Protein hydrolysates prepared by Alcalase using ultrasound and microwave pretreated almond meal and their characterization.

The study aimed to optimize ultrasonic (US: 40 kHz/200 W for 10, 20, 30, 40, and 50 min), and microwave (MW: 160 W for 45, 90, 125, 180, and 225 s) pretreatment conditions on protein extraction yield and degree of protein hydrolysis (DH) from almond de-oiled meal, an industrial by-product. First order model was used to describe the kinetics of almond protein hydrolysates obtained with Alcalase. The highest DH, 10.95% was recorded for the US-50 min and 8.87% for MW-45 s; while it was 5.76% for the untreated/control sample. At these optimized pretreatment conditions, a 1.16- and 1.18-fold increment in protein recovery was observed for the US and MW pretreatments, respectively in comparison to the conventional alkaline extraction. The molecular weight distribution recorded for pretreated samples disclosed a significant reduction in the band thickness in comparison with control. Both the pretreatments resulted in a significant increase (P < 0.05) in the antioxidant activity, and TCA solubility index when compared with the control. Results evinced that US and/or MW pretreatments before enzymatic hydrolysis can be a promising approach for the valorization of almond meal for its subsequent use as an ingredient for functional foods/nutraceuticals which otherwise fetches low value as an animal feed.

Biophotonics in Dentistry-An Overview.

Biophotonics, an interdisciplinary field merging biology with photonics, has transformed dentistry by offering innovative techniques and tools for diagnosis, treatment, and research. This overview explores the applications and benefits of biophotonics in dentistry, including early disease detection, precision in procedures, restorative dentistry assessment, real-time monitoring, and teeth whitening. We discuss how biophotonics improves patient care and the potential for future developments in personalized treatment, targeted therapy, enhanced imaging, and pain management. Biophotonics promises to continue revolutionizing oral healthcare, leading to better patient outcomes worldwide.

Synthesis and evaluation of highly potent HBV capsid assembly modulators (CAMs).

Chronic hepatitis B virus (HBV) infection remains a major global health burden. It affects more than 290 million individuals worldwide and is responsible for approximately 900,000 deaths annually. Anti-HBV treatment with a nucleoside analog in combination with pegylated interferon are considered first-line therapy for patients with chronic HBV infection and liver inflammation. However, because cure rates are low, most patients will require lifetime treatment. HBV Capsid Assembly Modulators (CAMs) have emerged as a promising new class of compounds as they can affect levels of HBV covalently closed-circular DNA (cccDNA) associated with viral persistence. SAR studies around the core structure of lead HBV CAM GLP-26 (Fig. 1B) was performed and led to the discovery of non-toxic compound 10a displaying sub-nanomolar anti-HBV activity. Advanced toxicity and cellular pharmacology profiles of compounds 10a were also established and the results are discussed herein.

Delayed adverse reaction in donors after whole blood donation: Is it a matter of concern?

BACKGROUND: Blood donation has a remarkable safety record and most of the donors have a good experience or only mild symptoms after blood donation were noted. Although even a very low rate of reactions may have gloomy effect diminishing their fondness to donate again. The main aim of our study was to determine the incidence and to analyze how various donor demographic factors tend to be associated with delayed adverse donor reactions (ADR). MATERIAL AND METHODS: The prospective observational study was conducted in Department of Immuno-hematology and Blood Transfusion of tertiary care hospital. All the whole blood donors, who gave consent to participate in the study were contacted telephonically after 24 hours and day 7 after donation. The donor who couldn't be contacted telephonically, was tried again at an interval of four hours in a day for two consecutive days before declaring the participant to be non-responder. RESULTS: A total of 2495 (92.4%) blood donors experienced delayed ADRs. The commonest delayed ADRs reported were generalized weakness (24.6%), bruises (24.2%) followed by painful arm (14.5%). Females, first-time blood donors, donors with low BMI and donors engaged in manual labor were more vulnerable to develop any adverse reactions. CONCLUSION: Blood donors can experience delayed ADRs. It is important to prevent these reactions especially in susceptible donors. Proper awareness and training to the staff was provided to minimize the incidence of ADR.

Effect of microfluidization on quality characteristics of sapodilla (Manilkara achras L.) juice.

Various oxidative enzymes account for the quality degradation of sapodilla (Manilkara achras L.) juice and need to be inactivated through emerging and continuous green pressure processing technology. In this study, pressurization of sapodilla juice was attempted via microfluidization (MF) at pressure range of 10,000-30,000 pound per square inch (psi) with 1-3 passes or cycles. The impact of microfluidization on the activity of polyphenol oxidase (PPO), peroxidase (POD), color, total soluble solid (TSS), viscosity, serum cloudiness along with particle size, and microbial load of sapodilla juice was assessed. Results showed that microfluidization (MF) decreased the residual PPO activity from 100 to 80.78 % and POD activity from 100 to 40.57%. However, TSS (18.81-19.01 %), viscosity (2.64-2.06 cP), serum cloudiness (2.19-1.22 %) and total color change (3.19-18.54) was also significantly affected. Most of these changes were observed due to particle size (PS) reduction that varied from 65.19 to 8.13 µm. Microfluidized juice revealed color improvement at particular MF pressure and pass due to enzyme inactivation. Moreover, lowest microbial load (2.89 Log CFU/ mL) was found at 30,000 psi/3 pass of MF as compared to control sample (unprocessed juice) (7.57 Log CFU/ mL). Consequently, MF can be potential candidate in processing of juices against spoilage.

Formulation and characterization of nano-curcumin fortified milk cream powder through microfluidization and spray drying.

In the present investigation, novel nano-curcumin enriched milk cream powder (CP) was formulated using microfluidization (at 100 MPa/2 passes) followed by spray drying (at three different temperatures: 150, 170 and 190 °C) with sodium caseinate (N) and gum arabic (G) as encapsulating materials. The effect of processing and encapsulating materials on the powder functionalities, particle size, encapsulation efficiency, morphology, fluorescence properties, bioaccessibility and cytotoxicity were studied. Results showed that NCP (spray dried at 190 °C) had significantly higher yield (68 %), encapsulation efficiency (EE) (93 %), and lower particle size (724 nm) than that of GCP. Fluorescence spectra of the powders revealed characteristic 'blue shift' phenomenon indicating better encapsulation and protection of the nano-curcumin corroborating EE results. SEM images showed distinctive features of NCP and GCP; wherein, NCP had shrivelled and irregular surface as compared to the GCP which exhibited round shape and smooth surface. TEM results confirmed that curcumin particles were in the nano-scale (50-250 nm) for both NCP and GCP. In vitro simulated digestion showed significantly high (88.48 %) curcumin bioaccessibility of NCP plus remarkable inhibition of HepG2 cells; whereas, no cytotoxicity was observed in Caco-2 cells by MTT assay. Formulation's applicability was shown by reconstituting powders as a 'cream spread' wherein high sensory acceptability observed. Sodium caseinate was found to be an excellent delivery vehicle for the fortification of nano-curcumin in the cream powder. To our knowledge, this is the first report of formulating a novel nano-curcumin fortified cream powder which has tremendous potential as a functional food ingredient.

Assessment of a Computational Approach to Predict Drug Resistance Mutations for HIV, HBV and SARS-CoV-2.

Viral resistance is a worldwide problem mitigating the effectiveness of antiviral drugs. Mutations in the drug-targeting proteins are the primary mechanism for the emergence of drug resistance. It is essential to identify the drug resistance mutations to elucidate the mechanism of resistance and to suggest promising treatment strategies to counter the drug resistance. However, experimental identification of drug resistance mutations is challenging, laborious and time-consuming. Hence, effective and time-saving computational structure-based approaches for predicting drug resistance mutations are essential and are of high interest in drug discovery research. However, these approaches are dependent on accurate estimation of binding free energies which indirectly correlate to the computational cost. Towards this goal, we developed a computational workflow to predict drug resistance mutations for any viral proteins where the structure is known. This approach can qualitatively predict the change in binding free energies due to mutations through residue scanning and Prime MM-GBSA calculations. To test the approach, we predicted resistance mutations in HIV-RT selected by (-)-FTC and demonstrated accurate identification of the clinical mutations. Furthermore, we predicted resistance mutations in HBV core protein for GLP-26 and in SARS-CoV-2 3CLpro for nirmatrelvir. Mutagenesis experiments were performed on two predicted resistance and three predicted sensitivity mutations in HBV core protein for GLP-26, corroborating the accuracy of the predictions.

In silico design of a novel nucleotide antiviral agent by free energy perturbation.

Nucleoside analogs are the backbone of antiviral therapies. Drugs from this class undergo processing by host or viral kinases to form the active nucleoside triphosphate species that selectively inhibits the viral polymerase. It is the central hypothesis that the nucleoside triphosphate analog must be a favorable substrate for the viral polymerase and the nucleoside precursor must be a satisfactory substrate for the host kinases to inhibit viral replication. Herein, free energy perturbation (FEP) was used to predict substrate affinity for both host and viral enzymes. Several uridine 5'-monophosphate prodrug analogs known to inhibit hepatitis C virus (HCV) were utilized in this study to validate the use of FEP. Binding free energies to the host monophosphate kinase and viral RNA-dependent RNA polymerase (RdRp) were calculated for methyl-substituted uridine analogs. The 2'-C-methyl-uridine and 4'-C-methyl-uridine scaffolds delivered favorable substrate binding to the host kinase and HCV RdRp that were consistent with results from cellular antiviral activity in support of our new approach. In a prospective evaluation, FEP results suggest that 2'-C-dimethyl-uridine scaffold delivered favorable monophosphate and triphosphate substrates for both host kinase and HCV RdRp, respectively. Novel 2'-C-dimethyl-uridine monophosphate prodrug was synthesized and exhibited sub-micromolar inhibition of HCV replication. Using this novel approach, we demonstrated for the first time that nucleoside analogs can be rationally designed that meet the multi-target requirements for antiviral activity.

Assessment of the Abbott BinaxNOW SARS-CoV-2 rapid antigen test against viral variants of concern.

As the emergence of SARS-CoV-2 variants brings the global pandemic to new levels, the performance of current rapid antigen tests against variants of concern and interest (VOC/I) is of significant public health concern. Here, we report assessment of the Abbot BinaxNOW COVID-19 Antigen Self-Test. Using genetically sequenced remnant clinical samples collected from individuals positive for SARS-CoV-2, we assessed the performance of BinaxNOW against the variants that currently pose public health threats. We measured the limit of detection of BinaxNOW against various VOC/I in a blinded manner. BinaxNOW successfully detected the Omicron (B.1.1.529), Mu (B.1.621), Delta (B.1.617.2), Lambda (C.37), Gamma (P.1), Alpha (B.1.1.7), Beta (B.1.351), Eta (B.1.525), and P.2 variants and at low viral concentrations. BinaxNOW also detected the Omicron variant in individual remnant clinical samples. Overall, these data indicate that this inexpensive and simple-to-use, FDA-authorized and broadly distributed rapid test can reliably detect Omicron, Delta, and other VOC/I.

Comparison of anti-SARS-CoV-2 activity and intracellular metabolism of remdesivir and its parent nucleoside.

Remdesivir, a monophosphate prodrug of nucleoside analog GS-441524, is widely used for the treatment of moderate to severe COVID-19. It has been suggested to use GS-441524 instead of remdesivir in the clinic and in new inhalation formulations. Thus, we compared the anti-SARS-CoV-2 activity of remdesivir and GS-441524 in Vero E6, Vero CCL-81, Calu-3, Caco-2 â€‹cells, and anti-HCoV-OC43 activity in Huh-7 â€‹cells. We also compared the cellular pharmacology of these two compounds in Vero E6, Vero CCL-81, Calu-3, Caco-2, Huh-7, 293T, BHK-21, 3T3 and human airway epithelial (HAE) cells. Overall, remdesivir exhibited greater potency and superior intracellular metabolism than GS-441524 except in Vero E6 and Vero CCL-81 â€‹cells.

Quantity and quality of complementary and alternative medicine recommendations in clinical practice guidelines for type 2 diabetes mellitus: A systematic review.

AIMS: Approximately 70% of Americans with diabetes have used complementary and alternative medicine (CAM) in the past year. Healthcare providers often receive minimal training on these therapies and subsequently rely on clinical practice guidelines (CPGs) to supplement their knowledge about the safe and effective use of CAM for the treatment/management of type 2 diabetes mellitus (T2DM). The purpose of this systematic review is to determine the quantity and assess the quality of CAM recommendations in CPGs for the treatment and/or management of T2DM. DATA SYNTHESIS: MEDLINE, EMBASE, and CINAHL were systematically searched from 2009 to 2020, in addition to the Guidelines International Network and the National Center for Complementary and Integrative Health websites. CPGs containing treatment and/or management recommendations for T2DM were eligible; those with CAM recommendations were quality-assessed with the AGREE II instrument twice, once for the overall CPG and once for the CAM sections. Twenty-seven CPGs were deemed eligible, of which 7 made CAM recommendations. Mean scaled domain percentages were (overall, CAM): scope and purpose (89.7%, 79.8%), clarity of presentation (85.7%, 48.4%), stakeholder involvement (67.9%, 28.2%), applicability (54.8%, 20.2%), rigour of development (49.7%, 35.7%), and editorial independence (44.1%, 44.1%). CONCLUSIONS: Quality varied within and across CPGs; domain scores across CAM sections generally scored lower than the overall CPG. Given that CAM therapies for T2DM are only represented in one-quarter of eligible CPGs and are of lower quality, a knowledge gap exists for healthcare providers who seek evidence-based information on this topic in order to effectively counsel inquiring patients.

Nanoencapsulated curcumin emulsion utilizing milk cream as a potential vehicle by microfluidization: Bioaccessibility, cytotoxicity and physico-functional properties.

Curcumin loaded milk cream emulsion was microfluidized at different pressures (50-200 MPa) and passes (1-4) using a full-factorial experimental design. Ultrasonicated and microfluidized emulsion was evaluated for particle size, morphological characteristics, antioxidant activity, rheological properties, bioaccessibility and cytotoxicity. Significant reduction was observed in the average particle size (358.2 nm) after microfluidization at 100 MPa/2nd pass. Transmission electron micrographs of the control (homogenized) and microfluidized (100 MPa/2nd pass) samples showed uniform distribution of fat globules in the microfluidized sample with partially dissolved curcumin particles (50-150 nm). Encapsulation efficiency of microfluidized emulsion was found to be significantly higher (97.88%) after processing as compared to control (91.21%). Two-fold (100%) increase in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and 25% increase in ferric-reducing antioxidant power (FRAP) was observed for microfluidized emulsions over control. Infrared spectrums of the emulsion exhibited shift in high intensity peaks indicating bond cleavage after microfluidization. After characterization, emulsions were subjected to in vitro digestion (oral, gastric and intestinal phase) to evaluate its bioaccessibility which was found to be remarkably increased by 30% after microfluidization. For assessing processing induced safety of the formulation, in vitro cytotoxicity of the microfluidized nanocurcumin emulsion was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay on HepG2 cells, wherein high % of cell viability (>93%) was seen even at a dose as high as 900 µg/mL revealing no toxic effect of the processing technique (microfluidization). This study highlights the efficacy of microfluidization as a technique and that of milk cream as an inexpensive, yet potential vehicle for generating stable and bio-accessible nano-curcumin emulsion.

Discovery and structure activity relationship of glyoxamide derivatives as anti-hepatitis B virus agents.

Chronic hepatitis B viral infection is a significant health problem world-wide, and currently available antiviral agents suppress HBV infections, but rarely cure this disease. It is presumed that antiviral agents that target the viral nuclear reservoir of transcriptionally active cccDNA may eliminate HBV infection. Through a series of chemical optimization, we identified a new series of glyoxamide derivatives affecting HBV nucleocapsid formation and cccDNA maintenance at low nanomolar levels. Among all the compounds synthesized, GLP-26 displays a major effect on HBV DNA, HBeAg secretion and cccDNA amplification. In addition, GLP-26 shows a promising pre-clinical profile and long-term effect on viral loads in a humanized mouse model.

Identifying the quantity and assessing the quality of clinical practice guidelines for the treatment and management of type 2 diabetes: A systematic review.

AIMS: Type 2 diabetes (T2D) is a condition where the body becomes insulin resistant and cannot use insulin made by the pancreas or is relatively insulin deficient causing high blood glucose levels. Assessing the quality of clinical practice guidelines (CPGs) for T2D is important to identify knowledge gaps and where improvements can be made. The purpose of this review was to identify the quantity and assess the quality of CPGs for the treatment and/or management of T2D. METHODS: A systematic review was conducted to identify T2D CPGs. MEDLINE, EMBASE, CINAHL and GIN were searched from 2008 to 2018. Eligible guidelines published on the treatment and/or management of T2D were assessed with the Appraisal of Guidelines for Research and Evaluation II (AGREE II) instrument. RESULTS: Seventeen CPGs were found to be eligible. Scaled domain percentages (highest to lowest) were: clarity of presentation (81.2%), scope and purpose (77.1%), stakeholder involvement (52.8%), applicability (42.9%), rigour of development (41.5%), and editorial independence (35.1%). CONCLUSIONS: CPGs that achieved higher AGREE II scores and favourable overall recommendations could be used by healthcare providers to facilitate informed discussions surrounding T2D therapies. CPGs that received lower scaled domain percentages or overall recommendations could be improved by using the AGREE II instrument.

Synthesis of 4'-Substituted-2'-Deoxy-2'-α-Fluoro Nucleoside Analogs as Potential Antiviral Agents.

Nucleoside analogs are widely used for the treatment of viral diseases (Hepatitis B/C, herpes and human immunodeficiency virus, HIV) and various malignancies. ALS-8176, a prodrug of the 4'-chloromethyl-2'-deoxy-2'-fluoro nucleoside ALS-8112, was evaluated in hospitalized infants for the treatment of respiratory syncytial virus (RSV), but was abandoned for unclear reasons. Based on the structure of ALS-8112, a series of novel 4'-modified-2'-deoxy-2'-fluoro nucleosides were synthesized. Newly prepared compounds were evaluated against RSV, but also against a panel of RNA viruses, including Dengue, West Nile, Chikungunya, and Zika viruses. Unfortunately, none of the compounds showed marked antiviral activity against these viruses.

Potent in vitro activity of ß-D-4'-chloromethyl-2'-deoxy-2'-fluorocytidine against Nipah virus.

Nipah virus (NiV) is a highly pathogenic zoonotic paramyxovirus that continues to cause outbreaks in humans characterized by high mortality and significant clinical sequelae in survivors. Currently, no therapeutics are approved for use in humans against NiV infection. Here, we report that 4'-chloromethyl-2'-deoxy-2'-fluorocytidine (ALS-8112) inhibits NiV. ALS-8112 is the parent nucleoside of lumicitabine, which has been evaluated in phase I and II clinical trials to treat pediatric and adult respiratory syncytial virus infection. In this study, we tested ALS-8112 against NiV and other major human respiratory pneumo- and paramyxoviruses in 2 human lung epithelial cell lines, and demonstrated the ability of ALS-8112 to reduce infectious wild-type NiV yield by over 6 orders of magnitude with no apparent cytotoxicity. However, further cytotoxicity testing in primary cells and bone marrow progenitor cells indicated cytotoxicity at higher concentrations of ALS-8112. Our results warrant the evaluation of lumicitabine against NiV infection in relevant animal models.

Novel Hepatitis B Virus Capsid Assembly Modulator Induces Potent Antiviral Responses In Vitro and in Humanized Mice.

Hepatitis B virus (HBV) affects an estimated 250 million chronic carriers worldwide. Though several vaccines exist, they are ineffective for those already infected. HBV persists due to the formation of covalently closed circular DNA (cccDNA)-the viral minichromosome-in the nucleus of hepatocytes. Current nucleoside analogs and interferon therapies rarely clear cccDNA, requiring lifelong treatment. Our group identified GLP-26, a novel glyoxamide derivative that alters HBV nucleocapsid assembly and prevents viral DNA replication. GLP-26 exhibited single-digit nanomolar anti-HBV activity, inhibition of HBV e antigen (HBeAg) secretion, and reduced cccDNA amplification, in addition to showing a promising preclinical profile. Strikingly, long term combination treatment with entecavir in a humanized mouse model induced a decrease in viral loads and viral antigens that was sustained for up to 12 weeks after treatment cessation.

Spontaneous Rupture of a Pseudoaneurysm of the Right Hepatic Artery Causing Massive Upper Gastrointestinal Bleeding.

We describe the case of an 84-year-old woman who presented with right lower chest pain, anaemia and newly deranged liver function which was followed by massive upper gastrointestinal (GI) bleeding with no source of bleeding found on upper GI endoscopy. CT angiography of the GI tract confirmed rupture of a pseudoaneurysm of the right hepatic artery (RHA) that was treated successfully with trans-arterial embolization of the RHA. LEARNING POINTS: If upper gastrointestinal (GI) endoscopy fails to identify the source of upper GI bleeding, CT angiography is required to search for rare causes such as pseudoaneurysm of the right hepatic artery (RHA) with fistula formation with the GI and biliary tract, along with other causes such as aorto-enteric fistula.Pseudoaneurysm of the RHA is commonly secondary to recent surgery or trauma and spontaneous occurrence is very rare.Endovascular repair using transcatheter arterial embolization is the treatment of choice but if it fails, emergency laparotomy should be considered.

Discovery of a Series of 2'-α-Fluoro,2'-ß-bromo-ribonucleosides and Their Phosphoramidate Prodrugs as Potent Pan-Genotypic Inhibitors of Hepatitis C Virus.

Hepatitis C virus (HCV) nucleoside inhibitors display pan-genotypic activity, a high barrier to the selection of resistant virus, and are some of the most potent direct-acting agents with durable sustained virologic response in humans. Herein, we report, the discovery of ß-d-2'-Br,2'-F-uridine phosphoramidate diastereomers 27 and 28, as nontoxic pan-genotypic anti-HCV agents. Extensive profiling of these two phosphorous diastereomers was performed to select one for in-depth preclinical profiling. The 5'-triphosphate formed from these phosphoramidates selectively inhibited HCV NS5B polymerase with no inhibition of human polymerases and cellular mitochondrial RNA polymerase up to 100 µM. Both are nontoxic by a variety of measures and display good stability in human blood and favorable metabolism in human intestinal microsomes and liver microsomes. Ultimately, a preliminary oral pharmacokinetics study in male beagles showed that 28 is superior to 27 and is an attractive candidate for further studies to establish its potential value as a new clinical anti-HCV agent.