Endocrine disruptors: Unravelling the link between chemical exposure and Women's reproductive health.

An Endocrine Disrupting Chemical (EDC) is any compound that disrupts the function of the endocrine system in humans and is ubiquitous in the environment either as a result of natural events or through anthropogenic activities. Bisphenol A, phthalates, parabens, pesticides, triclosan, polychlorinated biphenyls, and heavy metals, which are frequently found in the pharmaceutical, cosmetic, and packaging sectors, are some of the major sources of EDC pollutants. EDCs have been identified to have a deteriorating effect on the female reproductive system, as evidenced by the increasing number of reproductive disorders such as endometriosis, uterine fibroids, polycystic ovary syndrome, premature ovarian failure, menstrual irregularity, menarche, and infertility. Studying EDCs in relation to women's health is essential for understanding the complex interactions between environmental factors and health outcomes. It enables the development of strategies to mitigate risks, protect reproductive and overall health, and inform public policy decisions to safeguard women's well-being. Healthcare professionals must know the possible dangers of EDC exposure and ask about environmental exposures while evaluating patients. This may result in more precise diagnosis and personalized treatment regimens. This review summarises the existing understanding of prevalent EDCs that impact women's health and involvement in female reproductive dysfunction and underscores the need for more research. Further insights on potential mechanisms of action of EDCs on female has been emphasized in the article. We also discuss the role of nutritional intervention in reducing the effect of EDCs on women's reproductive health. EDC pollution can be further reduced by adhering to strict regulations prohibiting the release of estrogenic substances into the environment.

Pigmented skin exhibits accelerated wound healing compared to the nonpigmented skin in Guinea pig model.

This study investigated and compared the wound healing kinetics of pigmented (PG) and non-pigmented (NP) skin in guinea pigs, focusing on histological and transcriptional changes. Full-thickness wounds created on PG and NP skin were evaluated at various time points post-injury. Fontana-Masson staining and ultrastructural analysis suggested the presence of melanin and melanosomes in PG skin, which coincided with an upregulation of melanogenic genes cKIT, TYR, and DCT. On day 9 post-wound, PG skin exhibited a rapid transition from the inflammatory to proliferative phase, which correlated with the reappearance of epidermal pigmentation whereas the NP skin exhibited a delayed neo-epidermis formation. Furthermore, the study revealed that melanocyte-derived growth factors (conditioned media) positively regulated keratinocyte migration while inhibiting fibroblast differentiation. These effects were more prominent in tyrosine-treated (hyperpigmented) melanocyte-CM as was TGF- ß expression. These findings provide valuable insights into the mechanisms underlying skin repair and pigmentation.

Molecular mechanism of Enteroaggregative Escherichia coli induced apoptosis in cultured human intestinal epithelial cells.

Background. Enteroaggregative Escherichia coli (EAEC) is an evolving etiological agent of acute and persistent diarrhoea worldwide. The previous study from our laboratory has reported the apoptosis-inducing activity of EAEC in human small intestinal and colonic epithelial cell lines. In the present investigation, we have explored the underlying mechanism of EAEC-induced apoptosis in human intestinal epithelial cell lines.Methods. INT-407 and HCT-15 cells were infected with EAEC-T8 and EAEC-pT8 (plasmid cured strain of EAEC-T8) separately. Cells cultured in the absence of bacteria served as a negative control in all the experiments. For the subsequent experiments, the molecular mechanism(s) of epithelial cell aposptosis was measured in EAEC infecting both the cell lines by flow cytometry, real-time PCR and Western blotting.Results and conclusions. EAEC was found to activate the intrinsic/mitochondrial apoptotic pathway in both the cell lines through upregulation of pro-apoptotic Bax and Bak, un-alteration/reduction in the level of anti-apoptotic Bcl-2 and Bcl-XL, decrease in mitochondrial transmembrane potential, accumulation of cytosolic cytochrome c leading to activation of procaspase-9 and procaspase-3, which ultimately resulted in DNA fragmentation and apoptosis. Further, an increased expression of Fas, activation of procaspase-8 and upregulation of pro-apoptotic Bid in the EAEC-infected cells indicated the involvement of extrinsic apoptotic pathway too in this process. Our finding has undoubtedly led to an increased understanding of EAEC pathogenesis, which may be helpful to develop an improved strategy to combat the infection.

Cdk5-Mediated Brain Unfolded Protein Response Upregulation Associated with Cognitive Impairments in Type 2 Diabetes and Ameliorative Action of NAC.

The role of cyclin-dependent kinase 5 (Cdk5) in the normal functioning of the central nervous system and synaptic plasticity is well established. However, dysregulated kinase activity can have a significant impact on neurodegeneration and cognitive impairment. Cdk5 hyperactivation is linked to diabetes-associated neurodegeneration, but the underlying mechanism is not fully understood. Our study reveals that oxidative stress can lead to Cdk5 hyperactivity, which in turn is linked to neurodegeneration and cognitive impairment. Specifically, our experiments with N2A cells overexpressing Cdk5 and its activators p35 and p25 show ER stress, resulting in activation of the unfolded protein response (UPR) pathway. We identified Cdk5 as the epicenter of this regulatory process, leading to the activation of the CDK5-IRE1-XBP1 arm of UPR. Moreover, our study demonstrated that Cdk5 hyperactivation can lead to ER stress and activation of the UPR pathway, which may contribute to cognitive impairments associated with diabetes. Our findings also suggest that antioxidants such as NAC and GSH can decrease deregulated Cdk5 kinase activity and rescue cells from UPR-mediated ER stress. The accumulation of phosphorylated Tau protein in AD brain protein has been widely described earlier. Notably, we observed that oral treatment with NAC decreased Cdk5 kinase activity in the hippocampus, attenuated high levels of phospho-tau (ser396), and ameliorated memory and learning impairments in a type 2 diabetic (T2D) mouse model. Additionally, the high-fat-induced T2D model exhibits elevated phospho-tau levels, which are rescued by the NAC treatment. Taken together, these results suggest that targeting Cdk5 may be a promising therapeutic strategy for treating diabetes-associated cognitive impairments.

Mitochondrial calcium signaling mediated transcriptional regulation of keratin filaments is a critical determinant of melanogenesis.

Mitochondria are versatile organelles that regulate several physiological functions. Many mitochondria-controlled processes are driven by mitochondrial Ca2+ signaling. However, role of mitochondrial Ca2+ signaling in melanosome biology remains unknown. Here, we show that pigmentation requires mitochondrial Ca2+ uptake. In vitro gain and loss of function studies demonstrated that Mitochondrial Ca2+ Uniporter (MCU) is crucial for melanogenesis while the MCU rheostats, MCUb and MICU1 negatively control melanogenesis. Zebrafish and mouse models showed that MCU plays a vital role in pigmentation in vivo. Mechanistically, MCU controls activation of transcription factor NFAT2 to induce expression of three keratins (keratin 5, 7 and 8), which we report as positive regulators of melanogenesis. Interestingly, keratin 5 in turn modulates mitochondrial Ca2+ uptake thereby this signaling module acts as a negative feedback loop that fine-tunes both mitochondrial Ca2+ signaling and melanogenesis. Mitoxantrone, an FDA approved drug that inhibits MCU, decreases physiological melanogenesis. Collectively, our data demonstrates a critical role for mitochondrial Ca2+ signaling in vertebrate pigmentation and reveal the therapeutic potential of targeting MCU for clinical management of pigmentary disorders. Given the centrality of mitochondrial Ca2+ signaling and keratin filaments in cellular physiology, this feedback loop may be functional in a variety of other pathophysiological conditions.

Primary pulmonary lymphoepithelial-like carcinoma : A rare childhood malignancy.

Lymphoepithelial-like carcinoma (LELC) usually presents as a head and neck tumor with a close resemblance to nasopharyngeal carcinoma. We present an extremely rare case of Primary Pulmonary lymphoepithelioma in a 14-year-old female patient. The patient presented with a right-sided lung mass, which on biopsy revealed to be a lymphoepithelioma. There was no evidence of any mass elsewhere in the body, including the nasopharynx, as evidenced by PET CT. The IHC was positive for both cytokeratin and lymphoid cell markers. Hence, we conclude that lymphoepitheliomas can present as a primary lung mass in a young nonsmoking female, of which only two case reports are available from the Indian subcontinent till date.

Innovative strategies in algal biomass pretreatment for biohydrogen production.

Biohydrogen is one of the cleanest renewable energies with a high calorific value. Algal biomass can be utilized as a sustainable feedstock for biohydrogen production via dark fermentation. However, the recovery of fermentable sugar from algal biomass is challenging because of the diversity and complex cell wall composition and therefore, requires an additional pretreatment step. However, most of the conventional pretreatment strategies suffer from limited technological feasibility and poor economic viability. In this context, this review aims to present the structural complexities of the cell wall of algae and highlight the innovative approaches such as the use of hybrid technologies, biosurfactants, nanoparticles, and genetic engineering approaches for the hydrolysis of algal biomass and improved biohydrogen production. Additionally, a comprehensive discussion of the comparative evaluation of various pretreatment methods, and the techno-economic and life cycle assessment of algal biohydrogen production is also presented in this review.

Meckel's diverticulum causing acute intestinal obstruction: A case series.

Meckel's diverticulum (MD) is the most common congenital anomaly of the gastrointestinal tract. Most cases are asymptomatic and when symptomatic, preoperative diagnosis of MD is poor. Intestinal obstruction, Malena or hematochezia, and inflammation are major symptoms. We report three cases of 18-month, 2-year, and 9-year old male patients presenting with intestinal obstruction.

Fermentative bioconversion of food waste into biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) using Cupriavidus necator.

Dependency on plastic commodities has led to a recurrent increase in their global production every year. Conventionally, plastic products are derived from fossil fuels, leading to severe environmental concerns. The recent coronavirus disease 2019 pandemic has triggered an increase in medical waste. Conversely, it has disrupted the supply chain of personal protective equipment (PPE). Valorisation of food waste was performed to cultivate C. necator for fermentative production of biopolymer poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV). The increase in biomass, PHBV yield and molar 3-hydroxy valerate (3HV) content was estimated after feeding volatile fatty acids. The fed-batch fermentation strategy reported in this study produced 15.65 ± 0.14 g/L of biomass with 5.32 g/L of PHBV with 50% molar 3HV content. This is a crucial finding, as molar concentration of 3HV can be modulated to suit the specification of biopolymer (film or fabric). The strategy applied in this study addresses the issue of global food waste burden and subsequently generates biopolymer PHBV, turning waste to wealth.

Diclofenac induced anaphylaxis: A case report from Nepal.

Introduction: Diclofenac is considered a generally safe medication, and cause few side effects like dyspepsia, diarrhea/constipation, nausea, and vomiting, stomach bleeding, rash, urticaria, photosensitivity reactions, acute renal failure, analgesic nephropathy, bone marrow and liver diseases. Rarely, it causes anaphylactic reaction. Case presentation: We report a case of 40 years male who developed anaphylactic reaction after intravenous infusion with diclofenac. Discussion: Both the pulmonary embolism and anaphylaxis guidelines emphasize the importance of a thorough evaluation when making a diagnosis. Mast cells secrete tryptase, a neutral protease that is specifically concentrated in the secretory granules of human mast cells (but not basophils), along with histamine, as a marker of mast cell activation. Conclusion: Although intravenous diclofenac sodium is a safe and often used medicine, it can cause severe and perhaps fatal anaphylactic responses.

A case report on atypical spindle cell lipomatous tumor: A rare entity.

Introduction: Atypical spindle cell lipomatous tumor (ASPLT), a separate entity for the group of benign/low grade adipocytic neoplasm that is characterized by adipocyte adequacy and the presence of lipoblast-like cells and spindle cells at varying degrees. Case presentation: Here, we report a rare case of 60 years old male with atypical spindle cell lipomatous tumor in the anterior abdominal wall. Discussion: Histopathology is the gold standard for establishing the diagnosis and grade of soft tissue tumor and consistent radiology-pathology correlation is essential to avoid any diagnostic pitfalls [1]. Ultrasound is preferred as an initial investigation for superficially located lesions. Conclusion: ASPLT show a wide variety of microscopic features, and differential diagnosis is important and difficult. Recognition of morphologic clues and immunohistochemistry/molecular tests to confirm the diagnosis.

Effect of levulinic acid on production of polyhydroxyalkanoates from food waste by Haloferax mediterranei.

Polyhydroxyalkanoates (PHA), especially poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is considered as the most suitable candidate to replace petrochemical plastics. However, the high production cost and the composition of the monomers in the copolymer are the major constraints in production. The 3-hydroxyvalerate (3HV) rich copolymers are ideal for various applications due to their lower melting points, improved elasticity, and ductility. Haloferax mediterranei is a suitable microorganism for the production of biopolymer PHBV from biowaste. Nevertheless, the potential of H. mediterranei cultivated on food waste as sustainable substrate and levulinic acid as an inducer has not been explored for PHBV production. This study aims at the valorization of food waste as low-cost substrate and evaluation of effect of levulinic acid in the production and composition of PHBV using H. mediterranei. Shake-flask fermentations using different concentrations of salt, glucose and levulinic acid were first performed to optimize the cultivation conditions. The highest growth of the halophile was observed at salt concentration of 15% and glucose of concentration 10 g/L. Under optimized growth conditions, H. mediterranei was cultivated for PHBV production in fed-batch bioreactor with pulse fed levulinic acid. The maximum biomass of 3.19 ± 0.66 g/L was achieved after 140 h of cultivation with 3 g/L of levulinic acid. A decrease in H. mediterranei growth was noticed with the increase in levulinic acid concentration in the range of 3-10 g/L. The overall yield of PHBV at 3, 5, 7 and 10 g/L of levulinic acid were 18.23%, 56.70%, 31.54%, 21.29%, respectively. The optimum concentration of 5 g/L of levulinic acid was found to produce the maximum yield of 56.70% PHBV with 18.55 mol% 3HV content. A correlation between levulinic acid concentrations and PHBV production established in this study can serve as an important reference for future large-scale production.

Mitofusin-2 Negatively Regulates Melanogenesis by Modulating Mitochondrial ROS Generation.

Inter-organellar communication is emerging as one of the most crucial regulators of cellular physiology. One of the key regulators of inter-organellar communication is Mitofusin-2 (MFN2). MFN2 is also involved in mediating mitochondrial fusion-fission dynamics. Further, it facilitates mitochondrial crosstalk with the endoplasmic reticulum, lysosomes and melanosomes, which are lysosome-related organelles specialized in melanin synthesis within melanocytes. However, the role of MFN2 in regulating melanocyte-specific cellular function, i.e., melanogenesis, remains poorly understood. Here, using a B16 mouse melanoma cell line and primary human melanocytes, we report that MFN2 negatively regulates melanogenesis. Both the transient and stable knockdown of MFN2 leads to enhanced melanogenesis, which is associated with an increase in the number of mature (stage III and IV) melanosomes and the augmented expression of key melanogenic enzymes. Further, the ectopic expression of MFN2 in MFN2-silenced cells leads to the complete rescue of the phenotype at the cellular and molecular levels. Mechanistically, MFN2-silencing elevates mitochondrial reactive-oxygen-species (ROS) levels which in turn increases melanogenesis. ROS quenching with the antioxidant N-acetyl cysteine (NAC) reverses the MFN2-knockdown-mediated increase in melanogenesis. Moreover, MFN2 expression is significantly lower in the darkly pigmented primary human melanocytes in comparison to lightly pigmented melanocytes, highlighting a potential contribution of lower MFN2 levels to higher physiological pigmentation. Taken together, our work establishes MFN2 as a novel negative regulator of melanogenesis.

Perspective on Constructing Cellulose-Hydrogel-Based Gut-Like Bioreactors for Growth and Delivery of Multiple-Strain Probiotic Bacteria.

The current perspective presents an outlook on developing gut-like bioreactors with immobilized probiotic bacteria using cellulose hydrogels. The innovative concept of using hydrogels to simulate the human gut environment by generating and maintaining pH and oxygen gradients in the gut-like bioreactors is discussed. Fundamentally, this approach presents novel methods of production as well as delivery of multiple strains of probiotics using bioreactors. The relevant existing synthesis methods of cellulose hydrogels are discussed for producing porous hydrogels. Harvesting methods of multiple strains are discussed in the context of encapsulation of probiotic bacteria immobilized on cellulose hydrogels. Furthermore, we also discuss recent advances in using cellulose hydrogels for encapsulation of probiotic bacteria. This perspective also highlights the mechanism of probiotic protection by cellulose hydrogels. Such novel gut-like hydrogel bioreactors will have the potential to simulate the human gut ecosystem in the laboratory and stimulate new research on gut microbiota.

Lesional skin in vitiligo exhibits delayed in vivo reepithelialization compared to the nonlesional skin.

Vitiligo, a common skin disorder, is characterized by the loss of functional melanocytes resulting in the depigmentation of skin. Previous studies have demonstrated molecular and architectural alterations in the epidermal keratinocytes upon loss of melanocytes. The physiological implications of these "altered" keratinocytes are yet not known. We investigated the wound healing efficiency of lesional vs nonlesional skin in 12 subjects with stable nonsegmental vitiligo using histological and ultrastructural evaluation of partial-thickness wounds. The wounds were examined 12 days postinjury, coinciding with the reepithelialization phase of healing marked primarily by keratinocyte migration and proliferation. This study demonstrated a significant difference in the reepithelialization potential between the lesional and nonlesional skin. While all 12 nonlesional wounds demonstrated considerable neoepidermis formation on the 12th day post wound, only four of the corresponding lesional samples showed comparable reepithelialization; the rest remaining in the inflammatory phase. Ultrastructural studies using transmission electron microscopy as well as immunohistochemical staining revealed a reduced number of desmosomes, shorter keratin tonofilaments and an increase in myofibroblast population in the dermis of lesional reepithelialized tissue compared to the nonlesional reepithelialized samples. This study implicates gross functional perturbations in the lesional skin during physiological wound healing in vitiligo, suggesting that the breakdown of keratinocyte-melanocyte network results in delayed wound repair kinetics in the lesional skin when compared to patient-matched nonlesional skin.

ETS1 and PAX5 transcription factors recruit AID to Igh DNA.

B lymphocytes optimize antibody responses by class switch recombination (CSR), which changes the expressed constant region exon of the immunoglobulin heavy chain (IgH), and by somatic hypermutation (SH) that introduces point mutations in the variable regions of the antibody genes. Activation-induced cytidine deaminase (AID) is the key mutagenic enzyme that initiates both these antibody diversification processes by deaminating cytosine to uracil. Here we asked the question if transcription factors can mediate the specific targeting of the antibody diversification by recruiting AID. We have recently reported that AID is together with the transcription factors E2A, PAX5 and IRF4 in a complex on key sequences of the Igh locus. Here we report that also ETS1 is together with AID in this complex on key sequences of the Igh locus in splenic B cells of mice. Furthermore, we show that both ETS1 and PAX5 can directly recruit AID to DNA sequences from the Igh locus with the specific binding site for the transcription factor. Taken together, our findings support the notion of a targeting mechanism for the selective diversification of antibody genes with limited genome wide mutagenesis by recruitment of AID by PAX5 and ETS1 in a transcription factor complex.

Regulation of the DNA Repair Complex during Somatic Hypermutation and Class-Switch Recombination.

B lymphocytes optimize Ab responses by somatic hypermutation (SH), which introduces point mutations in the variable regions of the Ab genes and by class-switch recombination (CSR), which changes the expressed C region exon of the IgH. These Ab diversification processes are initiated by the deaminating enzyme activation-induced cytidine deaminase followed by many DNA repair enzymes, ultimately leading to deletions and a high mutation rate in the Ab genes, whereas DNA lesions made by activation-induced cytidine deaminase are repaired with low error rate on most other genes. This indicates an advanced regulation of DNA repair. In this study, we show that initiation of Ab diversification in B lymphocytes of mouse spleen leads to formation of a complex between many proteins in DNA repair. We show also that BCR activation, which signals the end of successful SH, reduces interactions between some proteins in the complex and increases other interactions in the complex with varying kinetics. Furthermore, we show increased localization of SH- and CSR-coupled proteins on switch regions of the Igh locus upon initiation of SH/CSR and differential changes in the localization upon BCR signaling, which terminates SH. These findings provide early evidence for a DNA repair complex or complexes that may be of functional significance for carrying out essential roles in SH and/or CSR in B cells.

Qualitative and quantitative metals liberation assessment for characterization of various waste printed circuit boards for recycling.

Metals liberation and composition are decisive attributes in characterization of e-waste for metal recycling. Though end-of-life printed circuit board (PCB) is an integral part of e-waste as secondary resource reservoir, yet no standardized procedure exists for metals liberation and dissolution for its characterization. Thus, the paper aims at assessment of metals liberation upon comminution employing scanning electron microscopy with energy dispersive X-ray spectroscopy (SEM-EDS) followed by comparative assessment of the existing United States Environmental Protection Agency (USEPA) digestion procedures, viz., USEPA 3050B, USEPA 3051A, and USEPA 3052, in effective dissolution of metals from comminuted particles of waste PCBs of computer, laptop, mobile phone, and television. Effect of comminution and digestion conditions was assessed to have significant role in metal liberation and dissolution from PCBs. The SEM-EDS analysis demonstrated partial release of metals from the silica matrix of PCBs. The USEPA digestion methods showed statistically significant (P < 0.05) difference with greater dissolution of metals complexed to PCB matrix by the USEPA 3052 method owing to use of strong acid like hydrofluoric acid. Base metals like Cu and Zn and toxic metals such as Pb and Cd were present in abundance in PCBs and in general exceeded the total threshold limit concentration (TTLC). The maximum contents of Cu (20.13 ± 0.04 wt.%) and Zn (1.89 ± 0.05 wt.%) in laptop PCBs, Pb (2.26 ± 0.08 wt.%) in TV PCBs, and Cd (0.0812 ± 0.0008 wt.%) in computer PCBs were observed.

Comparative assessment of metallurgical recovery of metals from electronic waste with special emphasis on bioleaching.

Waste electrical and electronic equipment (WEEE) or electronic waste (e-waste) is one of the fastest growing waste streams in the urban environment worldwide. The core component of printed circuit board (PCB) in e-waste contains a complex array of metals in rich quantity, some of which are toxic to the environment and all of which are valuable resources. Therefore, the recycling of e-waste is an important aspect not only from the point of waste treatment but also from the recovery of metals for economic growth. Conventional approaches for recovery of metals from e-waste, viz. pyrometallurgical and hydrometallurgical techniques, are rapid and efficient, but cause secondary pollution and economically unviable. Limitations of the conventional techniques have led to a shift towards biometallurgical technique involving microbiological leaching of metals from e-waste in eco-friendly manner. However, optimization of certain biotic and abiotic factors such as microbial species, pH, temperature, nutrients, and aeration rate affect the bioleaching process and can lead to profitable recovery of metals from e-waste. The present review provides a comprehensive assessment on the metallurgical techniques for recovery of metals from e-waste with special emphasis on bioleaching process and the associated factors.