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The goal of this study is to investigate the impact of the rs35829419 SNP on the serum level of NLRP3, and to assess the relationship between NLRP3 and its SNP and vulnerability to Pityriasis versicolor. Pityriasis versicolor (PV) is one of the most frequent skin conditions linked to skin pigmentation changes. Malassezia plays a key role in the pathogenesis of PV. A case-control study, 50 patients with pityriasis versicolor and 44 healthy controls. Real-time PCR was used to genotype NLRP3 (rs35829419) and ELISA assay of NLRP3 levels in tissue samples. There was a significantly higher median NLPR3 levels in PV patients than controls. A significant predominance of A allele of Q 705 K was in patients than controls. The risk of having the disease in the presence of A allele is nearly 10 times than having C allele. In PV patients, there was a significant relationship between NLPR3 levels and Q 705 K genotypes with higher NLPR3 levels in AA genotype. A potential correlation between PV and the Q705K polymorphism, pointing to evidence of NLRP3 alteration in PV patients. The NLRP3 inflammasome may be an appropriate therapeutic target for Malassezia-associated skin disorders. [ABSTRACT FROM AUTHOR]

This study aims to examine the genetic polymorphisms of the granulysin (GNLY) and pentraxin 3 (PTX3) genes and their association with acne in Egypt. Acne vulgaris is classified as a disorder of the pilosebaceous unit. Clinical, histological, and immunological findings indicate that inflammation is involved in every stage of acne development. GNLY and PTX3 are both involved in the body’s immune system and may play a role in the pathophysiology of acne. This case-control study included 180 participants who have acne and 180 healthy controls. Real-time PCR was used to genotype GNLY rs7908 and PTX3 rs2305619 polymorphisms. Genotype occurrence and allelic spreading for both single nucleotide polymorphisms (SNP) are in Hardy-Weinberg equilibrium. Regarding rs7908, no statistical difference was observed in the genotype and allele distributions between acne patients and controls. On the other hand, rs2305619 showed a statistical difference in the genotype and allele distributions between acne patients and controls, with a marked prevalence of the GG group and G allele in acne patients. Our study revealed a significant link between the PTX3 rs2305619 and acne susceptibility in Egypt, with the AG + GG genotype strongly predicting acne. In contrast, the GNYL rs7908 polymorphism was not associated with acne. These results highlight a genetic component to acne and suggest that PTX3 rs2305619 could be a key marker for understanding acne susceptibility. [ABSTRACT FROM AUTHOR]

Rickettsioses caused by Rickettsia and Orientia spp. are the re-emerging diseases in India, which are grossly underdiagnosed, particularly among children. They usually present as mild-febrile illness but may extend to severe life-threatening complications. Early diagnosis followed by proper treatment reduces the morbidity and mortality. Non-specific clinical symptoms and lack of point of care diagnosis may delay the treatment. Molecular assays like PCR may helpful in the early diagnosis and confirmation of rickettsial diseases. In this study, we used multiplex real-time PCR to detect Rickettsia spp. and Orientia spp. in febrile pediatric patients. Whole blood was collected from 239 clinically suspected febrile pediatric patients aged between 6 months to 12 years admitted in tertiary care hospital at Chennai, South India. Multiplex real-time PCR was used to target the gltA gene for Rickettsia spp. and the 47kDa gene for Orientia tsutsugamushi. To compare the sensitivity, nested PCR was performed on the 56kDa antigen gene of O. tsutsugamushi and the Rickettsia genus specific gltA gene. By multiplex real-time PCR, 15 samples were positive for O. tsutsugamushi and 3 were positive for Rickettsia spp. Nested PCR identified 35 positive samples for O. tsutsugamushi and 4 positive samples for Rickettsia spp. Even though multiplex real-time PCR had lower positivity than nPCR, it was effective in diagnosing O. tsutsugamushi and Rickettsia spp. in a single assay.

This study aims to examine whether the genetic variants in the genes for Granzyme B (GZMB) and Interferon Induced with Helicase C domain 1 (IFIH1) were associated with psoriasis. Psoriasis, a papulosquamous skin disease, was initially thought of as a disorder primarily of epidermal keratinocytes but is now recognized as one of the most common immune-mediated disorders. It is caused by the interplay between multiple genetic and environmental risk factors. This case-control study has 65 participants with psoriasis and 65 healthy controls. Real-time PCR was used to genotype GZMB (rs8192917) and IFIH1 (rs35667974). Genotype occurrence and allelic spreading for both SNPs are in Hardy – Weinberg equilibrium. The genotype and allele distributions of rs35667974 showed no differences between the studied groups. Regarding rs8192917, compared to Group II, there is a statistically significant rise in the CC genotype and C allele in Group I. Higher PASI scores are detected in the C/C and C/T genotypes more than the T/T genotype. Univariate and multivariate analyses revealed that BMI, catalase, MDA, and rs8192917 (C/C) are associated with psoriasis. GZMB rs8192917 was significantly related to psoriasis risk; its C allele is likewise associated with psoriasis vulnerability. However, our investigation found no link between rs35667974 and psoriasis. [ABSTRACT FROM AUTHOR]

Toxoplasmosis continues to be a prevalent parasitic zoonosis with a global distribution. This disease is caused by an intracellular parasite known as Toxoplasma gondii, and the development of effective novel drug targets to combat it is imperative. There is limited information available on the potential advantages of wheat germ oil (WGO) and propolis, both individually and in combination, against the acute phase of toxoplasmosis. In this study, acute toxoplasmosis was induced in Swiss albino mice, followed by the treatment of infected animals with WGO and propolis, either separately or in combination. After 10 days of experimental infection and treatment, mice from all groups were sacrificed, and their brains, uteri, and kidneys were excised for histopathological assessment. Additionally, the average parasite load in the brain was determined through parasitological assessment, and quantification of the parasite was performed using Real-Time Polymerase Chain Reaction targeting gene amplification. Remarkably, the study found that treating infected animals with wheat germ oil and propolis significantly reduced the parasite load compared to the control group that was infected but not treated. Moreover, the group treated with a combination of wheat germ oil and propolis exhibited a markedly greater reduction in parasitic load compared to the other groups. Similarly, the combination treatment effectively restored the histopathological changes observed in the brain, uterus, and kidney, and the scoring of these reported lesions confirmed these findings. In summary, the present results reveal intriguing insights into the potential therapeutic benefits of wheat germ oil and propolis in the treatment of acute toxoplasmosis. [ABSTRACT FROM AUTHOR]

Rickettsioses caused by Rickettsia and Orientia spp. are the re-emerging diseases in India, which are grossly underdiagnosed, particularly among children. They usually present as mild-febrile illness but may extend to severe life-threatening complications. Early diagnosis followed by proper treatment reduces the morbidity and mortality. Non-specific clinical symptoms and lack of point of care diagnosis may delay the treatment. Molecular assays like PCR may helpful in the early diagnosis and confirmation of rickettsial diseases. In this study, we used multiplex real-time PCR to detect Rickettsia spp. and Orientia spp. in febrile pediatric patients. Whole blood was collected from 239 clinically suspected febrile pediatric patients aged between 6 months to 12 years admitted in tertiary care hospital at Chennai, South India. Multiplex real-time PCR was used to target the gltA gene for Rickettsia spp. and the 47kDa gene for Orientia tsutsugamushi. To compare the sensitivity, nested PCR was performed on the 56kDa antigen gene of O. tsutsugamushi and the Rickettsia genus specific gltA gene. By multiplex real-time PCR, 15 samples were positive for O. tsutsugamushi and 3 were positive for Rickettsia spp. Nested PCR identified 35 positive samples for O. tsutsugamushi and 4 positive samples for Rickettsia spp. Even though multiplex real-time PCR had lower positivity than nPCR, it was effective in diagnosing O. tsutsugamushi and Rickettsia spp. in a single assay. [ABSTRACT FROM AUTHOR]

Marine fish commodities in Indonesia, both exported and imported, have experienced a significant increase. This increase opens up opportunities for the risk of entry and spread of fish pests and diseases, both on an export scale between countries and between areas within Indonesia. One of the pathogens that is acute that can cause major losses is the virus. The virus that often attacks sea water ornamental fish is Viral haemorrhagic septicemia. The purpose of this study was to detect the presence of VHSV that infects marine ornamental fish commodities using namely: necropsy, fixation, extraction, amplification, and reading of results. This research was conducted at Fish Quarantine Inspection Agency Denpasar, Bali. Based on the test results, VHS was found to attack six samples of marine fish species with codes namely NTC, BC 8.2, BC 9.2, BAR 8.2, 1252, NSR 8.2.

Background and purpose: The aim is to investigate the change in the expression level of miR-32 and PTEN genes in tumor samples with melanoma. Materials and methods: This study collected tumor samples from 50 patients and 50 samples from healthy marginal tissue around the tumor as a control sample. Following that, the RNA related to the samples was extracted and the expression of the desired gene was evaluated using real-time polymerase chain reaction. Appropriate statistical tests assessed differences in expression levels of the two groups. In statistical tests. Results: The expression of PTEN was lower in tumor samples than in healthy samples. In contrast, the expression of miR-32 was higher in tumor samples and the difference was significant in both cases (P

Copyright of Journal of Marine & Coastal Science is the property of Universitas Airlangga and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Fifty diabetic nephropathy (DN) children with type 1 diabetes mellitus (T1DM) and 50 healthy matched controls were included. Chromatographic assays of 14 amino acids, free carnitine and 27 carnitine esters using high‐performance liquid chromatography/electrospray ionization–mass spectroscopy, and genetic testing for JAK2v617f mutation using real‐time PCR were performed. Patients had significantly lower levels of tyrosine, branched‐chain amino acids (BCAAs), and BCAA/AAA (aromatic chain amino acids) ratios, glycine, arginine, ornithine, free carnitine and some carnitine esters (C5, 6, 12 and 16) and higher phenylalanine, phenylalanine/tyrosine ratio and C18 compared with the controls and in the macro‐albuminuria vs. the microalbuminuria group (p < 0.05 for all) except for free carnitine. Plasma carnitine was negatively correlated with eGFR (r = −0.488, p = 0.000). There were significant positive correlations between tyrosine with UACR ratio (r = 0.296, p = 0.037). The plasma BCAA/AAA ratio showed significant negative correlations with UACR (r = −0.484, p = 0.000). There was a significantly higher frequency of the JAK2V617F gene mutation in diabetic nephropathy patients compared with the control group and in macro‐albuminuria than the microalbuminuria group (p = 0.000) for both. When monitoring children with T1DM, plasma free amino acids and acylcarnitine profiles should be considered, especially if they have tested positive for JAK2V617F for the early diagnosis of DN. [ABSTRACT FROM AUTHOR]

Toxoplasmosis continues to be a prevalent parasitic zoonosis with a global distribution. This disease is caused by an intracellular parasite known as Toxoplasma gondii, and the development of effective novel drug targets to combat it is imperative. There is limited information available on the potential advantages of wheat germ oil (WGO) and propolis, both individually and in combination, against the acute phase of toxoplasmosis. In this study, acute toxoplasmosis was induced in Swiss albino mice, followed by the treatment of infected animals with WGO and propolis, either separately or in combination. After 10 days of experimental infection and treatment, mice from all groups were sacrificed, and their brains, uteri, and kidneys were excised for histopathological assessment. Additionally, the average parasite load in the brain was determined through parasitological assessment, and quantification of the parasite was performed using Real-Time Polymerase Chain Reaction targeting gene amplification. Remarkably, the study found that treating infected animals with wheat germ oil and propolis significantly reduced the parasite load compared to the control group that was infected but not treated. Moreover, the group treated with a combination of wheat germ oil and propolis exhibited a markedly greater reduction in parasitic load compared to the other groups. Similarly, the combination treatment effectively restored the histopathological changes observed in the brain, uterus, and kidney, and the scoring of these reported lesions confirmed these findings. In summary, the present results reveal intriguing insights into the potential therapeutic benefits of wheat germ oil and propolis in the treatment of acute toxoplasmosis.

Background: Peripheral artery disease (PAD) is an advanced atherosclerotic disease-causing augmented risks of hospitalization and death. So, several studies tried to detect novel biomarkers for PAD diagnosis. The study is aimed to explore the role of plasma miRNA 323b-5p and miRNA 486-5p levels to diagnose PAD. Methods: The research was performed in Medical Biochemistry & Cardiology Departments - Zagazig Faculty of Medicine. It included 100 volunteers categorized into 2 groups; Group (1): included 50 healthy volunteers served as control group with ankle brachial index (ABI) > 0.9. Group (2): included 50 PAD patients with ABI < 0.9. Plasma 323b-5p and miR 486-5p expression level were measured via real time PCR. Color Doppler was performed to calculate ABI, which was correlated with miRNA levels. Results: There was a highly significant difference between groups as regards miR 323b-5p. Furthermore, miR 486-5p expression level showed significant variance between groups. Regarding miRNA 323b-5p expression level, cutoff value was 2.55, sensitivity was 89.3% and specificity was 72.7%. Concerning miR 486-5p expression level, cutoff value was 1.92, sensitivity was 82.1% and specificity was 81.8%. As regards combined miR 323b-5p and miR 486-5p ROC curve, it was 96.4% sensitivity and 95.5% specificity. Each miRNA showed a negative correlation with ABI. Conclusion: miRNA 323B-5P and miRNA 486-5P can be a novel and potent biomarker for PAD diagnosis, particularly for risky patients. [ABSTRACT FROM AUTHOR]

Background: Curcumin is active ingredient of turmeric The main purpose of this study is evaluating impact of curcumin on suger, hypoalgesia and inflammatory factors in diabetic mice. Materials and methods: Male mice divided into six groups of 6. One group as a negative and the other five groups injected with Streptozotocin (STZ) (200 mg/kg). Diabetic mice in each group given different treatments for twenty-one days.After that, blood sugar and neuroathy studies have been done and tissue samples the liver were studied for gene expression. Result: Curcumin reduced blood sugar, but the rate of hypoglycemia was significantly lower than metformin group P > 0.05, and the comparison of the synergistic effect of curcumin and metformin with metformin was not significant P > 0.05. Also, in neuropathy studies, the groups which recieved curcumin and metformin have shown a significant difference with diabetic group P < 0.01, Also, by evaluating inflammatory factors, there was a significant difference in the expression of TNF-a, IL-6 and NfkB, but there is no significant difference in the expression of Sirt1 P < 0.05. Conclusions: The analgesic effect of curcumin was quite evident, probably due to the significant impact of this herbal drug in reducing the expression of inflammatory genes NF-kB, IL6, and TNF-α. [ABSTRACT FROM AUTHOR]

Chronic inflammation in the gallbladder leading to persistent epithelium damage promotes invasive cancer. The study aimed to assess the prognostic value of PDL1 and CA19-9 markers in cancer/inflammatory lesions of the gallbladder. A total of 29 cases (19 cancer and 10 inflammatory) were included. The PDL1 protein concentration level and mRNA expression were assessed in the tissues’ lysates by ELISA and real-time PCR, respectively. PDL1 and CA19-9 concentration levels were compared and statistically related with clinico-pathological variables. The PDL1 protein level and its relative mRNA expression were correlated. Kaplan–Meir survival and Cox regression analyses were conducted for predicting prognosis. This study investigated the PDL1 and CA19-9 marker expression in both cancer and inflammatory cases of the gallbladder (p = 0.48 and p = 0.17 respectively). PDL1 protein expression was significantly associated with the hormonal profile of the cases (p = 0.04) at an optimum cut-off value of 13 pg/mL, while the CA19-9 marker expression was correlated with the status of liver metastasis (p = 0.0043) and size of the tumor (p = 0.004). A low PDL1 concentration was found when compared to the CA19-9 level among cancer cases (p = 0.12) and proportional in the inflammatory lesions (p = 0.63). A significant positive correlation was found between the PDL1 protein and its relative mRNA expressions in the inflammatory lesions (p = 0.029) when compared to cancer cases (p = 0.069). Our results showed that a protein-based assay for PDL1 expression would be more sensitive compared to RNA based assays for GBC risk stratifications. Overall survival was predicted with CA19-9 and PDL1 levels (p = 0.0074, p = 0.23, respectively). PDL1 and CA19-9 may act as a probable predictor of a poor prognosis in gallbladder cancer (GBC) cases.

ObjectiveMedicinal plants have long been recognized as a rich source of essential oils and have been cultivated for a variety of uses since ancient times. White savory (Satureja mutica Fisch & CA mey) is one of the medicinal plants belonging to the mint family with high content of carvacrol in its essential oil. A large body of literature has suggested that abiotic stresses, especially drought stress, could considerably boost the production of essential oils in plants. This study aimed to evaluate the effect of drought stress on the expression of genes affecting the biosynthesis of the main component of white savory essential oil carvacrol.Materials and methodsFor this goal, 20 pot-planted seedlings of white savory at the 6-8 leaf stage were subjected in a greenhouse to two conditions of control and drought stress (10 pots for each condition) with irrigation regimes of 100% and 30% of field capacity, respectively. To ensure the implementation of stress on plants and to determine the appropriate time for sampling, leaf proline and relative water content of plants were measured.ResultsThe results of both proline and relative water content tests revealed a significant statistical difference between the control and stress conditions in white savory at 30% of field capacity. The KEGG (Kyoto Encyclopedia of Genes and Genomes) database was used to obtain information on the carvacrol biosynthesis pathway. The three key genes geranyl diphosphate synthase (gpps), terpene synthase (tps), and limonene hydroxylase (lh) were chosen for gene expression analysis. Due to the lack of genomic information on white savory, the sequence information of the genes in the closest genera within the family mint was used to design primers. The result of melting curve analysis from the real-time quantitative PCR (qPCR) assay confirmed the accuracy of the primer design. The raw qPCR data normalized to the housekeeping gene Actin, then differential expression analysis was performed using the delta-delta Ct method. qPCR result showed that expression of the genes gpps, tps, and lh were increased in stressed plants proximately 20, 18, and 50 times, compared to those in control, respectively.ConclusionsThe conclusion that must be drawn from the data available is that water stress leads to the process of plant response to stress and changes the expression of genes involved in the biosynthesis pathway of the active substance carvacrol.

Grass pea is a promising crop with the potential to provide food and fodder, but its genomics has not been adequately explored. Identifying genes for desirable traits, such as drought tolerance and disease resistance, is critical for improving the plant. Grass pea currently lacks known R-genes, including the nucleotide-binding site-leucine-rich repeat (NBS-LRR) gene family, which plays a key role in protecting the plant from biotic and abiotic stresses. In our study, we used the recently published grass pea genome and available transcriptomic data to identify 274 NBS-LRR genes. The evolutionary relationships between the classified genes on the reported plants and LsNBS revealed that 124 genes have TNL domains, while 150 genes have CNL domains. All genes contained exons, ranging from 1 to 7. Ten conserved motifs with lengths ranging from 16 to 30 amino acids were identified. We found TIR-domain-containing genes in 132 LsNBSs, with 63 TIR-1 and 69 TIR-2, and RX-CCLike in 84 LsNBSs. We also identified several popular motifs, including P-loop, Uup, kinase-GTPase, ABC, ChvD, CDC6, Rnase_H, Smc, CDC48, and SpoVK. According to the gene enrichment analysis, the identified genes undergo several biological processes such as plant defense, innate immunity, hydrolase activity, and DNA binding. In the upstream regions, 103 transcription factors were identified that govern the transcription of nearby genes affecting the plant excretion of salicylic acid, methyl jasmonate, ethylene, and abscisic acid. According to RNA-Seq expression analysis, 85% of the encoded genes have high expression levels. Nine LsNBS genes were selected for qPCR under salt stress conditions. The majority of the genes showed upregulation at 50 and 200 μM NaCl. However, LsNBS-D18, LsNBS-D204, and LsNBS-D180 showed reduced or drastic downregulation compared to their respective expression levels, providing further insights into the potential functions of LsNBSs under salt stress conditions. They provide valuable insights into the potential functions of LsNBSs under salt stress conditions. Our findings also shed light on the evolution and classification of NBS-LRR genes in legumes, highlighting the potential of grass pea. Further research could focus on the functional analysis of these genes, and their potential use in breeding programs to improve the salinity, drought, and disease resistance of this important crop.

Chronic inflammation in the gallbladder leading to persistent epithelium damage promotes invasive cancer. The study aimed to assess the prognostic value of PDL1 and CA19-9 markers in cancer/inflammatory lesions of the gallbladder. A total of 29 cases (19 cancer and 10 inflammatory) were included. The PDL1 protein concentration level and mRNA expression were assessed in the tissues' lysates by ELISA and real-time PCR, respectively. PDL1 and CA19-9 concentration levels were compared and statistically related with clinico-pathological variables. The PDL1 protein level and its relative mRNA expression were correlated. Kaplan–Meir survival and Cox regression analyses were conducted for predicting prognosis. This study investigated the PDL1 and CA19-9 marker expression in both cancer and inflammatory cases of the gallbladder (p = 0.48 and p = 0.17 respectively). PDL1 protein expression was significantly associated with the hormonal profile of the cases (p = 0.04) at an optimum cut-off value of 13 pg/mL, while the CA19-9 marker expression was correlated with the status of liver metastasis (p = 0.0043) and size of the tumor (p = 0.004). A low PDL1 concentration was found when compared to the CA19-9 level among cancer cases (p = 0.12) and proportional in the inflammatory lesions (p = 0.63). A significant positive correlation was found between the PDL1 protein and its relative mRNA expressions in the inflammatory lesions (p = 0.029) when compared to cancer cases (p = 0.069). Our results showed that a protein-based assay for PDL1 expression would be more sensitive compared to RNA based assays for GBC risk stratifications. Overall survival was predicted with CA19-9 and PDL1 levels (p = 0.0074, p = 0.23, respectively). PDL1 and CA19-9 may act as a probable predictor of a poor prognosis in gallbladder cancer (GBC) cases. [ABSTRACT FROM AUTHOR]

Introduction: Iranian Kakoti (.Ziziphora persica Bunge) which belongs to the mint genus is widely distributed in Iran. In some parts of Iran, it is used as a spice and condiment in food. Iranian Kakoti is also a medicinal plant with antioxidant and antimicrobial properties. Iranian Kakoti is used in traditional medicine as a stomach disinfectant and cold remedy. Since the effect of stresses on the increase of secondary metabolites of plants has been proven, the aim of this study was to investigate the effect of drought stress on the secondary metabolites of Iranian Kakoti in comparison with normal conditions by assessment of the expression of genes which are related to the most important secondary metabolites of the studied plant. In previous studies, it was found that pulegone has the largest share among the metabolites produced in this plant. Materials and Methods: Using the KEGG database, the metabolic pathway and genes involved in the production of the pulegone include the four key genes Isopiperitenol dehydrogenase. Limonene synthase, limonene hydroxylase and Isopiperitenone reductase were identified. Due to the lack of genomic information of Iranian Kakoti, the sequence of the four studied genes was predicted by the sequences of these genes in families close to this plant. The predicted sequences were used for primer design. The plants were cultivated under drought stress and normal conditions. The stress condition was achieved by reaching 30% Field Capacity. After observing phenotypic changes due to stress, the amount of proline and also Leaf relative water content (RWC) measured for the plants in both control and stress condition to confirm the accruing of the stress. Then leaf samples were collected. RNA extraction and cDNA synthesis were performed according to the manufacturer's instructions. The quantity and quality of the obtained RNA were evaluated by Nanodrop and gel electrophoresis. The qualified RNA samples were used for cDNA synthesis according to the manufacturer's protocols. Then the comparing the genes expression under drought stress with the control sample was performed. Results and Discussion: In response to drought stress, leaf RWC was decreased to 31% in the plants under stress condition and proline levels increased by 1.9-fold compared to control value. Analysis of real-time PCR data showed that. The expressions of limonene synthase, limonene hydroxylase, Isopiperitenon dehydrogenase, Isopiperitenon reductase were increased 115, 400, 50 and 120 folds compared to control respectively. Conclusion: The measurement of leaf RWC and proline were found to be beneficial methods to detection of incidence of drought stress in plant. The expression of genes involved in pulegone synthesis compared to the control plants changes with drought stress. The implement of drought stress is a useful method to increase essential oil in Iranian Kakoti. [ABSTRACT FROM AUTHOR]

Bovine brucellosis is a very important zoonotic disease of economic interest throughout the world. The current study was designed to evaluate the effects of phyto-chemo-immune therapeutic agents (antibiotics vs. herbal product) against bovine brucellosis in cattle. For this purpose, crossbred cows (n = 20) were divided into 4 groups (i.e., A, B, C and D; n = 5 in each group). The animals in group A, B and C included brucella positive cows, while group D had control negative (healthy) animals. Cows of group A were treated with antibiotics (oxytetracyclin and streptomycin), fluxin meglumin, vitamin E and selenium, and immunized with brucella RB-51 (Bivishot) and FMD oilbased vaccine; group B cows were treated with a herbal product Safi, instead of antiobiotics; group C cows were only treated with flunixin meglumin and vitamin E and selenium, and immunized with RB-51 and FMD vaccines, while cows in group D were only immunized with RB-51 and FMD vaccines. All cows were evaluated for brucella antibodies through ELISA on d 0, 30, 90 and 180 post-treatments, and for antigens through real time PCR on d 180 post-treatment. Moreover, the treated cows were also observed for retention of fetal membranes and birth weight of calves. According to the results, titers of antibodies (OD values) against brucella abortus on d 180 post-treatment were found to be less (P < 0.05) in groups A (0.83 ± 0.2) and D (0.60 ± 0.1) cows compared with group B (1.09 ± 0.2) and C (1.15 ± 0.2) cows. Moreover, no difference (P > 0.05) was found between OD values of group A and D cows. The Ct values, determined through Real time PCR, were greater in group A (37.2 ± 0.9) and D (> 40) cows compared with B (35.0 ± 2.5) and C (32.5 ± 1.2) cows. Furthermore, birth weight of calves was greater in A and D cows compared with B and C cows. Placenta dropping time was shorter in A and D animals compared with group B and C cows. In conclusion, combination of antibiotics, anti-inflammatory agents and immune boosters (Group-A) may be used as a therapeutic approach to overcome bovine Brucellosis. [ABSTRACT FROM AUTHOR]

Bacteria of the Borrelia burgdorferi sensu lato complex are the causative agents of Lyme borreliosis (LB). Even if the conventional diagnosis of LB does not rely on the species itself, an accurate species identification within the complex will provide a deepened epidemiological scenario, a better diagnosis leading to a more targeted therapeutic approach, as well as promote the general public's awareness. A comparative genomics approach based on the 210 Borrelia spp. genomes available in 2019 were used to set up three species-specific PCR protocols, able to detect and provide species typing of Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.) and Borrelia garinii, the three most common and important human pathogenic Lyme Borrelia species in Europe. The species-specificity of these protocols was confirmed on previously identified B. afzelii, B. burgdorferi s.s. and B. garinii specimens detected in Ixodes ricinus samples. In addition, the protocols were validated on 120 DNA samples from ticks collected in Sweden, showing 88% accuracy, 100% precision, 72% sensitivity and 100% specificity. The proposed approach represents an innovative tool in epidemiological studies focused on B. burgdorferi s.l. occurrence in ticks, and future studies could suggest its helpfulness in routine diagnostic tests for health care. [ABSTRACT FROM AUTHOR]

Newcastle disease is one of the worst avian illnesses, it was responsible for 32.7% of all deaths. The significant mortality rates from Newcastle disease illnesses necessitate that, they be given top care. The main purpose of the current study was Molecular diagnosis of Newcastle disease virus by RT-RCR and detection fusion gene using conventional PCR. With the discovery of Newcastle disease symptoms in 20 flocks, researchers tested 70 random samples and 40 samples as healthy control from all around the governorates of Babylon and Al-Najaf (loss of appetite, coughing, gasping, nasal discharge, watery eyes, bright green diarrhea and nervous signs such as paralysis and convulsions). In the beginning, an NDV kit test was utilized to quickly verify the infection. The best time to start raising chickens is between day 20 and day 35. The flocks were protected against NDV using LaSota vaccinations. The samples were collected between October 2021 and January 2022. To verify the tentative diagnosis based on clinical and pathological signs, 5 ml of blood were drawn. Each blood sample resulted in two separate test tubes (2.5ml putted in EDTA tube, to obtain of whole blood, and 2.5ml putted in gully tube to obtain serum). All samples were stored at -20C until analysis. Field examination was performed by an NDV rapid kit where it showed positive results for 63(90%) samples out of 70 samples were detected as ND according to clinical symptoms, out of 63 samples, 40(63.4%) samples give positive inhibition. Haemagglutination inhibition test were determination on 63 clinical samples that showed positive results of NDV by immunochromatography Rapid diagnostic test. The results showed that, out of 63 samples, 40(63.4%) samples give positive inhibition. There were significant difference between mean antibody titer of infected birds and healthy control group, the mean of AB titer was decrease (13.15±1.94) more than control (467.2±48.83). Results from an RT-PCR test revealed that, only 12(30%) of 40 isolates were positive. Seven of the twelve NDV isolates underwent RT-PCR to identify the F gene, and the findings indicated that, two of the samples provided a strong band, one gave a light band, one gave a weak band, and three did not exhibit any band of F gene. It was conclude that, in order to effectively develop a control strategy against the illness, rapid test detection and identification of the viruses are essential. As a result, the RT-PCR molecular technique may be used for quick and confirmatory detection of NDV from any kind of ND epidemic. The RT-PCR assay also had a higher detection rate of NDV than the HI test. Since this is the case, introducing the molecular approach (RT-PCR) for quick and confirmatory detection of NDV from any kind of ND outbreak at the field level of Iraq is feasible. [ABSTRACT FROM AUTHOR]

Objective: Medicinal plants have long been recognized as a rich source of essential oils and have been cultivated for a variety of uses since ancient times. White savory (Satureja mutica Fisch & CA mey) is one of the medicinal plants belonging to the mint family with high content of carvacrol in its essential oil. A large body of literature has suggested that abiotic stresses, especially drought stress, could considerably boost the production of essential oils in plants. This study aimed to evaluate the effect of drought stress on the expression of genes affecting the biosynthesis of the main component of white savory essential oil carvacrol. Materials and methods: For this goal, 20 pot-planted seedlings of white savory at the 6-8 leaf stage were subjected in a greenhouse to two conditions of control and drought stress (10 pots for each condition) with irrigation regimes of 100% and 30% of field capacity, respectively. To ensure the implementation of stress on plants and to determine the appropriate time for sampling, leaf proline and relative water content of plants were measured. Results: The results of both proline and relative water content tests revealed a significant statistical difference between the control and stress conditions in white savory at 30% of field capacity. The KEGG (Kyoto Encyclopedia of Genes and Genomes) database was used to obtain information on the carvacrol biosynthesis pathway. The three key genes geranyl diphosphate synthase (gpps), terpene synthase (tps), and limonene hydroxylase (lh) were chosen for gene expression analysis. Due to the lack of genomic information on white savory, the sequence information of the genes in the closest genera within the family mint was used to design primers. The result of melting curve analysis from the real-time quantitative PCR (qPCR) assay confirmed the accuracy of the primer design. The raw qPCR data normalized to the housekeeping gene Actin, then differential expression analysis was performed using the delta-delta Ct method. qPCR result showed that expression of the genes gpps, tps, and lh were increased in stressed plants proximately 20, 18, and 50 times, compared to those in control, respectively. Conclusions: The conclusion that must be drawn from the data available is that water stress leads to the process of plant response to stress and changes the expression of genes involved in the biosynthesis pathway of the active substance carvacrol. [ABSTRACT FROM AUTHOR]

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Hamdy N El-Tallawy,1 Sawsan Abuhamdah,2,3 Ahmed Y Nassar,4 Wafaa MA Farghaly,1 Tahia H Saleem,4 Sara A Atta,4 Ayat A Sayed,4 Amal M Tohamy,1 Mohammed H Hassan5 1Department of Neurology and Psychiatry, Faculty of Medicine, Assiut University, Assiut, Egypt; 2Department of Pharmaceutical Sciences, College of Pharmacy, Al Ain University, Abu Dhabi, United Arab Emirates; 3Department of Biopharmaceutics and Clinical Pharmacy, Faculty of Pharmacy, University of Jordan, Amman, Jordan; 4Department of Medical Biochemistry, Faculty of Medicine, Assiut University, Assiut, Egypt; 5Department of Medical Biochemistry, Faculty of Medicine, South Valley University, Qena, 83523, EgyptCorrespondence: Mohammed H Hassan Email mohammedhosnyhassaan@med.svu.edu.egPurpose: Gephyrin (GPHN) is an essential protein in the regulation of inhibitory postsynaptic density and polymorphism in the corresponding gene may have a role in the development of pharmacoresistant epilepsy (PRE). For the first time, we aimed to evaluate the association of rs928553T/C variants with PRE susceptibility. Moreover, we have analyzed the genetic polymorphism affecting CYP2C9 “rs12782374G/A” in the same population to detect the effect of SNP on the drug-metabolizing ability of patients with PRE.Patients and Methods: This case-control study enrolled 100 patients (group A) and 100 healthy, age and sex-matched controls, unrelated to patients (group B). TaqMan™ assays using real-time PCR were run for genotyping of rs928553T/C and rs12782374G/A in all participants.Results: GPHN T>C polymorphism revealed significant risk association with occurrence of PRE using dominant, recessive and codominant models as follows: TT vs (TC+CC): OR 0.23, 95%CI: 0.13– 0.43, P< 0.001. In addition, (TT+TC vs CC): OR 0.38, 95%CI: 0.18– 0.77, P< 0.001. Also, T vs C (OR 0.34, 95%CI: 0.22– 0.51, P=< 0.001). Similarly, CYP2C9 G>A polymorphism showed a significant increased risk of PRE (GG vs (GA+AA): OR 0.11, 95%CI: 0.05– 0.23, P< 0.001). Furthermore, (GG+GA vs AA): OR 0.18, 95%CI: 0.084– 0.39, P< 0.001. Also, G vs A (OR 0.24, 95%CI: 0.15– 0.366, P=< 0.001).Conclusion: Mutation of both GPHN (rs928553) and CYP2C9 (rs1278237) genes may be implicated as a genetic mediators of resistance in patients with PRE.Keywords: pharmacoresistant epilepsy, rs928553T/C, rs12782374G/A, real time-PCR

Background and Purpose: Black Cumin of Kerman (Bunium persicum) is an Iranian plant that is commonly used as an antispasmodic, carminative, and antimicrobial substance. The present study aimed to assess different components of the essence of B.persicum and its effect on antifungal activity, spore germination inhibition, and expressions of FUM1 and FUM14 genes in Fusarium verticillioides strains.Materials and Methods: The essence was extracted by hydrodistillation and analyzed through gas chromatography-mass spectroscopy. A broth microdilution method was used for the determination of the minimum inhibitory concentration (MIC). In addition, the expression of FUM1 and FUM14 genes of toxigenic F. verticillioides was assessed byusing the real-time polymerase chain reaction (RT-PCR) technique.Results: Based on the findings, most of the essence consisted of γ-terpinene (15.56%),propanal, and 2-methyl-3-phenyl (14.18%). The oil showed a good antifungal activity(mean MIC value: 2556.8 μg/ml) as well as the inhibition of spore germination and mycelial growth (P

INTRODUCTION: This study aimed to evaluate the test results of patients whose RT-PCR test result was negative, but thorax CT findings were compatible with COVID-19 and their infectiousness in terms of contacts. METHODS: The study was designed as cross-sectional, descriptive research. In the first group, thorax CT revealed findings compatible with COVID-19 pneumonia, and the RT-PCR test result was negative. In the second group, thorax CT revealed findings compatible with COVID-19 pneumonia, and the RT-PCR test result was positive. Data collected were assessed with SPSS 21.0 software program. The significance level was accepted as 5%. RESULTS: Of the participants, 54.41% were male, and 45.59% were female. There was no significant difference between the two groups in terms of contact environment and status of being a healthcare worker and exitus. There was no difference between the first and second groups in terms of the number of contacts. The rate of sample collection was higher in the second group. The number of people who had contact with the patients and positive RT-PCR test results was higher in the first group. DISCUSSION AND CONCLUSION: Even if patients have negative RT-PCR test results for COVID-19, they are still infectious.

Cotton fibers are the seed trichomes that are developed around the seed and are used to make clothes and yarn for the textile industry. Expression profiling of cotton fiber genes is very important to estimate the differential gene expression level at different fiber developmental stages. Expression analysis of fiber developing genes are very important to enhance the fiber length of cotton. The expression profiling of three gene families in five stages (0, 5, 10, 15 and 20 DPA) of cotton fiber tissues was carried out through real-time PCR. Expression analysis revealed that transcripts of GA-20 Oxidase, XTH, and PEPc were elevated from 5 to 20 days post-anthesis (DPA) fibers. Total RNA was extracted from various stages of cotton fiber development and was reverse transcribed to cDNA for PCR amplification. For data normalization, 18s rRNA was used as an internal control. The objective of this study was to explore the expression level of fiber developing genes at specific stages of fiber development. The results showed that most of the genes were expressed during the elongation phase in between 5 DPA to 15 DPA. Results obtained from this study may be helpful for the further identification of fiber genes and the improvement of fiber characteristics in cotton. PEPc and XTH genes that are expressed with a high rate during the fiber development may be used in breeding programmes for the improvement of fiber quality and quantity.

Objective: The aim of this study was to evaluate the QIAstat-Dx® Respiratory SARS-CoV-2 Panel (QIAstat-SARS-CoV-2), which is a closed, fully automated, multiplex polymerase chain reaction (PCR) assay that detects severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and 21 other pathogens that cause respiratory disease. Methods: Nasopharyngeal swabs from patients with or suspected of having coronavirus disease 2019 were collected and tested at Bichat–Claude Bernard Hospital, Paris, France. Using the World Health Organisation-approved real-time-PCR assay developed by the Charité Institute of Virology as the reference, positive percent agreement (PPA) and negative percent agreement (NPA) were calculated. Results: In total, 189 negative and 88 positive samples were analyzed. QIAstat-SARS-CoV-2 had an NPA of 90.48% (95% confidence interval (CI), 85.37%, 94.26%) and a PPA of 94.32% (95% CI, 87.24%, 98.13%). Co-infections were detected by QIAstat-SARS-CoV-2 in 4/277 specimens. The methods exhibited comparable failure rates (23/307 [7.5%] vs. 6/298 [2.0%] for QIAstat-SARS-CoV-2 and reference methods, respectively). The turnaround time was shorter for QIAstat-SARS-CoV-2 compared with the reference method (difference in mean –14:30 h [standard error, 0:03:23; 95% CI, –14:37, –14:24]; P < 0.001). Conclusions: QIAstat-SARS-CoV-2 shows good agreement with the reference assay, providing faster and accurate results for detecting SARS-CoV-2.

Background and Objective: Laboratory-based education is one of the basic pillars of change in the management of teaching and learning of students of basic medical sciences. In this study, we designed a workshop with a new approach to teach Real time –PCR technique to postgraduate students of basic medical sciences and to evaluate the practical skills learned in the workshop, we used the standard OSPE assessment. Methods: In this study, 18 graduate students of Babol university of medical sciences were included and they were divided into two groups of intervention and control. Pre-test was performed in both groups. The intervention group was provided with an electronic content (including theoretical and practical outlines of the workshop) before participating in the workshop. At the end of the workshop and before the OSPE assessment, a post-test was taken from all participants. On the third day of the workshop, the performance of students in practical and laboratory skills of Real time-PCR technique was evaluated using OSPE examination. Findings: Pre-test and post-test scores of the intervention group in both cognitive and skill domains were higher than the control group. High satisfaction of students in each of OSPE stations in both groups indicates their acceptance of this approach. Conclusion: Using the OSPE examination in practical and laboratory courses is a good way to both assess and learn skills.

Background & objectives In gastrointestinal stromal tumour (GIST), not only genetic abnormalities are responsible for adverse clinical events, but epigenetic modifications also play a crucial role. MicroRNA (miRNA) dysregulation plays a significant role in carcinogenesis as miRNAs serve as natural silencer for their targets. Our study aimed to explore the miRNAs expression and its association with molecular and histopathological characteristics of GIST. Methods Fifty GIST samples, including 45 formalin fixed paraffin embedded (FFPE) and fresh tissues were included. Peripheral non-tumour tissues were used as controls. All the cases were confirmed using immunohistochemistry. RNA was extracted using miRNA-specific kit, and the expression was performed using RT-qPCR. The data were evaluated using AriaMx software version 1.5 (Agilent, US). MiRNAs expression was analyzed by using the relative quantification method (ΔΔCT). Results miR-221, miR-222, miR-494 and miR-34a showed significant down-regulation in tumours relative to non-tumour tissues. The expression levels of these miRNAs were significantly down-regulated in c-KIT (proto-oncogene encoding the tyrosine kinase transmembrane receptor)-positive tumours compared to c-KIT-negative. Further analysis revealed that reduced expression was associated with spindle subtypes and gastric localization. However, there was no significant correlation with other histological features. Additionally, miR-221/222, and miR-494 were down-regulated in most of the KIT exon 11 mutant subtypes, while miRNA-34a was associated with platelet derived growth factor receptor alpha (PDGFRA) mutations. Interpretation & conclusions The present study showed that the down-regulation of these miRNAs may help better molecular classification and characterization of GISTs. Our results offer new insight into the association between miRNAs and histological features, enabling a more thorough understanding of GISTs at the molecular level.

Human Respiratory Syncytial virus (HRSV) was major cause of viral of lower respiratory tract infection among infants and young children less 2 years old. RSV was an enveloped virus with negative sense, single Stranded RNA, belonging to the family of pneumoviridae and genes Orthopneumovirus. Multiple methods were used for the laboratory diagnosis of HRSV infection, included enzyme linked immunosorbent assay ELISA and Real Time PCR technique for detection HRSV – antibodies and HRSV –RNA. This study was aimed to compare the efficiency of two diagnostic methods in detection of HRSV in infant and young children. This study included 144 samples of children suffering from respiratory infection and was divided into 84 blood samples with percentage of 58.33% to detect the seropositiveness of IgM, IgG Via the ELISA test, 40 swabs from the nasopharyngeal to detect the viral gene, with percentage 27.77% by using RT-PCR technique, and 10 control. Results of current study showed HRSV was found to be positive in 35%, 55.9% of samples by RT-PCR and ELISA technique respectively, and children under 4 years old were more likely to be infection with the virus. Autumn-winter was most common for HRSV infections. [ABSTRACT FROM AUTHOR]

Bacteria of the Borrelia burgdorferi sensu lato complex are the causative agents of Lyme borreliosis (LB). Even if the conventional diagnosis of LB does not rely on the species itself, an accurate species identification within the complex will provide a deepened epidemiological scenario, a better diagnosis leading to a more targeted therapeutic approach, as well as promote the general public’s awareness. A comparative genomics approach based on the 210 Borrelia spp. genomes available in 2019 were used to set up three species-specific PCR protocols, able to detect and provide species typing of Borrelia afzelii, Borrelia burgdorferi sensu stricto (s.s.) and Borrelia garinii, the three most common and important human pathogenic Lyme Borrelia species in Europe. The species-specificity of these protocols was confirmed on previously identified B. afzelii, B. burgdorferi s.s. and B. garinii specimens detected in Ixodes ricinus samples. In addition, the protocols were validated on 120 DNA samples from ticks collected in Sweden, showing 88% accuracy, 100% precision, 72% sensitivity and 100% specificity. The proposed approach represents an innovative tool in epidemiological studies focused on B. burgdorferi s.l. occurrence in ticks, and future studies could suggest its helpfulness in routine diagnostic tests for health care.

Background & Objective: Malignant melanoma is the fatal cutaneous neoplasm which is curable by the early diagnosis. The expression of occludin protein which is an integral membrane protein is altered in an epithelial-to-mesenchymal transition. Although, recent studies provide sufficient evidence supporting the functional importance of occludin in cancer, the prognostic significance of occludin expression levels in melanoma remains obscure. The aim of this study was to determine occludin expression level and itscorrelation with clinicopathological features of the patients with melanoma. Methods: The occludin mRNA level was compared between paraffin-embedded tissues of 40 patients with melanoma and 10 subjects with normal skin. The quality and quantity of the RNA was determined and occludin expression level was measured using Real-time PCR and ∆∆CT computational technique. Result: Theoccludin mRNA level reduced five-fold in the melanoma patients compared to the control group (P=0.000). No significant difference was observed between male and female cases (P=0.533). No significant correlation was observed between occludin mRNA level, mitotic count (P=0.252), and Breslow levels (P=0.171) Conclusion: We can conclude that down-regulation of occludin expression in the patients with melanoma is a hallmark of cancer progression and it might be used as a prognostic factor. No significant correlation was found between occludin gene expression and clinicopathological characteristics including Clark level, Breslow staging, mitotic count, age and gender (P

هدف: تقریباً سه ‌چهارم زنان در معرض واژینیت کاندیدیایی هستند. از جمله عوامل بیماری‌زا می ‌توان به قدرت اتصال کاندیدا به بافت میزبان از طریق عواملی مانند مولکول ‌های چسبنده و پروتئین‌ هایی که این اتصال را تسریع می‌کنند اشاره نمود. هدف از انجام مطالعه حاضر؛ بررسی فراوانی ژن‌ های مولد بیوفیلم در سویه‌ های کاندیدا آﻟﺒﯿﮑﻨﺲ جداشده از واژینوز (Vaginosis) و بررسی مقایسه‌ ای بیان ژن als در سویه‌ های تحت تیمار (Treatment) با نانوذره آهن در شرایط آزمایشگاهی به روش Real Time-PCR می ‌باشد. روش­ بررسی: در این مطالعه توصیفی- مقطعی، تعداد 100 نمونه کاندیدا که از نمونه ‌های واژینوز زنان به ‌دست ‌آمده بود مورد بررسی قرار گرفت. پس از شناسایی سویه ‌های کاندیدا آلبیکنس با استفاده از تست‌ های بیوشیمیایی و میکروبیولوژیکی استاندارد، استخراج DNA ژنوم با استفاده از کیت شرکت سیناژن انجام گردید. به‌ منظور ردیابی ژن ‌های Als، Hwp1 و Hyr آزمون Multiplex-PCR انجام گردید. همچنین به‌ منظور بررسی اثر نانوذره آهن ‌بر بیان ژن ALS، RNA ­ سویه های حاوی این ژن با استفاده از کیت Cinnapure استخراج گردید و پس از تبدیل به CDNA، آزمون Real-Time PCR انجام شد. یافته­ ها: نتایج آنالیز مولکولی نشان داد که از 60 ایزوله کاندیدا آلبیکنس 34 ایزوله (65/66%) واجد ژن hwp ، 46 ایزوله (76/66 %) واجد ژن Als و 33 ایزوله (55%) واجد ژن Hyr بودند. نتایج مربوط به منحنی ذوب ژن Als در سویه‌ های بالینی کاندیدا آلبیکنس و تیمار شده با نانو ذره آهن نشان داد که Peak اصلی برای این ژن در دمای 85/34 درجه سلسیوس رخ می ‌دهد. نتیجه ‌گیری: بیان ژن Als در ایزوله‌ های تحت مطالعه تیمار شده نسبت به ایزوله‌ های تیمار نشده پایین‌تر بدست آمد که نشان دهنده نقش مثبت تیمار نانوذره آهن در کاهش بیان ژنAls در ایزوله ‌ها می باشد.

Backgrounds: The human Parainfluenza and Influenza viruses are common causes of respiratory infections in humans. The main goals of this study was investigating the prevalence of these viruses in health centers of Mazandaran province and comparing the clinical manifestations. Materials and methods: The sample collection was done using Dacron swabs from the oropharynx of the patients, and the specimens were transferred to laboratory by Hanks medium. The Viral genomes were isolated by the extraction kits and the RT-PCR and Real time-PCR methods were used for detection of Influenza and Parainfluenza viruses’ prevalence in the specimens, respectively. Results: Out of 100 samples, 5 (5%) specimens were positive for HPIV and 24 (24%) samples were detected as Influenza virus containing specimens. Among the all patients, 50 (50%) of them were female. No HPIV positive patients had croup. While any significant correlation was detected between personality characteristics (sex and age) and the infection with Parainfluenza viruses, but there was a significant relationship between sex and the detection of Influenza virus (p-value

Objectives: This study aimed to evaluate the test results of patients whose RT-PCR test result was negative, but thorax CT findings were compatible with COVID-19 and their infectiousness in terms of contacts. Materials and Methods: The study was designed as cross-sectional, descriptive research. In the first group, thorax CT revealed findings compatible with COVID-19 pneumonia, and the RT-PCR test result was negative. In the second group, thorax CT revealed findings compatible with COVID-19 pneumonia, and the RT-PCR test result was positive. Data collected were assessed with SPSS 21.0 software program. The significance level was accepted as 5%. Results: Of the participants, 54.41% were male, and 45.59% were female. There was no significant difference between the two groups in terms of contact environment and status of being a healthcare worker and exitus. There was no difference between the first and second groups in terms of the number of contacts. The rate of sample collection was higher in the second group. The number of people who had contact with the patients and positive RT-PCR test results was higher in the first group. Conclusion: Even if patients have negative RT-PCR test results for COVID-19, they are still infectious. [ABSTRACT FROM AUTHOR]

Out of (100) clinical samples from teeth were collected from patients suffering from recurrent caries. Diagnosis of bacterial colonies was first done by bacteriological methods. 22 (53.65%) isolates were S. aureus and 16 (41.46%) showed S. mutans. To detect S. aureus and S. mutans by using PCR assay, we characterized the isolates based on the presence of virulence genes (SpaP, mecA) by using the real-time PCR technique. (20) of the 22 (53.65%) samples of patients observed carried the mecA gene and (16) of the 10 (41.46%) samples of patients observed carried the SpaP gene. [ABSTRACT FROM AUTHOR]

Background and Aims: Due to the increasing problems and side effects of the use of chemical antibacterial agents as well as antibiotic resistance, this study aimed to evaluate the effects of aloe vera gel on biofilm gene expression of sulfate-reducing bacteria (SRB) isolated from patients with periodontal infection by Real time-PCR method. Materials and Methods: For this study, 100 individuals including 50 patients and 50 healthy individuals were recruited and examined by a periodontist. After identifying sulfate-reducing bacteria by biochemical tests and specific media, the effect of aloe vera extract on them was investigated and the expression of BFR gene against housekeeping gene (16srDNA) was determined by Real time- PCR test via T-test analysis method. Results: The data showed that 12 strains of sulfate-reducing bacteria were isolated from the samples, 5 of which had BFR gene and their gene expression was significantly reduced by aloe vera gel (P<0.05). Conclusion: The data of this study in proving the anti-biofilm and antibacterial effects of aloe vera extract showed that the expression of the target gene is reduced. It seems that this substance can be used as an alternative to oral hygiene chemicals. [ABSTRACT FROM AUTHOR]

Sexually transmitted infections (STI) are a public health problem. Real-time PCR assays are the most sensitive test for screening and diagnosis of these infections. The aim of this study was to evaluate a new CT/NG/TV/MG Real-Time PCR (RT-PCR) kit (Vircell) for the detection of Chamydia trachomatis , Neisseria gonorrhoeae , Mycoplasma genitalium and Trichomonas vaginalis for the diagnosis of sexual transmitted infections using the Allplex STI Essential Assay (Seegene) as the reference's method. A total of 497 samples from different anatomical sites (endocervical, urethral, rectal, pharyngeal and urine) were analysed from October 2022 to February 2023. A total of 108 (21.73 %) and 106 (21.33 %) positive samples were found for any of the assays used. The most commonly detected pathogen was N. gonorrhoeae (52 samples; 10.46 %), and the least commonly detected was T. vaginalis (three samples; 0.60 %). The anatomical site with the highest prevalence of micro-organisms was a non-urogenital site, the pharynx (26 positive samples; 5.23 %). Using the Allplex STI Essential Assay (Seegene) as the reference method, the diagnosis performance showed that the average specificity of CT/NG/TV/MG RT-PCR Kit (Vircell) was 99.84 % and the sensitivity was 99.53 %. The overall concordance was k=0.98 (CI95 %; 0.96-1). In conclusion, the CT/NG/TV/MG RT-PCR Kit (Vircell) assay shows a good sensitivity and specificity and constitutes a promising and additional alternative to routine procedures for distinct types of clinical specimen in diagnosis STI.

To clone the first anion channel from Xenopus laevis (X. laevis), we isolated a calcium-activated chloride channel (CLCA)-like membrane protein 6 gene (CMP6) in X. laevis. As a first step in gene isolation, an expressed sequence tags database was screened to find the partial cDNA fragment. A putative partial cDNA sequence was obtained by comparison with rat CLCAs identified in our laboratory. First stranded cDNA was synthesized by reverse transcription polymerase-chain reaction (RT-PCR) using a specific primer designed for the target cDNA. Repeating the 5' and 3' rapid amplification of cDNA ends, full-length cDNA was constructed from the cDNA pool. The full-length CMP6 cDNA completed via 5'- and 3'-RACE was 2,940 bp long and had an open reading frame (ORF) of 940 amino acids. The predicted 940 polypeptides have four major transmembrane domains and showed about 50% identity with that of rat brain CLCAs in our previously published data. Semi-quantification analysis revealed that CMP6 was most abundantly expressed in small intestine, colon and liver. However, all tissues except small intestine, colon and liver had undetectable levels. This result became more credible after we did real-time PCR quantification for the target gene. In view of all CLCA studies focused on human or murine channels, this finding suggests a hypothetical protein as an ion channel, an X. laevis CLCA.

Introduction: Severe Early Childhood Caries (S-ECC) remains highly prevalent worldwide. Streptococcus mutans and Streptococcus sobrinus are causative pathogens of dental caries and strongly involved in plaque or oral biofilm formation. Streptococcus sanguinis has antagonistic relationship with S. mutans and it might delay the colonisation of S. mutans in the oral cavities. Aim: To quantify S. mutans, S. sobrinus and S. sanguinis between 2 groups {S-ECC and Caries Free (CF)} of Thai children and to analyse the association between these bacteria and caries-related factors. Materials and Methods: Supra-gingival overnight plaque samples were collected from 120 Thai children aged 2-5 years (S-ECC=CF=60) using sterile toothpicks and released in 1 mL of TrisBase and EDTA buffer from January 2015 to December 2017. They were asked to expectorate saliva into a cup, plaque and gingival indices and dmft scores were recorded, parent’s demographic and children’s oral hygiene care and diet were assessed using questionnaire. DNA extraction and quantitative real-time PCR was performed. Different amounts of each bacterium were analysed by Mann-Whitney U test (p

Abstract Background: The purpose of the present study is to investigate the effect of 8 weeks High-intensity Interval Training (HIIT) on the expression of two muscle growth regulating genes (myostatin and follistatin) in gastrocnemius muscle of healthy male rats. Materials and Methods: 16 male Wistar rats were randomly divided into two groups in the same number: control and HIIT. HIIT program was underwent 40 min each session, three sessions in a week for eight weeks. Each exercise training session consisted of 5 min warm-up and cool-down at 40-50 % VO2max, 30 min interval running including 4 min high-intensity (85-90% VO2max) and 2 min active recovery (at 50-60% VO2max). Rats in control group did not do any exercise training program. 48 h after the last training session, rats` gastrocnemius muscle was extracted and the expression of myostatin and follistatin genes was determined by Real Time-PCR. For statistical data analysis, independent t-test was used. Results: The expression of myostatin was significantly reduced 68% in HIIT group in comparison with the control group (p0.05). Gastrocnemius muscle weight was significantly increased 23% in the HIIT group compared to the control group (p

Introduction/Objective. The purpose of this study was to compare polypropylene and silk suture materials in terms of bacterial adherence and clinical features including the impact on soft tissue healing. Methods. Ten healthy patients were included in this study. Unilateral upper and lower wisdom teeth were extracted at the same time and wounds were sutured with different threads (one monofilament – polypropylene – and one multifilament – silk suture). Stitches were removed seven days postoperatively. Real-time polymerase chain reaction was used to analyze bacterial adherence. Intraoperative handling and ease of removal were assessed with the help of Visual Analogue Scale. Landry healing index was used for evaluation of soft tissue healing. Results. Significantly more pronounced bacterial adherence was found on silk compared to polypropylene sutures (p = 0.005). Superior intraoperative handling properties were registered suturing with polypropylene compared to silk (p = 0.005). Soft tissue healing was significantly better around polypropylene sutures, both on the third and the seventh postoperative day (p = 0.016). Patient discomfort was slightly higher for polypropylene sutures, but without statistical significance. Conclusion. Polypropylene suture material showed significantly lower bacterial adherence and superior clinical features compared to silk, including better soft tissue healing. [Project of the Serbian Ministry of Education, Science and Technological Development, Grant no. 175075]

Listeria monocytogenes, a foodborne pathogen and well known causative agent of listeriosis in humans that is also known to form biofilm on utensils used in the dairy industry is well studied for peptide-based Quorum sensing system (agr QS system). In the present study, we are reporting L. monocytogenes strain BN3 to respond to an acyl-homoserine lactone (AHL) by expression of virulence gene (hlyA) and the gene responsible for biofilm formation (srtA) using Real-Time PCR technology. It revealed that in L. monocytogenes, hlyA gene (encodes listeriolysin O) expression increased 0.7-fold in response to 500 nM AHL (C4-HSL treated) as compared to control. Also, in response to 500 nM AHL, (C4-HSL treated) strain BN3 showed 27-fold up-regulation of srtA gene (encodes enzyme sortase) as compared to control. Our study confirmed the cross-talk between Gram-positive and Gram-negative bacteria since L. monocytogenes strain BN3 responds to C4-HSL, which is normally produced by Gram-negative bacteria. Here, we suggest targeting both agr based QS and AHL based QS system together in L. monocytogenes to tackle biofilm formed on milk cans in dairy industries and when treating listeria infections. [ABSTRACT FROM AUTHOR]

Background: Stress triggers the release of corticotrophic release factor (CRF) from the hypothalamus which in turn stimulates the anterior pituitary to release adrenocorticotrophic hormone (ACTH). Methods: The design of this study is a quasi-experimental approach / quasi experimental and one group design pre and post test design with group control. The sample in this study amounted to 24 respondents Results: Based on the results of the study showed that the sample LA24 (7,935) was the highest sample fold fold change and the lowest in the sample LA15 (6,194). Whereas in the post test results it was found that the LB18 sample (10,580) was the highest sample in fold change and the lowest was in the LB15 sample (8,472). Conclusion: There is a significant relationship / correlation between CRHR1 levels and FKBP5 levels (p = 0.01 α). This is indicated by the high / strong correlation coefficient value which is 0.846. A positive value at 0.846 indicates that there is a positive relationship or a concomitant relationship, namely the influence of yoga exercises will result in a decrease in CRHR1 levels and will further reduce the effect of FKBP5 levels in third trimester pregnant women. [ABSTRACT FROM AUTHOR]

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Coronavirus disease COVID-19, caused by Severe Acute Respiratory Syndrome Corona Virus-2 (SARS-CoV-2), is highly contagious and has been a pandemic since March 2020. The SARS-CoV-2 is an enveloped, single-stranded, positive-sense RNA viruswhich spreadsthrough air droplets by sneezing and coughing from affected person. The diagnosis of the COVID-19 remains a challenge to the scientists since the genome of the SARS-CoV-2 was novel and varying. Various studies have reported the validated procedures for sampling and the detection method of SARS-CoV-2. This mini-review provides a brief introduction of the SARS-CoV-2 features and the current knowledge for the recommended COVID19 detection methods including sampling procedures and real time SARS-CoV-2 genome detection. [ABSTRACT FROM AUTHOR]

Introduction: Severe Early Childhood Caries (S-ECC) remains highly prevalent worldwide. Streptococcus mutans and Streptococcus sobrinus are causative pathogens of dental caries and strongly involved in plaque or oral biofilm formation. Streptococcus sanguinis has antagonistic relationship with S. mutans and it might delay the colonisation of S. mutans in the oral cavities. Aim: To quantify S. mutans, S. sobrinus and S. sanguinis between 2 groups {S-ECC and Caries Free (CF)} of Thai children and to analyse the association between these bacteria and caries-related factors. Materials and Methods: Supra-gingival overnight plaque samples were collected from 120 Thai children aged 2-5 years (S-ECC=CF=60) using sterile toothpicks and released in 1 mL of TrisBase and EDTA buffer from January 2015 to December 2017. They were asked to expectorate saliva into a cup, plaque and gingival indices and dmft scores were recorded, parent's demographic and children's oral hygiene care and diet were assessed using questionnaire. DNA extraction and quantitative real-time PCR was performed. Different amounts of each bacterium were analysed by Mann-Whitney U test (p<0.05). The correlation between amounts of each bacterium and other clinical factors were analysed by Spearman's correlation test (p<0.05). Results: The guardian's demographic data, habit of milk bottle and breast feedings, oral hygiene care and consumption of cariogenic snacks were different between the two groups. S.mutans and S.sobrinus in plaque were higher in S-ECC. S.sanguinis in saliva and total bacteria were higher in the CF group. Plaque and gingival indices in S-ECC were higher than in the CF group. S.mutans levels in dental plaque and saliva was not significantly different. Conclusion: S.mutans and S.sobrinus were associated with S-ECC while S.sanguinis was associated with caries-free. Socioeconomics and children's oral hygiene care and diet were important factors associated with S-ECC. [ABSTRACT FROM AUTHOR]