Estrogenic Effect of Bambusa arundinacea, Trichosanthes dioica and Punica granatum on Rats

Background: Phytoestrogens have recently become a hot topic among scientists. Phytoestrogens’ estrogen-like properties have led to their widespread use in the reproductive system. The aim of this research was to see whether the ethanolic extract of Bambusa arundinaceae, Trichosanthes dioica and Punica granatum had any estrogenic activity in female wistar rats. Methods: In female wistar rats, the estrogenic effect was studied using a uterotropic assay, vaginal cytology and vaginal opening. In ovariectomized immature and mature female wistar rats, a 400 mg/kg body weight (b.w.) dose of ethanolic extract of Bambusa arundinaceae, Trichosanthes dioica and Punica granatum was given. Result: When compared to ovariectomized control rats, the uterine wet weight increased significantly. The estrogen-treated rats had only cornified epithelial cells, indicating the existence of oestrogen, as well as 100% vaginal opening. At 400 mg/kg b.w., the ethanolic extract of Bambusa arundinaceae, Trichosanthes dioica and Punica granatum demonstrated promising estrogenic activity, as evidenced by uterotropic assays, vaginal opening measurements and histopathological changes. As a result of this research, it’s possible to infer that the ethanolic extract of Bambusa arundinaceae, Trichosanthes dioica and Punica granatum play an important role in estrogenic activity in female rats.

Targeted deletion of liver-expressed Choriogenin L results in the production of soft eggs and infertility in medaka, Oryzias latipes

Egg envelopes (chorions) in medaka, Oryzias latipes, are composed of three major glycoproteins: ZI-1, − 2, and − 3. These gene-encoded chorion glycoproteins are expressed in the liver and/or ovarian oocytes of sexually mature female fish. In medaka, the glycoproteins produced in the female liver are induced by estrogen as Choriogenin (Chg.) H and Chg. H minor (m), which correspond to the zona pellucida (ZP) B (ZPB) protein in mammals, and Chg. L, which corresponds to ZPC in mammals. Chg. H, Chg. Hm, and Chg. L, are then converted to ZI-1, − 2, and − 3, respectively, during oogenesis in medaka ovaries.In the present study, we established a medaka line in which the chg.l gene was inactivated using the transcription activator-like effector nuclease (TALEN) technique. Neither intact chg.l transcripts nor Chg. L proteins were detected in livers of sexually mature female homozygotes for the mutation (homozygous chg.l knockout: chg.l−/−). The chg.l−/− females spawned string-like materials containing “smashed eggs.” Closer examination revealed the oocytes in the ovaries of chg.l−/− females had thin chorions, particularly at the inner layer, despite a normal growth rate. In comparing chorions from normal (chg.l+/+) and chg.l−/− oocytes, the latter exhibited abnormal architecture in the chorion pore canals through which the oocyte microvilli pass. These microvilli mediate the nutritional exchange between the oocyte and surrounding spaces and promote sperm-egg interactions during fertilization. Thus, following in vitro fertilization, no embryos developed in the artificially inseminated oocytes isolated from chg.l−/− ovaries. These results demonstrated that medaka ZI-3 (Chg.L) is the major component of the inner layer of the chorion, as it supports and maintains the oocyte’s structural shape, enabling it to withstand the pressures exerted against the chorion during spawning, and is essential for successful fertilization. Therefore, gene products of oocyte-specific ZP genes that may be expressed in medaka oocytes cannot compensate for the loss Chg. L function to produce offspring for this species.

Vasoactive intestinal peptide indirectly elicits pituitary LH secretion independent of GnRH in female zebrafish

Vasoactive intestinal peptide (Vip) regulates luteinizing hormone (LH) release through the direct regulation of gonadotropin-releasing hormone (GnRH) neurons at the level of the brain in female rodents. However, little is known regarding the roles of Vip in teleost reproduction. Although GnRH is critical for fertility through the regulation of LH secretion in vertebrates, the exact role of the hypophysiotropic GnRH (GnRH3) in zebrafish is unclear since GnRH3 null fish are reproductively fertile. This phenomenon raises the possibility of a redundant regulatory pathway(s) for LH secretion in zebrafish. Here, we demonstrate that VipA (homologues of mammalian Vip) both inhibits and induces LH secretion in zebrafish. Despite the observation that VipA axons may reach the pituitary proximal pars distalis including LH cells, pituitary incubation with VipA in vitro, and intraperitoneal injection of VipA, did not induce LH secretion and lhβ mRNA expression in sexually mature females, respectively. On the other hand, intracerebroventricular administration of VipA augmented plasma LH levels in both wild type and gnrh3-/- females at 1 hour post-treatment, with no observed changes in pituitary GnRH2 and GnRH3 contents and gnrh3 mRNA levels in the brains. While VipA’s manner of inhibition of LH secretion has yet to be explored, the stimulation seems to occur via a different pathway than GnRH3, dopamine, and E2 in regulating LH secretion. The results indicate that VipA induces LH release possibly by acting with or through a non-GnRH factor(s), providing proof for the existence of functional redundancy of LH release in sexually mature female zebrafish.

Cortical Neurons and Astrocytes in Intrauterine Ethanol Exposure: The Role of Exposure Time and Postnatal Timeline

Background: The state of cortical neurons and astrocytes are pointers to the health of the brain. These cells are morphologically distorted by alcohol exposure. Intrauterine alcohol exposure remains a challenge with perinatal consequences. The role of exposure time and postnatal timeline on the degree of cortical cell derangement remains a subject of controversy till date. This study therefore examines alcohol exposure and postnatal changes on brain weight, cortical neurons and astrocytes at different developmental periods. Methods: Twenty mature female Wistar rats were time-mated and grouped into 4 groups. Group 1 (control) received distilled water (2 mL/kg), Groups 2, 3 and 4 were administered 2.5 mL/kg of 20% ethanol orally on the 4th, 11th and 18th days of gestation respectively. Rats produced litters and pups' brains were harvested and processed for H&E and Golgi Cox stains at the 3rd and 6th postnatal week. Neuronal and astrocytic densities in the cerebral cortex were evaluated. Results and Conclusion: There was statistically significant increase (p<0.05) in the density of degenerating neurons at the third postnatal week and sixth postnatal weeks in the experimental groups when compared with the control. There was also statistically significant increase (p<0.05) in astrocytic density in groups 2 and 4 at the 3rd postnatal week. There was no statistically significant difference (p>0.05) in the astrocytic densities across groups at the sixth postnatal week. It was concluded that intrauterine alcohol exposure in any of the developmental periods resulted in postnatal neuronal degeneration which persisted till the 6th week. However, increased astrocytic densities is a feature of 1st and 3rd trimester alcohol exposure noted in the 3rd but absent in the 6th postnatal week.

Abnormalities in the ovaries of sockeye salmon spawners in the Kamchatka River in 2019

Based on histological analysis of the ovaries of sockeye salmon spawners in the Kamchatka River, numerous abnormalities leading to resorption are shown in the structure of vitellogenic oocytes. Fertility of mature female sockeye salmon decreases due to the resorption of the vitellogenic oocytes. Presence of ovaries with abnormal oocytes implies oocyte resorption in fish being a phenomenon associated with mechanism of natural regulation of final fertility. Such mechanism is required in view of changing fish body length and weight in the sockeye salmon of the Kamchatka River. Numerous abnormalities in the ovaries indicate of a decreasing reproductive capacity of spawners and commercial quality of the eggs.

Enteric methane emission estimates for Kenyan cattle in a nighttime enclosure using a backward Lagrangian Stochastic dispersion technique

This study provides methane (CH4) emission estimates for mature female African beef cattle in a semi-arid region in Southern Kenya using open-path laser spectroscopy together with a backward Lagrangian Stochastic (bLS) dispersion modeling technique. We deployed two open-path lasers to determine 10-min averages of line-integrated CH4 measurements upwind and downwind of fenced enclosures (so-called bomas: a location where the cattle are gathered at night) during 14 nights in September/October 2019. The measurements were filtered for wind direction deviations and friction velocity before the model was applied. We compared the obtained emission factors (EFs) with the Intergovernmental Panel on Climate Change (IPCC) Tier 1 estimates for the Sub-Saharan African (SSA) countries, which were mostly derived from studies carried out in developed countries and adapted to the conditions in Africa. The resulting EF of 75.4 ± 15.99 kg year−1 and the EFs calculated from other studies carried out in Africa indicate the need for the further development of region-specific EFs depending on animal breed, livestock systems, feed quantity, and composition to improve the IPCC Tier 1 estimates.

α2-Macroglobulin-Rich Serum As a Master Inhibitor of Inflammatory Factors Attenuates Cartilage Degeneration in a Mini Pig Model of Osteoarthritis Induced By “Idealized” Anterior Cruciate Ligament Reconstruction

Background: α2-Macroglobulin (α2M) is important for chondral protection in post-traumatic osteoarthritis (PTOA). However, its injection into xenogeneic joint cavities has safety hazards, limiting clinical applications. Exploring serum α2M-enriching strategies and the therapeutic effect and mechanism of α2M-rich serum (α2MRS) autologous joint injection to treat PTOA has significant value.Methods: A unique filtration process was used to concentrate α2M from serum. Human osteoarthritic chondrocytes induced with interleukin (IL)-1β were used to evaluate catabolic enzymes, cell proliferation, apoptosis, and gene expression 24h after α2MRS treatment. Eighteen mature female mini pigs were randomized to three groups, sham (n = 6), “idealized” anterior cruciate ligament autograft reconstruction (IACL-R) (n = 6), and IACL-R+α2MRS (n = 6). Expression of inflammatory factors in the synovial fluid (SF) was measured using Luminex assays. Gait features were recorded using the Tekscan Walkway system. The extent of PTOA progression was evaluated using imaging, real-time PCR , and histology 3 months post-surgery.Results: The α2M concentration in α2MRS was higher than that in human and mini pig serum, respectively. In vitro, α2MRS significantly promoted human chondrocyte proliferation (p < 0.001) and reduced apoptosis (p < 0.001) and chondrocyte catabolic cytokine gene transcription (p < 0.001) and secretion (p < 0.001). In vivo, SF concentrations of all tested inflammatory factors were significantly lower in the IACL-R+α2MRS group than in the IACL-R group (p < 0.001). All gait parameters in the IACL-R+α2MRS group returned to normal significantly early compared to those in the IACL-R group (p < 0.05). Imaging , histology, and biochemistry data showed that cartilage degeneration in the IACL-R+α2MRS group was significantly diminished relative to that in the IACL-R group (p < 0.001).Conclusion: Injecting α2MRS into the joint cavity after IACL-R can significantly delay articular cartilage degeneration.

Molecular investigation and description of Iberozospeum n. gen., including the description of one new species (Eupulmonata, Ellobioidea, Carychiidae)

The subterranean realm of the Cantabrian-Pyrenean region of northern Spain harbours a rich diversity of Zospeum. Due to their tiny size and the difficulty of finding them alive, scarce animal material has been available for scientific investigation. Recent investigations of Zospeum shells have provided valuable, but limited insights towards our understanding of the evolutionary processes occurring within this taxon in northern Spain. In an integrative study, we investigate 57 populations of Zospeum from northern Spanish caves using two mitochondrial (COI and 16S) and two nuclear markers (H3 and 5.8 S rRNA + ITS2). Revealed is a separate radiation of the northern Spanish species for which the new genus, Iberozospeum, is proposed. The independent radiation of Dinaric Zospeum from that of northern Spain justifies the designation of Iberozospeum n. gen. Morphological evidence is provided via histological analysis of Iberozospeum vasconicum and SEM analyses of radulae of eastern Alpine, Dinaric and Iberian species. Important differences in morphological structure and character states are presented, including the first view of the sexually mature female and the presence of the giant albumen gland in an individual of the subterranean, troglobitic Carychiidae. Significant differences are revealed in superficial crystallographic structure of the columellar lamellae, the morphology of the columellar muscle and in the radula. Radular ribbon length, ribbon broadness, straightness of the ribbon base and cusp configuration are distinctive in the Iberian species. One new species is described corroborated by genetic and morphological characters.

Protandric Transcriptomes to Uncover Parts of the Crustacean Sex-Differentiation Puzzle

Hermaphrodite systems offer unique opportunities to study sexual differentiation, due to their high degree of sexual plasticity and to the fact that, unlike gonochoristic systems, the process is not confined to an early developmental stage. In protandric shrimp species, such as Hippolyte inermis and Pandalus platyceros, male differentiation is followed by transformation to femaleness during adulthood. The mechanisms controlling sexual differentiation have not been fully elucidated in crustaceans, but a key role has been attributed to the insulin-like hormone (IAG) produced by the androgenic gland (AG), a crustacean masculine endocrine organ. To uncover further transcriptomic toolkit elements affecting the sexual differentiation of H. inermis, we constructed eye and whole body RNA libraries of four representative stages during its protandric life cycle (immature, male, young female and mature female). The body libraries contained transcripts related to the reproductive system, among others, while the eye libraries contained transcripts related to the X-organ-sinus gland, a central endocrine complex that regulates crustacean reproduction. Binary pattern analysis, performed to mine for genes expressed differentially between the different life stages, yielded 19,605 and 6,175 transcripts with a specific expression pattern in the eye and body, respectively. Prominent sexually biased transcriptomic patterns were recorded for the IAG and vitellogenin genes, representing, respectively, a key factor within the masculine IAG-switch, and a precursor of the yolk protein, typical of feminine reproductive states. These patterns enabled the discovery of novel putative protein-coding transcripts exhibiting sexually biased expression in the H. inermis body and eye transcriptomes of males and females. Homologs to the above novel genes have been found in other decapod crustaceans, and a comparative study, using previously constructed transcriptomic libraries of another protandric shrimp, P. platyceros, showed similar sexually biased results, supporting the notion that such genes, mined from the H. inermis transcriptome, may be universal factors related to reproduction and sexual differentiation and their control in other crustaceans. This study thus demonstrates the potential of transcriptomic studies in protandric species to uncover unexplored layers of the complex crustacean sex-differentiation puzzle.