Alzheimer's disease-associated P460L variant of EphA1 dysregulates receptor activity and blood-brain barrier function.

INTRODUCTION: Genome-wide association studies link susceptibility to late-onset Alzheimer's disease (LOAD) with EphA1. Sequencing identified a non-synonymous substitution P460L as a LOAD risk variant. Other Ephs regulate vascular permeability and immune cell recruitment. We hypothesized that P460L dysregulates EphA1 receptor activity and impacts neuroinflammation. METHODS: EphA1/P460L receptor activity was assayed in isogenic Human Embryonic Kidney (HEK) cells. Soluble EphA1/P460L (sEphA1/sP460L) reverse signaling in brain endothelial cells was assessed by T-cell recruitment and barrier function assays. RESULTS: EphA1 and P460L were expressed in HEK cells, but membrane and soluble P460L were significantly reduced. Ligand engagement induced Y781 phosphorylation of EphA1 but not P460L. sEphA1 primed brain endothelial cells for increased T-cell recruitment; however, sP460L was less effective. sEphA1 decreased the integrity of the brain endothelial barrier, while sP460L had no effect. DISCUSSION: These findings suggest that P460L alters EphA1-dependent forward and reverse signaling, which may impact blood-brain barrier function in LOAD. HIGHLIGHTS: EphA1-dependent reverse signaling controls recruitment of T cells by brain endothelial cells. EphA1-dependent reverse signaling remodels brain endothelial cell contacts. LOAD-associated P460L variant of EphA1 shows reduced membrane expression and reduced ligand responses. LOAD-associated P460L variant of EphA1 fails to reverse signal to brain endothelial cells.

Increased Levels of Autoantibodies against ROS-Modified Proteins in Depressed Individuals with Decrease in Antibodies against SARS-CoV-2 Antigen (S1-RBD).

Coronavirus 2019 (COVID-19) disease management is highly dependent on the immune status of the infected individual. An increase in the incidence of depression has been observed during the ongoing COVID-19 pandemic. Autoantibodies against in vitro reactive oxygen species (ROS) modified BSA and Lys as well as antibodies against receptor binding domain subunit S1 (S1-RBD) (S1-RBD-Abs) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were estimated using direct binding and competition ELISA. Serum samples were also tested for fasting blood glucose (FBG), malondialdehyde (MDA), carbonyl content (CC), interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α). Significant structural changes were observed in ROS modified BSA and Lys. Female depressed subjects who were also smokers (F-D-S) showed the highest levels of oxidative stress (MDA and CC levels). Similarly, increased levels of autoantibodies against ROS modified proteins were detected in F-D-S subjects, in males who were depressed and in smokers (M-D-S) compared to the other subjects from the rest of the groups. However, contrary to this observation, levels of S1-RBD-Abs were found to be lowest in the F-D-S and M-D-S groups. During the pandemic, large numbers of individuals have experienced depression, which may induce excessive oxidative stress, causing modifications in circulatory proteins. Thus, the formation of neo-antigens is induced, which lead to the generation of autoantibodies. The concomitant effect of increased autoantibodies with elevated levels of IFN-γ and TNF-α possibly tilt the immune balance toward autoantibody generation rather than the formation of S1-RBD-Abs. Thus, it is important to identify individuals who are at risk of depression to determine immune status and facilitate the better management of COVID-19.

Biophysical, Biochemical, and Molecular Docking Investigations of Anti-Glycating, Antioxidant, and Protein Structural Stability Potential of Garlic.

Garlic has been reported to inhibit protein glycation, a process that underlies several disease processes, including chronic complications of diabetes mellitus. Biophysical, biochemical, and molecular docking investigations were conducted to assess anti-glycating, antioxidant, and protein structural protection activities of garlic. Results from spectral (UV and fluorescence) and circular dichroism (CD) analysis helped ascertain protein conformation and secondary structure protection against glycation to a significant extent. Further, garlic showed heat-induced protein denaturation inhibition activity (52.17%). It also inhibited glycation, advanced glycation end products (AGEs) formation as well as lent human serum albumin (HSA) protein structural stability, as revealed by reduction in browning intensity (65.23%), decrease in protein aggregation index (67.77%), and overall reduction in cross amyloid structure formation (33.26%) compared with positive controls (100%). The significant antioxidant nature of garlic was revealed by FRAP assay (58.23%) and DPPH assay (66.18%). Using molecular docking analysis, some of the important garlic metabolites were investigated for their interactions with the HSA molecule. Molecular docking analysis showed quercetin, a phenolic compound present in garlic, appears to be the most promising inhibitor of glucose interaction with the HSA molecule. Our findings show that garlic can prevent oxidative stress and glycation-induced biomolecular damage and that it can potentially be used in the treatment of several health conditions, including diabetes and other inflammatory diseases.

Insights into Multifunctional Nanoparticle-Based Drug Delivery Systems for Glioblastoma Treatment.

Glioblastoma (GB) is an aggressive cancer with high microvascular proliferation, resulting in accelerated invasion and diffused infiltration into the surrounding brain tissues with very low survival rates. Treatment options are often multimodal, such as surgical resection with concurrent radiotherapy and chemotherapy. The development of resistance of tumor cells to radiation in the areas of hypoxia decreases the efficiency of such treatments. Additionally, the difficulty of ensuring drugs effectively cross the natural blood-brain barrier (BBB) substantially reduces treatment efficiency. These conditions concomitantly limit the efficacy of standard chemotherapeutic agents available for GB. Indeed, there is an urgent need of a multifunctional drug vehicle system that has potential to transport anticancer drugs efficiently to the target and can successfully cross the BBB. In this review, we summarize some nanoparticle (NP)-based therapeutics attached to GB cells with antigens and membrane receptors for site-directed drug targeting. Such multicore drug delivery systems are potentially biodegradable, site-directed, nontoxic to normal cells and offer long-lasting therapeutic effects against brain cancer. These models could have better therapeutic potential for GB as well as efficient drug delivery reaching the tumor milieu. The goal of this article is to provide key considerations and a better understanding of the development of nanotherapeutics with good targetability and better tolerability in the fight against GB.

Glycation and Oxidative Stress Increase Autoantibodies in the Elderly.

Aging causes gradual changes in free radicals, antioxidants, and immune-imbalance in the elderly. This study aims to understand links among aging, gluco-oxidative stress, and autoantibodies in asymptomatic individuals. In vitro glycation of human serum albumin (Gly-HSA) induces appreciable biochemical changes. Significant inhibition of advanced glycation end products (AGEs) formation was achieved using garlic extract (53.75%) and epigallocatechin-3-gallate from green tea (72.5%). Increased amounts of serum carbonyl content (2.42 ± 0.5) and pentosidine (0.0321 ± 0.0029) were detected in IV-S (S represent smokers) vs. IV group individuals. Direct binding ELISA results exhibited significantly high autoantibodies against Gly-HSA in group IV-S (0.55 ± 0.054; p < 0.001) and III-S (0.40 ± 0.044; p < 0.01) individuals as compared to the age matched subjects who were non-smokers (group IV and III). Moreover, high average percent inhibition (51.3 ± 4.1%) was obtained against Gly-HSA in IV-S group individuals. Apparent association constant was found to be high for serum immunoglobulin-G (IgG) from group IV-S (1.18 × 10-6 M) vs. serum IgG from IV group (3.32 × 10-7 M). Aging induced gluco-oxidative stress and AGEs formation may generate neo-epitopes on blood-proteins, contributing to production of autoantibodies in the elderly, especially smokers. Use of anti-glycation natural products may reduce age-related pathophysiological changes.

Depression and Smoking Augment the Production of Circulating Autoantibodies against Glycated HSA in Rheumatoid Arthritis Patients.

BACKGROUND: Depression and smoking contribute to the prognosis of autoimmune rheumatoid arthritis (RA). Advanced glycation end products (AGEs) are also detected in RA patients. This study correlates RA in patients with various levels of depression and a history of smoking through the detection of antibodies against AGEs of proteins. METHODS: Sixty RA subjects were selected and divided into 4 groups based on their levels of depression and smoking habits. The division was as follows: group I consisted of RA patients classified as depressed (RA-D); group II consisted of RA patients with a history of smoking (RA-S); group III consisted of RA patients suffering from depression who were also smokers (RA-DS); and group IV consisted of patients with RA alone (RA-A), i.e., not depressed and non-smokers. In vitro human serum albumin (HSA) was modified by glucose, and the modifications were studied by biochemical and biophysical techniques. Glycated (G)-HSA was used as an antigen, and autoantibodies against G-HSA (G-HSA-Abs) were screened in serum samples of different groups of RA subjects. Oxidative stress levels in all patients and healthy individuals were analyzed by protein-bound carbonyl content estimations. RESULTS: Significant biochemical and biophysical changes were detected in G-HSA when compared to native (N)-HSA. All patients and control subjects were screened for circulating G-HSA-Abs and N-HSA-Abs. From the cohort of different samples, serum autoantibodies from RA-DS showed a high recognition of G-HSA-Abs titres compared to RA-D or RA-S. RA-A exhibited the least binding of circulating G-HSA-Abs of all the groups. The oxidative stress marker, the carbonyl content also exhibited highest levels in RA-DS, followed by RA-D and RA-S. Band shift assay showed the highest titres of immunoglobulin G in the serum samples of RA-DS. CONCLUSIONS: Smoking and concomitant depression in RA subjects may lead to enhanced oxidative stress levels responsible for the gradual formation and/or exposing of cryptic epitopes on HSA that induce the production of G-HSA-Abs. Hence, we postulate that by reducing depression and corresponding oxidative stress, it may be possible to control or limit the severity of the precipitation of RA disease activity and improve prognosis.

Prevalence and risk factors associated with multidrug-resistant bacteria in COVID-19 patients.

Bacterial coinfection among patients with confirmed coronavirus disease 2019 (COVID-19) is a critical medical concern that increases the disease severity and mortality rate. The current study is aimed at evaluating the effects of bacterial coinfections among COVID-19 patients, especially in relation to degree of severity and mortality. A retrospective study was conducted for patients with positive COVID-19 test, admitted to a regional COVID-19 hospital in Jeddah, Saudi Arabia, between May and August 2020. A specimen (e.g., blood, urine, or sputum) was collected from patients with confirmed COVID-19, and was cultured to determine bacterial coinfection caused by multidrug resistant (MDR) bacteria. COVID-19 patients were categorized into 2 groups based on the result of bacterial coinfection culture, as COVID-19 patients with coinfection and COVID-19 patients without coinfection. Independent sample t test or Mann-Whitney U test was used to compare age and hospitalization period between these groups. In addition, binominal logistic regression was applied to identify risk factors associated with mortality and bacterial coinfection. The study included 342 patients with laboratory confirmed COVID-19. Eighty (23.3%) patients were diagnosed with bacterial coinfection, while the remaining 262 (76.6%) patients did not test positive for bacterial coinfection. Length of hospital stay was prolonged among COVID-19 patients diagnosed with bacterial coinfection (16.01 ±â€…11.36 days) when compared with patients without bacterial coinfection (6.5 ±â€…6.12 days). Likewise, the mortality rate was significantly higher among COVID-19 patients with bacterial coinfection (90%) compared to those without bacterial coinfection (49.2%). Gram-negative bacteria were predominant compared to gram-positive, as Klebsiella pneumoniae (35 [43.8%]) and Acinetobacter baumanni (32 [40%]). On the other hand, Staphylococcus aureus (4 [5%]), Enterococcus faecalis (1 [1.3%]), and Enterococcus faecium (1 [1.3%]) were identified as gram-positive bacterial species from recruited patients. The findings of the current study showed that prolong hospitalization is the main risk factor associated with bacterial coinfection and death. Thus, health care providers should minimize hospitalization as well as following a continuous monitoring for bacterial coinfection among COVID-19 patients, to control the spread of infection and reducing the severity and mortality rate among COVID-19 patients.

NADP/H binding nearly doubles the stability of a Mycobacterium drug target: an unfolding study.

Mycobacterium Aspartate beta semialdehyde dehydrogenase (ASADH) was studied using various spectroscopic techniques and size exclusion chromatography to examine the unfolding of free (apo) and NADP/H-bound (holo) forms of ASADH. Non-cooperative guanidinium chloride (GdnHCl)-induced unfolding of the apo ASADH was discovered, and no partially folded intermediate structures were stabilized. On the other hand, it was observed that GdnHCl's unfolding of holoenzyme was a cooperative process without any stable intermediate structure. The native form of holoenzyme is found to be stable against the lower concentration of GdnHCl only (namely up to 1.25 M GdnHCl). The tryptophan environment appears to unfold cooperatively in case of the holoenzyme and is in well coordination with the overall unfolding of the holoenzyme. The presence of NADP/H shows a stabilizing effect on the tryptophan environment as well as on the native NADP/H-bound enzyme. ΔGSolvento values reveal nearly two-fold (∼1.9) conformationally more stable folded holoenzyme compared to its native apo state. The Cm for the apo and holo forms of ASADH are 1.3 and 1.9 M, respectively. Novel drug leads targeting the NADP/H binding domain of ASADH could offer promising drugs against extremely infective Mycobacterium tuberculosis.Communicated by Ramaswamy H. Sarma.

Directional preference for glioblastoma cancer cell membrane encapsulated nanoparticle population: A probabilistic approach for cancer therapeutics.

Background: Selective cancer cell recognition is the most challenging objective in the targeted delivery of anti-cancer agents. Extruded specific cancer cell membrane coated nanoparticles, exploiting the potential of homotypic binding along with certain protein-receptor interactions, have recently proven to be the method of choice for targeted delivery of anti-cancer drugs. Prediction of the selective targeting efficiency of the cancer cell membrane encapsulated nanoparticles (CCMEN) is the most critical aspect in selecting this strategy as a method of delivery. Materials and methods: A probabilistic model based on binding scores and differential expression levels of Glioblastoma cancer cells (GCC) membrane proteins (factors and receptors) was implemented on python 3.9.1. Conditional binding efficiency (CBE) was derived for each combination of protein involved in the interactions. Selective propensities and Odds ratios in favour of cancer cells interactions were determined for all the possible combination of surface proteins for 'k' degree of interaction. The model was experimentally validated by two types of Test cultures. Results: Several Glioblastoma cell surface antigens were identified from literature and databases. Those were screened based on the relevance, availability of expression levels and crystal structure in public databases. High priority eleven surface antigens were selected for probabilistic modelling. A new term, Break-even point (BEP) was defined as a characteristic of the typical cancer cell membrane encapsulated delivery agents. The model predictions lie within ±7% of the experimentally observed values for both experimental test culture types. Conclusion: The implemented probabilistic model efficiently predicted the directional preference of the exposed nanoparticle coated with cancer cell membrane (in this case GCC membrane). This model, however, is developed and validated for glioblastoma, can be easily tailored for any type of cancer involving CCMEN as delivery agents for potential cancer immunotherapy. This probabilistic model would help in the development of future cancer immunotherapeutic with greater specificity.

Increased inflammatory cytokines and oxidative stress enhanced antibody production in breast and prostate cancer patients with COVID-19 related depression.

Cancer management is highly dependent on the immune status of the patient. During the COVID-19 pandemic, a large number of people suffered from anxiety and depression, especially cancer patients. The effect of depression on breast cancer (BC) and prostate cancer (PC) patients, during the pandemic has been analyzed in this study. Levels of proinflammatory cytokines (IFN-γ, TNF-α, and IL-6) and oxidative stress markers malondialdehyde (MDA) and carbonyl content (CC) were estimated in patients' serum samples. Serum antibodies against in vitro hydroxyl radical (•OH) modified pDNA (•OH-pDNA-Abs) were estimated using direct binding and inhibition ELISA. Cancer patients showed increased levels of proinflammatory cytokines (IFN-γ, TNF-α, and IL-6) and oxidative stress markers (MDA and CC levels), which were further significantly enhanced in cancer patients with depression compared to normal healthy (NH) individuals. Increased levels of •OH-pDNA-Abs were detected in breast cancer (0.506 ± 0.063) and prostate cancer (0.441 ± 0.066) patients compared to NH subjects. Serum antibodies were found to be significantly elevated in BC patients with depression (BCD) (0.698 ± 0.078) and prostate cancer patients with depression (PCD) (0.636 ± 0.058). Inhibition ELISA also exhibited significantly high percent inhibition in BCD (68.8% ± 7.8%) and PCD (62.9% ± 8.3%) subjects compared to BC (48.9% ± 8.1%), and PC (43.4% ± 7.5%) subjects. Cancer is characterized by enhanced oxidative stress and increased inflammation, which may be exaggerated with COVID-19 related depression. High oxidative stress and compromised antioxidant homeostasis exerts alterations in DNA, leading to formation of neo-antigens, subsequently leading to the generation of antibodies. COVID-19 pandemic related depression needs to be addressed globally for improved cancer patient care and cancer disease management.

Assessment of key regulatory genes and identification of possible drug targets for Leprosy (Hansen's disease) using network-based approach.

Leprosy is a major health concern and continues to be a source of fear and stigma among people worldwide. Despite remarkable achievements in the treatment, understanding of pathogenesis and transmission, epidemiology of leprosy still remains inadequate. The prolonged incubation period, slow rates of occurrence in those exposed and deceptive clinical presentation pose challenges to develop reliable strategies to stop transmission. Hence, there is a need for improved diagnostics and therapies to prevent mortality caused by leprosy. The objectives of this study are to identify significant genes from protein-protein interactions (PPIs) network of leprosy and to choose the most effective therapeutic targets. Fifty genes related with leprosy were discovered by literature mining. These genes were used to construct a primary network. Leading Eigen Vector method was used to break down the primary network into various sub-networks or communities. It was found that the primary network was divided into many sub-networks at the 6 levels. Seed genes were traced at each level till key regulatory genes were identified. Three seed genes, namely, GNAI3, NOTCH1, and HIF1A, were able to make their way till the final motif stage. These genes along with their interacting partners were considered key regulators of the leprosy network. This study provides leprosy-associated key genes which can lead to improved diagnosis and therapies for leprosy patients.

Inhibitory effect of natural compounds on Dihydropteroate synthase of Mycobacterium leprae: Molecular dynamic study.

Leprosy is a chronic infectious disease caused by a bacillus, Mycobacterium leprae. According to official data from 139 countries in the 6 WHO Regions, there were 127558 new leprosy cases worldwide in 2020. Leprosy mainly affects the skin, the peripheral nerves, mucosa of the upper respiratory tract, and the eyes. If this disease is left untreated, can harm the skin, nerves, limbs, eyes, and skin permanently. The disease is curable with multidrug therapy. Over a period of time Mycobacterium leprae has become resistant to these drugs. Therefore, new therapeutic molecules are warranted. This study was aimed to carry out the in-silico analysis to determine the inhibitory effect of natural compounds on Dihydropteroate synthase (DHPS) of Mycobacterium leprae. The DHPS is a key enzyme in the folate biosynthesis pathway in M. leprae and acts as a competitive inhibitor of PABA. The 3D structure of DHPS protein was modeled using homology modeling and was validated. Molecular docking and simulation along with other in-silico methods were employed to determine the inhibitory effect of ligand molecules towards DHPS target protein. Results revealed ZINC03830554 molecule as a potential inhibitor of DHPS. Binding experiments and bioassays utilizing this strong inhibitor molecule against purified DHPS protein are necessary to validate these early findings.Communicated by Ramaswamy H. Sarma.

The effect of non-surgical periodontal therapy on salivary IL-6 levels in patients with moderate to severe generalized chronic periodontitis.

BACKGROUND: Gene regulation of IL-6 is characterized by the presence of inflammatory cytokines, bacterial products, viral infection, and activation of the diacylglycerol-, cyclic AMP-, or Ca + + -activated signal transduction pathways. AIM: Scaling and root planning (SRP), a non-surgical periodontal therapy, was studied in connection to several clinical parameters for its effect on salivary IL-6 levels in patients with generalized chronic periodontitis. METHODS: For this study, a total of 60 GCP patients were included. Plaque index (PI), gingival index (GI), pocket probing depth (PPD), bleeding on probing percentage (BOP%), and clinical attachment loss were among the clinical indicators covered (CAL). RESULTS: Following SRP, mean IL-6 levels in patients with GCP were significantly higher in the pre-treatment group (2.93 5.17 pg/ml; p 0.05) than in the posttreatment group (5.78 8.26 pg/ml; baseline). Pre- and post-treatment IL-6 levels for PI (pre), BOP percent (pre/post), GI (post), and PPD were found to be positively correlated (post). In patients with GCP, the study showed a statistically significant correlation between periodontal metrics and salivary IL-6. CONCLUSIONS: Changes in periodontal indices and IL-6 levels that are statistically significant over time indicate that non-surgical treatment is effective, and IL-6 can be regarded as a potent disease activity marker.

Comparison of Hematological Parameters Between Survivors and Non-Survivors COVID-19 Patients in Saudi Arabia.

Objective: Coronavirus disease 2019, caused by the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), is a communicable disease transmitted through the respiratory route and bodily contact. The severity of infection and mortality rate of COVID-19 cases was significantly high in the initial stages of the pandemic. This study aims to investigate the hematological profile of COVID-19 survivors and non-survivors. Methods: This is a single-center retrospective study. A total of 108 hospitalized patients with laboratory-confirmed COVID-19 at East Jeddah Hospital between April and August 2020 were categorized into two groups based on outcome as survivors (n = 54) and non-survivors (n = 54). Hematological parameters and clinical profiles were analyzed and compared between the two groups. Results: The mean age and standard deviation of the survived (30-71 years) and non-survived (33-83) groups was 53 ± 10.8 and 57.9 ± 12.2 years, respectively, with no statistically significant difference in age between groups (p = 0.0513). Non-survivors had a significantly longer median length of stay in the intensive care unit (ICU) (7 days, IQR: 4.24 to 12) compared to survivors COVID-19 patients (5 days, IQR: 0 to 11.75) (p = 0.0151). For the survivors group, the participant's age positively correlated with the length of hospital stay (r(52) = 0.21, p = 0.0005) and ICU length of stay r(52) = 0.18, p = 0.001). The median red blood cells (RBC) counts were significantly higher in the survived group (4.56x109/L, IQR: 4.02 to 5.11) in comparison with the non-survived (4.23x109/L, IQR: 3.75 to 4.23) group (p = 0.0011). All COVID-19 patients exhibited lymphocytopenia and a significant negative correlation was observed between the lymphocyte values and length of hospital stay among the survived group (p < 0.001) as well as length of ICU stay among the survived group (p < 0.0480). Disease-related mortality was significantly associated with reduced white blood cells (WBCs) (8.5×109/L, IQR: 6.1 to 11.7) and reduced basophils (0.09%, IQR: 0.02 to 0.19). Additionally, statistically significant differences were found between the survived and non-survived groups with respect to prothrombin time (PT) (12.5 sec. vs 14 sec., p < 0.0001) and partial thromboplastin time (PTT) (31.8 sec. vs 40 sec., p = 0.0008). Conclusion: Hematological parameters can serve as valuable indicators to identify patients with severe COVID-19 and expected poor-prognosis/outcomes upon hospital admission. Cell counts of lymphocytes, WBCs, basophils and parameters such as PT and PTT can serve as clinical indicators to assess disease severity and predict progression to critical illness.

Towards enhancement of fungal hydrolytic enzyme cocktail using waste algal biomass of Oscillatoria obscura and enzyme stability investigation under the influence of iron oxide nanoparticles.

Development of low-cost and economic cellulase production is among the key challenges due to its broad industrial applications. One of the main topics of research pertaining to sustainable biomass waste based biorefinaries is the development of economic cellulase production strategies. The main cause of the increase in cellulase production costs is the use of commercial substrates; as a result, the cost of any cellulase-based bioprocess can be decreased by employing a productive, low-cost substrate. The goal of the current study is to develop low-cost cellulase using the carbohydrate-rich, renewable, and widely accessible cyanobacteria algae Oscillatoria obscura as the production substrate. Maximum cellulase was produced utilising the fungus Rhizopus oryzae at substrate concentration of 7.0 g among various tested concentrations of algal biomass. Maximum production rates of 22 IU/gds FP, 105 IU/gds BGL, and 116 IU/gds EG in 72 h were possible under optimal conditions and substrate concentration. Further investigations on the crude enzyme's stability in the presence of iron oxide nanoparticles (IONPs) revealed that it was thermally stable at 60 °C for up to 8 h. Additionally, the crude enzyme demonstrated pH stability by maintaining its complete activity at pH 6.0 for 8 h in the presence of the optimal dose of 15 mg IONPs. The outcomes of this research may be used to investigate the possibility of producing such enzymes in large quantities at low cost for industrial use.

Bioremediation potential of Pseudomonas fluorescens strain isolated from the Ha'il region of Saudi Arabia.

Increased amounts of toxicants may cause sever health issues in humans as well as in aquatic life. Scientists are developing new technologies to combat these problems. Biological methods of detoxification are always beneficial for the environment. Pseudomonas fluorescens is known for its detoxification capacity. In this study Pseudomonas fluorescens stains were isolated from different locations of the Ha'il region, Saudia Arabia. The microbial strain AM-1 displayed resistance to heavy metals (Cr6+, Ni2+, Cd2+, Pb2+) and pesticides (BHC, 2,4-D, Mancozeb) at pollutant levels typical of highly contaminated areas. Additionally, AM-1 exhibited substantial detoxification potential, reducing toxicity by 40.67% for heavy metals and 47.4% for pesticides at 3x concentrations. These findings suggest that the AM-1 strain supports environmental remediation and pollution mitigation. Atomic absorption spectrometry (AAS) results exhibited bioremediation efficiency for metals Cr6+, Ni2+, and Pb2+ using immobilized cells of P. fluorescens AM-1 isolate, estimated to be 60.57%, 68.4%, and 53.93% respectively. These findings show that AM-1 strain has a potential role in bioremediation of water pollutants and may have future implications in wastewater treatment.

Seroprevalence of Anti-S1-RBD Antibodies in Pre-pandemic and Pandemic Subjects From Hail Region, KSA.

Background: Two years into the pandemic, yet the threat of new SARS-CoV-2 variants continues to loom large. Sustained efforts are required to fully understand the infection in asymptomatic individuals and those with complications. Identification, containment, care, and preventative strategies rely on understanding the varied humoral immune responses. Methods: An in-house ELISA was developed and standardized to screen for serum IgG antibodies against the SARS-CoV-2 S1-RBD protein as an antigen. This study aims to investigate the seroprevalence of serum antibodies against S1-RBD antigen in pre-pandemic (n = 120) and during the early pandemic period (n = 120) in subjects from the Hail region, KSA and to correlate it with clinical and demographic factors. Results: Samples collected from both male (n = 60) and female (n = 60) subjects during the pandemic in the age groups of 20-40 (0.31 ± 0.029 and 0.29 ± 0.024, respectively) and 41-60 years (0.35 ± 0.026 and 0.30 ± 0.025, respectively) showed significantly higher levels of serum antibodies against S-RBD antigen than the age-matched pre-pandemic samples [male (n = 60) and female (n = 60)]. Pandemic subjects exhibited significantly (p < 0.01) higher inhibition (80-88%) than age-matched pre-pandemic subjects (32-39%). Antibodies against S1-RBD antigen were detected in approximately 10% of the total pre-pandemic population (males and females). However, subjects > 60 years did not show antibodies. Conclusion: Antibody levels increased in samples collected during the pandemic, even though these subjects were not clinically COVID-19 positive. A small number of pre-pandemic subjects showed serum antibodies, suggesting prior exposure to other coronaviruses in the region. With dwindling neutralizing antibody levels and reduced vaccine efficacy against newer variants, it remains crucial to develop better assays for surveillance, management, and future research.

Awareness and Practice of COVID-19 Precautionary Measures Among Healthcare Professionals in Saudi Arabia.

OBJECTIVE: Healthcare workers (HCWs) are the backbone of the healthcare system and a skilled and healthy workforce is vital during a health crisis, such as the present coronavirus disease 2019 (COVID-19) pandemic. Healthcare workers are at higher risk of exposure to and transmission of the severe acute respiratory coronavirus-2 (SARS-CoV-2). Hence, HCWs should possess good knowledge and attitudes toward COVID-19 to protect not only themselves but also their colleagues, families and the larger community. Thus, the current study aims to assess the knowledge and practice of protective measures by HCWs in Saudi Arabia during the first wave of the COVID-19 pandemic to identify awareness of the disease, potential trends and associated predictors. METHODS: A cross-sectional, web-based study was conducted among HCWs about knowledge related to COVID-19 and practice of protective measures, such as social distancing and use of personal protective equipment (PPE) during the pandemic. A Fisher exact test and one-way analysis of variance (ANOVA) were used to investigate the level of association among variables. RESULTS: A total of 674 HCWs were recruited in the study (51.8% male, 42.7% aged 28-37 years old and 52% specialists). The Saudi Ministry of Health (MoH) was the main source of knowledge for most of the HCWs (89%) followed by the WHO (44.5%) and social media (42.3%). Washing hands before touching the face was the most selected choice (97.9%) as a precautionary method to limit SARS-CoV2 transmission. Most of the HCWs (74.6%) scored low for staying at home while the majority (71.2%) showed a high practice of personal protective methods, with pharmacists and general physicians scoring the highest. CONCLUSION: Saudi HCWs showed high knowledge and practice of protective measures for COVID-19. Good knowledge correlates with adoption of appropriate practices to prevent spread of infection. The current findings highlight the importance of interventions such as tailored education and training courses for those with low scores to improve overall knowledge and practice.

Biologically Active α-Amino Amide Analogs and γδ T Cells-A Unique Anticancer Approach for Leukemia.

Advanced stage cancers are aggressive and difficult to treat with mono-therapeutics, substantially decreasing patient survival rates. Hence, there is an urgent need to develop unique therapeutic approaches to treat cancer with superior potency and efficacy. This study investigates a new approach to develop a potent combinational therapy to treat advanced stage leukemia. Biologically active α-amino amide analogs (RS)-N-(2-(cyclohexylamino)-2-oxo-1-phenylethyl)-N-phenylpropiolamide (α-AAA-A) and (RS)-N-(2-(cyclohexylamino)-2-oxo-1-phenylethyl)-N-phenylbut2-enamide (α-AAA-B) were synthesized using linear Ugi multicomponent reaction. Cytotoxicities and IC50 values of α-AAA-A and α-AAA-B against leukemia cancer cell lines (HL-60 and K562) were analyzed though MTT assay. Cytotoxic assay analyzed percent killing of leukemia cell lines due to the effect of γδ T cells alone or in combination with α-AAA-A or α-AAA-B. Synthesized biologically active molecule α-AAA-A exhibited increased cytotoxicity of HL-60 (54%) and K562 (44%) compared with α-AAA-B (44% and 36% respectively). Similarly, α-AAA-A showed low IC50 values for HL-60 (1.61 ± 0.11 µM) and K562 (3.01 ± 0.14 µM) compared to α-AAA-B (3.12 ± 0.15 µM and 6.21 ± 0.17 µM respectively). Additive effect of amide analogs and γδ T cells showed significantly high leukemia cancer cell killing as compared to γδ T cells alone. A unique combinational therapy with γδ T cells and biologically active anti-cancer molecules (α-AAA-A/B), concomitantly may be a promising cancer therapy.

Optimization of methods for peripheral blood mononuclear cells isolation and expansion of human gamma delta T cells.

Human Vg9/Vδ2 T cells (γδ T cells) are immune surveillance cells both in innate and adaptive immunity and are a possible target for anticancer therapies, which can induce immune responses in a variety of cancers. Small non-peptide antigens such as zoledronate can do activation and expansion of T cells in vitro. It is evident that for adoptive cancer therapies, large numbers of functional cells are needed into cancer patients. Hence, optimization of methods needs to be carried out for the efficient expansion of these T cells. Standardization of peripheral blood mononuclear cells (PBMCs) isolation was devised. Cytokines (interleukin 2 (IL-2) and interleukin 15 (IL-15)) and zoledronate were also standardized for different concentrations. It was found that an increased number of PBMCs were recovered when washing was done at 1100 revolution per minute (rpm). Significantly high expansion fold was (2524 ± 787 expansion fold) achieved when stimulation of PBMCs was done with 1 µM of zoledronate and both cytokines IL-2 and IL-15 supported the expansion and survival of cells at the concentrations of 100 IU/ml and 10 ng/ml respectively. 14-day cultures showed highly pure (91.6 ± 5.1%) and live (96.5 ± 2.5%) expanded γδ T cells. This study aimed to standardize an easy to manipulate technique for the expansion of γδ T cells, giving a higher yield.