Glycosylation Effects on FSH-FSHR Interaction Dynamics: A Case Study of Different FSH Glycoforms by Molecular Dynamics Simulations

The gonadotropin known as follicle-stimulating hormone (FSH) plays a key role in regulating reproductive processes. Physiologically active FSH is a glycoprotein that can accommodate glycans on up to four asparagine residues, including two sites in the FSHα subunit that are critical for biochemical function, plus two sites in the β subunit, whose differential glycosylation states appear to correspond to physiologically distinct functions. Some degree of FSHβ hypo-glycosylation seems to confer advantages toward reproductive fertility of child-bearing females. In order to identify possible mechanistic underpinnings for this physiological difference we have pursued computationally intensive molecular dynamics simulations on complexes between the high affinity site of the gonadal FSH receptor (FSHR) and several FSH glycoforms including fully-glycosylated (FSH²⁴), hypo-glycosylated (e.g., FSH¹⁵), and completely deglycosylated FSH (dgFSH). These simulations suggest that deviations in FSH/FSHR binding profile as a function of glycosylation state are modest when FSH is adorned with only small glycans, such as single N-acetylglucosamine residues. However, substantial qualitative differences emerge between FSH¹⁵ and FSH²⁴ when FSH is decorated with a much larger, tetra-antennary glycan. Specifically, the FSHR complex with hypo-glycosylated FSH¹⁵ is observed to undergo a significant conformational shift after 5–10 ns of simulation, indicating that FSH¹⁵ has greater conformational flexibility than FSH²⁴ which may explain the more favorable FSH¹⁵ kinetic profile. FSH¹⁵ also exhibits a stronger binding free energy, due in large part to formation of closer and more persistent salt-bridges with FSHR.     

Fibroblasts Influence Survival and Therapeutic Response in a 3D Co-Culture Model

In recent years, evidence has indicated that the tumor microenvironment (TME) plays a significant role in tumor progression. Fibroblasts represent an abundant cell population in the TME and produce several growth factors and cytokines. Fibroblasts generate a suitable niche for tumor cell survival and metastasis under the influence of interactions between fibroblasts and tumor cells. Investigating these interactions requires suitable experimental systems to understand the cross-talk involved. Most in vitro experimental systems use 2D cell culture and trans-well assays to study these interactions even though these paradigms poorly represent the tumor, in which direct cell-cell contacts in 3D spaces naturally occur. Investigating these interactions in vivo is of limited value due to problems regarding the challenges caused by the species-specificity of many molecules. Thus, it is essential to use in vitro models in which human fibroblasts are co-cultured with tumor cells to understand their interactions. Here, we developed a 3D co-culture model that enables direct cell-cell contacts between pancreatic, breast and or lung tumor cells and human fibroblasts/ or tumor-associated fibroblasts (TAFs). We found that co-culturing with fibroblasts/TAFs increases the proliferation in of several types of cancer cells. We also observed that co-culture induces differential expression of soluble factors in a cancer type-specific manner. Treatment with blocking antibodies against selected factors or their receptors resulted in the inhibition of cancer cell proliferation in the co-cultures. Using our co-culture model, we further revealed that TAFs can influence the response to therapeutic agents in vitro. We suggest that this model can be reliably used as a tool to investigate the interactions between a tumor and the TME.     

Reproductive performance of carp, Catla catla (Ham.), reared on a formulated diet with PUFA supplementation

Effect of dietary supplementation of n‐3 and n‐6 polyunsaturated fatty acids (PUFA) on reproductive performances in both sexes of an Indian major carp, Catla catla (Ham.), were investigated in terms of sperm quality, spawning response, fecundity and spawn recovery over a consecutive 2‐year period. Sperm count and spermatocrit value were in the range of 3.2–3.8 × 10⁷ ml⁻¹ and 80–82%, respectively, in fish fed the PUFA‐enriched test diet, and significantly higher (P < 0.01) than in fish fed control diet (2.3–2.5 × 10⁷ ml⁻¹ and 64–70%). Spawning response in the test diet group was quite high (96%) compared to 76% in the control. Egg and larval quality were superior in the test diet group as evidenced by the significantly higher fertilization rate and larval survival in the range of 91–92% and 93.3%, respectively. Fatty acid profile of fertilized eggs and larvae showed that the quantities of 18:3 (n‐3), 20:5 (n‐3) and 22:6 (n‐3) were significantly (P < 0.01) increased in the test group while 18:1, 18:2 (n‐6) and 20:4 (n‐6) remained at higher levels in control. The results indicated that enrichment of n‐3 PUFA in the broodfish diet not only improved the quality of eggs and larvae in terms of recovery, but also enhanced the physiological response of broodfish to induced spawning. Oil enrichment in the broodfish diet should therefore be mandatory to ensure the optimum n‐3 PUFA content for warmwater carp for quality seed production.     

Expression profiling of the genes responding to zearalenone and its analogues using estrogen-responsive genes

To compare gene expression profiles in response to estrogen or 17β-estradiol (E₂) and a mycotoxin, zearalenone (ZEA), and its analogues (collectively termed ZEA compounds), breast cancer MCF-7 cells were treated with 10 nM of E₂ or ZEA compounds including ZEA, α-zearalenol, β-zearalenol, zearalanone, α-zearalanol and β-zearalanol. Expression profiles for 120 estrogen-responsive genes were subjected to cluster and statistical analyses using correlation coefficients or R-values. We found that all of the ZEA compounds stimulated the growth of MCF-7 cells, as much as E₂, and showed similar expression profiles to that of E₂ (R-values ranged from 0.82 to 0.96). The effect of ZEA compounds was likely mediated by estrogen-receptor-dependent Erk1/2-signaling. These results provide clues to understand the mechanism of their estrogen-like action.     

Isolation and characterization of microsatellite loci in giant freshwater prawn, Macrobrachium rosenbergii

This paper reports on isolation and characterization of 8 microsatellite loci from a partial genomic library of Macrobrachium rosenbergii using DIG labeled dinucleotides, (GT)₁₅ and (CT)₁₅ as probes. Primers were designed and PCR conditions optimized for repeat arrays. Polymorphism was studied using 24 individuals collected from the wild. All the loci were polymorphic with number of alleles ranging from 3 to 5 and observed heterozygosity 0.50–0.85. All the loci except one were in agreement with Hardy–Weinberg equilibrium. No significant pair wise linkage disequilibrium was found among the loci. These markers should prove useful for characterization of natural population as well as brood stock management of this species.     

Metabolic alternations of some amino acids,coenzymes, phytohormones and vitamins in chickpea crop grown from seeds soaked with defense stimulator

The metabolite profile of targeted amino acids, coenzymes, phytohormones and vitamins was evaluated in chickpea (Cicer arietinum L.) crop grown from seeds soaked with defense stimulator salicylic acid, benzothiadiazole or nicotinic acid (0.0, 10.0 or 20.0 µg/mL). The concentrations of analytes were determined at regular intervals covering five critical time points of crop growth. Liquid chromatography was used for the estimation of the test metabolites. In both leaf and root, the treatments progressively enhanced the biosynthesis of phenylalanine (Phe) and tyrosine (Tyr), oxidized nicotinamide adenine dinucleotide (NAD), reduced nicotinamide adenine dinucleotide phosphate (NADPH), pyridoxine, folic acid, riboflavin and rutin (vitamin P). The concentration of tryptophan (Trp), nicotinamide adenine dinucleotide phosphate (NADP), l-ascorbic acid (L-AA), niacin, thiamin (THI), β-carotene (vitamin A) and α-tocopherol (vitamin E) decreased. In leaf, the level of gibberellic acid (GA₃) was enhanced and of menadione (vitamin K₃) reduced. In root, the level of GA₃ was reduced and of vitamin K₃ increased. Consequent with the depletion of Trp level in plant, the levels of NAD increased whereas, those of master growth regulator indole-3-acetic acid and its precursor indole-3-butyric acid declined. With this, cytokinin level also reduced. NAD regulated the ratio of NAD: reduced form of NAD (NADH) which was less than that of NADP:NADPH. Tyr, Phe and Trp were the canonical variables for the observed metabolics. A strong correlation between the declining metabolite levels of vitamin E, L-AA, Trp, NADH, THI and vitamin K₃ in leaf; and vitamin E, GA₃, THI and vitamin A in root firstly attributed stress tolerance in chickpea agro-system.     

Pollution and Intimacy in a Transcendent Ethics of Care: A Case of Aged-care in India

In India, where children’s care of ageing parents is seen as practical and sacred, animated by notions of seva (selfless service), the outsourcing of elder care causes considerable concern. Meanwhile, carers’ work in old-age homes is treated as transactional, and their moral claims about this work are either overlooked or criticised. While gendered, socio-economic circumstances compel the women we discuss in this paper to care-work at an old-age home in Pune, they also understood this work as a register for the spiritual striving normally reserved for higher classes and castes. Accordingly, notions of polluting and non-polluting bodily waste inform the sense of kin-like intimacy through which they frame their labour. Attending to the institutional, spiritual, emotional and bodily registers of these carers’ work, we argue for a transcendent ethics of care, a conceptualisation that contributes to broader understandings of marginalisation and moral imagination in an ordinary ethics of care.     

Glyceraldehyde 3-P dehydrogenase, glycerate 3-P kinase and enolase mutants of Escherichia coli: Genetic studies

Summary The genetic loci for two enzymes of glycolysis have been established by transductional crosses. The eno locus, likely to be the structural gene for enolase maps in the 52-minute region of the Escherichia coli chromosome. A structural determinant for glycerate 3-P kinase (pgk) is located near serA. The map order is speB-pgk–serA-lysA-argA-eno-cysC. In the 35-minute region maps a locus affecting the structure of glyceraldehyde 3-P dehydrogenase.     

Oncocytic carcinoid tumor of the lung: a case report of diagnostic pitfall in filter membrane preparation of bronchial washings

BACKGROUND: Oncocytic carcinoid tumor of the lung is a rare variant of pulmonary carcinoid. This report describes the morphologic appearance of this rare tumor on filter membrane preparation along with potential pitfalls. CASE: A 49-year-old woman presented with cough and expectoration. On chest radiograph a mass lesion was seen in the upper zone of the right lung. Bronchial washings were sent for evaluation. On filter membrane (Millipore) preparation of bronchial washing the possibility of a non-small cell carcinoma, possibly squamous, was suggested. Right upper lobectomy was subsequently performed and a histologic diagnosis of oncocytic carcinoid given. The cytomorphologic features of this tumor on the Millipore preparation were reviewed. CONCLUSION: Differential diagnosis of oncocytic carcinoid should be kept in mind while assessing cytologic material when tumor cells show abundant granular cytoplasm and prominent nucleoli. Oncocytic carcinoid also must be differentiated from oncocytoma and granular cell tumor. Immunocytochemistry and electron microscopy are useful in confirming the diagnosis.