The structures of the colonic mucosa-associated and luminal microbial communities are distinct and differentially affected by a prolonged murine stressor

The commensal microbiota of the human gastrointestinal tract live in a largely stable community structure, assisting in host physiological and immunological functions. Changes to this structure can be injurious to the health of the host, a concept termed dysbiosis. Psychological stress is a factor that has been implicated in causing dysbiosis, and studies performed by our lab have shown that restraint stress can indeed shift the cecal microbiota structure as well as increase the severity of a colonic infection caused by Citrobacter rodentium. However, this study, like many others, have focused on fecal contents when examining the effect of dysbiosis-causing stimuli (e.g. psychological stress) upon the microbiota. Since the mucosa-associated microbiota have unique properties and functions that can act upon the host, it is important to understand how stressor exposure might affect this niche of bacteria. To begin to understand whether chronic restraint stress changes the mucosa-associated and/or luminal microbiota mice underwent 7 16-hour cycles of restraint stress, and the microbiota of both colonic tissue and fecal contents were analyzed by sequencing using next-gen bacterial tag-encoded FLX amplicon technology (bTEFAP) pyrosequencing. Both control and stress groups had significantly different mucosa-associated and luminal microbiota communities, highlighting the importance of focusing gastrointestinal community structure analysis by microbial niche. Furthermore, restraint stress was able to disrupt both the mucosa-associated and luminally-associated colonic microbiota by shifting the relative abundances of multiple groups of bacteria. Among these changes, there was a significant reduction in the immunomodulatory commensal genus Lactobacillus associated with colonic mucosa. The relative abundance of Lactobacillus spp. was not affected in the lumen. These results indicate that stressor-exposure can have distinct effects upon the colonic microbiota situated at the mucosal epithelium in comparison to the luminal-associated microbiota.     

Resurgence of therapeutically destitute tuberculosis: amalgamation of old and newer techniques

Most of thoracic surgery developed as a result of efforts to treat tuberculosis (TB). The role of surgical therapy has declined but the role of surgery in TB still remains in situations like diagnostic difficulties, persistent sputum positive state despite therapy and complications and sequel like haemoptysis, destroyed or bronchiectatic lungs or empyema with or without broncho-pleural fistula (BPF). Various procedures have a role according to the indication. Some of the procedures have become obsolete but lobectomy, pneumonectomy, thoracoplasty, decortication and open window thoracostomy continue to be relevant. Recent published series have demonstrated mortality ranging from 0% to 3.1%. Surgery for complications and sequel of pulmonary TB still remain an important intervention for alleviation of human misery.KEYWORDS : Thoracic surgery, tuberculosis (TB), surgery, pneumonectomy, thoracoplasty, decortication, window thoracostomy     

RIG-I–like receptor LGP2 protects tumor cells from ionizing radiation

An siRNA screen targeting 89 IFN stimulated genes in 14 different cancer cell lines pointed to the RIG-I (retinoic acid inducible gene I)–like receptor Laboratory of Genetics and Physiology 2 (LGP2) as playing a key role in conferring tumor cell survival following cytotoxic stress induced by ionizing radiation (IR). Studies on the role of LGP2 revealed the following: (i) Depletion of LGP2 in three cancer cell lines resulted in a significant increase in cell death following IR, (ii) ectopic expression of LGP2 in cells increased resistance to IR, and (iii) IR enhanced LGP2 expression in three cell lines tested. Studies designed to define the mechanism by which LGP2 acts point to its role in regulation of IFNβ. Specifically (i) suppression of LGP2 leads to enhanced IFNβ, (ii) cytotoxic effects following IR correlated with expression of IFNβ inasmuch as inhibition of IFNβ by neutralizing antibody conferred resistance to cell death, and (iii) mouse embryonic fibroblasts from IFN receptor 1 knockout mice are radioresistant compared with wild-type mouse embryonic fibroblasts. The role of LGP2 in cancer may be inferred from cumulative data showing elevated levels of LGP2 in cancer cells are associated with more adverse clinical outcomes. Our results indicate that cytotoxic stress exemplified by IR induces IFNβ and enhances the expression of LGP2. Enhanced expression of LGP2 suppresses the IFN stimulated genes associated with cytotoxic stress by turning off the expression of IFNβ.Several studies have shown that the response of tumor cells to ionizing radiation (IR) is associated with IFN-mediated signaling (1–6). IFN signaling leads to the induction of multiple IFN stimulated genes (ISGs) (7, 8) and activates growth arrest and cell death in exposed cell populations (9). The precise mechanism of IR-mediated induction of IFN signaling is unknown. Tumor cell clones that survive an initial cytotoxic insult are subsequently resistant to exposure to both IR and prodeath components of IFN signaling (10). These clones express IFN-dependent enhanced levels of constitutively expressed ISGs, which overlap in part with ISGs initially induced by cytotoxic stress. Many of these constitutively expressed ISGs have been characterized as antiviral genes (11). Recently, enhanced levels of constitutively expressed ISGs have been reported in advanced cancers and were often associated with a poor prognosis related to aggressive tumor growth, metastatic spread, resistance to IR/chemotherapy, or combinations of these factors (11–18). The studies presented in this report are based on the hypothesis that a specific set of constitutively expressed ISGs, whose enhanced expression is by cytotoxic stress, confers a selective advantage to individual tumor clones (5, 9, 10, 13, 19).To test this hypothesis, we designed a targeted siRNA screen against 89 ISGs selected from two sources. The first included ISGs identified in our earlier screen and designated the IFN-Related DNA Damage Signature (IRDS) (1, 13). The second set included related ISG signatures that have been reported in the literature (as described in Methods and Dataset S1). The 89 genes were individually targeted in 14 tumor cell lines derived from malignant gliomas, lung, breast, colon, head and neck, prostate, and bladder cancers.The most significant finding from this screen was that the RNA helicase Laboratory of Genetics and Physiology 2 (LGP2) encoded by DHX58 [DEXH (Asp-Glu-X-His) box polypeptide 58] confers survival and mediates the response to IR of multiple tumor cell lines. LGP2 acts as a suppressor of the RNA-activated cytoplasmic RIG-I RIG-I (retinoic acid inducible gene I)–like receptor’s pathway (20, 21). This pathway is a subtype of pattern recognition receptors responsible for primary recognition of pathogen and host-associated molecular patterns and the subsequent activation of type I IFN production that orchestrates an innate immune response (22–24). In addition to its role in inhibiting IFNβ expression, Suthar et al. recently demonstrated that LGP2 governs CD8+ T-cell fitness and survival by inhibiting death-receptor signaling (25). Here we demonstrate that suppression of LGP2 leads to an enhanced IFNβ expression and increased killing of tumor cells. Our results thereby provide a mechanistic connection between IR-induced cytotoxic response in tumor cells and the LGP2–IFNβ pathway.     

Diabetes and orthopaedic surgery: a review

Diabetes mellitus (DM) is common, estimated to affect 425 million people worldwide in 2017. It is a condition that is continually growing in prevalence and is often associated with multiple co-morbidities. Its multi-system effects on the body mean that its management can pose a challenge, even to more experienced clinicians. In orthopaedic practice, diabetic patients are commonly encountered owing to their increased fracture risk and complications of the disease such as diabetic foot. An appropriate knowledge of diabetes, its pathophysiology, immunology and the pharmacology of medications used in its treatment is essential, as the consequences of mismanagement can be grave. Optimal treatment of diabetics can often require the involvement of a wider multidisciplinary team. Complications that can be encountered in the perioperative and postoperative periods include, diabetic ketoacidosis, hyperosmolar hyperglycaemic state, surgical site infection and venous thromboembolism. This review outlines current concepts in the perioperative management of diabetes and its manifestations within orthopaedic surgery, with a focus on outcomes and complications. A review of the available literature reveals conflicting conclusions between studies, with no clear effect or consensus yet established for many issues. There is a need for a greater number of well-designed, high-quality, appropriately powered trials to establish the true effect of diabetes on outcomes in orthopaedic surgery.     

Case Report: Dropsy Outbreak in a Single Family in Punjab,India

Epidemic dropsy is caused by consumption of mustard oil contaminated with argemone oil. It usually occurs in outbreaks with acute manifestation of bilateral pitting edema, erythema, and local tenderness along with cardiac and respiratory problems in severe cases leading to death. We report an outbreak that is unusual because of its gradual onset, clustering in a single family, and with major manifestation of gastrointestinal illness mimicking acute gastroenteritis, hence leading to delayed diagnosis and high mortality. Thus, the diagnosis of epidemic dropsy should be considered as a strong possibility when there is clustering of cases in a single family with on and off gastrointestinal symptoms of vomiting and diarrhea in a mustard oil consuming belt.     

Redirecting intracellular trafficking and the secretion pattern of FSH dramatically enhances ovarian function in mice

FSH and luteinizing hormone (LH) are secreted constitutively or in pulses, respectively, from pituitary gonadotropes in many vertebrates, and regulate ovarian function. The molecular basis for this evolutionarily conserved gonadotropin-specific secretion pattern is not understood. Here, we show that the carboxyterminal heptapeptide in LH is a gonadotropin-sorting determinant in vivo that directs pulsatile secretion. FSH containing this heptapeptide enters the regulated pathway in gonadotropes of transgenic mice, and is released in response to gonadotropin-releasing hormone, similar to LH. FSH released from the LH secretory pathway rescued ovarian defects in Fshb-null mice as efficiently as constitutively secreted FSH. Interestingly, the rerouted FSH enhanced ovarian follicle survival, caused a dramatic increase in number of ovulations, and prolonged female reproductive lifespan. Furthermore, the rerouted FSH vastly improved the in vivo fertilization competency of eggs, their subsequent development in vitro and when transplanted, the ability to produce offspring. Our study demonstrates the feasibility to fine-tune the target tissue responses by modifying the intracellular trafficking and secretory fate of a pituitary trophic hormone. The approach to interconvert the secretory fate of proteins in vivo has pathophysiological significance, and could explain the etiology of several hormone hyperstimulation and resistance syndromes.During vertebrate evolution, the female reproductive pattern underwent a remarkable transition from spawning of large number of eggs in primitive species under favorable conditions to more tightly controlled ovarian cycles in higher vertebrates, such that only a limited number (rodents) or a single (human and nonhuman primates) egg is released per cycle (1, 2). Coincident with this event, the single pituitary gonadotropic hormone that exists in primitive vertebrates has given rise to two gonadotropins, FSH and luteinizing hormone (LH), which coordinate gametogenesis and steroidogenesis (3–7). FSH and LH are heterodimeric glycoproteins that contain a common α-subunit and a hormone-specific β-subunit (3). Although synthesized in the same cell, the gonadotrope, FSH is mostly constitutively released in many species, whereas LH is stored in dense core granules (DCGs) and secreted in pulses via the regulated pathway in response to gonadotropin-releasing hormone (GnRH) (8, 9). Although this pattern is evolutionarily conserved, how distinct modes of gonadotropin secretion affect ovarian development and target cell responses remain unclear. Although models in which variations in secretion of gonadotropins have been achieved in vivo (10) and in vitro, including basolateral and apically polarized secretion (11), the in vivo consequences of altered mode of gonadotropin trafficking and release (constitutive vs. regulated) from pituitary are untested. We sought to identify why the two gonadotropins, LH and FSH, expressed in the same pituitary cell have evolved to exit via different routes to regulate ovarian physiology.     

Biological activation of bone grafts using injectable platelet-rich fibrin

Platelet-rich fibrin (PRF) is gaining acceptance as a bioactive surgical additive in regenerative dentistry. However, PRF has only been available in gel or membrane form and is not suitable for injection. Recently, however, a liquid, injectable PRF has been introduced. This paper introduces the concept of injectable PRF and discusses its applications for biologic activation of bone grafts.     

Acute and subacute oral toxicity of copper oxide nanoparticles in female albino Wistar rats

The extensive use of copper oxide nanoparticles (CuO‐NPs) in various industries and their wide range of applications have led to their accumulation in different ecological niches of the environment. This excess exposure raises the concern about its potential toxic effects on various organisms including humans. However, the hazardous potential of CuO‐NPs in the literature is elusive, and it is essential to study its toxicity in different biological models. Hence, we have conducted single acute dose (2000 mg/kg) and multiple dose subacute (30, 300 and 1000 mg/kg daily for 28 days) oral toxicity studies of CuO‐NPs in female albino Wistar rats following OECD guidelines 420 and 407 respectively. Blood analysis, tissue aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and acetylcholinesterase, superoxide dismutase, catalase, lipid malondialdehyde and reduced glutathione assays, and histopathology of the tissues were carried out. The higher dose treatments of the acute and subacute study caused significant alterations in biochemical and antioxidant parameters of the liver, kidney and brain tissues of the rat. In addition, histopathological evaluation of these three organs of treated rats showed significantly high abnormalities in their histoarchitecture when compared to control rats. We infer from the results that the toxicity observed in the liver, kidney and brain of treated rats could be due to the increased generation of reactive oxygen species by CuO‐NPs.     

Effect of gestational exposure to perfluorononanoic acid on neonatal mice testes

Perfluoroalkyl acids (PFAAs) are widely used in commercial products and are found in many goods of daily use. Perfluorononanoic acid (PFNA) is one of the PFAAs that possesses endocrine disrupting properties and we have recently shown that PFNA affects testicular functions in Parkes mice. Exposure to environmental endocrine disruptors during fetal life is believed to affect gonadal development and they might produce reproductive abnormalities in males. Therefore, the present study examined the effect of gestational exposure to PFNA on the testes of neonatal mice offspring. Pregnant Parkes mice were orally administered PFNA (2 and 5 mg/kg body weight) or distilled water from gestational day 12 until parturition. Male pups were killed on postnatal day 3. PFNA treatment decreased testosterone biosynthesis by inhibiting expression of steroidogenic acute regulatory protein, cytochrome P450scc, and 3β‐ and 17β‐hydroxysteroid dehydrogenase; proliferation of testicular cells was also affected in treated mice. Furthermore, a marked decrease in expression of Wilms tumor 1, steroidogenic factor 1 and insulin‐like factor 3 was noted in neonatal mice testes, indicating that the PFNA treatment may affect the development of the testis. Moreover, observation of the dose‐related expression of anti‐müllerian hormone and c‐Kit in neonatal mice testes is also suggestive of an interference with gonadal development by PFNA exposure. In conclusion, the results suggest that the gestational exposure to PFNA decreased testosterone biosynthesis and altered the expression of critical factors involved in the development of the testis, thereby advocating a potential risk of PFNA to male reproductive health.     

Development,characterisation and efficacy evaluation of biochemical fungicidal formulations for postharvest control of anthracnose (Colletotrichum gloeosporioides Penz) disease in mango

The objectives of the study are to develop and characterise formulations with volatile molecules in an emulsifiable concentrate form, for their antimicrobial properties and to evaluate their efficacies against Colletotrichum gloeosporioides Penz., to control anthracnose in mangoes after harvest. Results showed EC39 and EC40 among formulations were characterised for their excellent emulsification properties, the droplet size of 192.34?±?0.48?nm and 227.4?±?0.71?nm and Zeta potential of ?52.5?±?2.76?mv and ?48.84?±?2.62?mv, respectively, with better storage stability at 10?±?20?°C and RH 80?±?5%. In vitro assay, 100% inhibition of visual spore germination by 0.15% and 0.2% MIC value for EC39 and EC40, respectively Studies on the efficacy of their fungicide properties also indicated the IC50 value of 0.161% and 0.162% for EC39 and EC40 respectively for mycelial growth inhibition. In vivo testing too, EC39 and EC40 effectively controlled anthracnose incidence in mango in a dosage-dependent manner.