2-deoxy-D-glucose as an adjunct to standard of care in the medical management of COVID-19: a proof-of-concept and dose-ranging randomised phase II clinical trial.

BACKGROUND: At present, no single efficacious therapeutic exists for acute COVID-19 management and a multimodal approach may be necessary. 2-deoxy-D-glucose (2-DG) is a metabolic inhibitor that has been shown to limit multiplication of SARS-CoV-2 in-vitro. We evaluated the efficacy and safety of 2-DG as adjunct to standard care in the treatment of moderate to severe COVID-19 patients. METHODS: We conducted a randomized, open-label, phase II, clinical study to evaluate the efficacy, safety, and tolerability of 2-DG administered as adjunct to standard of care (SOC). A total of 110 patients between the ages of 18 and 65 years with moderate to severe COVID-19 were included. Patients were randomized to receive 63, 90, or 126 mg/kg/day 2-DG in addition to SOC or SOC only. Times to maintaining SpO2 ≥ 94% on room air, discharge, clinical recovery, vital signs normalisation, improvement by 1 and 2 points on WHO clinical progression scale, negative conversion on RT-PCR, requirement for intensive care, and mortality were analyzed to assess the efficacy. RESULTS: Patients treated with 90 mg/kg/day 2-DG plus SOC showed better outcomes. Time to maintaining SpO2 ≥ 94% was significantly shorter in the 2-DG 90 mg compared to SOC (median 2.5 days vs. 5 days, Hazard ratio [95% confidence interval] = 2.3 [1.14, 4.64], p = 0.0201). Times to discharge from isolation ward, to clinical recovery, and to vital signs normalization were significantly shorter for the 2-DG 90 mg group. All three doses of 2-DG were well tolerated. Thirty-three (30.3%) patients reported 65 adverse events and were mostly (86%) mild. CONCLUSIONS: 2-DG 90 mg/kg/day as adjunct to SOC showed clinical benefit over SOC alone in the treatment of moderate to severe COVID-19. The promising trends observed in current phase II study is encouraging for confirmatory evaluation of the efficacy and safety of 2-DG in a larger phase III trial. TRIAL REGISTRATION: CTRI, CTRI/2020/06/025664. Registered 5th June 2020, http://ctri.nic.in/Clinicaltrials/pdf_generate.php?trialid=44369&EncHid=&modid=&compid=%27,%2744369det%27 .

Prevalence of chronic respiratory diseases from a rural area in Kerala, southern India.

BACKGROUND: Chronic lung diseases are one of the leading causes of morbidity in developing countries. A community based survey was undertaken with an objective to estimate the prevalence of chronic respiratory diseases and to describe the profile of people with CRDs in the rural area Nilamel health block in Kollam district, Kerala, southern India. METHODS: A household information sheet and a translated respiratory symptom questionnaire based on International Union against Tuberculosis and Lung Disease (IUATLD) bronchial symptoms questionnaire was administered to 12,556 people above 15 years, selected randomly from Nilamel health block. RESULTS: Prevalence of self reported asthma was 2.82% (95% CI 2.52-3.12) and that of chronic bronchitis was 6.19% (95% CI 5.76-6.62) while other CRDs which did not fit to either constitute 1.89%. Prevalence of asthma among males was 2.44% (95% CI 2.05-2.85) while that of females was 3.14% (95% CI 2.71-3.57). Chronic bronchitis prevalence was 6.73% and 5.67% among males and females respectively. CONCLUSION: Although India has devised a programme to combat cancer, diabetes, cardio vascular disease and stroke, none have been devised for chronic respiratory illness till date. Considering high prevalence and its contributions to morbidity and mortality, a comprehensive programme to tackle chronic respiratory diseases is needed.

Biocompatible amphiphilic pentablock copolymeric nanoparticles for anti-cancer drug delivery.

A pentablock copolymer of Poly(Lactide-co-Glycolide) and Pluronic F68 was synthesized using ring-opening polymerization and characterized by NMR and FTIR for confirming the structure of the block copolymer. TG-DTA studies showed PLGA:Pluronic ratio to be 4:1. As the PLGA-PEO-PPO-PEO-PLGA Pentablock Copolymer (PPPC) prepared is amphiphilic, its Critical Vesicular Concentration, was measured, which was lower at 37 degrees C than at 25 degrees C, which could provide better stability to the system at physiological temperature. The nanoparticles of PPPC vary in topographyand range from 150 to 500 nm in size, according to the synthesis route used viz Emulsion Solvent Evaporation and simple dialysis. Pentablock copolymer nanoparticles were found to entrap about 84% of hydrophobic drug, docetaxel. Drug release profile of docetaxel showed about 50% release in first 2 hours at pH 4.6 and about 80% docetaxel was released at pH 7.4, at the end of 2 days. The PPPC nanoparticles was found to be biocompatible to L929 cell lines up to 1 mg/ml concentration. Preliminary in vitro cytotoxic effect of docetaxel loaded PPPC nanoparticles against four different cancer cell lines showed 50% inhibitory concentration of 6 nM in A431 (Squamous cell carcinoma), 250 nM in HeLa (Cervical carcinoma), 800 nM in PC3 (Prostate carcinoma) and 1 microM in KB (Epidermoid carcinoma) cells.

Polyelectrolyte coated polymeric nanoparticles for controlled release of docetaxel.

Polyelectrolyte coatings are effective means of minimizing the rate of release of small molecules from a nanoparticle system. The current investigation aim at developing biodegradable drug delivery carriers composed of three types of polymers viz poly(lactide-co-glycolic) acid [PLGA], poly(lactide-co-glycolic) acid-polyethylene imine [PLGA-PEI] and poly lactic acid [PLA]. The PLGA and PLGA-PEI nanoparticles were in the size range approximately 150 nm while PLA nanoparticles were approximately 80 nm in size respectively. The nanoparticles were found to encapsulate 66%, 62% and 65% of the hydrophobic drug, Docetaxel (DOCE) respectively. "Layer by Layer (LbL)" self assembly technique was then performed to coat these particles with polyelectrolyte thin films (PEs). DLS studies showed hydrodynamic diameter of 330 nm, 350 nm and 310 nm for the coated PLGA, PLGA-PEI and PLA nanoparticles respectively, while SEM and TEM studies demonstrated that after coating the particle sizes were approximately 200 nm for all the three nanoparticles. In vitro drug release studies demonstrated that at the end of 24 hours, about 83%, 90% and 88% of drug was released from uncoated PLA, PLGA-PEI and PLGA nanoparticles respectively. Using LbL coating, the same amount of the drug was found to be released in a sustained way for up to 7 days for PLGA and up to 6 days for PLGA-PEI and PLA nanoparticles. In vitro cytocompatibility studies were carried out with each system and the cell viability at end of 48 hours was found to be in the range of 70% to 100%. Current approach of sustained drug delivery can help in improving the therapeutic efficiency of the breast cancer drug, DOCE.

Juvenile hormone stimulated tyrosine kinase-mediated protein phosphorylation in the CNS of the silk worm, Bombyx mori.

In vitro studies with the larval CNS of the silkworm, Bombyx mori revealed the phosphorylation of a 48-kDa protein, which was not dependent on cyclic nucleotides. Studies also revealed modest phosphorylation of this protein by a calcium-dependent but calmodulin-independent mechanism. However, phosphorylation of this protein was greatly enhanced in the presence of juvenile hormone (JH) I by a calcium-independent mechanism. This stimulatory effect of JH was seen in both homogenates as well as in intact CNS of Bombyx. Immunoblotting studies revealed the cross-reaction of this 48-kDa protein with phosphotyrosine monoclonal antibody and the phosphorylation of this protein was inhibited by genistein. This study suggests that the 48-kDa protein is a substrate for tyrosine kinase. The phosphorylation of this protein was also observed in other larval tissues such as salivary gland, fat body, and epidermis of Bombyx.

tau kinases in the rat heat shock model: possible implications for Alzheimer disease.

We have previously shown, by using the phosphate-dependent anti-tau antibodies Tau-1 and PHF-1, that heat shock induces rapid dephosphorylation of tau followed by hyperphosphorylation in female rats. In this study, we analyzed in forebrain homogenates from female Sprague-Dawley rats the activities of extracellular signal regulated kinase 1/2 (ERK1/2), c-Jun NH(2)-terminal kinase (JNK), glycogen synthase kinase-3beta (GSK-3beta), cyclin-dependent kinase 5 (Cdk5), cAMP-dependent protein kinase A (PKA), and Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) at 0 (n = 5), 3 (n = 4), 6 (n = 5), and 12 (n = 5) h after heat shock and in non-heat-shocked controls (n = 5). Immunoprecipitation kinase assays at 0 h showed suppression of the activities of all kinases except of GSK-3beta, which showed increased activity. At 3-6 h, the activities of ERK1/2, JNK, Cdk5, and GSK-3beta toward selective substrates were increased; however, only JNK, Cdk5, and GSK-3beta but not ERK1/2 were overactivated toward purified bovine tau. At 3-6 h, kinase assays specific for PKA and CaMKII showed no increased activity toward either tau or selective substrates. All of eight anti-tau antibodies tested showed dephosphorylation at 0 h and hyperphosphorylation at 3-6 h, except for 12E8, which showed hyperphosphorylation also at 0 h. Immunoblot analysis using activity-dependent antibodies against ERK1/2, JNK, and GSK-3beta confirmed the above data. Increased activation and inhibition of kinases after heat shock were statistically significant in comparison with controls. Because tau is hyperphosphorylated in Alzheimer disease these findings suggest that JNK, GSK-3beta, and Cdk5 may play a role in its pathogenesis.

Non-Hodgkin's lymphoma in children: disease pattern and survival.

The use of intensive chemotherapy and incorporation of prophylactic treatment of the central nervous system have dramatically improved the outcome of children with non-Hodgkin's lymphoma (NHL). The authors analyzed retrospectively the disease characteristics and survival data of 34 children with NHL during a 7-year period. There were 26 boys and 8 girls with a median age of 8 years. The primary sites were the abdomen (41%) and peripheral node (41%). Histopathologically lymphoblastic and undifferentiated lymphoma (small nonclaved cell lymphoma) were equally distributed (41%). Thirteen patients had localized disease (stage I and II) and 21 patients had advanced disease (stage III and IV). Surgical removal of the primary tumor was done in 6 patients with localized gastrointestinal lesions. All 34 patients received chemotherapy, either cyclophosphamide, vincristine, methotrexate, and prednisolone (COMP) or adriamycin, cyclophosphamide, vincristine, and prednisolone (ACOP). Thirty patients achieved complete remission (88.2%). The 5-year event-free survival rate was 64%. The results indicate that most children with localized disease can be cured by COMP chemotherapy, but more aggressive chemotherapy is necessary to improve survival in advanced-stage disease.

Identification, characterization, immunocytochemical localization, and developmental changes in the activity of calcium/calmodulin-dependent protein kinase II in the CNS of Bombyx mori during postembryonic development.

In the present investigation, in vitro phosphorylation of CNS proteins of the silkworm Bombyx mori during the postembryonic development have been studied. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography of phosphorylated proteins revealed the presence of major phosphoproteins of 59/60 kDa. Based on molecular mass, calcium/calmodulin-dependent autophosphorylation, substrate specificity, KN-62 inhibition, apparent Km for ATP and syntide-2, these proteins were identified as calcium/calmodulin-dependent protein kinase II (CaM kinase II). Anti-rat CaM kinase II monoclonal antibody showed immunoreactivity with Bombyx CaM kinase II isoforms. This kinase showed a high degree of autophosphorylation in neural tissue. During postembryonic development of Bombyx, two distinct peaks of enzyme activity could be noticed, one at the late-larval and another at the late-pupal stage, which were associated with an increase in amount of the enzyme. These results suggested that the expression of CaM kinase II in the CNS of Bombyx was developmentally regulated.

Ecdysteroid mediated muscle actin synthesis during the larval development of rice moth, Corcyra cephalonica.

Protein fractions from the whole body homogenates were separated on SDS-PAGE and the muscle actin band was identified by western blotting, using commercial antibody raised against chick back muscle actin (Sigma Chemical Co., USA). The antibody was found to be specific for muscle actin, as it did not cross react with cytoplasmic actin of the nervous system. The molecular weight of the muscle actin in Corcyra was found to be around 46 kDa. The concentration of actin was found to be more or less the same in penultimate and early-last instar larvae and it increased during mid and late-last instar development. Thereafter, it has dropped to its minimum during the pupal stage. But, once again it increased to its maximum in adults. Thorax-ligation in early and late-last instar larvae resulted in decreased synthesis of muscle actin and this was more pronounced after 48 h of ligation. Injection of 20-hydroxyecdysone to insects which were thorax-ligated for 24 and 48 h was found to significantly increase the concentration of actin, suggesting that the ecdysteroids mediate the muscle actin synthesis during the larval development of Corcyra.