Microfluidic sperm sorting selects a subpopulation of high-quality sperm with a higher potential for fertilization.

STUDY QUESTION: Is a microfluidic sperm sorter (MSS) able to select higher quality sperm compared to conventional methods? SUMMARY ANSWER: The MSS selects sperm with improved parameters, lower DNA fragmentation, and higher fertilizing potential. WHAT IS KNOWN ALREADY: To date, the few studies that have compared microfluidics sperm selection with conventional methods have used heterogeneous study population and have lacked molecular investigations. STUDY DESIGN, SIZE, DURATION: The efficiency of a newly designed MSS in isolating high-quality sperm was compared to the density-gradient centrifugation (DGC) and swim-up (SU) methods, using 100 semen samples in two groups, during 2023-2024. PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen specimens from 50 normozoospermic and 50 non-normozoospermic men were sorted using MSS, DGC, and SU methods to compare parameters related to the quality and fertilizing potential of sperm. The fertilizing potential of sperm was determined by measurement of phospholipase C zeta (PLCζ) and post-acrosomal sheath WW domain-binding protein (PAWP) expression using flow cytometry, and the chromatin dispersion test was used to assess sperm DNA damage. MAIN RESULTS AND THE ROLE OF CHANCE: In both normozoospermic and non-normozoospermic groups, the MSS-selected sperm with the highest progressive motility, PLCζ positive expression and PLCζ and PAWP fluorescence intensity the lowest non-progressive motility, and minimal DNA fragmentation, compared to sperm selected by DGC and SU methods (P < 0.05). LIMITATION, REASONS FOR CAUTION: The major limitations of our study were the low yield of sperm in the MSS chips and intentional exclusion of severe male factor infertility to yield a sufficient sperm count for molecular experiments; thus testing with severe oligozoospermic semen and samples with low count and motility is still required. In addition, due to ethical considerations, at present, it was impossible to use the sperm achieved from MSS in the clinic to assess the fertilization rate and further outcomes. WIDER IMPLICATIONS OF THE FINDINGS: Our research presents new evidence that microfluidic sperm sorting may result in the selection of high-quality sperm from raw semen. This novel technology might be a key to improving clinical outcomes of assisted reproduction in infertile patients. STUDY FUNDING/COMPETING INTEREST(S): The study is funded by the Iran University of Medical Sciences and no competing interest exists. TRIAL REGISTRATION NUMBER: N/A.

How sperm protects itself: A journey in the female reproductive system.

Sperm must pass a complex route in the female reproductive tract (FRT) to reach the fertilization site and join the oocyte. Thus, it should employ several mechanisms to survive against the female immune system, fertilize the oocyte, and successfully transmit paternal genes to the next generation. In addition to self-protection, sperm may be involved in the immune tolerance to the developing embryo and regulating the FRT for embryo implantation and subsequent pregnancy. Hence, this review intends to summarize the mechanisms that protect sperm in the FRT: including immunomodulatory factors that are carried by seminal plasma, cell-to-cell and molecular interaction of sperm with epithelial and immune cells of the FRT, high regulated secretions of inflammatory factors such as cytokines, chemokines, and growth factors, inducing immune tolerance to paternal antigens, and specialized expression of cell receptors and binding proteins. In most of these events sperm induces the FRT to protect itself by modulating immune responses for its own benefit. However, not all sperm in the semen are able to trigger the survival mechanisms and only high-quality sperm will overcome this challenge. A clear understanding of the molecular mechanisms that maintain sperm viability and function in the FRT can lead to new knowledge about infertility etiology and a new approach in assisted reproductive technologies for the preparation and selection of the best sperm based on the criteria that physiologically happen in-vivo.

Downregulation of miR-211 expression in the blood plasma of infertile men compared to the fertile controls.

OBJECTIVES: Infertility is inability to conceive after 12 months of regular unprotected sex. MiRNA expression changes can serve as potential biomarkers for infertility in males due to impaired spermatogenesis. This research was conducted to measure the expression level of miR-211 in plasma samples as a factor identifying infertility in comparison with the control group. METHODS: In this study, blood plasma were taken from the infertile men (n = 103) nonobstructive azoospermia (NOA) or severe oligozoospermia (SO) and the control group (n = 121). The expression of circulating miR-211 in plasma was assessed by qRT-PCR. A relative quantification strategy was adopted using the 2-ΔΔCT method to calculate the target miR-211 expression level in both study groups. RESULTS: Plasma miR-211 levels were significantly lower in infertile men compared to the control group (0.544 ± 0.028 and 1.203 ± 0.035, respectively, p < 0.001). Pearson's correlation analysis showed that miR-211 expression level has a positive and significant correlation with sperm parameters, including sperm concentration, sperm total motility, progressive motility, and normal morphology (p < 0.001). CONCLUSIONS: Decreased expression of miR-211 in blood plasma seems to be associated with male infertility. This experiment showed that miR-211 can be considered as a biomarker for evaluation, diagnosis, and confirmation of the results of semen analysis in male infertility.

Prevalence of congenital anomalies and related factors in live births in Zahedan, Southeast of Iran: A cross-sectional study.

Background: The term congenital anomalies (CAs) refers to structural or functional abnormalities at the time of conception. Approximately 12 deaths related to congenital disabilities occur in every 10,000 babies born. Objective: This study aimed to evaluate the prevalence and associated factors of single and multiple CAs in live births in Zahedan, Southeast Iran. Materials and Methods: This cross-sectional study was conducted on 59,087 live births in a referral hospital in Zahedan located in the southeast of Iran from 2009 to 2019. All live births were examined by pediatricians and the CAs and categorized based on the international classification of diseases. Results: Of 59,085 live births, at least 883 had a significant anomaly, and the prevalence rate of CAs was about 149 per 10,000. Anomalies of the nervous (24.1%) and cardiovascular systems (21.10%) were the most frequent, occurring in 213 and 187 of the live births, respectively. Spina bifida is the most common anomaly of the central nervous system. The most common anomalies in the cardiovascular system were unspecified heart malformations (17.1%), cardiovascular malformations (18.7%), and patent ductus arteriosus (11.7%). Significant correlations were found between the parent's consanguinity marriage, the mother's age, an existing anomaly in the family, and relatives in single and multiple CAs (p = 0.02, p = 0.02, p < 0.001, p = 0.01, respectively). Conclusion: The prevalence of CAs was 149 per 10,000 live births. The highest prevalence of CAs was related to the central nervous system. Increasing the public's knowledge about fetal defects can reduce the prevalence of CAs.

The possible impact of DNA-binding chromodomain-helicase 5 polymorphisms on male infertility: A case-control study.

BACKGROUND: Chromodomain-helicase 5 (CHD5) is a conventional tumor-suppressing gene in many tumors. The CHD5 gene, as a key factor in the chromatin density process during sperm maturation, can affect the risk of infertility. This study aimed to determine whether CHD5 variants contribute to the risk of male infertility. METHODS: Gene variants were identified using tetra primer-ARMS-PCR method on nonobstructive azoospermia and severe oligozoospermia in a case-control study. SPSS software 20 (SPSS Inc. Chicago, IL, USA) was used for data recording and statistical analysis. RESULTS: In the codominant pattern, the rs12067480 TT variant versus CC significantly increased the risk of disease, and also, in the recessive pattern, TT variant versus CC + CT and T allele versus C. The rs2273032 variant was associated with the risk of infertility in codominant pattern AA versus GG and recessive pattern AA versus GG + GA and allele. We discovered that the rs12067480 T and rs2273032 A alleles increase the risk of male infertility. Also, the interaction of the CT/GA, CT/AA, TT/GA genotypes and rs12067480T/rs2273032A and rs12067480T/rs2273032G haplotypes significantly increased the risk of infertility. CONCLUSIONS: Our results suggest that the CHD5 gene polymorphisms contribute to the risk of male infertility. Our findings can be valuable in improving the diagnosis and treatment of infertility.

Immunoexpression of interferon-gamma in the interdental gingiva of chronic periodontitis patients with interferon-gamma (+874A/T) rs62559044 polymorphism.

Interferon-gamma (IFNγ), which is involved in inflammation, has an essential role in the pathogenesis of chronic periodontitis (CP). The association of IFNγ +874A/T gene polymorphisms with a higher incidence of CP has been reported previously. We investigated the immunoexpression of IFNγ in human gingiva from CP patients with IFNγ +874A/T gene polymorphisms compared to a healthy group. Biopsies from interdental gingiva (n = 60) were obtained from CP patients (n = 38) and individuals who had clinically healthy gingiva (n = 22). The specimens were classified according to genotypes of IFNγ +874A/T gene polymorphism using amplification-refractory mutation system-polymerase chain reaction (ARMS-PCR) (10AA, 18AT, and 10 TT). After tissue processing and slide preparation, we evaluated the expression levels of IFNγ in the interdental gingiva using the immunohistochemistry technique. Values were compared between two study groups by Mann-Whitney and Kruskal-Wallis analysis. P < 0.05 was considered the level of significance. There was a noticeable increase in the immunoexpression of IFNγ in the gingival epithelium, fibroblasts, and capillaries of gingival tissue from CP patients compared to the control group. We found different levels of IFNγ expression in different parts of the gingival epithelium. The highest levels were found in the middle regions of the gingival epithelium in patients with TT genotypes. Immune cells and vascular endothelium of gingiva in the AA group also showed increased expression of IFNγ. Results suggest that IFNγ +874A/T gene polymorphism may change the expression level of IFNγ and subsequently the progression of CP.

The role of myo-inositol supplement in assisted reproductive techniques.

Assisted reproductive techniques can help many infertile couples conceive. Therefore, there is a need for an effective method to overcome the widespread problems of infertile men and women. Oocyte and sperm quality can increase the chances of successful in vitro fertilisation. The maturation environment in which gametes are present can affect their competency for fertilisation. It is well established that myo-inositol (MI) plays a pivotal role in reproductive physiology. It participates in cell membrane formation, lipid synthesis, cell proliferation, cardiac regulation, metabolic alterations, and fertility. This molecule also acts as a direct messenger of insulin and improves glucose uptake in various reproductive tissues. Evidence suggests that MI regulates events such as gamete maturation, fertilisation, and embryo growth through intracellular Ca2 + release and various signalling pathways. In addition to the in-vivo production of MI from glucose in the reproductive organs, its synthesis by in vitro-cultured sperm and follicles has also been reported. Therefore, MI is suggested as a therapeutic approach to maintain sperm and oocyte health in men and women with reproductive disorders and individuals of reproductive age.

Microfluidic chips as a method for sperm selection improve fertilization rate in couples with fertilization failure.

PURPOSE: Sperm quality plays a vital role in successful fertilization and pregnancy. Patients with fertilization failure (total failure or low-fertilization rate) despite having normal semen parameters are a challenging group whose sperm cannot fertilize the oocyte via the intracytoplasmic sperm injection (ICSI) technique. Microfluidics is offered as a new method for proper sperm sorting. METHODS: This study aimed to evaluate sperm parameters, DNA fragmentation index (DFI), expression of phospholipase C zeta 1 (PLCZ1), and transition nuclear proteins 1 (TNP1) mRNAs in sperm selected by microfluidic sperm sorting (MSS) chip compared with conventional density gradient centrifugation technique in patients with fertilization failure following ICSI. Subsequence fertilization rate and embryo quality were assayed. RESULTS: Normal morphology and total motility were significantly higher, and DFI was significantly lower in sperm selected by the MSS chip in fertilization failure and control groups. The RT-PCR results demonstrated a significant increase in the expression of PLCZ1 and TNP1 genes in sperm of both groups selected by MSS chips compared to the DGC method. In addition, with the selected sperm by MSS chip, an increase in fertilization rate and improvement of embryo quality was obtained. CONCLUSION: The present study findings show that sperm sorting by the microfluidic method improves fertilization rate in patients with poor fertilization outcomes following ICSI.

Sperm-oviduct interaction: Differential gene expression of growth factors induced by sperm DNA fragmentation.

The present study investigated the effects of DNA fragmentation of spermatozoa on the growth factors expression by a human oviduct epithelial cell line (OE-E6/E7). Two separate groups were examined in this study. The cell line was cultured in the presence of spermatozoa with normal DNA fragmentation index (DFI) or abnormal DFI. Total RNA from the cell line in each group was isolated, and relative expression of objective genes was analysed using PCR array. Also, the concentration of VEGF, BMP-2, BMP-7 and MSTN in the supernatant of cell culture was analysed by the ELISA method. The PCR array analysis revealed that most of the growth factors had been upregulated in the abnormal group. However, the differences between groups were statistically significant (p < 0.05) for five genes, including VEGF-A, BMP-2, BMP-6, BMP-7 and OSM. Furthermore, MSTN was the only gene that down-regulated significantly under the influence of the spermatozoa with abnormal DFI. Moreover, the results of ELISA analysis were in agreement with the data of the PCR array. It has been concluded that DNA fragmentation in human spermatozoa can probably change regular events throughout the oviducts. Consequently, the genes of interest may change sperm function and probably its fate in the female reproductive tract.

Reduced volumetric parameters of the placenta and extravillous trophoblastic cells in complicated pregnancies may lead to intrauterine growth restriction and small for gestational age birth.

AIM: We undertook this study to determine quantitative changes of the placenta, focusing on extravillous trophoblastic cells (EVTs) in pregnancies with intrauterine growth restriction (IUGR) and small for gestational age (SGA) compared to the control group. METHODS: Placentas from pregnancies complicated with SGA-IUGR (n = 10) and control group (n = 10) were obtained after cesarean surgery and evaluated using stereological assays after routine tissue processing and Masson's trichrome staining. Mann-Whitney U-test was employed, and the level of statistical significance was set at p <0.05. RESULTS: Our results showed that the volumetric parameters, including the total volume and volume density of chorionic villi, intervillous spaces, blood vessels in chorionic villi, and syncytiotrophoblast, decreased significantly in the SGA-IUGR group compared to control placentas (p <0.05). Also, total volume, number of EVTs, volume, the diameter of cytoplasm, and diameter of the nucleus in these cells were significantly lower in the SGA-IUGR group (p <0.05). In addition, the nucleus to cytoplasm ratio of EVTs was also higher in the SGA-IUGR group (p <0.05). CONCLUSIONS: There are several significant histological and stereological differences in the placenta, particularly its EVTs from the SGA-IUGR group compared to the control group. It seems that histological changes in the placental tissues could be helpful for the retrospective explanations of pregnancy complications.

Interleukin 22 Expression During the Implantation Window in the Endometrium of Women with Unexplained Recurrent Pregnancy Loss and Unexplained Infertility Compared to Healthy Parturient Individuals.

We evaluated the expression of interleukin-22 (IL-22) in the endometrium of women with unexplained recurrent pregnancy loss (uRPL) and unexplained infertility (UI) compared to the women with normal pregnancies. Endometrial tissues were collected from 20 women with UI, 20 women with uRPL, and 24 healthy women as a control group. Immunohistochemical expression and gene expression of IL-22 were analyzed by immunohistochemistry (IHC) and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) methods. The controls showed lower IL-22 expression than the uRPL group (P > 0.05) using PCR. It was also found that patients with UI had lower levels of IL-22 expression compared to the uRPL group (P > 0.05). Although IL-22 expression in the endometrium of patients with UI was higher than the control group, this difference was not statistically significant (P < 0.05). IL-22 immunoreactivity was observed in the endometrial glands and stromal tissues using IHC. We found the lowest IL-22 expression in the control group and the highest in uRPL samples (P < 0.05). Our findings suggest that a significant increase in IL-22 expression in uRPL patients may affect fertility and pregnancy outcomes or even have a considerable impact on immune function deficits. Further studies on the critical function of IL-22 during pregnancy are suggested.

Cannabinoid receptor type-1 and its correlation with CB1 gene polymorphism-1359G/A in ectopic pregnancy compared to the control group.

AIM: Ectopic pregnancy (EP) is one of the most important causes of maternal mortality. This study aimed to evaluate the immunohistochemical (IHC) expression of the cannabinoid receptor type-1 (CB1) and its association with CB1-1359G/A gene polymorphism (rs1049353) in the fallopian tubes in EP compared to controls. METHODS: In this case-control study, 100 women with EP (cases) and 100 women that underwent abdominal surgery due to the hysterectomy or uterine tubal ligation (healthy controls) were included. Genotyping of CB1-1359G/A polymorphism, tissue expression of CB1 at the protein and mRNA levels were studied using restriction fragment length polymorphism, IHC method, and quantitative real-time polymerase chain reaction (qRT-PCR) analysis. RESULTS: Genotyping showed that in EP, the frequency of AA, AA+AG genotypes, and A allele was significantly higher than healthy control subjects (p = 0.001). Also, patients with EP had significantly increased IHC expression of CB-1 compared to the control samples (p = 0.016). Patients with AA and AG genotypes had a significantly higher IHC expression of CB-1 compared to the GG genotype. qRT-PCR analysis showed that patients with EP had significantly increased expression of CB-1 compared to the control samples (p < 0.001). Patients with AA and AG genotypes had higher significant mRNA expression of CB-1 compared to the GG genotype. CONCLUSIONS: CB1 is likely to be effective in creating innate immunity in humans and can affect the process of EP in the fallopian tube. CB1 is also a pathological valuable factor in identifying the pathway of inflammation during ectopic implantation.

Elevated immunoexpression of interferon-gamma in placenta tissue samples from pregnancies complicated with preeclampsia compared to the placenta previa.

AIM: The present study aimed to compare the immunohistochemical expression of interferon-gamma (IFN-γ) in placentas from pregnancies complicated with preeclampsia (PE) and placenta previa (PP) and normal healthy placentas. METHODS: Placentas were collected from cases of PE, PP and normal pregnancies as a control group (10 placentas in each group). All the deliveries were at full-term (37-42 weeks) by cesarean section and newborns were without any complications or diseases. Expression of IFN-γ in the placenta was determined using immunohistochemical methods and findings were compared. Statistical analysis was performed by Mann-Whitney and Kruskal-Wallis tests for comparing the mean values of IFN-γ expression in the placentas from PE, PP and control groups. Our results showed that the immunoexpression of IFN-γ in syncytiotrophoblast cells, extravillous trophoblast cells, vascular endothelium and basal plate of the placenta from PE group were more than control and PP groups (P < 0.05) and in PP group were more than the control group (P < 0.05). CONCLUSION: We concluded that the immunoexpression of IFN-γ was increased significantly in placenta tissue samples of the PE group compared to the PP group and normal pregnancies. It is proposed that IFN-γ has an important role in the different mechanisms of PE and PP progression.

Upregulation of elafin expression in the fallopian tube of ectopic tubal pregnancies compared to the normal tubes.

BACKGROUND: Ectopic pregnancy is one of the most important causes of maternal deaths and fallopian tubes are the location of 95% of ectopic pregnancies. Elafin is a natural antimicrobial molecule that plays an important role as an anti-inflammatory agent in mucosal surfaces and has been found in the female reproductive tract. OBJECTIVES: The aim of this study was to investigate elafin expression, in the fallopian tube mucosa of ectopic pregnancies compared to the normal tubes using immunohistochemistry (IHC) techniques and quantitative reverse transcription (qRT)-PCR. METHODS: In this case-control study, uterine tube samples were obtained from patients with an indication for surgical removal of the tubes. The case group (n = 20) consisted of patients who were undergoing salpingectomy due to an ectopic pregnancy, the control group (n = 20) included patients who had a salpingectomy and hysterectomy. Using qRT-PCR and IHC, the expression of elafin was investigated in both study groups. RESULTS: Immunohistochemical expression of elafin in the epithelium and connective tissue was significantly increased in the implantation site of the patients in comparison with the control group (P < 0.001). The level of elafin mRNA increased in the mucous membrane of the fallopian tube from patients with the ectopic pregnancy compared to the normal mucosa (P < 0.001). CONCLUSION: Increasing expression of elafin during an ectopic pregnancy may be a mechanism for enhancing innate immune response and be involved in related pathological conditions such as infection and ectopic implantation.

Effects of gestational diabetes mellitus on stereological parameters and extravillous trophoblast cells of placenta compared to the control group.

The quantitative changes of extravillous trophoblast cells (EVTs) in placenta of gestational diabetes mellitus (GDM) patients were investigated compared to healthy controls using stereological methods. The volumetric parameters of the placenta and EVTs were estimated using Cavalieri's principle and Physical Disector stereological methods. The placental volume and weight in the GDM group increased compared to the control group (p < .05). The difference in the total volume of intervillous space and blood vessels of the placenta between GDM and control groups was statistically significant (p < .05). In addition, there was a significant difference in the volume density of blood vessels and syncytiotrophoblast between the GDM group and the control group (p < .05). The total volume of the EVTs, nucleus and cytoplasm diameter, volume of the nucleus and cytoplasm, nucleus to cytoplasm ratio (N/C) of EVTs and the total number of EVTs per unit volume of the placenta in the GDM group showed a significant increase compared to the controls (p < .05). Impact statement What is already known on this subject? It is reported that extravillous trophoblast cells (EVTs) played an important role in pregnancy complications. There are limited studies on the quantitative changes of EVTs in the placental bed of GDM patients. What do the results of this study add? The results showed that volumetric parameters and number of EVTs were significantly altered in GDM placentas. These changes can be associated with disturbances in trophoblastic invasion in GDM pregnancies and may affect the development and survival of the embryo. What are the implications of these findings for clinical practice and/or further research? In the present study, there is a new insight to placenta structure that probably could be useful to understanding possible mechanisms of pregnancy complications and the achievement of new therapeutic strategies. Further investigation on the molecular biology of these cells in pregnancy complications will be needed to clarify this hypothesis.

Placenta structural changes in heavy smoking mothers: a stereological aspect.

OBJECTIVE: Smoking during pregnancy is able to alter the structure and function of the placenta. In the present study, quantitative changes of the placenta in smoking mothers were investigated compared to healthy controls by Cavalieri's point counting method. METHODS: Twenty placentas from heavy smoking mothers and non-smoker controls (n = 10 in each group) were selected. Systematic uniform random sampling (SURS) was used for sample selection and tissue sectioning. Quantitative parameters of the placenta in the selected sections were estimated after Masson's trichrome staining. Differences between the two groups were determined by the Mann Whitney U test and the significance level was set at p < .05. RESULTS: Results showed that there was a significant difference in the placental weight, total volume of placenta, intervillous space, fibrin and syncytiotrophoblast between the heavy smoker group and the control group (p < .05). The differences in the volume density of fibrin and blood vessels between the smoker and control groups were statistically significant (p < .05). CONCLUSIONS: Our findings suggested that quantitative parameters of the placenta significantly changed in placentas from the smoker group compared to controls. These changes can probably be associated with pregnancy complications in smoking mothers and may affect the development and survival of the fetus and even its future life.

Quantitative changes of extravillous trophoblast cells in heavy smoker mothers compared with healthy controls.

Maternal smoking during pregnancy can induce structural and functional changes in the placenta. Placentas from heavy smoker (>20 cigarettes per day) mothers and non-smoker healthy controls (n=10 in each group) were enrolled in the present case-control study. Sample selection and sectioning were performed by systematic uniform random sampling (SURS). Selected sections were stained using Masson's trichrome to estimate quantitative parameters of placental extravillous trophoblast cells (EVTs) and the number of EVTs. Differences between groups were evaluated using the Mann-Whitney U-test, with significance set at P<0.05. There was a significant difference in placental weight and the total volume of the placenta between the heavy smoker and control groups (P<0.05). The total volume of EVTs, nucleus diameter, cytoplasm diameter, the volume of the nucleus and cytoplasm and the nucleus to cytoplasm ratio of EVTs were significantly greater in the heavy smoker compared with control group (P<0.05 for all). In placentas from heavy smokers, the total number of EVTs per unit volume of placental bed were significantly greater than in the control group (P<0.05 for both). In conclusion, the findings suggest that maternal smoking could affect fetal health by changing the quantitative parameters of the placenta, and likely the invasive properties of EVTs.

Association of macrophage migration inhibitory factor gene polymorphisms with chronic periodontitis in a South Eastern Iranian population.

BACKGROUND: Macrophage migration inhibitory factor (MIF) is a key proinflammatory mediator. It plays a vital role in immune response against the oral disease. MIF is a regulator of innate immunity, and bacterial antigens can stimulate serum level of this protein. In experimental gingivitis, the expression level of MIF increases and this increment positively correlates with oral plaque index. The single nucleotide polymorphisms in the gene encoding the MIF protein can control the function of MIF. The aim of the present study was a clarification of the associations between MIF-173 G/C, MIF 95 bp, and 189 bp insertion/deletion (I/D) polymorphisms and chronic periodontitis (CP) compared with healthy controls. MATERIALS AND METHODS: This case-control study was carried out on 210 CP patients and 100 normal subjects. MIF-173 G/C and MIF 95 bp and 189 bp I/D polymorphisms were genotyped, using polymerase chain reaction-restriction fragment-length polymorphism (PCR-RFLP) and PCR, respectively. Allele and genotype frequencies of the variants were compared between patients and controls using Chi-square. test. The value of P < 0.05 was considered statistically significant. RESULTS: The study findings showed that MIF-173 G/C polymorphism, especially the C allele increased the risk of CP. The 95-bp I/D polymorphism was not associated with CP and the 185-bp I/D variant was not polymorphic in our population. CONCLUSION: Therefore, MIF-137 G/C variant increased the risk of CP in the South East of the Iranian population. In other words, polymorphisms in MIF gene influence clinical outcome of CP infection and influence the susceptibility to disease. Further studies with larger sample sizes and different ethnicities are required to validate our findings.

Quantitative changes of extravillous trophoblast cells in placentas of systemic lupus erythematosus patients.

In the present study, quantitative changes of extravillous trophoblast cells (EVTs) in the placentas of SLE patients were investigated compared to healthy controls using stereological methods. Volumetric parameters and number of EVTs per unit volume of the placenta were estimated respectively, using Cavalieri's principle and Physical Disector stereological methods. Placental volume in the SLE group was increased compared to the control group, but this increase was not statistically significant (p > .05). Placental weight in the patient group showed a significant decrease compared to controls (p < .05). Total volume of EVTs, diameter and volume of the nucleus and cytoplasm and the N/C ratio of EVTs in the SLE group showed a significant increase compared to the controls (p < .05). In SLE placentas the total number of EVTs per unit volume of the placenta was increased significantly compared to the control group (p < .05). Impact statement In the present study, there is a new insight to placenta structure that may be useful in understanding possible mechanisms of pregnancy complications and the achievement of new therapeutic strategies. In the present study, for the first time quantitative changes of extravillous trophoblast cells (EVTs) in the placental bed of SLE patients were investigated compared to healthy controls using stereological methods. Results showed that volumetric parameters and number of EVTs were significantly altered in SLE placentas. These changes can be associated with disturbances in trophoblastic invasion in SLE pregnancies and may affect the development and survival of the embryo. Further investigation on the molecular biology of these cells in pregnancy complications will be needed to clarify this hypothesis.

Quantitative Parameters of Interdental Gingiva in Chronic Periodontitis Patients with IFN-γ Gene Polymorphism.

Chronic periodontitis (CP), an infectious disease resulting in inflammation within the periodontal tissue, is the main cause of adult tooth loss. CP is a multi-factorial disorder and the interaction between multiple genetic and environmental factors results in the manifestation of this disease. Recent researches in periodontitis has focused on cytokine gene polymorphisms that play important role in periodontal inflammation, but few studies investigated histological change that occur during CP in the supporting tissue of teeth. The aims of this study were to investigate the association of IFN-γ +874 A/T polymorphisms and quantitative parameters of interdental gingiva in CP patients. The study samples were interdental gingiva biopsies from 60 individuals including 38 patients and 22 healthy subjects. After determination of IFN-γ +874 A/T gene polymorphism by amplification refractory mutation system-polymerase chain reaction (ARMS-PCR), patients were divided in three subgroups: 10 AA, 18 AT and 10 TT. After slides preparation, quantitative parameters were estimated by Cavalieri's point-counting method. Statistical analyses were performed using Mann-Whitney and Kruskal-Wallis test to compare differences between groups. The volume density (Vv) of epithelium, connective tissue and its components were significantly different between the control and CP groups (P<0.05). Statistically significant differences in the Vv of collagenous and non-collagenous matrix of interdental gingiva between AA, AT and TT groups were found (P<0.05). Result of present study shows that IFN-γ +874 A/T is strongly associated with some quantitative parameters of connective tissue constituents of interdental papilla in CP patients.