46 females with PCOS and 46 controls. OUTCOMES: VDR FokI (rs10735810) CC,CT,TT,C and T genotypic/allelic frequencies were 22(47.83), 20(43.48), 4(8.696), 64(69.57) and 28(30.43) in cases and 29(63.04), 15(32.61), 2(4.348), 73(79.35) and 19(20.65) in controls, respectively. The frequencies of VDR FokI C and T alleles were 0.7 and 0.3 in cases, and 0.79 and 0.21 in controls, respectively. VDR BsmI (rs1544410) Intron 8 (A/G) AA,AG,GG,A and G genotypic/allelic frequencies were 15(32.6), 27(58.7), 4(8.7), 57(62), and 35(38) in cases and 20(43.5), 24(52.2), 2(4.35), 64(69.6), and 28(30.4) in controls, respectively. The frequencies of VDR BsmI (rs1544410) Intron 8 A and G alleles were 0.7 and 0.3 in cases, and 0.62 and 0.38 in controls, respectively. Statistical analysis showed that the differences in genotypic/allelic frequencies between the cases and controls were not statistically significant regarding of VDR FokI(rs10735810) Exon 2 (C/T) and VDR BsmI (rs1544410) Intron 8 (A>G) (p >0.05). CONCLUSIONS: It can be concluded that FokI (rs10735810) Exon 2 (C/T) and VDR BsmI (rs1544410) Intron 8 (A>G) were not associated with PCOS susceptibility in studied group. Present investigation is the first study in its own kind in Iranian Azeri Turkish women.

Synergistic effects of glutamine and ciprofloxacin in reduction of Pseudomonas aeruginosa-induced septic shock severity.

BACKGROUND: Systemic inflammatory response induced by over expressing inflammatory mediators is the main pathogenic mechanism of septic shock. Glutamine (Gln) has been demonstrated to inhibit pro-inflammatory cytokine release through enhanced heat shock protein (HSP) expression. OBJECTIVE: To assess the effect of co-administration of Gln and antibiotic ciprofloxacin in reduction of septic shock severity caused by Pseudomonas aeruginosa in mice. METHODS: Six- to eight-week old male BALB/c mice were used. At first, P. aeruginosa susceptibility to ciprofloxacin was determined. Then, 75% lethal dose (LD 75) of P. aeruginosa in a 10-day period was assessed. For determining survival rate, fifty mice were divided into 5 groups which included control (+), control (-), Gln, ciprofloxacin, and "glutamine+ciprofloxacin" group. All mice, except for control (-), were given an LD75 dose of P. aeruginosa and after 30 min each group received its special treatment: control (-) and control (+) groups received only 500λ phosphate buffer saline (PBS). Gln group received 500λ Ala-Gln, Cip group received 500λ ciprofloxacin. The Cip+Gln group received 500λ Gln and ciprofloxacin. Finally serum TNF-α, IL-10 and HSP-70 concentrations were measured and the severity of liver necrosis was examined. RESULTS: Glutamine in combination with ciprofloxacin significantly increased survival rate and serum HSP-70 and IL-10 concentration and significantly decreased serum TNF-α concentration and the liver necrosis severity in comparison to control (+) group. CONCLUSION: Gln has synergistic effects with ciprofloxacin in reduction of P. aeruginosa-induced septic shock.

A novel adjuvant, a mixture of alum and the general opioid antagonist naloxone, elicits both humoral and cellular immune responses for heat-killed Salmonella typhimurium vaccine.

In the current study, we tested the efficacy of the mixture of naloxone, an opioid receptor antagonist, and alum, as a new adjuvant, in the induction of humoral and cellular immunity in response to heat-killed Salmonella typhimurium (HKST) as a model vaccine. BALB/c mice were divided into five groups. Mice in the experimental groups received either the HKST vaccine alone or in combination with the adjuvant alum, naloxone or the alum-naloxone mixture. Mice in the negative control group received phosphate-buffered saline. All mice were immunized two times on days 0 and 14. Two weeks after the last immunization, immune responses to S. typhimurium were assessed. Our results indicated that the administration of the alum-naloxone mixture as an adjuvant increased the ability of the HKST vaccine to enhance lymphocyte proliferation, shifted the immune response towards a T-helper 1 (Th1) pattern and increased S. typhimurium-specific immunoglobulin G (IgG), IgG2a, IgG1 and the ratio of IgG2a to IgG1. This resulted in improved protective immunity against S. typhimurium. In conclusion, the administration of the alum-naloxone mixture as an adjuvant, in combination with the HKST vaccine, can enhance both humoral and cellular immunity and shift the immune responses to a Th1 pattern.

Naloxone and alum synergistically augment adjuvant activities of each other in a mouse vaccine model of Salmonella typhimurium infection.

Alum is the most commonly used adjuvant for human vaccination but is a poor inducer of cell mediated immunity and T helper 1 (Th1) responses. We have previously shown that naloxone (NLX), which is a general opioid antagonist, acts as an effective adjuvant in enhancing vaccine-induced cellular immunity and Th1 immune responses. Here, we tested the efficacy of an alum-NLX mixture, as a new adjuvant, in the induction of humoral and cellular immunity in response to endotoxin-removed lysate (ERL) of Salmonella typhimurium (S. typhimurium) as a model vaccine. BALB/c mice were divided into five vaccination groups. Mice in the experimental groups received either the ERL vaccine alone or in combination with the adjuvant alum, NLX or the alum-NLX mixture. Mice in the negative control group received phosphate-buffered saline. All mice were immunized on days 0 and 7. Two weeks after the last immunization, immune responses to S. typhimurium were assessed. Our results indicate that including the alum-NLX mixture as an adjuvant during vaccination increased the ability of the ERL vaccine to enhance lymphocyte proliferation, shifted the immune response toward a Th1 profile and increased S. typhimurium-specific IgG, IgG2a and the ratio of IgG2a to IgG1. This resulted in improved protective immunity against S. typhimurium. In conclusion, administering an alum-NLX mixture adjuvant in combination with the ERL vaccine enhances both humoral and cellular immunity, and shifts the immune response to a Th1 pattern.

Detection of metallo-ß-lactamase-encoding genes among clinical isolates of Pseudomonas aeruginosa in northwest of Iran.

The prevalence of metallo-ß-lactamase (MBL) production among 104 clinical isolates of Pseudomonas aeruginosa from northwest of Iran was investigated by phenotypic and polymerase chain reaction (PCR) methods. Thirty-nine (37.50%) of isolates were MBL positive by double-disk synergy test. Results of PCR revealed that 18 (17.31%) and 6 (5.77%) imipenem nonsusceptible isolates of P. aeruginosa carried bla(VIM) and bla(IMP) genes respectively, while 92.4% (62/67) of isolates contained class 1 integron gene. This is the first report of MBL-producing P. aeruginosa from northwest of Iran.

Evaluation of the adjuvant activity of naloxone, an opioid receptor antagonist, in combination with heat-killed Listeria monocytogenes vaccine.

We have previously demonstrated the adjuvant activity of naloxone (NLX), a general opioid antagonist, using a DNA vaccine for herpes simplex virus type 1. Here, the adjuvant activity of NLX has been evaluated using a heat-killed Listeria monocytogenes (HKLM) vaccine as a model for general immunization against intracellular bacteria. BALB/c mice were divided into three groups: the Vac group received the HKLM vaccine alone; the NLX-Vac group received the HKLM vaccine in combination with the adjuvant NLX; and the control group received phosphate buffered saline (PBS). Our results indicate that the administration of NLX as an adjuvant enhances the ability of the HKLM vaccine to increase lymphocyte proliferation, delayed type hypersensitivity, and skewing of the immune response toward a T-helper 1 (Th1) pattern. Additionally, combination of NLX with the HKLM vaccine improves protective immunity against L. monocytogenes. In conclusion, administration of NLX as an adjuvant for the HKLM vaccine can enhance cell-mediated immunity and shift the immune response to Th1.

Post heat shock tolerance: a neuroimmunological anti-inflammatory phenomenon.

We previously showed that the progression of burn-induced injury was inhibited by exposing the peripheral area of injured skin to sublethal hyperthermia following the burn. We called this phenomenon post-heat shock tolerance. Here we suggest a mechanism for this phenomenon. Exposure of the peripheral primary hyperalgesic/allodynic area of burned skin to local hyperthermia (45 degrees C, 30 seconds), which is a non-painful stimulus for normal skin, results in a painful sensation transmitted by nociceptors. This hyperthermia is too mild to induce any tissue injury, but it does result in pain due to burn-induced hyperalgesia/allodynia. This mild painful stimulus can result in the induction of descending anti-nociceptive mechanisms, especially in the adjacent burned area. Some of these inhibitory mechanisms, such as alterations of sympathetic outflow and the production of endogenous opioids, can modify peripheral tissue inflammation. This decrease in burn-induced inflammation can diminish the progression of burn injury.

Protective effects of Helicobacter pylori against gastroesophageal reflux disease may be due to a neuroimmunological anti-inflammatory mechanism.

There is some evidence that Helicobacter pylori infection has a protective effect against gastroesophageal reflux disease (GORD) and its complications such as Barrett's oesophagus and oesophageal adenocarcinoma. In this paper, we propose that a neuroimmunological mechanism is responsible for the protective effect of H. pylori on GORD. H. pylori infection of the gastric mucosa induces a T helper1-like immune response and production of pro-inflammatory cytokines. These cytokines can inhibit local sympathetic tone, whereas they increase systemic sympathetic tone. Increased sympathetic tone can induce an anti-inflammatory milieu, which in turn can inhibit inflammation in the oesophagus and lower oesophageal sphincter (LOS). Furthermore, H. pylori infection may stimulate the cholinergic anti-inflammatory pathway. It has been suggested that reflux-induced oesophageal inflammation plays an important role in the pathogenesis of reflux oesophagitis. Reduction of oesophageal inflammation by increased systemic sympathetic tone and vagal activity may lead to a decrease in reflux-induced oesophageal injury and LOS dysfunction in GORD.

The effect of post-burn local hyperthermia on the reducing burn injury: the possible role of opioids.

PURPOSE: This paper studied the effect of post-burn local hyperthermia on burn induced injury. METHODS: A second-degree burn injury was induced on the right and left flanks of Balb/c mice. Thirty-two burn wounds were divided into four groups. Opioid receptor blocking was done for groups 3 and 4 by intra-peritoneal administration of Naloxone (NLX) 30 min before the thermal injury. Local hyperthermia (45 degrees C, 30 s) was applied only for the burn wounds of groups 2 and 4. Twenty-four hours after burn injury, the burned wounds were assessed for the level of iNOS (by immunohistochemistry) and the number of hair follicles (as an indicator of tissue injury). RESULTS: The wounds that received hyperthermia (group 2) had significantly more hair follicles (p < 0.001) compared to the control wounds (group 1). There was no significant difference between the number of hair follicles and acute inflammation of group 1 and group 3 (NLX + burn). Group 4 (NLX + burn + hyperthermia) had significantly fewer hair follicles compared to group 1 (p < 0.001), group 2 (p < 0.001) and group 3 (p < 0.001). The level of iNOS in groups 1, 3 and 4 was not significantly different but significantly more than group 2 (p < 0.001, p < 0.001 and p < 0.001, respectively). CONCLUSIONS: The results showed that local hyperthermia after second degree burn decreased the tissue injury and iNOS expression. It is also concluded that endogenous opioid response may have a key role in the above mentioned effects of post-burn local hyperthermia.

Sympathetic nervous system plays an important role in the relationship between immune mediated diseases.

UNLABELLED: T helper (Th) lymphocytes have been classified into distinct subsets, Th1 and Th2 on the basis of the cytokines they produce. According to the cross-regulatory properties of Th1 and Th2 cells, one would assume that to be affected by a Th1 type disease increases susceptibility to a Th1 type disease and inhibits a Th2 type disease and vice versa about being affected by a Th2 type disease. However, the pattern of related diseases does not necessarily follow the conventional pattern of inhibitory effects of Th1 and Th2 immune responses on each other. For example, Mycobacteria including BCG, that induce Th1 immune responses; can modulate some Th1 type autoimmune diseases including MS, experimental autoimmune encephalomyelitis (EAE; an animal model for Multiple Sclerosis) and insulin-dependent diabetes mellitus (IDDM) thereby leading to an alleviation of their symptoms. Also BCG precipitates a syndrome similar to systemic lupus erythematosus (SLE), a Th2 type disease; in NOD mice. The coexistence of the major Th2-mediated atopic diseases such as asthma, eczema and allergic rhinitis with the Th1-mediated autoimmune conditions including; coeliac disease (CD), IDDM, rheumatoid arthritis (RA) and psoriasis is another example that is in apparent disagreement with counter-regulatory effects of Th1/Th2 phenotypes. HYPOTHESIS: SNS can be stimulated by pro-inflammatory cytokines, production of which is induced by mycobacteria including BCG. Although these cytokines can inhibit SNS activity in the site of inflammation and secondary lymphoid organs, they increase sympathetic tone in other places. Increased sympathetic tone can induce an anti-inflammatory and Th2 type milieu. This milieu can inhibit MS and IDDM and provide a susceptible environment for starting of SLE. Atopic diseases are Th2 type immune mediated diseases; therefore, they increase the production of Th2 type cytokine and decrease production of pro-inflammatory cytokines in the site of allergic reaction and also in secondary lymphoid organs. Therefore, atopic diseases decrease sympathetic tone in all tissues except in the sites of allergic reaction and secondary lymphoid organs. Decreased sympathetic tone results in a pro-inflammatory milieu and in such an environment, Th1 type autoimmune diseases can affect tissues.

Any beneficial effects of mycobacteria on multiple sclerosis and experimental autoimmune encephalitis may include stimulation of the sympathetic nervous system.

UNLABELLED: The inhibitory effects of mycobacterial infection and mycobacterium components on multiple sclerosis (MS) and experimental autoimmune encephalitis (EAE; an animal model for MS) have been known for years. However, this effect seems like a paradox that both mycobacterial infection and MS induce type I immune responses. Some mechanisms have been proposed or even proven for this effect in different studies, but among them there is no hint of a possible role for the nervous system (NS). Regarding the close relations between sympathetic nervous system (SNS) and MS disease course, it can be hypothesized that SNS may have a role in the effects of mycobacterium on MS. HYPOTHESIS: SNS can be stimulated by pro-inflammatory cytokines such as TNF-alpha and IL1-beta, production of which are induced by mycobacterial infection or mycobacterium components. Although these cytokines can inhibit SNS in the site of inflammation caused by mycobacterium, they increase sympathetic tone in other places. The beneficial role of SNS in inhibiting or attenuating the course of MS and EAE has been suggested. Inhibitory effects of stimulated SNS on MS may occur via different ways such as inhibiting the production of pro-inflammatory cytokines and inducing the synthesis of anti-inflammatory cytokines, in other words, shifting the immune responses from type 1 toward type 2, as well as, induction of suppressor/regulator T lymphocytes, induction of heat shock proteins in brain and increasing the expression of Fas and Fas-ligand. Therefore, it seems that stimulation of SNS by mycobacterial infection or mycobacterium components is a key step in the mechanism of beneficial effects of mycobacterium on MS.