Effect of metabolic status on response to SIV infection and antiretroviral therapy in nonhuman primates.

Current antiretroviral therapy (ART) regimens efficiently limit HIV replication, thereby improving life expectancy of people living with HIV, but also cause metabolic side effects. The ongoing obesity epidemic has resulted in more people with metabolic comorbidities at the time of HIV infection, yet the impact of pre-existing metabolic dysregulation on infection sequelae and response to ART is unclear. Here, to investigate the impact of preexisting obesity and insulin resistance on acute infection and subsequent long-term ART, we infected a cohort of lean and obese adult male macaques with SIV and administered ART. The responses of lean and obese macaques to SIV and ART were similar with respect to plasma and cell-associated viral loads, ART drug levels in plasma and tissues, SIV-specific immune responses, adipose tissue and islet morphology, and colon inflammation, with baseline differences between lean and obese groups largely maintained. Both groups exhibited a striking depletion of CD4+ T cells from adipose tissue that did not recover with ART. However, differential responses to SIV and ART were observed for body weight, omental adipocyte size, and the adiponectin/leptin ratio, a marker of cardiometabolic risk. Thus, obesity and insulin resistance had limited effects on multiple responses to acute SIV infection and ART, while several factors that underlie long-term metabolic comorbidities were influenced by prior obesity and insulin resistance. These studies provide the foundation for future investigations into the efficacy of adjunct therapies such as metformin and glucagon-like peptide-1 receptor agonists in the prevention of metabolic comorbidities in people living with HIV.

Safety and Efficacy of SAB-185 for Non-hospitalized Adults with COVID-19: A Randomized Clinical Trial.

BACKGROUND: To address the need for novel COVID-19 therapies, we evaluated the fully-human polyclonal antibody product SAB-185 in a phase 3 clinical trial. METHODS: Non-hospitalized high-risk adults within 7 days of COVID-19 symptom onset were randomized 1:1 to open-label SAB-185 3,840 units/kg or casirivimab/imdevimab 1200 mg. Non-inferiority comparison was undertaken for the pre-Omicron population (casirivimab/imdevimab expected to be fully active) and superiority comparison for the Omicron population (casirivimab/imdevimab not expected to be active). Primary outcomes were the composite of all-cause hospitalizations/deaths and grade ≥3 treatment-emergent adverse events (TEAEs) through day 28. Secondary outcomes included time to sustained symptom improvement and resolution. RESULTS: Enrollment was terminated early due to low hospitalization/death rates upon Omicron emergence. 733 adults were randomized, 255 included in pre-Omicron and 392 in Omicron analysis populations. Hospitalizations/deaths occurred in 6 (5.0%) and 3 (2.2%) of pre-Omicron SAB-185 and casirivimab/imdevimab arms, respectively (absolute difference [95% CI] 2.7% [-2.3%, 8.6%]), inconclusive for non-inferiority; and 5 (2.5%) versus 3 (1.5%) (absolute difference 1.0% [-2.3%, 4.5%]) for Omicron. Risk ratios for grade ≥3 TEAEs were 0.94 [0.52, 1.71] (pre-Omicron) and 1.71 [0.96, 3.07] (Omicron). Time to symptom improvement and resolution were shorter for SAB-185, median 11 vs 14 (pre-Omicron) and 11 vs 13 days (Omicron) (symptom improvement), and 16 vs 24 days and 18 vs >25 days (symptom resolution), p<0.05 for symptom resolution for Omicron only. CONCLUSIONS: SAB-185 had an acceptable safety profile with faster symptom resolution in the Omicron population. Additional studies are needed to characterize its efficacy for COVID-19.

Establishment of a Four-Cell In Vitro Blood-Brain Barrier Model With Human Primary Brain Cells.

The blood-brain barrier (BBB) constitutes a crucial protective anatomical layer with a microenvironment that tightly controls material transit. Constructing an in vitro BBB model to replicate in vivo features requires the sequential layering of constituent cell types. Maintaining heightened integrity in the observed tight junctions during both the establishment and post-experiment phases is crucial to the success of these models. We have developed an in vitro BBB model that replicates the cellular composition and spatial orientation of in vivo BBB observed in humans. The experiment includes comprehensive procedures and steps aimed at enhancing the integration of the four-cell model. Departing from conventional in vitro BBB models, our methodology eliminates the necessity for pre-coated plates to facilitate cell adhesion, thereby improving cell visualization throughout the procedure. An in-house coating strategy and a simple yet effective approach significantly reduce costs and provides superior imaging of cells and corresponding tight junction protein expression. Also, our BBB model includes all four primary cell types that are structural parts of the human BBB. With its innovative and user-friendly features, our in-house optimized in vitro four-cell-based BBB model showcases novel methodology and provides a promising experimental platform for drug screening processes. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Coating and culture system Basic Protocol 2: Cell seeding and Transwell insert handling Basic Protocol 3: Assessment of model functionality.

In-vitro and in-vivo assessment of nirmatrelvir penetration into CSF, central nervous system cells, tissues, and peripheral blood mononuclear cells.

Three years after SARS-CoV-2 emerged as a global infectious threat, the virus has become endemic. The neurological complications such as depression, anxiety, and other CNS complications after COVID-19 disease are increasing. The brain, and CSF have been shown as viral reservoirs for SARS-CoV-2, yielding a potential hypothesis for CNS effects. Thus, we investigated the CNS pharmacology of orally dosed nirmatrelvir/ritonavir (NMR/RTV). Using both an in vitro and an in vivo rodent model, we investigated CNS penetration and potential pharmacodynamic activity of NMR. Through pharmacokinetic modeling, we estimated the median CSF penetration of NMR to be low at 18.11% of plasma with very low accumulation in rodent brain tissue. Based on the multiples of the 90% maximal effective concentration (EC90) for SARS-CoV-2, NMR concentrations in the CSF and brain do not achieve an exposure level similar to that of plasma. A median of only 16% of all the predicted CSF concentrations in rats were > 3xEC90 (unadjusted for protein binding). This may have implications for viral persistence and neurologic post-acute sequelae of COVID-19 if increased NMR penetration in the CNS leads to decreased CNS viral loads and decreased CNS inflammation.

Safety, Efficacy, and Pharmacokinetics of Combination SARS-CoV-2 Neutralizing Monoclonal Antibodies BMS-986414 (C135-LS) and BMS-986413 (C144-LS) Administered Subcutaneously in Non-Hospitalized Persons with COVID-19 in a Phase 2 Trial.

Background: Outpatient COVID-19 monoclonal antibody (mAb) treatment via subcutaneous delivery, if effective, overcomes the logistical burdens of intravenous administration. Methods: ACTIV-2/A5401 was a randomized, masked placebo-controlled platform trial where participants with COVID-19 at low risk for progression were randomized 1:1 to subcutaneously administered BMS-986414 (C135-LS) 200 mg, plus BMS-986413 (C144-LS) 200 mg, (BMS mAbs), or placebo. Coprimary outcomes were time to symptom improvement through 28 days; nasopharyngeal SARS-CoV-2 RNA below the lower limit of quantification (LLoQ) on days 3, 7, or 14; and treatment-emergent grade 3 or higher adverse events (TEAEs) through 28 days. Results: A total of 211 participants (105 BMS mAbs and 106 placebo) initiated study product. Time to symptom improvement favored the active therapy but was not significant (median 8 vs 10 days, P=0.19). There was no significant difference in the proportion with SARS-CoV-2 RNA

Human bronchopulmonary disposition and plasma pharmacokinetics of oral bemnifosbuvir (AT-527), an experimental guanosine nucleotide prodrug for COVID-19.

BACKGROUND: Bemnifosbuvir (AT-527) is a novel oral guanosine nucleotide antiviral drug for the treatment of persons with COVID-19. Direct assessment of drug disposition in the lungs, via bronchoalveolar lavage, is necessary to ensure antiviral drug levels at the primary site of SARS-CoV-2 infection are achieved. OBJECTIVES: This Phase 1 study in healthy subjects aimed to assess the bronchopulmonary pharmacokinetics, safety and tolerability of repeated doses of bemnifosbuvir. METHODS: A total of 24 subjects were assigned to receive bemnifosbuvir twice daily at doses of 275, 550 or 825 mg for up to 3.5 days. RESULTS: AT-511, the free base of bemnifosbuvir, was largely eliminated from the plasma within 6 h post dose in all dosing groups. Antiviral drug levels of bemnifosbuvir were consistently achieved in the lungs with bemnifosbuvir 550 mg twice daily. The mean level of the guanosine nucleoside metabolite AT-273, the surrogate of the active triphosphate metabolite of the drug, measured in the epithelial lining fluid of the lungs was 0.62 µM at 4-5 h post dose. This exceeded the target in vitro 90% effective concentration (EC90) of 0.5 µM for antiviral drug exposure against SARS-CoV-2 replication in human airway epithelial cells. Bemnifosbuvir was well tolerated across all doses tested, and most treatment-emergent adverse events reported were mild in severity and resolved. CONCLUSIONS: The favourable pharmacokinetics and safety profile of bemnifosbuvir demonstrates its potential as an oral antiviral treatment for COVID-19, with 550 mg bemnifosbuvir twice daily currently under further clinical evaluation in persons with COVID-19.

An overview of current research on the biological effects of extracts from the unripe ackee (Blighia sapida L.) - abstract

It has been demonstrated that subacute intraperitoneal administration of high doses of the lipid fraction of the unripe ackee resulted in marked neutropaenia, pulmonary toxicity and changes in the blood chemistry in rats. This study has been carried out to further isolate the neutropaenic principle by testing both aqueous and lipid fractions and to see if this effect could be demonstrated in a different animal species. In mice, intraperitoneal administration of aqueous and lipid extracts of the ackee at a dose regimen of 300mg/kg thrice weekly for six weeks showed an initial rise, followed by a lowering of the neutrophil count, red blood cell count, haemoglobin, platelet count, serum albumin and alkaline phosphatase also decreased. It is concluded that in addition to it's hypoglycaemic principle, both aqueous and lipid extracts of unripe ackee, when adminstered separately, have neutropaenic activity. Further research involving characterization and testing is in progress. (AU)

Extraction of an hyperglycaemic principle from the annatto (Bixa orellana)

The red powdery extract from the seeds of the annatto, Bixa orellana, is a well known food colouring. In an oil suspension it is used as a folk remedy (bush tea) in the West Indies, for diabetes mellitus. Detailed investigations on this extract, yielded a methyl ester, trans-bixin, molecular weight 394 and molecular formula C24 H30 O4. This purified substance was demonstrated, in anaaesthetised mongrel dogs, to cause hyperglycaemia. Concomitant electron microscopy of tissues biopsies, revealed damage to mitochrondria and endoplasmic reticulum mainly in liver and pancreas. When dogs were fed on a diet fortified with riboflavin, there was neither demonstratable tissue damage nor associated hyperglycaemia. These findings point to: (i) the potential dangers of informal medications such as 'bush teas'; (ii) the possible role of plant extracts/food additives in the development of diabetes mellitus especially in the undernourished state. (AU)

Renal subcapsular islet cell transplantation in the dog: a preliminary report

During the last two decades, islet cell transplantation has been pursued both experimentally and clinically in an effort to ameliorate diabetes mellitus. At present, however, islet cell transplantation still remains at the experimental stages as far as the treatment of diabetes is concerned. Also, culture of islet cells has proved to be rather frustrating and difficult. No consistent techniques have been developed, and simplified methods for islet cell preparation and adequate sites for islet cell placement would allow for further progress in this area. Ultimately, rejection remains the greatest obstacle to success. We report a simplified technique for enriching dog pancreatic islet cells. This preparation was injected into the renal subcapsular space in both homograft (3 experiments) and heterograft (3 experiments) situations. After six weeks, nephrectomy was performed, and histochemical techniques demonstrated many groups of live islets in betweeen the tubules in the renal cortex. No acinar cells were observed. Blood samples from the renal artery and renal vein at the time of nephrectomy revealed an average 36.9 percent increase in insulin concentration in the renal veins, supporting an active secretory role of these transported islet cells. This technique points to (i) the possible role of a "renal factor" in promoting growth of islet cells and (ii) the feasibility of successful transplantation of enriched islet cells as a potential approach to the curative treatment of diabetes mellitus. (AU)

Isolation, purification and identification of the hyperglycaemic principle of the annatto (Bixa orellana) - abstract

The crude extract eluted from the seed of Bixa orellana was seen to cause hyperglycaemia in anaesthetized dogs. Further investigation was carried out to find the biologically active component. Column chromatography, PLC and recrystallizations yielded a pure residue with m.p. 220. The experimental dogs in quantities below one gram, it caused sustained hyperglycaemia and severe damage to tissues of the liver, kidney and pancreas. The level of toxicity associated with this plant is of interest, since its extract is widley used in various goods (AU)

Toxicity of the hyperglycaemic-inducing extract of the Annatto (Bixa orellana) in the dog

The Annatto seed coat extract, which is commonly used in folklore in treating diabetes mellitus, has been investigated for its effects on blood sugar levels in the dog. An alkali-soluble hyperglycaemia-inducing fraction was detected, and detailed studies revealed toxic effects in the pancreas and liver accompanied by hyperglycaemia and apparent tendency to increased insulin levels. The toxicity was diminished by feeding riboflavin. The relevance of this finding points to a possible aetiological role in the development of malnutrition-related diabetes mellitus and also to the potential toxicity inherent in the widespread use of folklore medicines (AU)