Immunodiagnosis of groundnut and watermelon bud necrosis viruses using polyclonal antiserum to recombinant nucleocapsid protein of Groundnut bud necrosis virus

In vitro gene expression strategy was used for the production of polyclonal antiserum to the nucleocapsid protein (NP) of Groundnut bud necrosis virus (GBNV). The GBNV NP gene from cowpea isolate was cloned into 6x His-tagged UA cloning vector and expressed in Escherichia coli [M15] cells. The fusion protein was detected in insoluble fraction and was purified by using Ni-NTA agarose resin. The purified 6x His-fusion protein (32 kDa) was used for immunisation to produce a high titre polyclonal antiserum. The antiserum to the NP of GBNV at 1:4000 dilution detected successfully natural infection of GBNV and Watermelon bud necrosis virus in a wide range of cucurbitaceous, leguminous and solanaceous hosts from different locations.     

Serial Dexamethasone Suppression Tests and Clinical Outcome in ECT

Serial dexamethasone suppression tests (DST), obtained during a course of electroconvulsive therapy in 43 severely depressed patients, did not exhibit relationships between the initial DST, final DST, or the change in DST with clinical outcome measures at the time of discharge. In 37 patients reviewed six months after discharge, no relationship with the continuation of improvement, rehospitalization, or suicide was found. We are unable to confirm a clinical application for the DST in the management of patients during a course of convulsive therapy.     

UPLC-QTOF-MS fingerprinting combined with chemometrics to assess the solvent extraction efficiency,phytochemical variation,and antioxidant activities of Beta vulgaris L.

Accurate assays of plant antioxidants and other phytochemicals require efficient extraction conditions and enable rigorous assessments of crop varieties and production systems. This study assessed the extraction of phytochemicals and antioxidants from conventionally or organically grown red and golden beets (Beta vulgaris L.), using twenty solvent (S1–S20) mixtures containing water, methanol, and ethanol alone or with acids (ascorbic, formic, acetic). Red beetroot extracted with methanol with or without acid had the highest betanin content (2791.0 μg/g and 8222.3 μg/g of fresh weight [FW], respectively) and golden beetroot extracted with methanol/ascorbic acid/water had the highest vulgaxanthin I (193.7 μg/g and 15.0 μg/g of FW, respectively). The radical-scavenging activity and total phenolics in beetroot extracts reflected the different extraction efficiency of each solvent. UHPLC-QTOF-MS was used to identify twenty-seven phytochemicals, including 23 betalains, 2 amino acids, and 2 phenolic acids. Chemometric approaches discriminated the beet varieties and different extracts within one variety based on the composition and abundance of the key phytochemicals. The red beetroot extracted with aqueous ethanol with or without acid (S5, S7, S8, S9), and golden beetroot extracted with methanol-containing solvents (S15 for conventionally and S20 for organically) had the highest levels of phytochemicals, suggesting that these conditions efficiently extract key phytochemicals.     

Microbiome as mediator: Do systemic infections start in the gut?

The intestinal microbiome is emerging as a crucial mediator between external insults and systemic infections. New research suggests that our intestinal microorganisms contribute to critical illness and the development of non-gastrointestinal infectious diseases.Common pathways include a loss of fecal intestinal bacterial diversity and a disproportionate increase in toxogenic bacterial species. Therapeutic interventions targeting the microbiome- primarily probiotics- have yielded limited results to date. However, knowledge in this area is rapidly expanding and microbiome-based therapy such as short-chain fatty acids may eventually become a standard strategy for preventing systemic infections in the context of critical illness.     

Electroconvulsive Therapy and Neuroleptic Medication in Therapy-Resistant Positive-Symptom Psychosis

Eight patients with persistent psychosis and positive symptoms, who had not improved with at least two courses of neuroleptic medications, were treated with electroconvulsive therapy (ECT) combined with an antipsychotic medication. Seven of the eight patients improved; the improvement was sustained in five patients after 6 and 12 months. ECT and neuroleptic drugs appear to be synergistic in antipsychotic activity. This enhancement may result from greater brain levels of neuroleptic drugs secondary to increased permeability of the blood-brain barrier accompanying seizures. Our experience is consistent with other recent clinical reports, and encourages trials of ECT combined with neuroleptic medications in therapy-resistant psychotic patients, including those classified as having schizophrenic disorders.     

When combination therapy isn’t working: Emerging therapies for the management of inflammatory bowel disease

Although antagonists of tumor necrosis factor have resulted in major therapeutic benefits in inflammatory bowel disease, the magnitude and durability of response are variable. Similar to previously available drugs such as 5-aminosalicylates and immunomodulators, the therapeutic effect is not universal leaving many people searching for options. The development of newer agents has benefited from advances in the understanding of the pathophysiology of the disease. Uncontrolled activation of the acquired immune system has an important role, and lymphocytes, cytokines, and adhesion molecules are broadly targeted for therapeutic intervention. There is increasing evidence of an important role of the innate immune system and the intestinal epithelium, and the therapeutic paradigm is also shifting from immunosuppression to the reinforcement of the intestinal barrier, and modification of the disease process. In this review, we explore the limitation of current therapy as well as mechanisms of actions of new drugs and the efficacy and adverse events from data from clinical trials.     

Standardization of Regeneration,Agrobacterium-Mediated Transformation,and Introduction of Nucleocapsid Gene of Watermelon Bud Necrosis Virus in Watermelon

Proceedings of the National Academy of Sciences, India Section B: Biological Sciences - Seven types of explants (proximal cotyledon, distal cotyledon, distal cotyledon leaves, proximal cotyledon...     

Cox-2 is regulated by Toll-like receptor-4 (TLR4) signaling: Role in proliferation and apoptosis in the intestine

BACKGROUND & AIMS: We recently showed that mice deficient in Toll-like receptor 4 (TLR4) or its adapter molecule MyD88 have increased signs of colitis compared with wild-type (WT) mice after dextran sodium sulfate (DSS)-induced injury. We wished to test the hypothesis that cyclooxygenase 2 (Cox-2)-derived prostaglandin E2 (PGE2) is important in TLR4-related mucosal repair. METHODS: Cox-2 expression was analyzed by real-time polymerase chain reaction, immunohistochemistry, Western blotting, and luciferase reporter constructs. Small interfering RNA was used to inhibit expression of MyD88. TLR4-/- or WT mice were given 2.5% DSS for 7 days. Proliferation and apoptosis were assessed using bromodeoxyuridine staining and terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling assays, respectively. PGE2 was given orally to DSS-treated mice. RESULTS: Intestinal epithelial cell lines up-regulated Cox-2 expression in a TLR4- and MyD88-dependent fashion. Lipopolysaccharide-mediated stimulation of PGE2 production was blocked by a selective Cox-2 inhibitor or small interfering RNA against MyD88. After DSS injury, Cox-2 expression increased only in WT mice. TLR4-/- mice have significantly reduced proliferation and increased apoptosis after DSS injury compared with WT mice. PGE2 supplementation of TLR4-/- mice resulted in improvement in clinical signs of colitis and restoration of proliferation and apoptosis to WT values. The mechanism for improved epithelial repair may be through PGE2-dependent activation of the epidermal growth factor receptor. CONCLUSIONS: We describe an important link between TLR4 signaling and Cox-2 expression in the gut. TLR4 and MyD88 signaling are required for optimal proliferation and protection against apoptosis in the injured intestine. Although TLR4 signaling is beneficial in the short term, chronic signaling through TLR4 may lower the threshold for colitis-associated cancer.     

Long term non-invasive ventilation in the community for patients with musculoskeletal disorders: 46 year experience and review

BACKGROUND—A study wasundertaken to assess the long term physiological and clinical outcomein 79 patients with musculoskeletal disorders (73 neuromuscular, six ofthe chest wall) who received non-invasive ventilation for chronicrespiratory failure over a period of 46years.
METHODS—Vital capacity(VC) and carbon dioxide tension (PCO₂) beforeand after initiation of ventilation, type and duration of ventilatory assistance, the need for tracheostomy, and mortality wereretrospectively studied in 48 patients who were managed withmouth/nasal intermittent positive pressure ventilation (M/NIPPV) and 31 who received body ventilation. The two largest groups analysed were 45 patients with poliomyelitis and 15 with Duchenne's muscular dystrophy. Twenty five patients with poliomyelitis received body ventilation (fora mean of 290 months) and 20 were supported by M/NIPPV (mean 38 months). All 15 patients with Duchenne's muscular dystrophy wereventilated by NIPPV (mean 22months).
RESULTS—Fourteenpatients with poliomyelitis on body ventilation (56%) but only one onM/NIPPV, and 10 of 15 patients (67%) with Duchenne's musculardystrophy eventually received tracheostomies for ventilatory support.Five patients with other neuromuscular disorders required tracheostomies. Twenty of 29 tracheostomies (69%) were provided because of progressive disease and hypercarbia which could not becontrolled by non-invasive ventilation; the remaining nine were placedbecause of bulbar dysfunction and aspiration related complications.Nine of 10 deaths occurred in patients on body ventilation (six withpoliomyelitis), although the causes of death were varied and notnecessarily related to respiratory complications. A proportionatelygreater number of patients on M/NIPPV (67%) reported positive outcomes(improved sense of wellbeing and independence) than did those on bodyventilation (29%, p<0.01). However, other than tracheostomies anddeaths, negative outcomes in the form of machine/interface discomfortand self-discontinuation of ventilation also occurred at a rate 2.3 times higher than in the group who received body ventilation. None ofthe six patients with chest wall disorders (all on M/NIPPV) requiredtracheostomy or died. Hospital admission rates increased nearlyeightfold in patients receiving body ventilation (all poliomyelitispatients) compared with before ventilation (p<0.01) while in thosesupported by M/NIPPV they were reduced by 36%.
CONCLUSIONS—Non-invasiveventilation (NIV) in the community over prolonged periods is a feasiblealthough variably tolerated form of management in patients withneuromuscular disorders. While patients who received body ventilationwere followed the longest (mean 24 years), the need for tracheostomyand deaths occurred more often in this group (most commonly in thepoliomyelitis patients). Despite a number of discomforts associatedwith M/NIPPV, a larger proportion of patients experienced improvedwellbeing, independence, and ability to perform daily activities.