Assay Method Research Articles

The relationship between oxLDL, sLOX-1, PCSK9 and carotid intima-media thickness in patients with prediabetes and type 2 diabetes

Affection of vascular structures is a known complication in diabetes and prediabetes. Subclinical atherosclerosis begins to develop years before both cardiovascular and cerebrovascular diseases become symptomatic. The aim of this study was to evaluate the relationship between circulating oxidized LDL (oxLDL), soluble lectin-like oxidized LDL (sLOX)-1 and proprotein convertase subtilisin/kexin type 9 (PCSK9) levels and carotid intima-media thickness (CIMT) in patients with prediabetes and T2DM. We recruited 50 T2DM patients with macrovascular complications, 50 T2DM patients with uncomplicated, 50 prediabetes patients, and 50 healthy participants. The common carotid arteries were visualized by high-resolution B-mode carotid artery ultrasonography. Measurement of serum oxLDL, sLOX-1 and PCSK9 levels were assessed by using a commercially available human enzyme-linked immunosorbent assay (ELISA) method. Homeostasis model assessment for insulin resistance (HOMA-IR) was also calculated. Circulating LDL-C, oxLDL, sLOX-1 and PCSK9 levels were significantly higher in T2DM patients with macrovascular complications compared to control group, prediabetes and uncomplicated diabetes. There was significant difference, especially between the controls and the T2DM patients with macrovascular complications. The CIMT increased progressively from control through to T2DM. Both right and left CIMT also showed significant differences between the groups of prediabetes versus uncomplicated diabetes at p < 0.001 showing progressive increase in vascular involvement with progression of disease. There was a positive correlation between PCSK9 levels with LDL-C, oxLDL, sLOX-1, and CIMT. ROC curve analyses showed that PCSK9, as well as oxLDL and LOX-1, was a strong predictor of T2DM with high sensitivity and specificity. In this study, we found that oxLDL, sLOX-1 and PCSK9 levels were significantly associated with carotid intima-media thickness. Our findings suggest that these biomarkers may be used as potential markers for the assessment of cardiovascular risk in patients with prediabetes and type 2 diabetes.

The effect of probiotic-fortified kefir on depression, appetite, oxidative stress, and inflammatory parameters in Iranian overweight and obese elderly: a randomized, double-blind, placebo-controlled clinical trial

BackgroundIt has been shown that the microflora of the gastrointestinal tract undergoes changes in obese individuals. The present study aimed to investigate the effect of kefir fortified with two strains, Lactobacillus helveticus and Bifidobacterium longum, on depression, appetite, oxidative stress, and inflammatory parameters in overweight and obese elderly individuals.MethodsThis study was a double-blind, randomized, and placebo-controlled clinical trial conducted on 67 elderly men aged over 65, who were randomly divided into two groups. One group (n = 35) received one bottle (240 cc) of regular kefir as a placebo, while the intervention group (n = 32) received one bottle of probiotic-fortified kefir for eight weeks. Depression and appetite were evaluated using the Geriatric Depression Scale-15 (GDS-15) and a validated Visual Analogue Scale (VAS), respectively. Oxidative stress parameters were assessed using the standard calorimetric method, and inflammatory parameters were measured via the enzyme-linked immunosorbent assay method (ELISA). The differences between the two groups were compared using the independent samples T-test.ResultsThe median age of participant in both groups was 65 years. A significant difference in depression scores and the mean change between the two groups was observed after eight weeks (p = 0.001 and p = 0.042, respectively). Within-group comparison revealed a significant increase in appetite scores in both groups (p < 0.05 for both). Moreover, a significant difference in the changes in total antioxidant capacity (TAC) was noted (p = 0.009). However, no significant differences were observed in other oxidative and inflammatory parameters between the two groups (p˃0.05 for all).ConclusionsThe results demonstrated the positive impact of two specific strains of Bifidobacterium and Lactobacillus on improving depression in the elderly. However, when comparing the two groups, no significant effects were observed on appetite, inflammation, and oxidative stress parameters, except for TAC.

Anticariogenic potential of selected medicinal plants from Dayak Benuaq tribe, Indonesia

Several selected medicinal plants from the indigenous Dayak Benuaq tribe of Indonesia were analyzed for their potential as anticariogenic. The plant parts of leaves, stems, roots, and tubers were extracted using methanol. Streptococcus sobrinus and Streptococcus mutans are used to determine antibacterial and anticariogenic activities. Toxicity activity was evaluated using Artemia salina with the brine shrimp lethality assay (BSLT) method. This study also calculated the content of total tannins, total flavonoids and total phenols. The results of this study indicate that four selected medicinal plants, namely Cratoxylum sumatranum (Jack) Blume., Areca catechu L., Syzygium aromaticum (L.) Merr. &amp; L.M. Perry., and Lepisanthes amoena (Hassk.) Leenh., have the potential to inhibit the growth of S. sobrinus bacteria with inhibition values of 89.75%, 80.17%, 71.37%, and 74.69% in concentrations of 100 ppm. The highest content of total tannins, total phenols and total flavonoids was in Uncaria gambir (W. Hunter) Roxb, with respective values of 205.94 g/g, 478.52 mg GAE/g, and 0.725 g CE/g. Anticariogenic activity showed that U. gambir, Helminthostachys. zeylanica, S. aromaticum, L. amoena, and Eurycoma longifolia (Jack) had the potential to inhibit acid production in bacteria with a pH of 6.380, 6.563, 6.140, 5.987 and 5.933, respectively. Besides that, it can also inhibit the attachment of bacterial cells with values of 31%, 50%, 43%, 53%, and 40%. The results of this study indicate the potential of several selected medicinal plants from the Dayak Benuaq tribe in Temula village as natural antibacterial, anticancer, and anticariogenic agents.

Chemical Analysis of Euphorbia denticulata Lam. and Study of extracted oil on Cytotoxicity and anti-tumor potential on AMGM-5 and HBL 100 cell lines

Study have been conducted on the cytotoxicity of Euphorbia denticulate oil. In The normal human breast cell line HBL 100 and human brain cancer cell line AMGM-5 Depending on the MTT(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide assay method Following the application of various oil concentrations (0.01, 0.05, 0.1, 0.5, 1, 5) μg \ ml to the cells, the current study was able to determine the The Half maximal inhibitory concentration( IC50) value in AMGM-5 to be 0.21505%, while the normal cell line HBL100 revealed an IC50 value of 0.18688%). the MTT assay test findings suggested a suppression of the proliferation of cancer cells. The AMGM-5cell lines changed cellularly as a result of the oil Following a 48-hour course of therapy, the degeneration showed varying degrees of atrophy, swelling, and breakdown. Ultimately, the cells completely disintegrated, resulting in the formation of necrotic regions devoid of cells. Cultures of cells: When the cells were exposed to high oil concentrations, these alterations worsened

Development and validation of an LC-MS/MS multiplex assay for the quantification of bedaquiline, n-desmethyl bedaquiline, linezolid, levofloxacin, and clofazimine in dried blood spots.

Dried blood spot (DBS) assays to quantify novel and repurposed drugs for the treatment of rifampicin-resistant tuberculosis (RR-TB) would facilitate pharmacokinetic studies and therapeutic drug monitoring in low-middle income settings, considering their ease of application and simple sample storage requirements. We describe a DBS method for the simultaneous quantification of bedaquiline and metabolite N-desmethyl bedaquiline, linezolid, levofloxacin, and clofazimine. The analytes were extracted from the matrix and isolated by solid-phase extraction. Two LC-MS/MS systems were used, optimized for the separate analysis of the more polar compounds (linezolid and levofloxacin), and less polar compounds (bedaquiline, N-desmethyl bedaquiline, and clofazimine), employing gradient elution. Electrospray ionization and multiple reaction monitoring were used to quantify the analytes on a Sciex API3200 and an API5500 triple quadrupole mass spectrometer, for the more polar and less polar analytes, respectively. Isotopically labelled internal standards were used to compensate for variability in the quantification of each analyte. The method was validated according to international guidelines and applied to samples from a clinical trial. We performed correlation and agreement analysis of the DBS assay and in-house plasma methods using Deming regressions and Bland-Altman plots. Coefficients of correlation between measured plasma and DBS concentrations ranged from 0.866 (95% CI: 0.817-0.902) to 0.989 (95% CI: 0.985-0.992). More than 67% of the samples showed a difference between the observed and estimated plasma concentrations within 20% of their means, meeting EMA requirements for method reproducibility and demonstrating the interchangeability of our DBS and plasma LC-MS/MS methods.

Mitochondria-rich extracellular vesicles derived from the culture supernatant of human synovial Fluid-derived mesenchymal stem cells Inhibited senescence of Stressed/inflammatory Licensed chondrocytes and Delayed Osteoarthritis progression.

Mitochondrial dysfunction induces chondrocyte senescence, thereby precipitating articular cartilage (AC) degeneration in the pathogenesis of osteoarthritis (OA). Although the transfer of mitochondria from mesenchymal stem cells (MSCs) to host cells and their potential protective role have been demonstrated, whether MSCs can alleviate chondrocyte mitochondrial dysfunction or reverse OA progression remains unclear. A mitochondrial tracer was used to investigate the transfer of mitochondria-rich extracellular vesicles (MEV) derived from the culture supernatant of human synovial fluid-derived mesenchymal stem cells (hSF-MSCs). Human articular chondrocytes (hACs) impaired by oxidative stress co-incubated with MEV were used for experimental research in vitro. Healthy hACs and stressed hACs were cultured separately acting as the control groups. The MEV was injected into the OA rats' knee joint serving as experimental group. Healthy and OA rats were served as the control groups. Quantitative reverse transcription polymerase chain reaction (qRT-PCR), western blot (WB), enzyme- linked immunosorbent assay (ELISA), flow cytometry (FC), immunofluorescence (IF), fluorescence spectrophotometer (FS), immunohistochemistry (IHC) and other methods are used to analyze the effect of MEV on hACs and OA progression. MEV derived from hSF-MSCs could transfer into hACs. Compared to the negative control group, co-incubation with MEV resulted in a significant down-regulation of oxidative stress markers and senescence-associated proteins in hACs, while improved mitochondrial function of hACs. Moreover, the MEV could traverse the dense interstitial layer and migrate towards the deeper cartilage, while intra-articular injection of MEV could effectively attenuate AC degeneration. The transfer of MEV derived from hSF-MSCs represents a promising strategy for safeguarding AC, thereby offering a potential avenue and mechanism for the treatment of OA.

Novel solid-phase extraction promotes simultaneous colloidal gold immunochromatographic assay of malachite green, leuco-malachite green, chloramphenicol, and semi-carbazone metabolites in aquatic products.

Solid phase extraction (SPE) using traditional column materials offers several advantages, but its ability to simultaneously extract and purify commonly used veterinary drugs, such as chloramphenicol, malachite green, leuco-malachite green and semi-carbazone, remains unproven.Therefore, in the present study, a rapid analytical method based on ultrasound-assisted SPE combined with colloidal gold test strips was developed and applied herein to determine the residues of these veterinary drugs in common aquatic products. Under optimized conditions, the method achieved a limit of detection in the range of 0.01-0.5μg/kg, inter-day RSDs of the spiked samples in the ranges of 0.99-5.58% and 1.6-7.2%, and absolute recoveries in the range of 84.2-112.9% using the colloidal gold immunoassay and enzyme-linked immunosorbent assay methods, respectively. The SPE column efficiently enriched the four veterinary drugs simultaneously, showing considerable potential for determining common veterinary drug residues in aquatic products.

Extracellular hydrolytic enzyme activities and biofilm formation in Candida species isolated from people living with human immunodeficiency virus with oropharyngeal candidiasis at HIV/AIDS clinics in Uganda.

Commensal oral Candida species can become opportunistic and transition to pathogenic causes of oropharyngeal candidiasis (OPC) in individuals with impaired immunity through ecological cues and the expression of extracellular hydrolytic enzyme activities and biofilm formation. We evaluated phospholipase, proteinase, hemolysin, esterase, and coagulase enzymatic activities and biofilm formation in Candida species isolated from people living with human immunodeficiency virus (PLHIV) with OPC. Thirty-five Candida isolates from PLHIV with OPC were retrieved from a sample repository and evaluated for phospholipase activity using the egg yolk agar method, proteinase activity using the bovine serum albumin agar method, hemolysin activity using the blood agar plate method, esterase activity using the Tween 80 opacity test medium method, coagulase activity using the classical tube method, and biofilm formation using the microtiter plate assay method in vitro. A total of 35 Candida isolates obtained from PLHIV with OPC were included in this study, and phospholipase and proteinase activities were detected in 33/35 (94.3%) and 31/35 (88.6%) Candida isolates, respectively. Up to 25/35 (71.4%) of the Candida isolates exhibited biofilm formation, whereas esterase activity was demonstrated in 23/35 (65.7%) of the Candida isolates. Fewer isolates (21/35, 60%) produced hemolysin, and coagulase production was the least common virulence activity detected in 18/35 (51.4%) of the Candida isolates. Phospholipase and proteinase activities were the strongest in oropharyngeal Candida species.

Assessment of serum levels of 8-hydroxy-2’ deoxyguanosine and f2-isoprostanes in newly diagnosed adult hypertensive with clinical depression in NAUTH, Nnewi, Nigeria

Despite the high prevalence of depression and hypertension, the relationship between the two diseases has received little attention. The potential roles of some biomarkes of oxidative stress in disease progression and management is very crucial. Aim: The present study therefore, evaluated the serum levels of 8-hydroxy-2’ deoxyguanosine (8-OHdG) and F-2 isoprostanes in newly diagnosed hypertensive individuals with and without depression at Nnamdi Azikiwe University Teaching Hospital, Nnewi. Method: This was a cross sectional study carried out on randomly selected 121 consented adult male and female participants within the ages of 18-65 years which comprises 30 newly diagnosed hypertensive individuals without depression (Hypertensive), 31 hypertensive individuals with depression selected from the department of internal medicine. 30 non hypertensive with depression (Depressive) and 30 apparently healthy non-hypertensive individuals without depression which served as control selected among the hospital staff. Their bio-data was obtained from their hospital records. 3 ml of venous blood were collected from each of the participants, dispensed into a plain container, centrifuged and serum separated into another container and stored at -20 0C for the assessment of serum 8-OHdG using Enzyme Linked Immunosorbent Assay (ELISA) Method. Result: The result showed that Hypertensive individuals with and without clinical depression (107.08±22.05, 129.16±82.49) had significantly higher serum level of 8-OHdG when compared with control group (53.95±28.25) (P&lt;0.05 respectively). Hypertensive individuals (129.16±82.49) also had significantly higher 8-OHdG level when compared with non-hypertensive with depression (63.31±37.30) (P&lt;0.05). Hypertensive individuals with depression (9.94±7.14) and depressive individuals (8.99±5.84) also had significantly higher F2-Isoprostanes level when compared with control (5.15±1.47) (P&lt;0.05 respectively). There was a moderately strong positive correlation between 8-OHdG level and body mass index in both hypertensive and depressive individuals (r=0.682, P&lt;0.05). Conclusion: The study observed higher 8-OHdG and F2-Isoprostanes in individuals with hypertension with and without clinical depression. In addition, there is evidence that oxidative stress is related to inflammation and endothelial activation in individuals with both conditions leading to disease severity.

Verification of analytical performance of Creatine Kinase isoenzyme MB (CK-MB) on the Alinity c® Experience from the Biochemistry Laboratory of Mohammed VI University Hospital in Oujda

The verification of analytical methods is a requirement of the NF EN ISO 15189 standard. It involves evaluating the performance of an analytical method according to a well-defined protocol and comparing it to pre-established analytical objectives. In this study, we present the results of the verification protocol for the measurement method of creatine kinase isoenzyme MB (CK-MB) on the Alinity c® analyzer, which is employed for functional exploration of the heart. Our study is conducted according to the criteria of Scope A detailed in the Verification/Validation Guide for Methods in Medical Biology, following the SH GTA 04 recommendations from COFRAC. The verification focused on the CK-MB assay on the Alinity c® analyzer using International Federation of Clinical Chemistry (IFCC) method based on immunoinhibition principle for an assessment of analytical performance in terms of repeatability and intermediate precision. these results were aligned with the manufacturer's specifications. The results obtained for various CK-MB assay verification criteria on the Alinity c® analyzer demonstrate satisfactory repeatability across one level with CV = 1.29%. Intra-laboratory reproducibility was satisfactory for one level with CV = 2.58%. The coefficient of variation (CV) values obtained in our study were compared with manufacturer's specifications, no CV reference values were established by French Society of Clinical Biology SFBC. The findings from this study have allowed us to verify the performance of the CK-MB assay method and to compare them with the objectives set in the accreditation process to which our laboratory is committed.

Verification of the analytical performance of the urinary creatinine assay on Abbott Architect ci8200

The urinary creatinine assay is a pivotal diagnostic tool for assessing kidney function, hydration status, and muscle metabolism. This study evaluated the analytical performance of the urinary creatinine assay method using an Abbott kit on the Architect ci8200® automated system in the biochemistry laboratory of CHU Mohammed VI, Oujda. Method verification was conducted in two phases: intermediate fidelity testing using internal quality controls and repeatability testing on patient urine samples across low and high creatinine concentration levels. The results demonstrated excellent analytical performance, with coefficients of variation (CV) for intermediate fidelity (CV1 = 1,95% and CV2 = 1.46%) and repeatability (CV1 = 0.78% and CV2 = 0.87%) well within the limits established by the French Society of Clinical Biology (SFBC). Levey-Jennings charts illustrated consistent and robust performance. This method also underwent a comparative analysis using the Bland-Altman diagram, which confirmed its accuracy and precision relative to established standards. These findings underscore the reliability of the urinary creatinine assay method, ensuring high-quality diagnostic outcomes.

In vitro Antioxidant and Antibacterial Activities of Ethanolic Extract of Mentha viridis

Mentha viridis has been used as folk medicine traditionally for diverse pharmacological activity and used worldwide for its aroma for different purposes in particular with tea. This research was investigated to examine the antioxidant and antibacterial activities of ethanolic extract of M. viridis. The antioxidant capacity was assessed by several assays namely DPPH free radical scavenging assay, total antioxidant assay, total phenolics and total flavonoids content assay. Determination of total phenolics and total flavonoids were performed spectrophotometrically using Folin-Ciocalteu reagent and aluminium chloride colorimetric assay methods respectively, while the free radical scavenging activity was estimated by DPPH (2, 2-diphenyl-2-picrylhydrazyl) method. The antibacterial activity was estimated by agar disk diffusion method against a number of bacterial strains. The results exhibited that ethanolic extract M. viridis is the abundant source of phenolics and flavonoids but the total antioxidant activity was detected to be the highest in comparison to other phytochemical constituents. Extract showed robust free radical scavenging activity at a concentration of 500 μg/ml. The ethanolic extract of M. viridis displayed moderate inhibitory activity against the microorganisms S. aureus and E.coli. The interpretation stipulates that ethanolic extract of M. viridis is a potential origin of natural antioxidants due to presence of mainly polyphenolic components in it. Bangladesh Pharmaceutical Journal 28(1): 1-10, 2025 (January)

Verification of analytical performance of glycosuria assay on the Abbott Architect® analyzer: Experience of the Biochemistry Laboratory, Mohammed VI University Hospital of Oujda

The aim of our study was to evaluate the analytical performance of the glycosuria assay method using the ARCHITECT c Systems automated analyzer in the biochemistry laboratory of the Mohammed VI University Hospital in Oujda. Glycosuria, defined as the presence of glucose in urine, is a key biomarker in the diagnosis and monitoring of diabetes mellitus, reflecting disturbances in glucose metabolism. We conducted a performance study of the ARCHITECT c Systems analyzer, focusing on repeatability and reproducibility, in accordance with the COFRAC GTA 04 accreditation technical guide, which aligns with the quality requirements of ISO 15189 standards. The results of our study demonstrate satisfactory repeatability for two levels of glycosuria (Low, High), with CV1 = 1.70%, CV2 = 1.68%, Similarly, reproducibility was satisfactory with coefficients of variation (CV) for the same levels, showing CV1 = 3.66%, CV2 = 1.55%. The reliability of our glycosuria assay results is confirmed by the satisfactory analytical performance obtained, which meets the quality standards required for clinical laboratory diagnostics. These findings underline the robustness of the ARCHITECT c Systems analyzer and its compliance with ISO 15189 accreditation criteria.

89. First-in-human study of a novel half-life extended monoclonal antibody (GB-0669) against SARS-CoV2 and related sarbecoviruses

Abstract Background GB-0669 is a novel half-life extended monoclonal antibody for the prophylaxis of SARS-CoV2 infection and is one of the first biologics designed using artificial intelligence/machine learning to reach clinical development and the first against SARS-CoV2. GB-0669 targets the previously undruggable S2 domain of the SARS-CoV2 spike protein, which contains the fusion peptide and stem-helix peptides, and was selected because the S2 domain is conserved across all SARS-CoV2 variants to-date. The S2 domain is not immunodominant and is therefore not subject to selective pressure from natural infection or vaccine induced immunity. This suggesting GB-0669 will maintain its activity, unlike monoclonal antibodies directed against the SARS-CoV2 receptor-binding domain. GB-0669 is also able to neutralize other sarbecoviruses, such as SARS-CoV2 and WIV1 (Table).Table:GB-0669 EC50 against SARS-CoV2 variants and other sarbecoviruses in a VSV pseudoneutralisation assay Methods In this single-ascending dose study, healthy volunteers aged 18–55 years were administered either GB-0669 or placebo intravenously. There were five sequential dosing cohorts 100 mg, 300 mg, 600 mg, 1200 mg, 2400 mg and each cohort was randomized to receive either GB-0669 or placebo in ratios of 3:3, 3:3, 10:3, 10:3, 10:3 respectively. Total follow-up will be 43 weeks; although dosing is now complete, follow-up is ongoing.Figure:Simulated GB-0669 drug concentrations over 43 weeks (plasma and lung) following a single dose of 1200 mg Results No dose-limiting toxicity has been observed to date and all treatment-related adverse reactions observed were mild (grade 1 or 2) at all doses. Preliminary pharmacokinetic data shows dose-proportionality up to 2400 mg the highest dose tested, with a half-life of approximately 70 days. An exploratory analysis of live virus neutralization shows a clear dose response and separation from placebo at 600 and 1200 mg. Limited anti-drug antibodies were observed in a few participants, but had no observable impact on PK or viral neutralization titers. Simulated PK of GB-0669 confirmed with observed data (assuming 10% lung penetration) are at levels anticipated to protect against COVID-19 for at least 150 days (Figure). Conclusion The preliminary data support the hypothesis that GB-0669 will be able to provide a six-month duration of prophylactic efficacy against currently circulating viral variants when administered as part of a combination. Disclosures Dinesh De Alwis, PhD, Generate Biomedicines: Stocks/Bonds (Private Company) Iñaki F. Troconiz, Pharmacy, Generate Biomedicines: Advisor/Consultant Daria Hazuda, PhD, Generate Biomedicines: Stocks/Bonds (Private Company) Gavin CKW Koh, PhD, AstraZeneca: Stocks/Bonds (Public Company)|Generate:Biomedicines: Stocks/Bonds (Private Company)|GlaxoSmithKline: Stocks/Bonds (Public Company) Alexandra Snyder, MD, Generate Biomedicines: employee

Development of new powdery mildew resistant lines in garden pea (Pisum sativum L.) using induced mutagenesis and validation of resistance for the er1 and er2 gene through molecular markers

Powdery mildew (PM) caused by Erysiphie pisi Syd. is the most devastating disease of pea, affecting fresh pea production as well as the quality of the marketable harvest worldwide. The efforts were made to develop PM-resistant mutants of popular pea varieties “Lincoln” and “Azad P-1” through induced mutations by following gamma irradiation (300, 400, 500, and 600 Gy) and chemical mutagenesis, i.e., ethyl methane sulfonate (EMS) (0.3% and 0.4%). The screening of 13,868 M2 progenies at Kukumseri (summer season) followed by M3 generation at Palampur (winter season) resulted in the isolation of six putative PM-resistant mutants. The rigorous evaluation of these progenies under in vivo (field screening) and in vitro (artificial screening under greenhouse conditions and using the detached leaf assay method) conditions over the years resulted in the isolation of three PM-resistant mutants, viz., L-40-1014, L-0.3-139, and AP-0.3-129. SSR markers “PSMPSAD60 d” and “PSMPA5 c” linked to the er-1 gene indicated the presence of the “er1” gene in the mutant L-0.3-139 while the er-2 gene-linked SCAR marker “ScX171400” and SSR marker “AD141” indicated the probability of the “er-2” gene in mutant L-40-1014. The known markers linked to PM resistance genes could not be validated in the mutant AP-0.3-129, suggested to identify new markers linked to PM resistance. These PM-resistant mutants can be promising candidates as the new source of resistance for future pea breeding programs.

Comparative Assessment of Acute Pulmonary Effects Induced by Heat-Not-Burn Tobacco Aerosol Inhalation in a Murine Model

Tobacco smoking remains a major global health concern, causing preventable deaths and economic strain. Although new tobacco products such as heat-not-burn (HnB) are safer alternatives to traditional cigarettes, research on their associated risks remains limited. This study aimed to investigate the effects of HnB smoke exposure on the lungs compared to those of traditional cigarettes and the combined use of HnB and cigarettes using experiments with a mouse model. We quantitatively analyzed changes in the levels of 92 blood plasma proteins using the proximity extension assay method and observed significant changes in their levels in mice exposed to different smoke conditions; specifically, the levels of certain proteins, including Ccl20, Cxcl1, and Pdgfb, increased in the HnB smoke-exposed group, suggesting activation of nicotine pathways. Comparative analysis with traditional cigarette smoke-exposed mice further highlighted similarities and differences in their protein expression profiles. This study contributes to an improved understanding of the biological mechanisms underlying the harmful effects of alternative nicotine delivery systems and identifies potential biomarkers associated with the harmful effects of HnB smoke exposure. However, the precise impact of nicotine on the immune system may be influenced by various factors, necessitating further research.

Proximate and mineral analysis of the mushroom biscuits

Mushroom is a rich source of nutrients and is widely consumed in Asian countries. Biscuits can be easily fortified with mushroom-rich flour to provide convenient foods to supplement protein in the diet. The research study evaluated the proximate and mineral analysis of the most widely used mushroom cultivated in Chattogram and developed mushroom biscuits. The mushroom varieties (Pleurotus ostreatus, Pleurotus sajor-caju, Agaricus bitorquis, and Ganoderma lucidum) were collected and dried, then taken to the laboratory for proximate and mineral analysis by following standard assay methods. Results demonstrated that the moisture, total solids, protein, fat, carbohydrate, fibre, ash, and energy content of different types of mushrooms ranged from 53.10 to 90.10%, 9.90 to 47.00%, 3.23 to 19.70%, 0.47 to 4.20%, 4.21 to13.02%, 0.51 to 3.90%, 1.28 to 6.18 and 34.78 to 172.37 (kcal/100 g), respectively. The highest sodium, phosphorus, and copper content value was found in Agaricus bitorquis. Magnesium content was found to be the highest in Pleurotus ostreatus. Potassium, calcium, chloride, iron, and zinc were the most elevated in Ganoderma lucidum. In mushroom biscuits, the moisture, ash, protein, fat, fibre, carbohydrate, and energy value ranged from 4.03 to 3.89%, 3.47 to 4.11%, 12.15 to 13.43%, 5.43 to 5.83%, 4.71 to 5.09%, 67.03 to 70.35% and 374.15 to 378.87 (Kcal/100 g), respectively. From the sensory and microbiological point of view, mushroom biscuits exhibited significant acceptability and stability. Mushrooms and mushroom-based products are valuable food sources for human needs, able to meet day-to-day nutritional requirements as a portion of food and help the country's economy

Relationship Between Systemic Inflammatory Response Exponents, Levels of ADAM10, ADAM17 Proteins and Selected Clinical Parameters in Patients with Colorectal Cancer: Original Research Study

Chronic inflammation is a confirmed risk factor for colorectal cancer (CRC). Indicators of systemic inflammatory response (SIR), such as neutrophil-to-lymphocyte ratio (NLR) or platelet-to-lymphocyte ratio (PLR), are easily accessible indicators of the generalized inflammatory response. At the molecular level, inflammation-related carcinogenesis involves proteins from the adamalysin family: ADAM10 and ADAM17. The aim of the study was to assess NLR and PLR and their relationship with selected clinical parameters in CRC patients, as well as the correlation between ADAM10 and ADAM17 in tumor tissue and matched surgical margins with NLR and PLR values. Tumor tissue material matched surgical margins, and blood was collected from 66 patients who underwent surgery because of CRC. The concentrations of ADAM10 and ADAM17 in the collected material were tested using the enzyme-linked immunosorbent assay (ELISA) method. SIR parameters (NLR, PLR) were also determined. The results were statistically analyzed and compared with selected clinical parameters. Results: The study showed that PLR was lower in patients with comorbid cardiovascular diseases (CVD). In patients who underwent preoperative treatment, both the NLR and PLR values were higher than in patients who underwent primary surgery. There was also a negative correlation between ADAM17 concentrations in the surgical margin and PLR values. In conclusion, the presence of additional diseases such as CVD or diabetes mellitus type 2 (DMT2) or the use of preoperative treatment should be taken into account when assessing SIR parameters in CRC patients. Moreover, no clear correlations have been found between ADAM10 and ADAM17 and SIR parameters.

From Diversity to Standardization: Unveiling Key Parameters in Chick Ileum Bioassays for Enhanced Pharmacology Education and Research

Background As traditional laboratory animal use faces ethical and logistical constraints, the chick ileum has been experimented with as an alternate tissue for bioassays, which is crucial for drug discovery and pharmacology education. However, the absence of uniform bioassay protocols for chick ileum presents a significant challenge, impacting the accuracy and reliability of results in pharmacological studies. Purpose The objective of this review was to compile existing data on chick ileum bioassays from published sources, aiming to enhance future experiments and contribute to the establishment of standardized methods for conducting these bioassays. Methods We conducted a systematic literature review, utilizing keywords like “Chick ileum,” “Bioassay,” “Acetylcholine,” and “Histamine” across databases such as PubMed, Google Scholar, and ScienceDirect. The study selection adhered to specified inclusion and exclusion criteria, focusing on chick ileum research. Data extraction and subsequent analysis were performed to retrieve data for the experiment. Results Eleven articles were selected after applying strict inclusion criteria. Studies ranged from 1967 to 2019, with diverse physiological salt solutions and varied responses to essential chemicals like acetylcholine and histamine. Assay methods varied, but three-point assays were common. Commonly employed tissue lengths and temperatures were identified, and additional factors like organ bath volume and aeration were crucial for consistent results. Conclusion The review highlights the feasibility and ethical advantages of using chick ileum for bioassays. Despite challenges like variable tissue responses and methodological diversity, the review suggests these can be navigated with careful optimization, making chick ileum a valuable model for pharmacological research and education.

Occurrence of Alternaria Toxins, Ochratoxin A and Aflatoxins B1 in Green Arabic Coffee (Coffea arabica) in Saudi Arabia: Risk Assessment on HepG-2 Cell Line and Male Albino Rats

Arabica coffee (Coffee arabica L.) is among the most popular and widely consumed drinks globally today. The current study aimed to detect and quantify of potential mycotoxins in coffee beans and to investigate their potential risk on HepG-2 human liver cancer cells and male albino rats. Eighteen fully matured coffee beans samples were collected from different geographical locations in Saudi Arabia. Mycotoxins detected and quantified utilizing high-performance liquid chromatography (HPLC). Cytotoxicity of the detected toxins was investigated utilizing the MTT assay method toward HepG-2 cells and male albino rats. Cytotoxic properties of the detected mycotoxins were investigated at doses of 31.25, 62.50, 125, 250 and 500 µg/ml following exposure durations of 24 and 48 h in comparison with control. HPLC analysis revealed the detection of four Alternaria toxins (ALT-Ts) namely Altenuene (ALT), Tenuazonic acid (TA), Alternariol (AOH), and Altenuisol being AOH reported as the frequently detected mycotoxin in 18 coffee beans samples representing 75 % with quantity ranged 0.98 to 8.97 μg/g. OTA was frequently detected (10 samples, 55.5%) in concentrations varied from 0.75 to 9.87 μg/g. In addition, Aflatoxin B1 was also detected with low quantity ranged 0.89 to 3.22 μg/g. Treatment of HepG-2 cell line with combined Alternaria toxins (ALT, TeA, AOH, and Altenuisol), Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) as well as their combination triggered a decline in cell viability that was proportional to the dose administered. The elevated doses (62.50, 125, 250 and 500 µg/ml) notably reduced cell viability and induced cellular damages when contrasted with the lowest dose (31.25 µg/ml) and control. The IC50 for the tested mycotoxins registered for ALT-Ts, AFB1 and OTA as well as their combination were 115.95, 56.68, 59.86 and 31.05 µg/ml after 48 h of exposure. Secreting inflammatory markers (Interleukin-6 (IL-6), interleukin-1β (IL-1β) and COX-2), and oxidative markers (total antioxidant capacity (TAC), glutathione (GSH) and nitrite (NO) were downregulated by all mycotoxins with the highest decrease induced by their mixture. Malonaldehydes (MDA) and TAC were significantly increased by all toxins in treated rats. Hematological results revealed that AFB1, OTA and ALT-Ts-treated rats exhibited a notable decline in hemoglobin (Hb), red blood cells (RBCs), white blood cells (WBCs), and packed cell volume % (PCV). However, single and double dose-treated rats with OTA, Mix1 and Mix2 exhibited a marked increase in WBCs. The results demonstrated a notable decline in liver and kidney functions in all mycotoxins-treated rats. The studied organs (liver, kidney and spleen) exhibited significant damage, with the most severe alterations observed in the Mix1 and Mix2-treated group in comparison with the groups receiving individual treatment. This indicates that the mixture of AFB1, ALT-Ts and OTA had an enhanced toxic consequence against treated HepG-2 cell line and rats. Our results indicated the synergistic effect of the combined different mycotoxins against HepG-2 human liver cancer cells and male albino rats. This work could help fill in data gaps and enhance risk assessment for human health by addressing some of the possible risks associated with Alternaria toxins