Novel insights on the probable mechanism associated with histamine oral model-inducing neuropathological and behavioral toxicity in rats.

Histamine (HIS) is an important chemical mediator that causes vasodilation and contributes to anaphylactic reactions. Recently, HIS is an understudied neurotransmitter in the central nervous system, and its potential role in neuroinflammation and neurodegeneration is a critical area of research. So, the study's goal is to investigate the consequences of repeated oral intake of HIS on the rat's brain and explore the mechanistic way of its neurotoxicity. Thirty male rats were divided into three groups (n = 10). The following treatments were administered orally to all rats every day for 14 days. Group (1) was given distilled water, whereas groups (2 & 3) were given HIS at dosage levels 250 and 500 mg/kg body weight (BWT), respectively. Brain tissue samples were collected at 7- and 14-days from the beginning of the experiment. Our results revealed that continuous oral administration of HIS at both doses for 14 days significantly reduced the BWT and induced severe neurobehavioral changes, including depression, dullness, lethargy, tremors, abnormal walking, and loss of spatial learning and memory in rats. In all HIS receiving groups, HPLC data showed a considerable raise in the HIS contents of the brain. Additionally, the daily consumption of HIS causes oxidative stress that is dose- and time-dependent which is characterized by elevation of malondialdehyde levels along with reduction of catalase activity and reduced glutathione levels. The neuropathological lesions were commonly observed in the cerebrum, striatum, and cerebellum and confirmed by the immunohistochemistry staining that demonstrating moderate to strong caspase-3 and inducible nitric oxide synthase expressions in all HIS receiving groups, mainly those receiving 500 mg/kg HIS. NF-κB, TNF-α, and IL-1ß gene levels were also upregulated at 7- and 14-days in all HIS groups, particularly in those getting 500 mg/kg. We concluded that ROS-induced apoptosis and inflammation was the essential mechanism involved in HIS-mediated neurobehavioral toxicity and histopathology.

Polymorphisms and expressions of ADSL, MC4R and CAPN1 genes and their effects on economic traits in Egyptian chicken breeds.

In recent years, strategic plans for poultry production have emphasized quantitative traits, particularly body weight and carcass traits (meat yield), in response to overpopulation challenges. Candidate genes such as adenylosuccinate lyase (ADSL), melanocortin-4-receptor (MC4R), and calpain 1 (CAPN1) have played vital roles in this context due to their associations with muscle growth and body composition. This study aims to investigate the influence of polymorphisms and gene expressions of the aforementioned genes on body weight (BW), growth rate (GR), breast weight (BrW), and thigh weight (TW) across four distinct chicken breeds: Fayoumi, Matrouh, Mamourah, and Leghorn. The use of PCR-SSCP analysis revealed genetic polymorphisms through the identification of various patterns (genotypes) within the three examined genes. The ADSL, MC4R, and CAPN1 genes exhibited five, three, and two different genotypes, respectively. These polymorphisms displayed promising connections with enhancing economically significant production traits, particularly BW, BrW and TW. Furthermore, gene expression analyses were conducted on breast and thigh tissues obtained from the chicken breeds at 60 days of age, where ADSL and MC4R exhibited a noteworthy up-regulation in Fayoumi and Matrouh breeds, and down-regulation in Mamourah and Leghorn. In contrast, CAPN1 expression decreased across most breeds with a slight increase noted in Fayoumi breed. In conclusion, this investigation underscores the substantial impact of ADSL, MC4R, and CAPN1 genes on economically important production traits within Egyptian domestic chicken breeds. Consequently, these genes emerge as significant molecular markers, holding potential utility in avian selection and breeding programs aimed at enhancing productive performance.

Colorimetric Polymerase Chain Reaction Enabled by a Fast Light-Activated Substrate Chromogenic Detection Platform.

Miniaturization of nucleic acid tests (NATs) into portable, inexpensive detection platforms may aid disease diagnosis in point-of-care (POC) settings. Colorimetric signals are ideal readouts for portable NATs, and it remains of high demand to develop color readouts that are simple, quantitative, and versatile. Thus motivated, we report a fast light-activated substrate chromogenic polymerase chain reaction (FLASH PCR) that uses DNA intercalating dyes (DIDs) to enable colorimetric nucleic acid detection and quantification. The FLASH system is established on our finding that DID-DNA intercalation can promote the rapid photooxidation of chromogenic substrates through light-induced production of singlet oxygen. Using this principle, we have successfully converted DID-based fluorescent PCR assays into colorimetric FLASH PCR. To demonstrate the practical applicability of FLASH PCR to POC diagnosis, we also fabricated two readout platforms, including a portable electronic FLASH reader and a paper-based FLASH strip. Using the FLASH reader, we were able to detect as low as 60 copies of DNA standards, a limit of detection (LOD) comparable with commercial quantitative PCR. The FLASH strip further enables the reader-free detection of PCR amplicons by converting the colorimetric signal into the visual measurement of distance as a readout. Finally, the practical applicability of the FLASH PCR was demonstrated by the detection and/or quantification of nucleic acid markers in diverse clinical and biological samples.

Naked-Eye Detection of Grapevine Red-Blotch Viral Infection Using a Plasmonic CRISPR Cas12a Assay.

Herein, we described a novel plasmonic CRISPR Cas12a assay for the visual, colorimetric detection of grapevine viral infections. Our assay generates rapid and specific colorimetric signals for nucleic acid amplicons by combining the unique target-induced incriminate single-stranded DNase activity of Cas12a with plasmon coupling of DNA functionalized gold nanoparticles. The practical applicability of our plasmonic assay was successfully demonstrated through the detection of emerging red-blotch viral infections in grapevine samples collected from commercial vineyards.

Histamine-Producing Bacteria and Histamine Induction in Retail Sardine and Mackerel from Fish Markets in Egypt.

This study examined the occurrence of histamine-producing bacteria (HPB) and histamine induction in retail sardine and mackerel in Egypt; and whether the fish vendors play a role in the transmission of HPB. Fish were collected from the fish markets, additionally; hand swab samples were taken from the fish vendors. All samples were cultured on modified Niven's medium (MNM); the positive colonies were subcultured on Violet Red Bile Glucose (VRBG) agar, followed by biochemical identification and histidine decarboxylase (hdc)-gene-PCR of the VRBG-positive isolates. The hdc-gene-positive fish and human isolates were subjected to partial hdc-gene-sequencing and phylogenetic analysis. Production of histamine in the fish muscles was measured by high-performance liquid chromatography. A higher percentage of sardine showed the presence of MNM-positive bacteria (84%) than mackerel (53%). Enterobacteriaceae was the dominant family; the most frequent species were Enterobacter cloacae, Raoultella planticola, Citrobacter freundii, and Enterobacter aerogenes. Higher proportion of the R. planticola isolates were hdc positive as compared with the other species. Only 32% sardine and 17% mackerel of the MNM-positive isolates carried the hdc gene. Fish muscles that contain hdc-positive bacteria exhibit higher levels of histamine (median 86; IQR 80-1112 mg/kg) than those with hdc-negative bacteria (48; 75-223 mg/kg). The level of histamine was significantly higher in sardine (109; 104-1094 mg/kg) than in mackerel (40; 49-106 mg/kg). The 20 fish vendor samples were MNM positive, 2 of them were hdc-gene positive. The close genetic relatedness between the human and fish strains isolated from the same markets suggests a possible bidirectional transmission of the HPB. This warns for the presence of HPB carrying hdc gene in retail sardine and mackerel, which is associated with a relatively high level of histamine. Regular inspection of the fish markets is required, including accurate determination of HPB by using a combination of the MNM culture, hdc-gene PCR, and measurement of histamine level.

Two versus three doses of COVID-19 vaccine and post-vaccination COVID-19 infection in hemodialysis patients.

Background and aim: Patients with chronic kidney disease including those undergoing hemodialysis (HD) constitute a particularly challenging group regarding COVID-19 vaccination. The present study aimed to compare the rate of reinfection after two and three doses of Sinopharm COVID-19 vaccine in HD patients. Patients and methods: The study included 80 HD patients who received three doses of Sinopharm COVID-19 vaccine. In addition, there were another 80 patients who received only two doses of the vaccine. Patients in the latter group were selected based on propensity matching score with 1:1 ratio. Patients were monitored for post-vaccination COVID-19 infection using PCR examination of nasopharyngeal swabs. Patients were also monitored for post-vaccination complications including general complaints (headache, fever, fatigue), injection site complaints (arm pain, swelling, itching, rash), musculoskeletal complaints (muscle spasm or pain, joint pain) and others. All patients were followed for six months. Results: The present study included 80 patients submitted to COVID-19 vaccination with two doses of Sinopharm vaccine (GI) and other 80 patients who received three doses of the same vaccine (GII). At the end of follow up, 11 patients (13.8 %) in GI caught COVID-19 infection. In contrast, no patient in GII had infection (P<0.001). Comparison between patients who had COVID-19 infection and those without infection revealed that the former subgroup had significantly lower BMI (23.3 ± 2.3 versus 27.5 ± 8.1 Kg/m2), higher frequency of associated Hepatitis C (HCV) infection (54.6 % versus 2.9 %, P<0.001) and higher serum ferritin levels [median (IQR): 1101.0 (836.0-1564.0) versus 675.0 (467.0-767.7) ng/mL, P=0.01]. Binary logistic regression analysis identified high serum ferritin levels [OR (95% CI): 0.014 (0.001-0.15), P<0.001] and associated HCV infection [OR (95% CI): 0.99 (0.98-1.01), P=0.02] as significant predictors of post-vaccination COVID-19 infection in multivariate analysis. Conclusions: A three dose regime of Sinopharm COVID-19 vaccine associated with significantly lower rate of reinfection COVID-19 infection in HD patients. Infected patients had significantly lower BMI, higher frequency of HCV and higher ferritin levels.

The potential mechanism of histamine-inducing cardiopulmonary inflammation and apoptosis in a novel oral model of rat intoxication.

Histamine (HIS) is a potent vasodilator that contributes to anaphylactic reactions. Our investigation aims to study the possible toxic impact of repeated oral administration of histamine on the target organs of HIS poisoning (lung & heart) in rats as a model of scombroid poisoning. We used 15 rats that were separated into three groups with 5 rats in each. All rats received the treatments orally for 14 days as follows; (1): distilled water, (2) HIS at a dosage level of 250 mg/kg BWT daily and (3) HIS at a dosage level of 1750 mg/kg BWT weekly. Our results revealed that the consumption of HIS either daily or weekly could cause marked cardiopulmonary toxicity in rats. HIS can trigger inflammatory reactions in the cardiopulmonary tissues and induce oxidative stress damage along with apoptosis of such organs. HIS was markedly increase the MDA levels and decrease the CAT and GSH activity in both lung and heart tissues. The main pathological lesion observed is inflammation which was confirmed by immunohistochemistry and demonstrated strong iNOS and TNF-α protein expressions. Cardiac muscles showed extensive degeneration and necrosis and displayed strong casp-3 protein expression. Additionally, all HIS receiving groups noticed marked elevation of the pulmonary transcription levels of Cox2, TNF-α, and IL1ß along with substantial elevation of casp-3 and bax genes and downregulation of Bcl2 gene in the cardiac tissue. We concluded that the oral administration of HIS either daily or weekly can induce cardiopulmonary toxicity via the upregulation of proinflammatory cytokines resulting in ROS overgeneration and inducing both oxidative stress and apoptosis.

The effects of novel alginate-lauric arginate coatings with temperature on bacterial quality, oxidative stability, and organoleptic characteristics of frozen stored chicken drumsticks.

Although alginate has been reported to be used as an edible film and coating in food, to the best of our knowledge, this study is the first to investigate the individual effects of coatings, temperatures, storage times, as well as their interactions, on the bacterial quality, deterioration criteria, and sensory attributes of chicken drumsticks. To accomplish this, six groups of chicken drumsticks were treated with different coatings, temperatures, and storage conditions. The groups included 2 % alginate at 25 °C (Alg25) and 50 °C (Alg50), a mixture of 2 % alginate and 2 % LAE at 25 °C (M25) and 50 °C (M50), besides control untreated samples in distilled water at 25 °C (C25) and 50 °C (C50). The chicken drumsticks were stored at -18 °C for 3 months. The results showed that all treated chicken drumsticks induced a significant reduction in all bacterial counts, as well as a significant decrease in pH and thiobarbituric acid values, with an improvement in all sensory attributes, particularly in the M25 and M50 coated samples compared to the C25 and C50. Furthermore, exposing alginate and LAE to a temperature of 50 °C can increase their antimicrobial activity. In conclusion, the innovative combinations of LAE and alginate can be used successfully to decontaminate chicken carcasses in poultry processing plants.

Amplified detection of nucleic acids and proteins using an isothermal proximity CRISPR Cas12a assay.

Herein, we describe an isothermal proximity CRISPR Cas12a assay that harnesses the target-induced indiscrimitive single-stranded DNase activity of Cas12a for the quantitative profiling of gene expression at the mRNA level and detection of proteins with high sensitivity and specificity. The target recognition is achieved through proximity binding rather than recognition by CRISPR RNA (crRNA), which allows for flexible assay design. A binding-induced primer extension reaction is used to generate a predesigned CRISPR-targetable sequence as a barcode for further signal amplification. Through this dual amplification protocol, we were able to detect as low as 1 fM target nucleic acid and 100 fM target protein isothermally. The practical applicability of this assay was successfully demonstrated for the temporal profiling of interleukin-6 gene expression during allergen-mediated mast cell activation.

Expanding detection windows for discriminating single nucleotide variants using rationally designed DNA equalizer probes.

Combining experimental and simulation strategies to facilitate the design and operation of nucleic acid hybridization probes are highly important to both fundamental DNA nanotechnology and diverse biological/biomedical applications. Herein, we introduce a DNA equalizer gate (DEG) approach, a class of simulation-guided nucleic acid hybridization probes that drastically expand detection windows for discriminating single nucleotide variants in double-stranded DNA (dsDNA) via the user-definable transformation of the quantitative relationship between the detection signal and target concentrations. A thermodynamic-driven theoretical model was also developed, which quantitatively simulates and predicts the performance of DEG. The effectiveness of DEG for expanding detection windows and improving sequence selectivity was demonstrated both in silico and experimentally. As DEG acts directly on dsDNA, it is readily adaptable to nucleic acid amplification techniques, such as polymerase chain reaction (PCR). The practical usefulness of DEG was demonstrated through the simultaneous detection of infections and the screening of drug-resistance in clinical parasitic worm samples collected from rural areas of Honduras.

Paper-Based DNA Reader for Visualized Quantification of Soil-Transmitted Helminth Infections.

Soil-transmitted helminth (STH) infections are a global health issue affecting nearly one-third of the world's population. As most endemic areas of STH are impoverished countries or regions with limited healthcare resources, the accurate diagnosis of STH requires analytical tools that are not only quantitative, but also portable, inexpensive, and with no or minimal demand for external instrument. Herein, we introduce a novel paper-based diagnostic device, termed quantitative paper-based DNA reader (qPDR), capable of quantifying STH at the molecular level by measuring distance as readout, thus eliminating the need for external readers. On the basis of the unique interfacial interaction of a DNA intercalating dye, SYBR Green I, with native cellulose on a chromatographic paper, qPDR allows the distance-based quantification of minute amounts of double-stranded DNA as short as 6 min. By integrating qPDR with polymerase chain reactions that were performed using a smartphone-controlled portable thermal cycler, we were able to quantify minute amount of genetic markers from adult worms of an STH (Trichuris trichiura) that were expelled post-treatment by infected children living in the rural areas of Honduras.

Salivary changes in type 2 diabetic patients.

OBJECTIVE: this study was conducted to determine the effect of type 2 diabetes mellitus on salivary secretion of glucose, amylase and immunoglobulin A levels and also to fined out if saliva could be used as anon invasive method to monitor glycaemic control in type 2 diabbetes. This study was conducted on 40 human subjectes, They were 20 males and 20 females, their ages ranged from 35 years to 64 years,and they were divided into two groups, the first one is the patient group which contains 20 diabetic paient. (10 males and 10 females,aged between 38 years to 64 years). the second one is the control group which contains 20 healthy adult (10 males and 10 females,aged between 35 years to 60 years) they were age and sex matched. All studied group were subjected to clinical and laboratory investigation which includes post prandial blood glucose, HA1C, salivary glucose, salivary amylase, and salivary immunoglobulin A. RESULTS: there was a highly significant increase in the level of post prandial blood glucose, HBA1C; Salivary glucose, Salivary amylase & Salivary immunoglobulin A in diabetic group compared with control group and There was a significant positive correlation between post prandial blood glucose and salivary glucose in diabetic group. CONCLUSIONS: These results suggest that diabetes influences the composition of saliva and that saliva can be used as less painful,non invasive biomarker for monitoring the blood glucose concentration in the patients with diabetes mellitus.

The use of natural herbal extracts for improving the lipid stability and sensory characteristics of irradiated ground beef.

Ground Longissimus dorsi of beef were treated with herbal extracts of marjoram, rosemary and sage at concentration of 0.04% (v/w), radiation (2 or 4.5 kGy) or their combination. Treated samples were stored at 5°C and analyzed periodically for thiobarbituric acid reactive substances (TBARS), sensory characteristics and psychrotrophic bacterial counts during storage for 41 and 48 days for samples treated at 2 and 4.5 kGy respectively. Results demonstrated a significant benefit of the addition of herbal extracts to the ground beef prior to irradiation. All three extracts significantly (P<0.05) lowered the TBARS values and off-odor scores and significantly (P<0.05) increased color and acceptability scores in all samples with marjoram being the most effective. The combination treatment with herbal extracts plus irradiation resulted in extension of the shelf life of samples treated with 2 kGy by one week and samples treated with 4.5 kGy by two weeks, over that treated with irradiation alone. In conclusion, the addition of herbal extracts can minimize lipid oxidation, improve color and decrease off-odor production in irradiated ground beef.