Expression，co-localization and interaction analysis of mitochondrial membrane proteins prohibitin and stomatin-like protein 2 during spermatogenesis in Larimichthys polyactis.

Hexavalent chromium Cr (VI), an endocrine disruptor, has been a possible risk factor for male-oriented infertility, due to its damaging effect on testes. Thus, we sought to analyze the protective effect of Aloe vera (A. vera) and their mediated silver nanoparticles on testes in mice. Sixty male mice were exposed to Cr (VI) and A. vera extract (AV) and their mediated silver nanoparticles (AV+NP) as protective agents for 60 days. The organ index, hormone analysis, enzyme analysis, metal estimation, computer assisted sperm analysis (CASA), histopathology of testes and micrometric data were assessed using one-way ANOVA. A significant reduction was found in the level of catalase (87.1 ± 7.02 mmol/mL), superoxide dismutase (61.8 ± 2.5 mmol/mL), glutathione (1.51 ± 0.9 µmol/mL), luteinizing hormone (LH) (1.6 ± 0.3 ng/mL), and testosterone (2.3 ± 0.2 ng/mL) in Cr exposed group. However, a significant increase was found in Cr exposed mice in follicle-stimulating hormone (FSH) (159.9 ± 4.98 ng/mL) level. With administration of AV and AV+NP, we found a significant increase in level of catalase (143.4 ± 2.6 mmol/mL; 134.8 ± 9.1 mmol/mL), superoxide dismutase (205.8 ± 13.0 mmol/mL; 187.3 ± 9.9 mmol/mL), glutathione (2.98 ± 0.2 µmol/mL; 3.06 ± 0.2 µmol/mL), luteinizing hormone (LH) (2.9 ± 0.2 ng/mL; 3.1 ± 0.2 ng/mL), and testosterone (3.9 ± 0.1 ng/mL; 4.58 ± 0.2 ng/mL) respectively, whereas, FSH level was decreased (84.7 ± 7.2 ng/mL; 70.4 ± 4 ng/mL). The CASA showed a noticeable decrease in sperm quantity and kinematics after Cr exposure. In AV and AV+NP exposed groups, the sperm kinematics were improved significantly. The histopathological studies showed necrosis of Leydig cells, absence of sperm tails, and destruction in the zone of mitosis and meiosis in Cr-exposed groups. The micro morphometric analysis showed a significant reduction in the size of spermatogonia (38.9 ± 0.7 µm), size of spermatocytes (26.91 ± 1.4 µm), head breadth (5.4 ± 0.7 µm), head length (19.98 ± 1.2 µm), middle piece, and tail length (0.00 ± 0.0 µm; 0.00 ± 0.0 µm) in Cr exposed mice. However, with the AV and AV+NP administration, the testis architecture was improved, exhibiting an improved zone of mitosis and meiosis. A. vera exposure induced a significant decrease in the size of spermatogonia (24.1 ± 2.4 µm), and size of spermatocytes (18.7 ± 1.2 µm), whereas a significant increase in sperm head breadth (15.1 ± 0.8 µm), and head length (34.4 ± 2.8 µm) was found. We found that AV and AV+NP can potentially reduce oxidative damage induced by Cr (VI) in testes and promote fertility.

Spermatogenesis and ultrastructure of the spermatozoa at three ploidy levels in the bivalve species, Corbicula fluminea.

The CMTM family plays pivotal roles in various physiological and pathological processes, including those associated with the male reproductive system. Among them, CMTM2 exhibits specific expression in testicular tissue; however, its relationship with spermatogenesis, sperm function, and the underlying molecular mechanisms remains unclear. This study systematically investigated the localization of CMTM2 in human testis and sperm, as well as its correlation with spermatogenesis and sperm quality. The results revealed that CMTM2 expression was specifically localized in human testis, predominantly in germ cells, with significantly lower levels observed in sperm from patients with asthenozoospermia and teratozoospermia compared to normospermic individuals. Immunofluorescence analysis revealed that CMTM2 was predominantly localized in the middle piece of sperm, with significant reductions in infertile samples. Western blot analysis confirmed decreased CMTM2 levels in sperm from asthenozoospermia, oligozoospermia, and teratozoospermia patients, indicating a strong correlation between CMTM2 expression and sperm quality. Furthermore, CMTM2 expression positively correlated with sperm progressive motility, but not with sperm concentration, suggesting its potential role in regulating sperm motility in both normal and asthenozoospermia patients. The findings suggested a significant correlation between the CMTM2 expression and spermatogenesis as well as sperm quality, thereby providing valuable insights for further investigations into the molecular mechanisms involved. These results lay a foundational basis for exploring the potential roles of other members within the CMTM family in male reproduction.

Background: The Asian warty newt is a rare amphibian species endemic to China and is listed as a second-class national protected wild animal. This study observes the spermiogenesis and sperm ultrastructure of the Asian warty newt and explores the relationship between structure and function, elucidating the male reproductive biology of this rare amphibian at the cellular and molecular levels. Methods: The collected samples were fixed in 2.5% glutaraldehyde and stored at 4°C. The corresponding electron microscope samples were prepared respectively. The spermiogenesis and ultrastructure changes of spermatids were observed by transmission electron microscopy (TEM) and the morphology and structure of sperms were studied by scanning electron microscopy (SEM). Result: During spermiogenesis, the morphology and structure of spermatids undergo significant changes, ultimately forming mature sperm with highly condensed chromatin, an elongated nucleus and structures such as the acrosome and tail. The sperm ultrastructure of the Asian warty newt is divided into the head, neck and tail: the head is slender with an acrosome hook; the neck is short and not prominent; and the tail is divided into the middle piece, principal piece and end piece. The study found that certain structural features of the sperm are related to its fertilization method and reproductive habits and its morphological structure shares some common characteristics with the sperm of the Salamandridae family.

In the present study, we developed an unsolvable behavioral metacognitive task for kindergarten children. The task was designed to gain insight into how children’s metacognitive processes, measured as monitoring (e.g., checking the plan) and control behavior (e.g., seeking a piece), operate in a problem-solving task that mimics real-life scenarios. Five to six-year-old kindergarten children (N = 72) were asked to build a wooden snake according to a plan. The middle piece of the snake (fourth out of seven pieces) was missing, making the task unsolvable. Other than expected, metacognitive behavior was not related to teacher ratings of metacognitive self-regulation. However, we found age differences. Children in kindergarten year two (M = 5.85 years old) showed more control behavior than children in kindergarten year one (M = 5.05 years old). Surprisingly, we did not find age differences in monitoring behavior. Lastly, we found that metacognitive behavior differed between the solvable part (before the missing piece is reached) and the unsolvable part (after the missing piece is reached). Children showed more monitoring and less control behavior in the solvable part than in the unsolvable part. The current study contributes to the metacognitive research methodology by capturing children’s metacognitive processes in action using an ecological-valid, unsolvable behavioral task.

Tubulin polymerization-promoting protein2 (TPPP2) is one of the three paralogs of mammalian TPPP proteins. Its possible role in spermatogenesis is described in this narrative review. TPPP2 is expressed specifically in the male reproductive system, mainly in testes and sperm, and also in the epididymis. In testes, TPPP2 is exclusively expressed in elongating spermatids; in the epididymis, it is located in the middle piece of the sperm tail. TPPP2 is involved in spermiogenesis, in steps which are determinative for the formation and morphology of spermatids. The inhibition of TPPP2 decreases sperm motility (the curvilinear velocity of sperms), probably due to influencing mitochondrial energy production since TPPP2 knockout mice possess an impaired mitochondrial structure. There are data on the role of TPPP2 in various mammalian species: human, mouse, swine, and various ruminants; there is a significant homology among TPPP2s from different species. Experiments with Tppp2-/--mice show that the absence of TPPP2 results in decreased sperm count and serious dysfunction of sperm, including decreased motility; however, the in vitro capacitation and acrosome reaction are not influenced. The symptoms show that Tppp2-/--mice may be considered as a model for oligoasthenozoospermia.

Abstract Plasmaspheric hiss waves are important to shape the Earth’s electron radiation belt. These waves are commonly envisioned to have a long lifetime which allows them to permeate the global plasmasphere from a spatially restricted source. However, this hypothesis has not been experimentally confirmed yet, because of the challenging observational requirements in terms of location and timing. With wave and particle measurements from five magnetospheric satellites and detailed modeling, we present the first report of long lifetime (∼42 s) hiss rays in the substorm‐disturbed plasmasphere. The low‐frequency hiss waves are found to originate from the middle piece of the plasmaspheric plume, bounce between two hemispheres, and eventually drift into the plasmaspheric core. These hiss rays can travel through ∼3 hr magnetic local time and ∼4 magnetic shell. Such a long‐time and large‐scale permeation of hiss rays could benefit from the ducting process by plasmaspheric field‐aligned density irregularities.

Chromium (Cr) is a vital micronutrient for sugar metabolism while its unauthorized use may agonize health fatalities. This study was conducted about the novelty about exposure of hexavalent chromium (Cr+6) histopatholgical potentials along with Mulberry Fruit Extract and Jamun Fruit Extract to define the ameliorative aptitude. The histological finding in Cr exposure specify; cessation of testicular seminiferous tubules (ST), annihilation of smooth muscles in basement membrane, Leydig’s cell and spermatids. The ST have halted spermatogenesis, with irregular boundary and dead spermatogonia and Leydig’s cells are disorganized. The sperm head cross sectional area (CSA), tail length and middle piece diameter (p ≤ 0.001) significantly reduced while there is significant elevation of hepatocytes nuclei size, central vein CSA and mean width of Sinusoidal Spaces as compared to control. The protuberant marks of steatosis, fibrosis, dehydration and atrophy were nullified by designated fruit extracts and their possible bio-chelating effects was obvious due to the presence of anthocyanin, β-sitosterol and phytochemicals. The given results specify that hexavalent Cr induce andro-hepatic anomalies when it is freely used without scientific authentication. Cr+6 as food additives in coloring rice and local sweets should be prohibited while bio-products of Morus and Jamun can be recommended and sponsored for traditional medicines.

György Harag worked several times as a guest director in Yugoslavia and between 1978 and 1982 staged three of Chekhov’s plays at the Novi Sad Theatre, referred to by the critics as his Chekhov trilogy. Harag’s production of The Cherry Orchard, which premiered in 1979 as the middle piece of the trilogy – between Three sisters and Uncle Vanya – showed a modern, tragicomic interpretation of the dramatic text in a non-traditional visual setting. The production, which was delayed for several years, received an overwhelmingly positive critical reception from critics both in Yugoslavia and in Hungary and is regarded as one of the most defying productions in the history of the Novi Sad Theatre that helped to shape the profile of the institution. Using the Philther method, this study aims to analyse and reconstruct the production.

Simple SummaryBull fertility greatly impacts the profitability of the cattle production system. Therefore, in vivo semen analysis is an option to predict the potential fertility of the sire. The results showed an effect of the mating period on bull sperm morphometric and displayed spermatozoon with a more linear and faster progressiveness. In addition, we demonstrated an overall decrease in overtime on sperm morphometrical traits of the sperm head and midpiece. Therefore, the results obtained help us understand how the mating period affects a bull’s sperm quality while opening the possibility for future research in other species.The objective of the present study was to determine the effects that the reproductive season has on the motility, kinematics, morphology, and sperm morphometry of Brahman bulls evaluated with a commercial CASA system. The experiment was carried out at the Costa Rica Institute of Technology from March to August 2021. A total of eight Brahman bulls were used. A total of 28 ejaculates were collected in the pre-mating period (PMP), during it (DMP), and after it (AMP) using an electroejaculator. The sperm concentration was measured with the Accuread photometer. The motility was measured using a Spermtrack® counting chamber. The analyses were performed with the CASA-Mot ISAS®v1 system. The morphology was analyzed using a microscope with a negative phase contrast objective. Morphometry was evaluated with the CASA-Morph. The sperm concentration did not present differences between the PMP and AMP; however, it was significantly higher than DMP (p > 0.05). Regarding the progressiveness variables, linearity on forward progression (LIN), straightness (STR), and wobble (WOB) were higher (p < 0.05) DMP. A kinematic principal component analysis grouped all the variables into three factors and an effect on the reproductive period was found (p < 0.05) in the parameters of the head and middle part of the sperm, such as width and perimeter, which were greater in the PMP. The length of the sperm head in the PMP and DMP did not show differences; however, both were larger (p < 0.05) than AMP. The insertion distance of the middle piece of the sperm was significantly greater than DMP. Finally, the PMP contained cells with a larger insertion angle (p < 0.05) than AMP. These findings are important to understand the implications of reproductive status on sperm quality and to consider them in andrological evaluations.

We present a new technique that allows an intraocular lens to be explanted through the small incisions used in modern cataract surgery. The intraocular lens optic is cut into three connected pieces at the 1-mm-wide end with scissors. Then, with the stabilizing counterforce provided by a pair of vitreoretinal forceps through a paracentesis, the middle piece is removed first, followed by the two side pieces connected with haptics flipped over at the connected part. These two parts overlap each other when passing through the incision, eventually resulting in the explantation of the intraocular lens, as an intact piece. We believe this method provides a simple and effective way to remove intraocular lens through very small incisions, which could also reduce complications and hasten patient's recovery.

The aim of the research was to determine the morphometry comparison of spermatozoa of simental cattle and ongole cattle such as head length, head width, head area, middle piece length, tail length and overall body length. This research was carried out at the UPTD Laboratory of the Livestock Breeding Center, Horticulture and Animal Husbandry Food Crops Service, Jambi Province. This methodology uses exploration and observation of spermatozoa, using a microscope with a magnification of 400 times. The samples measured were 480 spermatozoa, taken from frozen semen randomly at the Department of Horticultural Food Crops and Livestock, Jambi Province, then smear preparations were made with eosin dye. The variables observed were head length (PK), head width (LK), head area (LKP), middle piece length (PMD), tail length (PE) and overall body length (PTK). The research results showed that the morphometry of simental cattle spermatozoa PK (9.46 µm), LK (5.11 µm), LKP (41.96 µm), MD (13.77 µm), PE (47.68 µm) and PTK ( 70.92 µm). Meanwhile, the size of the spermatozoa of Ongole cattle PK (9.48 µm), LK (5.17 µm), LKP (42.57 µm), MD (13.83 µm), PE (47.96 µm) and PTK (71.28 µm); The statistical results obtained were not significantly different in the morphology of the two cow spermatozoa. The relationship between PTK parameters of simental cattle spermatozoa and PE was very significant (r=0.870) and likewise for Ongole Cow Spermatozoa, the relationship between PTK and PE was very significant (r=0.953). This situation shows that there is a very close relationship between body length and tail length. The percentage of X chromosome simental cattle spermatozoa was (44.38%) and Y chromosome spermatozoa was (55.62%), while the percentage of X chromosome Ongole cattle spermatozoa was (45.83) and Y chromosome carrying spermatozoa was (54.17).

Sperm morphology is considered a species-specific character and has been used as a tool in the classification of numerous mammalian taxa. Neotropical bats have been poorly studied, and important aspects on sperm morphology have not been elucidated. The aim of the present study was to describe and compare the sperm morphology and morphometry of Molossus molossus and Molossops temminckii. A total of 14 adults specimens were analyzed from the Colección Mamíferos Lillo, Universidad Nacional de Tucumán: five M. molossus and nine M. temminckii. The epididymis were extracted and macerated in Farmer's solution, followed by a coloration with different stains. To carry out the description and morphometric analysis, microphotographs were taken under an optical, epifluorescence, and scanning electron microscope (SEM). A total of 50 sperm from each individual were measured for morphometric analysis. The length and width of the head, midpiece and tail were taken as variables. Sperm from M. molossus and M. temminckii were practically identical, both morphologically and morphometrically. In both species, a distal bulge was observed at the end of the intermediate piece in a percentage greater than 85%. The main characteristics shared between the species were: presence of acrosomal blebs in the upper half of the head of the spermatozoa; cephalic equatorial segment with filiform ornamentations; intermembrane space of head apex wedge-shaped; helical middle piece and annulus at the end of middle piece. In the present study, SEM allowed us to visualize structures, such as acrosomal vesicles, that were not detected with other types of microscopy. RESEARCH HIGHLIGHTS: The similarities in the sperm morphology between M. molossus and M. temminckii were observed with three types of microscopy: optical, epifluorescence and scanning electron, and supported by morphometric and statistical analyses.

Objective: To verify that urine collectors are not toxic to sperm. Methods: The toxicity of glass (control) and plastic (urine collector) containers with semen samples of good concentration and sperm motility was evaluated by studying sperm motility at intervals of 1 hour, at room temperature, for 4 hours, following guidelines of the fifth edition of the World Health Organization Manual for the processing of semen samples. The progressive motility between both types of containers (glass and plastic) was compared and the morphotintorial characteristics of the spermatozoa were evaluated with the modified fluorescence differential stain. Results: No statistically significant differences (p = 0.334) were found between the groups. Spermatozoa that emitted a green fluorescence were categorized as normal and presented a strong correlation (r = 1,000; p = 0.000); Whereas, yellow (r = -0.838, p = 0.009) and orange (r = 0.940, p = 0.000) stains were highly correlated with sperm abnormalities, highlighting that orange fluorescence had a high correlation (r = 0.940, p = 0.000) with head abnormalities. Conclusion: The plastic collector has no toxic effects on sperm, it satisfies the needs of collection of the semen sample. With modified fluorescence differential staining, green staining is associated with morphologically normal spermatozoa and yellow and orange stains are associated with abnormalities of the head, middle piece, and main piece of sperm.

ABSTRACT Nesfatin-1, a polypeptide hormone derived from the nucleobindin 2 (NUCB2) precursor protein, is known to regulate appetite and energy metabolism. Recent studies have also shown that NUCB2/nesfatin-1 is expressed in the reproductive organs of mice. However, the expression and potential role of NUCB2/nesfatin-1 in the mouse epididymis remain unclear. Therefore, we investigated the expression of NUCB2/nesfatin-1 in the mouse epididymis and its potential function. NUCB2/nesfatin-1 was detected in the epididymis by qRT-PCR and western blotting, and high expression levels were observed in epididymal epithelial cells by immunohistochemical staining. Pregnant mare’s serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) injections significantly increased NUCB2/nesfatin-1 expression in the epididymis. After castration, NUCB2/nesfatin-1 expression in the epididymis decreased, but was significantly increased by testosterone injection. Nesfatin-1-binding sites were found in the middle piece of testicular sperm, but were scarcely detected in the sperm head. By contrast, nesfatin-1 binding sites were identified on the sperm head within the epididymis. Furthermore, nesfatin-1 treatment inhibited the acrosome reaction in epididymal sperm. These results suggest that the nesfatin-1 protein produced in the epididymis binds to nesfatin-1 binding sites on the sperm head and plays a role in suppressing the acrosome reaction before ejaculation.

Pharmacological intervention of biosynthesized Nigella sativa silver nanoparticles against hexavalent chromium induced toxicity in male albino mice

Methamphetamine (METH) can induce spermatogenesis impairment, testicular apoptosis, and abnormal sperm quality. It also promotes changes in the expression of receptors for sex hormones and neurotransmitters, including GABA receptors in the testis. Proteomic assessment focusing on proteins involved in the calcium signalling pathway in the testis can facilitate diagnostic factors contributing to testicular and sperm functions, especially those related to spermatogenesis and fertilisation. In this study, we proposed to determine the localisation and differential expression of GABA A receptor alpha 1 subunit (GABA A-α1) in the spermatozoa of METH-administered rats. The differential proteomic profile of the testis was also observed by focusing on proteins in the KEGG pathways belonging to the calcium signalling pathway. There were 212 differentially expressed proteins in the rat testis, based on the cut-off value of 1.2-fold change. Most of those proteins, 13 proteins, were classified in the calcium signalling pathway, including 4 down-regulated and 9 up-regulated proteins. An immunolocalisation study of the GABA A-α1 receptor and calbindin revealed their localisation in the equatorial segment of the head in the rat spermatozoa. The expression of calbindin is also found in the middle piece of sperm. An increase in GABA A-α1 receptor in rat spermatozoa was correlated with an increase in abnormal sperm motility and morphology after methamphetamine exposure. Moreover, calbindin expression in sperm decreased in METH-administered rats. All our findings demonstrate that METH influences intracellular calcium homeostasis by acting through the calcium signalling pathway-associated proteins. Moreover, it might disrupt ion homeostasis in sperm through the GABA A-α1 receptor and calbindin, triggering a change in intracellular calcium and chloride ions. These changes may cause abnormalities in spermatogenesis, testicular apoptosis, and sperm quality impairment.

Background: Sperm morphology is the major benchmark in assessment of male infertility. There are limited studies available on frequency of qualitative defects of sperm in patients with normal sperm count. Hence, the present study was done to assess the morphological defects in patients with normal sperm counts who came for infertility work up. Aims and Objectives: The aim of the study was to evaluate the correlation of qualitative defects of sperm in patients having normal sperm counts. Materials and Methods: We have collected 70 samples with normal sperm count (≥15 million/mL). After routine sperm count and motility, Smears were fixed in ethanol and stained with Papanicolaou stain. Minimum of 200 spermatozoa were analyzed per sample. We have recorded normal sperm morphology and abnormal morphology and we have further categorized the data. Morphology was categorized as defect in head, middle piece and tail. Results: Most common morphological abnormalities detected in head region were double head (60%) followed by tapering head (42%). Most common defect observed in middle piece region was thick middle piece (57%) followed by bent neck (48%). Most frequent defect in tail region was coiled tail (14%). We have recorded 40 cases (57%) out of 70 showed multiple morphological defects. Conclusion: Qualitative defects in sperm play a key role in infertility workup. We must have to acknowledge the importance of morphological assessment of semen.

Spermatogenesis is a complex process that includes spermatogonia self-renewal, spermatocyte meiosis and spermatozoa assembly. Recent studies have revealed that WD40-repeat domain-containing (WDR) proteins play important roles in spermatocyte division, spermatozoa flagella assembly and head shaping. In this study, we investigated the expression pattern of WDR87 and found that it was highly expressed in the testis of both humans and mice. Immunofluorescence staining revealed that mouse WDR87 was distributed in the perinuclear cytoplasm of primary spermatocytes, secondary spermatocytes and round spermatids. In the spermiogenesis stage, with extension of the nucleus, WDR87 migrated to the manchette and finally localized to the middle piece of the spermatozoa tail. Furthermore, we identified a cilia- and flagella-associated protein, CFAP47, which interacted with WDR87 in the flagellar midpiece of the spermatozoa, suggesting that WDR87 may be associated with multiple morphological abnormalities of the flagella (MMAF). Subsequently, we screened gene mutations in seven MMAF individuals and found two novel mutations in CFAP47 (c.706G>A, Val236Met; c.1337C>T, Thr446Met) in one case. Immunoblotting and immunofluorescence revealed that CFAP47 was dramatically reduced in spermatozoa from the CFAP47-mutated man. Meanwhile, the expression of WDR87 was also significantly decreased, and weak signals were detected adjacent to the spermatozoa nuclei, indicating that CFAP47 was necessary for WDR87 transportation during spermatozoa flagella biogenesis. These data indicate that WDR87 is located in the middle piece of the sperm tail and interacts with CFAP47 to form a complex which is involved in spermatozoa tail assembly.