Genetic Variation in Toll-Like Receptors (TLRs) 2, 4, and 9 Influences HIV Disease Progression Toward Active TB and AIDS.

Background: Toll-like receptors (TLRs) are identified as one of the key components of the innate immune system. The objective of this study was to explore the influence of genetic variability in these TLRs on human immunodeficiency virus (HIV) disease progression with and without tuberculosis (TB) co-infection. Materials and Methods: This prospective, cross-sectional, and longitudinal study included 373 HIV-positive patients without TB infection. This study aimed to examine the genetic variation in TLRs (TLR2, TLR4, and TLR9) between patients with HIV-1 infection and those who progressed to active TB during the two years of follow-up. Results: During the two year follow-up of 373 positive patients, 98 patients progressed to active TB/AIDS (acquired immunodeficiency syndrome). When comparing 98 HIV patients who developed active TB/AIDS to 275 HIV patients who did not, it was discovered that the frequency of the A allele in TLR9 was considerably higher (p <0.001) in HIV patients progressed to active TB/AIDS. Ninety eight HIV individuals who advanced to active TB/AIDS showed a significantly higher frequency of the AA genotype in TLR9 than did in HIV patients who had no TB/AIDS (p <0.001). Conclusion: The increased association of the AA genotype of TLR9 in HIV patients who progressed to active TB during follow-up suggests that HIV-positive patients with the AA genotype of TLR9 have increased susceptibility towards TB during the disease progression.

Flap Closure in Neonatal Neural Tube Defects: Challenges in Flap Design and Lessons Learnt.

Background: Management of neural tube defects (NTDs) is challenging and the outcome is demanding. Aims: To analyze the outcomes in operated cases of NTDs closed using various types of flaps. Materials and Methods: The data between June 2017 and May 2023 were analyzed. The mode of presentation, timing of intervention, type of flap, neurological status after closure, status of the wound, presence of hydrocephalous, flap blackening, flap necrosis, features of sepsis, and the outcome were recorded and analyzed. Covered NTD; closure done using primary closure or 'Z' Plasty (everywhere); incomplete data; lost to follow-up; and not giving consent were excluded from the study. Results: Out of 92 cases, 35 were operated using the rhomboid flap, 33 using dufourmentel modification of limberg flap, and 24 using keystone island flap. The mean age at presentation was 4 days (range: 0-28 days). The mean duration of surgery after presentation was 2 days (range: 1-3 days). Mean operating time was 1.15 h (range: 0.45-3.15 h). A ventriculoperitoneal shunt was required in 62 cases at various stages. The preoperative and the postoperative power were nearly the same in all. Wound infection was seen in 2, 3, and 1 cases in each group. Blackening of the flap was seen in 3, 2, and 1 cases in three groups. Cerebrospinal fluid (CSF) leak was seen in 2, 2, and 0 cases. Wound dehiscence was present in one case in each group and sepsis was present in 2, 3, and 2, respectively. Conclusion: The management of open NTD requires adequate planning. CSF shunting and flap closure are often required.

Glans Anthropometry for Preputial Reconstruction in Hypospadias.

Background: Cases of hypospadias present for poor stream or cosmetic appearance. The main aim is to provide a visibly normal phallus. Preputial reconstruction is technical. A properly planned reconstruction based on anthropometry may improve the result. We are presenting our experience of reconstruction based on glans anthropometry. Aim: The aim of the study was to evaluate the importance of glans anthropometry in preputial reconstruction in cases of hypospadias. Materials and Methods: All cases of hypospadias operated between June 2014 and March 2022 were included. Glans width was measured at the base. The marking sutures for preputial reconstruction were taken at distance thrice the glans width at base. Those requiring religious circumcision along with repair, associated significant chordee, catheter came out before 2 weeks, or history of any previous penile surgery were excluded. All the cases were subjected to urethroplasty, meatoplasty, and preputioplasty. The results obtained were analyzed. Results: One hundred and forty-eight out of 159 cases formed the study group. There were 31 glanular, 42 distal penile, 58 mid-penile, and 17 proximal penile hypospadias. Mean glans width at base was 16 mm (range: 11-21 mm). Mean distance of marking suture at prepuce was 38 mm (range: 33-63 mm). Mean follow-up was 12 months (range: 1-36 months). Mean age at presentation was 23 months (range: 14-72 months). Mean operating time was 50 min (range: 45-60 min). Fistula at the base of preputioplasty was seen in four. Dehiscence of preputioplasty was seen in six. Meatal stenosis was seen in three cases. Conclusion: Preputial reconstruction improves the cosmetic appearance of the hypospadiac penis. Reconstruction based on glans anthropometry improves the result and avoids complications.

Comparison of the Efficacy of Amniotic Membrane Versus Buccal Fat Pad in Treatment of Oral Submucous Fibrosis.

Background: Oral and maxillofacial surgery deals with wide range of oral defects, wound closure, tissue resection, and tissue reconstruction. The purpose of our study is to use amniotic membrane for closure of post surgery defect in patient of oral submucous fibrosis to utilize its growth factor and scaffold nature for effective healing and to evaluate effectiveness of amniotic membrane in treatment outcome. The objectives are to compare post-operative mouth opening, healing of amniotic membrane and buccal fat pad. Material and Method: Diagnosed patients with OSMF are divided into two surgical site Group I (n = 5patients)-Left side buccal mucosa in which resection of fibrous band with coronoidectomy followed by reconstruction of the mucosal defect with BFP. Group II-Right side buccal mucosa in which resection of fibrous band with coronoidectomy followed by reconstruction of the mucosal defect with freeze dried irradiated amniotic membrane. Result: This study suggested that in comparison to buccal fat pad flap, the HAM graft is a better option for oral reconstruction in terms of infection, graft failure, MMO, inflammation, pain. Outcome indicated that the HAM is biologically ideal graft for oral wounds and could be used as clinical alternative for various repair surgery for oral defects. Conclusion: The amniotic membrane was found easy to handle and easy to use with inherent hemostatic property which is observed in all patients. No patients had shown any evidence of any complications. Good pain control observed in patients throughout postoperative period.

Identification and characterization of the Cyamopsis tetragonoloba transcription factor MYC (CtMYC) under drought stress.

The MYC transcription factor (TF) has a variety of roles in abiotic stress responses of plants. In the present work, MYC TF named CtMYC (Cymopsis tetragonoloba) from guar plant, which is induced by drought stress, was identified. The mature leaves of guar were employed to detect the full-length CtMYC TF on the 8th day of drought stress. The CtMYC gene showed tissue-specific expression and up regulated under drought stress conditions as compared to the control and maximum expression was observed in mature leaves. Additionally, CtMYC TF was cloned and expressed in E. coli Rosetta cells and CtMYC protein was purified. The circular dichroism (CD) analysis revealed the presence of helical content and beta sheets and in the presence of genomic DNA the conformational changes were observed in secondary structure, which showed DNA binding potential of CtMYC. These results were analyzed by CD and fluorescence studies. In silico studies reveal the presence of conserved bHLH domain and DNA-binding amino acid residues His, Glu and Arg in CtMYC. This is first report on CtMYC TF with DNA binding potential that is responsive to drought. This study provides the structure and characterization of CtMYC TF and DNA binding ability in drought tolerance mechanism in guar.

Adverse childhood experiences and maternal self-efficacy: Examining the mediating role of intimate partner violence and the moderating role of caste membership in rural India.

Maternal self-efficacy (MSE) is associated with healthy functioning in mothers and children globally. Maternal exposure to adverse childhood experiences (ACEs) and intimate partner violence (IPV) is known to negatively impact MSE in high-income countries; however, the association has not been examined in low-and-middle-income countries, such as India, which face socioeconomic risks including poverty, illiteracy, and discrimination based on caste membership. The present study examines the mediating role of IPV in the association between ACEs (specifically-emotional, physical, and sexual abuse, neglect, household dysfunction, and discrimination) and MSE and tests caste membership as a moderator. A community-based, cross-sectional survey was performed with 316 mothers with at least one child between 0 and 24 months in a rural area in the North Indian state of Uttar Pradesh. A structural equation framework was used to test the moderated-mediation model. Results from the moderated-mediation model indicate that greater ACEs exposure was associated with lower MSE and this association was mediated by IPV exposure for low-caste but not high-caste mothers, even after controlling for wealth and literacy. These findings add to existing evidence on ACEs exposure as a significant burden for rural Indian mothers, negatively impacting parenting outcomes such as MSE. The critical role of caste membership is also highlighted, providing implications for future research.

La autoeficacia materna (MSE) se asocia con el saludable funcionamiento en las madres y niños globalmente. Se conoce que el hecho de que la madre haya estado expuesta a experiencias adversas en la niñez (ACE) y a la violencia de la pareja íntima (IPV) tiene un negativo impacto en MSE en países de altas entradas económicas; sin embargo, esta asociación no se ha examinado en países donde las entradas económicas son bajas o medias, como India, que enfrenta riesgos socioeconómicos entre los que se incluyen la pobreza, el analfabetismo, así como la discriminación basada en la pertenencia a una casta. El presente estudio examina el papel mediador de IPV en la asociación entre ACE (específicamente - el abuso emocional, físico y sexual, negligencia, disfuncionalidad en el hogar y discriminación) y MSE, y pone a prueba la pertenencia a una casta como aspecto moderador. Se llevó a cabo una encuesta de base comunitaria e inter-seccional con 316 madres con por lo menos un niño entre 0 y 24 meses de edad en un área rural en el estado de Uttar Pradesh en el norte de India. Se usó un marco de trabajo de ecuación estructural para examinar el modelo de moderación y mediación. Los resultados del modelo de moderación y mediación indican que una mayor exposición a ACE estaba asociada con una más baja MSE y que la exposición a IPV mediaba esa asociación para madres de castas bajas, pero no para madres de castas altas, aun después del factor control de recursos económicos y alfabetismo. Estos resultados contribuyen a la existente evidencia de que el haber estado expuesta a ACE es una carga significativa para las madres en la India rural, la cual tiene un impacto negativo en los resultados de crianza tales como MSE. También se subraya el papel esencial de la pertenencia a una casta, lo cual aporta implicaciones para la investigación futura.

L'auto-efficacité maternelle (MSE en anglais) est globalement liée à un fonctionnement sain chez les mères et les enfants. L'exposition maternelle à des expériences de l'enfance adverses (ACE en anglais) et à la violence intime ou conjugale (IPV) est connue comme impactant de manière négative l'auto-efficacité maternelle dans les pays à revenus élevés. Cependant ce lien n'a pas toujours été examiné dans les pays à faibles ou moyens revenus, tels que l'Inde qui fait face à des risques socioéconomiques qui comportent la pauvreté, l'illettrisme, la discrimination en fonction de l'appartenance à une caste. Cette étude examine le rôle médiateur de la violence conjugale (ou violence entre partenaires intimes) dans le lien entre les ACE (plus spécifiquement l'abus émotionnel, physiques, sexuel, la négligence, la dysfonction au sein du foyer et la discrimination) et l'auto-efficacité maternelle et les tests d'appartenance à une caste en tant que modérateurs. Un questionnaire communautaire, en coupe transversale, a été présenté à 116 mères ayant au moins un enfant entre l'âge de 0-24 mois dans une région rurale de l'état du nord de l'Inde, Uttar Pradesh. Un cadre d'équation structurelle a été utilisé pour tester le modèle de modération-médiation. Les résultats de ce modèle de modération-médiation indiquent que plus l'exposition aux ACE est grande, plus l'auto-efficacité maternelle est basse et cette association est affectée par l'exposition à la violence conjugale pour les castes moins élevées mais pas pour les mères des castes plus élevées, même en effectuant un contrôle pour la richesse et l'alphabétisation. Ces résultats s'ajoutent aux preuves existantes sur l'exposition aux ACE en tant que poids important pour les mères indienne de milieux ruraux, ce qui impacte de manière négative les résultats de parentage telles que l'auto-efficacité maternelle. Le rôle critique de l'appartenance à une caste est également mis en lumière, offrant des implications pour les recherches à venir.

MMP modulated differentiation of mouse embryonic stem cells on engineered cell derived matrices.

Stem cell differentiation is dictated by the dynamic crosstalk between cells and their underlying extracellular matrix. While the importance of matrix degradation mediated by enzymes such as matrix metalloproteinases (MMPs) in the context of cancer invasion is well established, the role of MMPs in stem cell differentiation remains relatively unexplored. Here we address this question by assaying MMP expression and activity during differentiation of mouse embryonic stem cells (mESCs) on mouse embryonic fibroblast (MEF) derived matrices (MEFDMs) of varying stiffness and composition. We show that mESC differentiation into different germ layers is associated with expression of several MMPs including MMP-11, 2, 17, 25 and 9, with MMP-9 detected in cell secreted media. Different extents of softening of the different MEFDMs led to altered integrin expression, activated distinct mechanotransduction and metabolic pathways, and induced expression of germ layer-specific markers. Inhibition of MMP proteolytic activity by the broad spectrum MMP inhibitor GM6001 led to alterations in germ layer commitment of the differentiating mESCs. Together, our results illustrate the effect of MMPs in regulating mESC differentiation on engineered cell derived matrices and establish MEFDMs as suitable substrates for understanding molecular mechanisms regulating stem cell development and for regenerative medicine applications.

Modulation of E-Cadherin and N-Cadherin by ovarian steroids and embryonic stimuli.

Endometrium is a dynamic tissue that undergoes extensive remodelling to attain a receptive state which is further modulated in presence of an embryo for successful initiation of pregnancy. Cadherins are the proteins of the junctional complex of which E-cadherin (E-Cad) is crucial for maintaining epithelial cell state and integrity of the epithelial barrier; gain of N-cadherin (N-Cad) in epithelial cells leads to epithelial to mesenchymal transition (EMT). In the present study, we investigated the expression of E-Cad and N-Cad in the mouse endometrial luminal epithelium and its modulation by estrogen, progesterone, and embryonic stimuli. We observed that E-Cad is diffusely expressed in the luminal epithelium of mouse endometrium during the estrus stage and upon estrogen treatment. It is apico-laterally and basolaterally sorted at the diestrus stage and in response to the combined treatment of estrogen and progesterone. In 3D spheroids of human endometrial epithelial cells, combined treatment with estrogen and progesterone led to lateral sorting of E-Cad without any effects on its mRNA levels. at the time of embryo implantation, there is loss of E-Cad along with the gain of N-Cad and SNAIL expression suggestive of EMT in the luminal epithelium. This EMT is possibly driven by embryonic stimuli as treatment with estrogen and progesterone did not lead to the gain of N-Cad expression in the mouse endometrium in vivo or in human endometrial epithelial cells in vitro. In conclusion, the present study demonstrates that steroid hormones directly affect E-Cad sorting in the endometrial epithelium which undergo EMT in response to embryonic stimuli.

Immunoinformatics approach for a novel multi-epitope subunit vaccine design against various subtypes of Influenza A virus.

Vaccination is the best strategy for the control and prevention of contagious diseases caused by Influenza A viruses. Extraordinary genetic variability and continual evolvability are responsible for the virus having survival and adaptation to host cell immune response, thus rendering the current influenza vaccines with suboptimal effectiveness.Therefore, in the present study, using a novel immunoinformatics approach, we have designed a universal influenza subunit vaccine based on the highly conserved epitopic sequences of rapidly evolving (HA), a moderately evolving (NP) and slow evolving (M1) proteins of the virus. The vaccine design includes 2 peptide adjuvants, 26 CTL epitopes, 9 HTL epitopes, and 7 linear BCL epitopes to induce innate, cellular, and humoral immune responses against Influenza A viruses. We also analyzed the physicochemical properties of the designed construct to validate its thermodynamic stability, hydrophilicity, PI, antigenicity, and allergenicity. Furthermore, we predicted a highly stable tertiary model of the designed subunit vaccine, wherein additional disulfide bonds were incorporated to enhance its stability. The molecular docking and molecular dynamics simulations of the refined vaccine model with TLR3, TLR7, TLR8, MHC-I and MHC-II showed stable vaccine and receptors complexes, thus confirming the immunogenicity of the designed vaccine. Collectively, these findings suggest that our multi-epitope vaccine construct may confer protection against various strains of influenza A virus subtypes, which could prevent the need for annual reformulation of vaccine and alleviate disease burden.

TLR-2 expression and dysregulated human Treg/Th17 phenotype in Aspergillus flavus infected patients of chronic rhinosinusitis with nasal polyposis.

BACKGROUND: T helper (Th)17 and regulatory T (Treg) cells with toll-like receptor (TLR)-2 have been acknowledged to play a critical role in chronic rhinosinusitis with nasal polyposis (CRSwNP). However, its pathogenesis has been perplexed by conflicting reports on the role of Th17/Treg cells in patients of distinct ethnicities. We attempted to understand the role of Th responses induced during host defense against Aspergillus flavus. RESULTS: The percentages of Th17 (CD4+CD161+IL23R+) and Treg (CD4+CD25+FoxP3+) cell populations and various cytokine profiles in peripheral blood mononuclear cells (PBMCs) challenged by A. flavus antigens were characterized from 50 CRSwNP cases, before and after treatment, and in 50 healthy controls. TLR-2 expression was analyzed in tissues of cases and controls for disease co-relation. The major pathogen identified in our study was A. flavus by mycological investigations. A marked immune imbalance was noted with elevated Th17 and decreased Tregs in PBMCs of CRSwNP patients after A. flavus stimulation. Comparatively, interleukin (IL)-17 and IL-10 levels were increased, with low transforming growth factor (TGF)-ß levels in A. flavus stimulated PBMC supernatants of patients. The mRNA expression of TLR-2 in polyps of CRSwNP patients indicated significant (p = 0.001) upregulation in comparison to the controls. CONCLUSIONS: Our data highlights the excessive expression of TLR-2 in nasal polyps contributing to the imbalance in Th17/Tregs population in patients. After therapy, recovery of Tregs cells indicates restoration and tissue homeostasis, though high circulating CD4+CD161+ Th17 cells may continue to be a threat to patients predisposed to future recurrences. The constant exposure and tendency of A. flavus to colonize nasal cavities can lead to a Th17 driven airway inflammation. Dysregulated Th17 with TLR-2 promote resistance to treatment and progression to the chronicity of the disease.

Upregulation of miR-101 during Influenza A Virus Infection Abrogates Viral Life Cycle by Targeting mTOR Pathway.

Micro RNAs (miRNAs) are a class of small non-coding single-stranded RNA, which play an important role in modulating host-Influenza A virus (IAV) crosstalk. The interplay between influenza and miRNA interaction is defined by a plethora of complex mechanisms, which are not fully understood yet. Here, we demonstrate that in IAV infected A549 cells, a synchronous increase was observed in the expression of mTOR up to 24 hpi and significant downregulation at 48 hpi. Additionally, NP of IAV interacts with mTOR and modulates the levels of mTOR mRNA and protein, thus regulating the translation of host cell. RNA sequencing and qPCR analysis of IAV-infected A549 cells and NP transfected cells revealed that miR-101 downregulates mTOR transcripts at later stages of infection. Ectopic expression of miR-101 mimic led to a decrease in expression of NP, a reduction in IAV titer and replication. Moreover, treatment of the cells with Everolimus, a potent inhibitor of mTOR, resulted in an increase of miR-101 transcript levels, which further suppressed the viral protein synthesis. Collectively, the data suggest a novel mechanism that IAV stimulates mTOR pathway at early stages of infection; however, at a later time-point, positive regulation of miR-101 restrains the mTOR expression, and hence, the viral propagation.

Improved uptake and therapeutic intervention of curcumin via designing binary lipid nanoparticulate formulation for oral delivery in inflammatory bowel disorder.

This study was focussed on development of curcumin loaded solid binary lipid nanoparticles (C-SBLNs) to ameliorate stability, uptake and therapeutic potential of curcumin during inflammatory bowel disease (IBD). C-SBLNs with nano-size range (210.56 ± 41.22 nm) and high entrapment efficiency (83.12 ± 6.57%) were prepared by solvent emulsification evaporation method using binary lipids i.e. stearic acid and tristearin after optimizing various formulation and process variables. Physicochemical characterization of C-SBLNs by ATR-FTIR confirmed drug entrapment whereas thermal and pXRD study corroborated loss of crystallinity of drug into C-SBLNs. Lyophilized C-SBLNs were found to be spherical shaped with good gastrointestinal stability and prolonged drug release up to 24 h. Optimized C-SBLNs formulation displayed significantly enhanced cellular uptake and localization in inflamed tissues during IBD. Oral administration of C-SBLNs in DSS induced colitis model revealed significant reduction in leucocyte infiltration, oxidative stress, pro-inflammatory cytokine (TNF-α) secretion and maintenance of colonic structure similar to healthy animal group compared to curcumin. Thus, in vitro and preclinical findings of study clearly confirmed that C-SBLNs could be a stable and efficacious alternative platform for curcumin delivery with strong competence in IBD chemotherapy.

Transanastomotic tube in intestinal atresia: How beneficial are they?

INTRODUCTION: Intestinal atresia requires multiple surgeries and long hospital stay. We tried managing these cases by primary anastomosis with transanastomotic tube (TAT) for early feeding. AIMS: The aim of the study was to analyse the outcomes in patients of intestinal atresia who underwent primary anastomosis with a TAT. MATERIALS AND METHODS: The records between June 2014 and November 2017 were analysed. Those with incomplete data or unclear final outcome were excluded. Patients managed by primary anastomosis with TAT (Group A) or without TAT (Group B) were included. The TAT was kept for 6 weeks. Oral feeds were started after 2 weeks in all the cases. P < 0.05 was considered as statistically significant. RESULTS: Forty-eight cases were included. There were two duodenal atresia, 29 jejunal atresia and 17 ileal atresia. The mean age at surgery was 2 days (range: 1-16 days). There were 42 cases in Group A (with TAT) and six in Group B (without TAT). The average duration of start of feeds was 78 h (range: 72-96 h) in Group A and 402 h (range: 360-504 h) in Group B (P = 0.01). The mean duration of hospital stay was 7 days (range: 5-15 days) and 27 days (range: 19-48 days) in Group A and B, respectively (P = 0.02). The overall survival was 38 (91%) and 3 (50%) in Group A and B, respectively (P = 0.01). Reexploration was required in 2/42 and 2/6 cases in Group A and B, respectively (P = 0.4). Total parental nutrition was required in 2/42 and all cases in Group A and B, respectively. CONCLUSION: Primary repair in intestinal atresia with a TAT is a practical option. The overall outcome is better.

Identification of a novel homozygous mutation in transmembrane channel like 1 (TMC1) gene, one of the second-tier hearing loss genes after GJB2 in India.

BACKGROUND & OBJECTIVES: Hearing impairment is a common and heterogeneous sensory disorder in humans. Among about 90 genes, which are known to be associated with hearing impairment, mutations in the GJB2 (gap junction protein beta 2) gene are the most prevalent in individuals with hereditary hearing loss. Contribution of the other deafness-causing genes is relatively poorly understood. Here, we present our findings on two families with transmembrane channel like 1 (TMC1) gene variants of the 47 families with nonsyndromic hearing loss (NSHL) studied. METHODS: Forty seven families including 26 consanguineous families with at least two hearing impaired children and one normal hearing child and 21 non-consanguineous families having at least three hearing impaired children and one normal hearing child were enrolled for this study. Genetic linkage studies were carried out in 41 families that were GJB2 (Connexin 26) negative. Seven polymorphic short tandem repeat markers at the DFNB7/11 locus were studied employing fluorescently labelled markers. RESULTS: A novel homozygous missense mutation c.1283C>A (p.Ala428Asp) was identified co-segregating with hearing loss. This change results in substitution of a highly conserved polar alanine to a charged aspartic acid and is predicted to be deleterious. In addition, a previously reported nonsense mutation, p.R34X in TMC1, was found. INTERPRETATION & CONCLUSIONS: While mutations in TMC1 are not as common a cause of NSHL as those in GJB2, TMC1 should be considered for diagnostic investigations in cases of NSHL in GJB2-negative families.

Decidual Control of Trophoblast Invasion.

At the time of implantation, the trophoblast cells of the embryo adhere and then invade into the maternal endometrium and eventually establish placentation. The endometrium at the same time undergoes decidualization, which is essential for successful pregnancy. While the NK cells of the decidua have been implicated to play a key role in trophoblast invasion, few evidence are now available to demonstrate a pro-invasive property of decidual stromal cells. Secretions from decidualized endometrial stromal cells promote invasion of primary trophoblasts and model cell lines by activating proteases and altering expression of adhesion-related molecules. The decidual secretions contain high amounts of pro-invasive factors that include IL-1ß, IL-5, IL-6, IL-7, IL-8, IL-9, IL-13, IL-15, Eotaxin CCL11, IP-10 and RANTES, and anti-invasive factors IL-10, IL-12 and VEGF. It appears that these decidual factors promote invasion by regulating the protease pathways and integrin expression utilizing the STAT pathways in the trophoblast cells. At the same time the decidua also seem to secrete some anti-invasive factors that are antagonist to the matrix metalloproteinases and/or are activators of tissue inhibitors of metalloproteinases. This might be essential to neutralize the effects of the invasion-promoting factors and restrain overinvasion. It is tempting to propose that during the course of pregnancy, the decidua must balance the production of these pro and anti-invasive molecules and such harmonizing production would allow a timely and regulated invasion.

Flow behaviour of gellan sol with selected cations.

An understanding of the flow behaviour of the sols before gel formation is important for developing nutrient enriched gels. The influence of cations like CaCl2 (0.05 and 0.1 %, w/w) and FeSO4 (0.05 and 0.1 %, w/w) on the rheological properties of 1 % gellan sol (w/w) prior to gelling was investigated. The apparent viscosity, reported at a shear-rate of 100 s(-1), indicated that the gellan dispersion without any cation possessed lower values compared to other samples containing different cations. The Cross model provided the best fit (0.97 ≤ r ≤ 0.99, p ≤ 0.01) compared to moderate fitting to power law model (0.94 ≤ r ≤ 0.98). Among the different Cross model parameters, the zero-shear viscosity (ηo) increased with the addition of CaCl2 and FeSO4, and with an increase in their concentrations. Zero-shear viscosity values were 0.46 Pas for gellan sol, 0.79 Pas for gellan with 0.05 % (w/w) CaCl2, 1.41 Pas for gellan with 0.1 % CaCl2, 3.85 Pas for gellan with 0.05 % FeSO4 and 4.33 Pas for gellan with 0.1 % FeSO4. An increase in cation concentration from 0.05 to 0.10 % (w/w) marginally increased the relaxation time (λ) values indicating the development of more solid characteristics in the sol.

Influenza A viral nucleoprotein interacts with cytoskeleton scaffolding protein α-actinin-4 for viral replication.

Influenza A virus (IAV), similar to other viruses, exploits the machinery of human host cells for its survival and replication. We identified α-actinin-4, a host cytoskeletal protein, as an interacting partner of IAV nucleoprotein (NP). We confirmed this interaction using co-immunoprecipitation studies, first in a coupled in vitro transcription-translation assay and then in cells either transiently co-expressing the two proteins or infected with whole IAV. Importantly, the NP-actinin-4 interaction was observed in several IAV subtypes, including the 2009 H1N1 pandemic virus. Moreover, immunofluorescence studies revealed that both NP and actinin-4 co-localized largely around the nucleus and also in the cytoplasmic region of virus-infected A549 cells. Silencing of actinin-4 expression resulted in not only a significant decrease in NP, M2 and NS1 viral protein expression, but also a reduction of both NP mRNA and viral RNA levels, as well as viral titers, 24 h post-infection with IAV, suggesting that actinin-4 was critical for viral replication. Furthermore, actinin-4 depletion reduced the amount of NP localized in the nucleus. Treatment of infected cells with wortmannin, a known inhibitor of actinin-4, led to a decrease in NP mRNA levels and also caused the nuclear retention of NP, further strengthening our previous observations. Taken together, the results of the present study indicate that actinin-4, a novel interacting partner of IAV NP, plays a crucial role in viral replication and this interaction may participate in nuclear localization of NP and/or viral ribonucleoproteins.

LIMD1 antagonizes E2F1 activity and cell cycle progression by enhancing Rb function in cancer cells.

Tumour suppressor genes restrain inappropriate cell growth and division, as well as stimulate cell death to maintain tissue homeostasis. Loss of function leads to abnormal cellular behaviour, including hyperproliferation of cell and perturbation of cell cycle regulation. LIMD1 is a tumour suppressor gene located at chromosome 3p21.3, a region commonly deleted in many solid malignancies. LIMD1 interacts with retinoblastoma (Rb) and is involved in Rb-mediated downregulation of E2F1-target genes. However, the role of LIMD1 in cell cycle regulation remains unclear. We propose that LIMD1 induces cell cycle arrest, utilising Rb-E2F1 axis, and show that ectopic expression of LIMD1 in A549 cells results in hypo-phosphorylation that potentiates Rb function, which correlates with downregulation of E2F1. In agreement with these observations, LIMD1 overexpression retards cell cycle progression and blocks S-phase entry, as cells accumulate in G0/G1 phase and have reduced incorporation of BrdU. Most significantly, LIMD1-dependent effects on Rb function and cell cycle are reversed on depletion of endogenous LIMD1, underscoring its centrality in Rb-mediated cell cycle regulation. Hence, our findings provide new insight into cell cycle control by Rb-LIMD1 nexus.

Norrie disease: first mutation report and prenatal diagnosis in an Indian family.

Norrie Disease (ND) is a rare X-linked recessive disorder characterised by congenital blindness due to severe retinal dysgenesis. Hearing loss and intellectual disability is present in 30-50 % cases. ND is caused by mutations in the NDP gene, located at Xp11.3. The authors describe mutation analysis of a proband with ND and subsequently prenatal diagnosis. Sequence analysis of the NDP gene revealed a hemizygous missense mutation arginine to serine in codon 41 (p.Arg41Ser) in the affected child. Mother was carrier for the mutation. In a subsequent di-chorionic di-amniotic pregnancy, the authors performed prenatal diagnosis by mutation analysis on chorionic villi sample at 11 wk of gestation. The fetuses were unaffected. This is a first mutation report and prenatal diagnosis of a familial case of Norrie disease from India. The importance of genetic testing of Norrie disease for confirmation, carrier testing, prenatal diagnosis and genetic counseling is emphasized.