TLR-1, TLR-2, and TLR-6 MYD88–dependent signaling pathway: A potential factor in the interaction of high-DNA fragmentation human sperm with fallopian tube epithelial cells / 대한생식의학회지

Objective@#The DNA integrity of spermatozoa that attach to fallopian tube (FT) cells is higher than spermatozoa that do not attach. FT epithelial cells can distinguish normal and abnormal sperm chromatin. This study investigated the effects of sperm with a high-DNA fragmentation index (DFI) from men with unexplained repeated implantation failure (RIF) on the Toll-like receptor (TLR) signaling pathway in human FT cells in vitro. @*Methods@#Ten men with a RIF history and high-DFI and 10 healthy donors with low-DFI comprised the high-DFI (>30%) and control (<30%) groups, respectively. After fresh semen preparation, sperm were co-cultured with a human FT epithelial cell line (OE-E6/E7) for 24 hours. RNA was extracted from the cell line and the human innate and adaptive immune responses were tested using an RT2 profiler polymerase chain reaction (PCR) array. @*Results@#The PCR array data showed significantly higher TLR-1, TLR-2, TLR-3, TLR-6, interleukin 1α (IL-1α), IL-1β, IL-6, IL-12, interferon α (IFN-α), IFN-β, tumor necrosis factor α (TNF-α), CXCL8, GM-CSF, G-CSF, CD14, ELK1, IRAK1, IRAK2, IRAK4, IRF1, IRF3, LY96, MAP2K3, MAP2K4, MAP3K7, MAP4K4, MAPK8, MAPK8IP3, MYD88, NFKB1, NFKB2, REL, TIRAP, and TRAF6 expression in the high-DFI group than in the control group. These factors are all involved in the TLR-MyD88 signaling pathway. @*Conclusion@#The MyD88-dependent pathway through TLR-1, TLR-2, and TLR-6 activation may be one of the main inflammatory pathways activated by high-DFI sperm from men with RIF. Following activation of this pathway, epithelial cells produce inflammatory cytokines, resulting in neutrophil infiltration, activation, phagocytosis, neutrophil extracellular trap formation, and apoptosis.

Evaluation of polyglycolic acid as an animal-free biomaterial for three-dimensional culture of human endometrial cells / 대한생식의학회지

Objective@#Animal-free scaffolds have emerged as a potential foundation for consistent, chemically defined, and low-cost materials. Because of its good potential for high biocompatibility with reproductive tissues and well-characterized scaffold design, we investigated whether polyglycolic acid (PGA) could be used as an animal-free scaffold instead of natural fibrin-agarose, which has been used successfully for three-dimensional human endometrial cell culture. @*Methods@#Isolated primary endometrial cells was cultured on fibrin-agarose and PGA polymers and evaluated various design parameters, such as scaffold porosity and mean fiber diameter. Cytotoxicity, scanning electron microscopy (SEM), and immunostaining experiments were conducted to examine cell activity on fabricated scaffolds. @*Results@#The MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay and SEM results showed that endometrial cells grew and proliferated on both scaffolds. Immunostaining showed cytokeratin and vimentin expression in seeded cells after 7 days of culture. On both scaffolds, an epithelial arrangement of cultured cells was found on the top layer and stromal arrangement matrix on the bottom layer of the scaffolds. Therefore, fibrin-agarose and PGA scaffolds successfully mimicked the human endometrium in a way suitable for in vitro analysis. @*Conclusion@#Both fibrin-agarose and PGA scaffolds could be used to simulate endometrial structures. However, because of environmental and ethical concerns and the low cost of synthetic polymers, we recommend using PGA as a synthetic polymer for scaffolding in research instead of natural biomaterials.

Diversity of Phlebotomine sand flies (Diptera: Psychodidae) in mountainous and plain areas of an endemic focus of anthroponotic cutaneous leishmaniasis in Iran

Objective: To determine the diversity of sand flies in different biotopes of mountainous and plain areas of Bam County as the most infected focus of anthroponotic cutaneous leishmaniasis in southeast Iran, and synanthropic index of Phlebotomus sergenti Parrot, and Phlebotomus papatasi Scopoli as the main vectors of cutaneous leishmaniasis in Iran. Methods: Sand flies were captured once a month using sticky traps in domestic, peri-domestic, agricultural, and sylvatic biotopes in the plain and mountainous areas. Alpha diversity indices, including richness, evenness, Shannon-Wiener; beta diversity indices (Jaccard's and Sorensen's similarity indices) and synanthropic index were calculated. Results: A total of 2 664 specimens of 9 sand fly species were collected from mountainous (47%) and plain (53%) areas. Species richness, species evenness, and Shannon-Wiener indices were obtained as 9, 0.637, and 1.399, respectively in the mountainous area. Phlebotomus sergenti and Phlebotomus papatasi were constant species with the synanthropic index of-18.463 and-29.412, respectively. In addition, species richness, species evenness, and Shannon-Wiener indices were 4, 0.690, and 0.956, respectively in the plain area. Phlebotomus sergenti and Phlebotomus papatasi were dominant species with the synanthropic index of +9.695 and +36.207, respectively. Similarity indices were low among different biotopes of plain and mountainous areas. Conclusions: A basic knowledge about the diversity of sand flies in various biotopes is essential to design sound control programs. Biodiversity and synanthropic indices of sand flies are different in plain and mountainous areas due to the difference in biotic and abiotic factors between the two areas.

Effect of the estrus cycle stage on the establishment of murine endometriosis lesions

Background: Establishment of a standardized animal endometriosis model is necessary for evaluation of new drug effects and for explaining different ethological aspects of this disease. For this purpose, we need a model which has more similarity to human endometriosis

Objective: Our objective was to establish an autologous endometriosis mouse model based on endogenous estrogen level and analyze the influence of estrus cycle on the maintenance of endometriotic lesions

Materials and Methods: In this experimental study, endometriotic lesions were induced in 52 female NMRI mice by suturing uterine tissue samples to the abdominal wall. The transplantation was either performed at proestrus/estrus or at metestrus/diestrus cycles. Urine-soaked beddings from males and also male vasectomized mice were transferred to the cages to synchronize and maintenance of estrus cycle in female mice. The mice were sacrificed after different transplantation periods [2, 4, 6 or 8 wk]. The lesions size, macroscopic growth, model success rate, histological and immune-histochemical analyses were assessed at the end

Results: From a total of 200 tissue samples sutured into the peritoneal cavity, 83 endometriotic lesions were confirmed by histopathology [41.5%]. Model success rate for proestrus/estrus mice was 60.7% vs. 79.2% for metestrus/diestrus mice. The endometriotic lesions had similar growth in both groups. Number of caspase-3, Ki67-positive cells and CD31-positive micro vessels were also similar in endometriotic lesions of two groups

Conclusion: If we maintain the endogenous estrogen levels in mice, we can induce endometriosis mouse model in both proestrus/estrus and metestrus/diestrus cycle without any significant difference

Altered expression of 3´paralogus HOX A-D clusters in endometriosis disease: a case-control study

Background: Endometriosis is a prevalent gynecological disease, with limited known etiology and more researches are required to identify its etiology. In this manner, there is no evidence for expression and function of 3and acute;HOX genes in 4 clusters in the limb and pelvic organs such as the uterus and its disorders [Genes in the HOXA-D clusters are subdivided into 13 paralogous groups]

Objective: This study designed to investigate the expression profile of 5 paralogous [1-5] in four clusters of HOX genes [A, B, C, and D] in ectopic and eutopic tissues of women with endometriosis compared to the normal endometrium

Materials and Methods: Samples were obtained from thirty patients [15 with and 15 without endometriosis] of reproductive age with normal menstrual cycles. The same patient provided both eutopic and ectopic tissues and control women were laparoscopically checked for the absence of endometriosis. The expression profile of these HOX genes was investigated by quantitative real-time polymerase chain reaction technique

Results: We observed significant up-regulation of some members of HOXC and D clusters [HOXD1, HOXD3, HOXC4 and HOXC5] in ectopic and eutopic tissues vs. control. Also, our data showed significant down-regulation of all of HOXA and HOXB paralogous except HOXA1 in ectopic tissues versus control

Conclusion: Our data showed specific cluster dependent modulation of the HOX genes expression in endometriosis [over-expression of some HOX genes in cluster C and D and down-regulation of HOX genes in cluster A and B] in ectopic and eutopic tissues compare to control group. Therefore, it is possible that change of expression level of these genes in endometrium plays a role in the pathogenesis of endometriosis

Sex hormones alter the response of Toll-like receptor 3 to its specific ligand in fallopian tube epithelial cells / 대한생식의학회지

OBJECTIVE: The fallopian tubes play a critical role in the early events of fertilization. The rapid innate immune defense is an important part of the fallopian tubes. Toll-like receptor 3 (TLR3), as a part of the innate immune system, plays an important role in detecting viral infections. In this basic and experimental study, the effect of sex hormones on the function of TLR3 in the OE-E6/E7 cell line was investigated. METHODS: The functionality of TLR3 in this cell line was evaluated by cytokine measurements (interleukin [IL]-6 and IL-1b) and the effects of sex hormones on TLR3 were tested by an enzyme-linked immunosorbent assay kit. Additionally, TLR3 small interfering RNA (siRNA) and a TLR3 function-blocking antibody were used to confirm our findings. RESULTS: The production of IL-6 significantly increased in the presence of polyinosinic-polycytidylic acid (poly(I:C)) as the TLR3 ligand. Using a TLR3-siRNA-ransfected OE-E6/E7 cell line and function-blocking antibody confirmed that cytokine production was due to TLR3. In addition, 17-β estradiol and progesterone suppressed the production of IL-6 in the presence and absence of poly(I:C). CONCLUSION: These results imply that sex hormones exerted a suppressive effect on the function of TLR3 in the fallopian tube cell line when different concentrations of sex hormones were present. The current results also suggest that estrogen receptor beta and nuclear progesterone receptor B are likely to mediate the hormonal regulation of TLR3, as these two receptors are the main estrogen and progesterone receptors in OE-E6/E7 cell line.

Variable localization of Toll-like receptors in human fallopian tube epithelial cells / 대한생식의학회지

OBJECTIVE: To determine the localization, expression, and function of Toll-like receptors (TLRs) in fallopian tube epithelial cells. METHODS: The localization of TLRs in fallopian tube epithelial cells was investigated by immunostaining. Surprisingly, the intensity of staining was not equal in the secretory and ciliated cells. After primary cell culture of fallopian tube epithelial cells, ring cloning was used to isolate colonies of ciliated epithelial cells, distinct from non-ciliated epithelial cells. The expression of TLRs 1–10 was examined by quantitative real-time polymerase chain reaction, and protein localization was confirmed by immunostaining. The function of the TLRs was determined by interleukin (IL)-6 and IL-8 production in response to TLR2, TLR3, TLR5, TLR7, and TLR9 ligands. RESULTS: Fallopian tube epithelial cells expressed TLRs 1–10 in a cell-type-specific manner. Exposing fallopian tube epithelial cells to TLR2, TLR3, TLR5, TLR7, and TLR9 agonists induced the secretion of proinflammatory cytokines such as IL-6 and IL-8. CONCLUSION: Our findings suggest that TLR expression in the fallopian tubes is cell-type-specific. According to our results, ciliated cells may play more effective role than non-ciliated cells in the innate immune defense of the fallopian tubes, and in interactions with gametes and embryos.

Isolation, culture and characterization of human sertoli cells by flow cytometry: development of procedure

Background: the sertoli cells in the testis create unique and safe environment to protect seminiferous tubules from auto antigens and invading pathogens. These cells produce the survival factor of the blood-testis barrier and produce special materials such as androgen binding proteins and contribute to the coordinated action of spermatogenesis. Given that the sertoli cells play an essential role in spermatogenesis and the lack of these cells leads to the disruption of spermatogenesis, it is necessary to investigate the behavior and performance of these cells. To achieve this goal, the cells must first be extracted. The aim of this study was to develop a procedure to isolate, culture, and characterize human sertoli cells

Methods: in order to isolate the sertoli cells of azoospermia patients who underwent [testicular sperm extraction] TESE surgery, washing up and multi_stage enzyme digestion of single cells, culture on petri dishes impregnated with datura stramonium lectin agglutinin [DSA] were done and then the cells were passaged for several times and isolated. For more purification, flow cytometry method with FSH receptor antibody was used. Immunocytochemistry assays and Elisa test for identification of these cells were employed

Results: the purification method resulted in more than 97% purity. The nature of sertoli cells was confirmed by morphology evaluation, detecting anti-mullerian hormone in sertoli cell culture media and the presence of FSH receptor on sertoli cells

Conclusion: this study introduced and applied a method to isolate, culture, and purify human sertoli cells with high purity which made possible further researches on these cells

expression of TLR2 and TLR3 in sertoli cells of azoospermic patients

Objective: Toll-like receptors [TLRs] on Sertoli cells are thought to have essential roles in sperm protection. This study was conducted to investigate the expression of TLR2 and TLR3 in Sertoli cells of men with azoospermia

Materials and Methods: In this experimental study, testicular biopsies were taken from ten azoospermic men. Following enzymatic dissociation, the samples were moved to lectin coated petri dishes. After a few passages, all cells were cultivated and Seroli cells were sorted by flow cytometry. To confirm Sertoli cell purification, alkaline phosphatase activity [ALP] and immunohistochemistry assays were employed. The expression of TLR2 and TLR3 at the transcript and protein levels was examined with real-time quantitative reverse transcription-polymerase chain reaction [RT-QPCR] and western blot, respectively

Results: Isolation, purification and cultivation of human Sertoli cells were performed successfully. Efficacy of purification of Sertoli cells by fluorescence-activated cell sorting [FACS] sorter was 97%. The type of cultured cells was confirmed by vimentin and follicle-stimulating hormone [FSH] receptor markers. Furthermore, the existence of anti- Mullerian hormone in culture was confirmed. RT-PCR showed that both genes were expressed in Sertoli cells. Consistently, proteins of both were also expressed in Sertoli cells. Moreover, QPCR showed that the relative expression of TLR3 transcripts was significantly higher than TLR2 in Sertoli cells. Although both genes are expressed in fibroblast cells, their level of expression was significantly lower than in Sertoli cells

Conclusion: This study confirmed expression of TLR2 and TLR3 in human Sertoli cells. This may be an indicator of their roles in developing immunity against pathogens as well as allo- and auto-antigens or viral antigens in seminiferous tubules

comparative study on the results of agonist and antagonist protocols based on serum AMH levels in patients undergoing intracytoplasmic sperm injection

Background: Serum concentrations of antimullerian hormone [AMH] correlate with ovarian response during assisted reproduction treatment [ART] cycles

Objective: This retrospective study attempted to evaluate the selection of ovarian stimulation protocols based on serum AMH levels in patients and its impact on the results of ART

Materials and Methods: Based on AMH levels, the patients with tubal factor infertility were divided in three groups of normal, low and high AMH levels. Oocyte, good embryo number and pregnancy rate in each group were analyzed

Results: Using agonist and antagonist protocols, an increase in serum AMH led to higher number of oocytes and better quality embryos. At all low, normal and high AMH levels, the agonist protocol led to a more significant increase in the number of oocytes than the antagonist protocol [p<0.05]. The number of high quality embryos significantly increased by the agonist protocol than antagonist protocol in women with normal AMH levels of 1.3-2.6 ng/ml [p=0.00]. Moreover, the results for the number of high quality embryos at AMH >2.6 ng/ml was in favor of the antagonist protocol [p=0.00]. The results showed the lowest pregnancy rate at AMH <1.3 ng/ml. At AMH >2.6 ng/ml, there was a significant increase in pregnancy rate through the antagonist protocol [p=0.04]

Conclusion: Findings of this study suggested that the ART results are predictable, taking into account the AMH levels. The protocol specific to each patient can be used given the AMH level in each individual. This is because the results of each protocol depend on individual conditions

Effect of estradiol and progesterone on toll like receptor gene expression in a human fallopian tube epithelial cell line

Objective: Toll like receptors [TLRs] are one of the main components of the innate im-mune system. It has been reported that expression of these receptors are altered in the female reproductive tract [FRT] during menstrual cycle. Here we used a fallopian tube epithelial cell line [OE-E6/E7] to evaluate the effect of two sex hormones in modulating TLR expression

Materials and Methods: In this experimental study, initially TLR gene expression in OE-E6/E7 cells was evaluated and compared with that of fallopian tube tissue using quantitative real time-polymerase chain reaction [qRT-PCR] and immunostaining. Thereafter, OE-E6/E7 cells were cultured with different concentrations of estradiol and progesterone, and combination of both. qRT-PCR was performed to reveal any changes in expression of TLR genes as a result of hormonal treatment

Results: TLR1-10 genes were expressed in human fallopian tube tissue. TLR1-6 genes and their respective proteins were expressed in the OE-E6/E7 cell line. Although estradiol and progesterone separately had no significant effect on TLR expression, their combined treatment altered the expression of TLRs in this cell line. Also, the pattern of TLR expression in preovulation [P], mensturation [M] and window of implantation [W] were the same for all TLRs with no significant differences between P, M and W groups

Conclusion: These data show the significant involvement of the combination of estradiol and progesterone in modulation of TLR gene expression in this human fallopian tube cell line. Further experiments may reveal the regulatory mechanism and signalling pathway behind the effect of sex hormones in modulating TLRs in the human FRT

MRNA expression of VEGF and its receptors in fallopian tubes of women with ectopic pregnancies

Establishment of viable pregnancy requires embryo implantation and placentation. Ectopic pregnancy [EP] is a pregnancy complication which occurs when an embryo implants outside of the uterine cavity, most often in a fallopian tube. On the other hand, an important aspect of successful implantation is angiogenesis. Vascular endothelial growth factor [VEGF] is a potent angiogenic factor responsible for vascular development that acts through its receptors, VEGF receptor 1 [VEGFR1] and VEGFR2. This study aims to investigate mRNA expression of VEGF and its receptors in fallopian tubes of women who have EP compared with fallopian tubes of pseudo-pregnant women. We hypothesize that expression of VEGF and its receptors in human fallopian tubes may change during EP. This was a case-control study. The case group consisted of women who underwent salpingectomy because of EP. The control group consisted of women with normal fallopian tubes that underwent hysterectomy. Prior to tubal sampling, each control subject received an injection of human chorionic gonadotropin [hCG] to produce a state of pseudo-pregnancy. Fallopian tubes from both groups were procured. We investigated VEGF, VEGFR1 and VEGFR2 mRNA expressions in different sections of these tubes [infundibulum, ampulla and isthmus] by reverse transcription polymerase chain reaction [RT-PCR] and quantitative PCR [Q-PCR]. RT-PCR showed expressions of these genes in all sections of the fallopian tubes in both groups. Q-PCR analysis revealed that expressions of VEGF, VEGFR1 and VEGFR2 were lower in all sections of the fallopian tubes from the case group compared to the controls. Only VEGFR2 had higher expression in the ampulla of the case group. Decreased expressions of VEGF, VEGFR1 and VEGFR2 in the EP group may have a role in the pathogenesis of embryo implantation in fallopian tubes

Phylogenetic groups, extended-spectrum beta-lactamases and metallo- beta-lactamase in Escherichia coli isolated from fecal samples of patients with diarrhea in Iran

The aims of this study were to investigate the phenotypic and genotypic of extended-spectrum beta-lactamase [ESBL] and metallo-beta-lactamases [MBLs] and determine phylogenetic background E. coli isolates from fecal samples of patients with diarrhea in Kerman, southeast of Iran The emergence of ESBLs and MBLs-producing E. coli caused problems in antibiotic treatments. E. coli strains can be assigned to four main phylog-groups, including: A, B1, B2 and D. E. coli isolates [n=216] were obtained from fecal samples of patients with diarrhea between June and December 2013. ESBLs and MBLs were confirmed by disk-diffusion and broth micro-dilution methods. Using PCR, the ESBL-positive isolates were screened to determine the phylo-groups and the presence of bla[CTX-M-15], bla[OXA-1], bla[PER-1], bla[VIM]and bla[IMP] genes. ESBL-positive isolates [n= 56] were detected. Among ESBL-positive isolates, 51 isolates were positive for bla[CTX-M15] and one isolate was positive for both bla[CTX-M-15] and bla[OXA-1] genes. None of the isolates were positive for bla[PER-1], bla[VIM]and bla[IMP] genes. PCR assay for phylotyping of isolates indicated that the isolates were belonged to groups A [54.16%], B1 [11.11%], B2 [12.96%] and D [21.75%]. The isolates possessed bla[CTX-M-15] gene were belonged to A [35 isolates], B1 [5], B2 [3] and D [8] phylo-groups. Our results indicate that bla[CTX-M-15] gene is widespread among diarrheagenic E. coli isolates. ESBLproducing E. coli isolates were disseminated among a diversity of phylo-groups. Further studies are necessary to identify the ESBL genes in relation to phylogenetic groups

Gene expression analysis of VEGF and its receptors and assessment of its serum level in unexplained recurrent spontaneous abortion

Unexplained recurrent spontaneous abortion [URSA] is one of the main complications of pregnancy which is usually defined as three or more consecutive pregnancy losses before the 20[th] week of gestation without a known cause. Vascular endothelial growth factor [VEGF] is a potent angiogenic factor and shown, along with its receptors [VEGFR1, 2], to play important roles in several physiologic processes including reproduction. The aim of the present study was to analyze gene expression of VEGF and VEGF receptors in endometrium of patients with a history of URSA compared with normal fertile women. In addition, serum VEGF concentration was assessed and compared between the two groups at the same time. In this case control study, endometrial and blood samples were obtained between day 19[th] and 24[th] of menstrual cycle [window of implantation] from 10 women with a history of URSA [case group] and 6 fertile women who had at least one successful pregnancy [control group]. Expression of VEGF and VEGFRs was studied by reverse transcription- polymerase chain reaction [RT-PCR] and then quantified by real time PCR. Normalization of expression levels was done by comparison with beta-actin expression level as an internal control. Relative VEGF, VEGFR1 and VEGFR2 expression quantities were compared between the two groups. Enzyme linked immunosorbent assay [ELISA] was used for serum VEGF assay. VEGF, VEGFR1 and VEGFR2 gene expression was detected in endometrial samples of both groups. The mean relative expression of VEGF gene was lower in the case group compared with control women, however, both VEGF receptors were expressed higher in endometrium of the case group. In addition, the serum level of VEGF was significantly higher in the case group compared with the controls. Alteration in gene expression of VEGF and its receptors in endometrium and changes of serum VEGF might play important roles in pathogenesis of unexplained RSA

[Body mass index status in secondary school students in Kerman, Iran, in 2012]

Assessment of physical growth is one of the most important methods of determining nutrition and health status. Body mass index [BMI] is a suitable index for growth monitoring and helps in the identification of growth disorders and malnutrition in teenagers. The aim of this study was to determine BMI status in secondary school students in Kerman, Iran, in 2012. This was a descriptive-analytic study in which data were collected cross-sectionally and compared with the results of the study in 1997. After gaining permission from the Department of Education of Kerman, the samples were selected randomly from the secondary schools in Kerman. Height and weight were measured by standard tools. Data were analyzed through SPSS software and using Students' independent t-test. Mean BMI of the students [n = 424] was 19 +/- 4.2 and 44.8% of students had normal BMI. Mean BMI of boys was significantly lower than girls. In total, 6% of the students were excessively thin and they were mainly from public schools in the suburb of the city. Over 50% of students had weight problems; underweight was the main problem in the studied population and only 9.2% of our subjects were overweight. Since the pubertal height and weight growth spurt occurs earlier in girls [10-13 years] than boys, the absolute comparison of BMI based on sex is not wise; if necessary it should to be performed after this period. Malnutrition or lack of access to food, or cultural factors and body shape care could explain our findings in regard to the distribution of BMI in the student population of Kerman

Expression of Toll-like receptors 7-10 in human fallopian tubes

The human female reproductive tract [FRT] is constantly deal with the invading pathogens. Recognition of these pathogens is attributed to the family of Toll like receptors [TLR] as a major part of the innate immune system. We and others have previously revealed that TLRs1-6 express in the female reproductive tract. However, more studies should be done to detect TLRs 7-10 in the female reproductive tract, especially in the fallopian tubes. To examine the expression of TLRs7-10 in human fallopian tube tissue. Using immunostaining techniques, distribution of TLR7-10 was studied in surgical sections from the uterine tubes, obtained from patients undergoing tubal ligation and hysterectomy for benign gynecological conditions. RT-PCR was used to show the existence of TLR7-10 genes in fallopian tube tissue. TLR7-10 proteins were detected in the fallopian tube epithelium, although the intensity of staining was not equal in cases. TLR7-10 genes were expressed in human fallopian tube tissue. This study indicates that TLR7-10 is expressed in fallopian tubes tissues, and may play an important role in microbial recognition, and in host defense against ascending infection

Canine visceral leishmaniasis in Kerman, southeast of Iran: a seroepidemiological, histopathological and molecular study

Canine visceral leishmaniasis [CVL] is a systemic disease with a high mortality rate, caused by a diphasic protozoan parasite, Leishmania infantum/chagasi in the world. The objective of the present study was to determine the presence of CVL in the city and suburbs of Kerman, using a range of serological, histopathological and molecular methods. Blood samples were taken from 80 clinically symptomatic stray dogs All the collected blood samples were tested by direct agglutination test [DAT] to detect the anti-Leishmania antibodies in dogs, using a cut-off value of >/= 1:320. Pathological specimens including spleen, liver and lymph nodes were prepared for paraffin blocks, sectioning, staining and final microscopic examination in the pathology laboratory. PCR amplification of kDNA from 9 samples of DAT positive stray dogs was studied. The anti-Leishmania antibody was detected in 9 dogs [11.25%] of the total 80 studied dogs. No significant difference was found between VL infection and gender. In contrast, there was a significant difference between seropositivity and age [P<0.05]. Pathological samples showed changes including hyperplasia of infected macrophages and inflammatory cells that occupied sinusoids and splenic cords. Among the samples which was characterized by PCR, only one specimen revealed to be mixed infection between L. infantum and L. tropica. The results revealed a high prevalence of L. infantum infection in stray dogs in Kerman. This kind of information is needed for implementation of future control programs

[ frequency of IgM anti-Coxiella burnetii [Q fever] antibodies among slaughterhouse workers in Kerman city, 2012]

Q fever is an important zoonotic disease caused by infection with Coxiella burnetii, a Gram negative, obligate intracellular bacterium classified within the order Legionellales. Farmers, veterinarians, abattoir workers and laboratory personnel are among persons at risk of Q fever. The aim of this study was to determine frequency of IgM anti-Coxiella burnetii antibodies in slaughterhouse workers in Kerman city/ Iran. In this survey, 64 sera samples were gathered during May - June 2011 from slaughterhouse workers to evaluate the presence of phase II IgM antibodies against Q fever, using a commercial indirect ELISA test [Virion/Sermon, Germany]. Among all sera samples tested, only 5 samples [7.8%] were positive for the presence of IgM antibodies. Since chronic Q fever leads to more complex conditions like endocarditics, chronic fatigue syndrome and recurrent abortion, preventive measures like using mask or available vaccines are recommended. Moreover, early diagnosis of Q fever followed by appropriate treatment is necessary

Toll-like receptors expression in follicular cells of patients with poor ovarian response

Background: Poor ovarian response [POR] to gonadotropin stimulation has led to a significant decline in success rate of fertility treatment. The immune system may play an important role in pathophysiology of POR by dysfunctions of cytokines and the growth factor network, and the presence of ovarian auto-antibodies. The aim of this study is to investigate the expression of toll-like receptors [TLR] 1, 2, 4, 5, 6 and cyclooxygenase [COX] 2 genes in follicular cells and concentration of interleukin [IL]-6, IL-8 and macrophage migration inhibitory factor [MIF], as major parts of innate immunity, in follicular fluid [FF] obtained from POR women in comparison with normal women

Materials and Methods: In this case-control study, 20 infertile POR patients and 20 normal women took part in this study and underwent controlled ovarian stimulation. The FF was obtained from the largest follicle [>18 mm]. The FF was centrifuged and cellular pellet was then used for evaluation of expression of TLRs and COX2 genes by real-time PCR. FF was used for quantitative analysis for IL-6, IL-8 and MIF by enzyme-linked immunosorbent assay [ELISA]

Results: TLR1, 2, 4, 5, 6 and COX2 gene expression were significantly higher in POR [p<0.05]. Concentration of IL-6, IL-8 and MIF proteins was significantly increased in POR compared with normal women [p<0.05]

Conclusion: These findings support the hypothesis that the immune system may be involved in pathophysiology of POR through TLRs

Seroprevalence of bovine leptospiral antibodies by microscopic agglutination test in Southeast of Iran

<p><b>OBJECTIVE</b>To evaluate serological findings of bovine leptospirosis which is a zoonotic disease with worldwide distribution caused by Leptospira interrogans.</p><p><b>METHODS</b>One hundred and sixty seven sera were collected from 9 commercial dairy herds in jiroft suburbs, from July to October 2011. Microscopic agglutination test (MAT) was used to evaluates serological findings of bovine leptospirosis in Jiroft suburb dairy farms, Kerman province, Iran.</p><p><b>RESULTS</b>Antibodies were found by MAT at least against one serovar of Leptospira interrogans in 29 samples (17.36%) among 167 sera at a dilution 1:100 or higher, and Leptospira pomona was the most prevalent serovar. Positive titers against more than one serovar were detected in 6 sera of the positive samples.</p><p><b>CONCLUSION</b>This study is the first report of leptospirosis in Southeast Iran and showed that Leptospira pomona was the most and Leptospira icterohaemorrhagiae the least prevalent serovars in Southeast Iran.</p>