Aquaporin expression in cryopreserved human sperm: exploring the capabilities of natural deep eutectic solvents (NADEs).

BACKGROUND: Aquaporins (AQPs) are essential proteins that facilitate the rapid movement of water and cryoprotective agents (CPAs) during the cryopreservation process, and ensure the cryo-tolerance of sperm cells. OBJECTIVE: This study evaluated the preservation of aquaporin levels in human sperm after undergoing freezing using natural deep eutectic solvents (NADES) as CPAs for cryoprotection. MATERIALS AND METHODS: From June 2021 to October 2022, 35 semen samples with normal sperm parameters were acquired from the Mehr Infertility Treatment Institute in Rasht, Iran. The samples were divided into several groups for analysis: control group (not frozen), group frozen with SpermFreeze Solution, and groups frozen with different NADESs, including ChS, ChX, ChU, ChG, GlyP, and EtP. After thawing, various aspects for each group were assessed, including the integrity and condensation of sperm chromatin, viability, motility, integrity of acrosome, and the expression of AQP1, AQP3, AQP7, AQP8, and AQP9 genes. RESULTS: The analysis of gene expression revealed that freezing with ChS and GlyP preserved the expression of the AQP1 and AQP3 genes compared to the control group. Regarding AQP7 and AQP8, significant differences were not observed in expression levels between certain NADES groups (e.g., ChS, ChU, and GlyP) and the control group. Additionally, samples frozen with specific NADESs, such as ChS, ChG, EtP, and GlyP, exhibited preserved levels of AQP9 expression when compared to the control group. CONCLUSION: These findings emphasize the importance of NADES in preserving the expression of aquaporins in cryopreserved human sperm and their important fertility parameters. Doi.org/10.54680/fr24310110512.

Brucellosis as a neglected disease in a neglected population: a seroepidemiological study of migratory nomads in the Fars province of Iran.

This study assessed the seroprevalence of brucellosis and its risk factors in migratory nomads in the Fars province of Iran. Active brucellosis was defined as the combination of clinical symptoms, including fever, chills, night sweats, headache, low back pain, arthralgia, or myalgia, and positive laboratory testing, including either a serum agglutination test (SAT) ⩾1:80 with a 2-mercaptoethanol (2-ME) test ⩾1:40, or a SAT <1:80 combined with a positive Coombs Wright test (CWT) at a titre of at least threefold higher than SAT titre results. For the 536 participants, the female (316, 59%) to male (220, 41%) ratio was 1·4 and the participants' mean age was 32·4 ± 18·9 (range 1-96) years. Of all participants, 325 (60·6%) showed clinical symptoms; in symptomatic participants, the Rose Bengal plate test was positive in 33 (6·1%) cases, the SAT was positive in 18 (3·3%) cases, and the 2-ME test was positive in 30 (5·5%) cases. Positive SAT and 2-ME results were seen in 18 (3·3%) cases, but a negative SAT and a positive CWT were found in 36 (6·7%) cases. As a result, active brucellosis was detected in 54 cases, indicating a prevalence of 10% (95% confidence interval 8-12). In conclusion, we determined that brucellosis is a prevalent yet neglected disease in this nomadic population. Brucellosis control is not possible as long as these high-risk populations remain neglected.

Tacrolimus Dose Requirement in Iranian Kidney Transplant Recipients within the First Three Weeks after Transplantation.

BACKGROUND: Tacrolimus is the main immunosuppressive agent in many kidney transplant protocols with an initial recommended daily dose of 0.2 mg/kg of ideal body weight (IBW). However, due to the high inter- and intra-patient variability in its pharmacokinetics, the required tacrolimus doses may differ markedly from patient to patient. OBJECTIVE: To assess the required tacrolimus dose to achieve the desired whole blood concentration within the first three weeks after kidney transplantation among Iranian patients. METHODS: This cross-sectional study was performed at kidney transplantation ward of Imam Khomeini Hospital Complex where almost all patients receive thymoglobulin induction therapy and a calcineurin inhibitor, mainly tacrolimus, plus mycophenolate, and prednisolone as maintenance immnosuppressive drugs with the target tacrolimus whole blood concentration of 8-12 ng/mL for the first month after transplantation. RESULTS: The mean±SD administered daily dose of tacrolimus during the first three weeks after transplantation was 0.085±0.024 mg/kg of IBW that resulted in a mean±SD whole blood concentration of 10.34±5.44 ng/mL. The required mean±SD dose of the drug to achieve the desired whole blood level of 8-10 ng/mL was 0.08±0.02 mg/kg. Only 27.4% of the assessed tacrolimus blood levels were within the desired range. Compared with males, females needed 19% more daily dose of tacrolimus to reach similar whole blood levels. Tacrolimus blood levels were significantly correlated with daily tacrolimus doses (r=0.307, p=0.001) and patients' age (r=0.283, p=0.003). CONCLUSION: It seems that Iranian kidney transplant recipients need lower daily doses of tacrolimus to achieve the desired whole blood levels; compared with males, females need a higher dose.

Spontaneous abortion and functional polymorphism (Val16Ala) in the manganese SOD gene.

Spontaneous abortion is the most common complication of early pregnancy. Genetic factors have been hypothesised to play a role in spontaneous abortion. Since it is possible that the balance of oxidants and antioxidants can be affected by different genetic variants, gene polymorphisms have been proposed as a susceptibility factor that increases the chance of miscarriage. Manganese superoxide dismutase is an important antioxidant enzyme encoded by manganese superoxide dismutase (MnSOD) gene. The aim of this experiment was to assess whether Val16Ala polymorphism of MnSOD gene is associated with miscarriage in northern Iran. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used for genotyping. Statistical analyses were conducted using the χ(2)-test. The genetic distributions did not differ significantly between cases and controls, however slightly more Val/Val genotypes were found among the patients compared with control subjects (p = 0.059). No correlation was observed between susceptibility to abortion and MnSOD Val16Ala polymorphism. Larger population-based studies are needed for clarifying the relationship between abortion and MnSOD genotypes.

Polymorphism of catalase gene (CAT C-262T) in women with endometriosis.

Endometriosis is defined as the presence of ectopic endometrial glands and stroma outside of the uterine cavity. Recent studies have shown that the oxidative stress causes irreparable damage, which leads to oxidative enzymopathies. Catalase gene encodes an antioxidant enzyme, detoxifying hydrogen peroxide to H2O and O2. The aim of this study was to determine whether the polymorphism at position -262 in the promoter region of catalase gene (C-262T), which alters the expression and enzyme blood levels, could have an impact on the risk of endometriosis. Extracted DNA from peripheral blood leucocytes was genotyped using allele-specific PCR (AS-PCR). The χ(2)-test was used for statistical analyses. In endometriosis subjects, the frequencies of the CAT CC/CT/TT were 67.5%, 32.5% and 0%, respectively, while in healthy women, they were 12%, 68% and 20%, respectively. Significant differences in allele and genotype distribution among controls and patients were found (OR, 178.76 95% CI, 10.11-3159.1202; p = 0.0004). This study indicates that catalase C-262T polymorphism is associated with the endometriosis. Randomised multicentre trials with greater sample sizes are still needed to clarify our results.