Seroprevalence and Molecular Detection of Influenza A Virus (H1N1) in Sulaimani Governorate-Iraq

Influenza A (H1N1) virus is now rapidly scattering across the world. Early detection is one of the most effective measures to stop the further spread of the virus. The current study was aimed to detect influenza A (H1N1) serologically and by polymerase chain reaction (PCR) techniques. From September 2020 to June 2021, three hundred nasopharyngeal swabs and blood samples were collected from Hiwa and Shahid Tahir Hospitals in Sulaimani city. Obtained results revealed that 23.3% of the tested patients were seropositive anti-IgG for Influenza A, while 13.3% showed anti-IgM seropositive results although 10% of the tested cases were with both anti-IgG and anti-IgM seropositive results. Gender, residency, and flu symptoms showed no significant relations with seropositive results (p<0.05) whereas valuable relations were found between seropositive observations and smoking, the previous history of chronic diseases as well as employment status (p<0.05). It was concluded that hematologic investigations (CBC) were not dependable if H1N1 diagnosis and detection. Only 1% of the tested samples showed positive results for influenza A (H1N1) RNA using reverse transcription-PCR.

Avirulence gene based RFLP and rep-PCR distinguish the genetic variation of Xanthomonas oryzae pv. oryzae pathotypes in Bangladesh

Bacterial blight (BB) caused by X. oryzae pv. oryzae is one of the devastating diseases of rice mostly in Asia. Genomes of X. oryzae pv. oryzae is highly variable due to rearrangement of the large contents of transposable elements and dynamic changes of X. oryzae pv. oryzae population regulated efficiency of the control measures used for BB management of rice worldwide. In this study, genetic variation of X. oryzae pv. oryzae pathotypes of Bangladesh was studied using aviruelnce gene based RFLP and rep-PCR techniques aimed to formulate pathogen targeted effective control measures against BB of rice. Eight pathotypes of X. oryzae pv. oryzae field isolates were identified based on their reactions against 10 Near Isogenic Lines (NILs). Among eight pathotypes, pathotypes IV and V contained higher number of isolates which were 30.13% and 23.01% respectively while pathotype VIII revealed as minimum containing only 2.51% of total isolates. These eight pathotypes were studied for their genetic variation by RFLP using avrBs3 repeat domain as probe. The results conceded that Bangladeshi X. oryzae pv. oryzae strains seem carrying a minimum of two and maximum of nine avrBs3 family genes homologs. The resistance phenotype on IRBB7 and IRBB10 NILs also indicated presence of two major avrBs3 family genes viz. avrxa7 and avrXa10 in some pathotypes. Relationship of phylogenicity exhibited that X. oryzae pv. oryzae pathotypes assorted into two RFLP haplotypes as well as these haplotypes are largely distributed in Bangladesh. Phylogenetic analyses carried out by (REP, ERIC), rep-PCR and BOX depicted the presence of two main molecular haplotypes of X. oryzae pv. oryzae pathotypes. The relationship between pathotypes and molecular haplotypes of X. oryzae pv. oryzae in Bangladesh indicated that the same lineage possesses different pathotypes and different lineage possesses different pathotypes. The results indicated that eight different pathotypes might have originated from common inherited haplotypes with a wide genetic variation.

Early determination of fetal sex in goat a comparison between real time PCR and ultrasonography

The point of the current study is to assess the productivity of the real time PCR and ultrasound techniques in early determination of fetal sex in Iraqi singleton pregnant goats. Our investigation has been led in Iraq, Al-Diwanya city from 10/8/2020 – 15/1/2021. The examination incorporates 45 singleton pregnant Iraqi goats, which initially inspected by ultrasound to affirm pregnancy and to decide the fetal sex depending on the restriction of the genital tubercle of the goat fetuses, after that, blood specimens had been gathered from the jugular vein of all examined does to detect fetal sex by discovery of AMLX and SRY genes in the circling cells free fetal DNA (ccffDNA) in these maternal blood specimens by utilizing real time PCR. Our outcomes showed an exceptionally high level of accuracy in real time PCR in contrast with the ultrasound strategy. The outcomes were affirmed by the true fetal sex after parturition in the inspected does. The complete symptomatic rate were 51.11% (23/45) and 97.78% (44/45) for ultrasound and PCR strategies separately. The exactness level of genuine analyzed female and male caprine kidding were 58.33% (7/12), 48.48% (16/33), and 100% (12/12), 96.97% (32/33) for ultrasound and real time PCR techniques separately. While the exactness rates of the two techniques utilized in this investigation for early caprine fetal sexing in respect to early pregnancies periods analyzed uncovered 100% (13/13), 96.3% (26/27), 100% (5/5), and 61.54% (8/13), 40.74% (11/27), 80% (4/5) in early pregnancy periods (58-62, 63-67, 68-73) days for real time PCR and ultrasound strategies individually. In conclusion our outcomes revealed a huge predominant exactness and productivity in fetal sexing in Iraqi singleton pregnant does in early development periods, with very high accuracy in real time PCR in compare to ultrasound techniques.

PCR-Based Analytical Methods for Quantification and Quality Control of Recombinant Adeno-Associated Viral Vector Preparations

Recombinant adeno-associated viral vectors (rAAV) represent a gene therapy tool of ever-increasing importance. Their utilization as a delivery vehicle for gene replacement, silencing and editing, among other purposes, demonstrate considerable versatility. Emerging vector utilization in various experimental, preclinical and clinical applications establishes the necessity of producing and characterizing a wide variety of rAAV preparations. Critically important characteristics concerning quality control are rAAV titer quantification and the detection of impurities. Differences in rAAV constructs necessitate the development of highly standardized quantification assays to make direct comparisons of different preparations in terms of assembly or purification efficiency, as well as experimental or therapeutic dosages. The development of universal methods for impurities quantification is rather complicated, since variable production platforms are utilized for rAAV assembly. However, general agreements also should be achieved to address this issue. The majority of methods for rAAV quantification and quality control are based on PCR techniques. Despite the progress made, increasing evidence concerning high variability in titration assays indicates poor standardization of the methods undertaken to date. This review summarizes successes in the field of rAAV quality control and emphasizes ongoing challenges in PCR applications for rAAV characterization. General considerations regarding possible solutions are also provided.

Genetic Markers as Risk Factors for the Development of Impulsive-Compulsive Behaviors in Patients with Parkinson’s Disease Receiving Dopaminergic Therapy

Impulsive–compulsive and related behavioral disorders (ICD) are drug-induced non-motor symptoms of Parkinson’s disease (PD). Recently research has focused on evaluating whether ICD could be predicted and managed using a pharmacogenetic approach based on dopaminergic therapies, which are the main risk factors. The aim of our study was to evaluate the role of candidate genes such as DBH, DRD2, MAOA, BDNF, COMT, SLC6A4, SLC6A3, ACE, DRD1 gene polymorphisms in the pathogenesis of ICD in PD. We compared patients with PD and ICD (n = 49), patients with PD without ICD (n = 36) and a healthy control group (n = 365). ICD was diagnosed using the QUIP questionnaires and specific diagnostic criteria for subtypes of ICD. Genotyping was conducted using a number of PCR techniques and SNaPshot. Statistical analysis was performed using WinPepi and APSampler v3.6 software. PCA testing was conducted using RStudio software v1.4.1106-5. The following substitutions showed statistically significant correlations with PD and ICD: DBH (rs2097629, rs1611115), DRD2 (rs6275, rs12364283, rs1076560), ACE (rs4646994), DRD1 (rs686), BDNF (rs6265), these associations are novel in Russian PD patients. Our findings suggest that polymorphisms in DBH, BDNF, DRD2, ACE genes in Russian subjects are associated with an increased risk of ICD development.

Effect of Imipenem and Amikacin Combination against Multi-Drug Resistant Pseudomonas aeruginosa

Pseudomonas aeruginosa is an opportunistic nosocomial pathogen associated with high morbidity and mortality rates. Combination of antibiotics has been found to combat multi-drug resistant or extensively drug resistance P. aeruginosa. In this study we investigate the in vitro and in vivo effect of amikacin and imipenem combination against resistant P. aeruginosa. The checkerboard technique and time-killing curve have been performed for in vitro studies showed synergistic effect for combination. A peritonitis mouse model has been used for evaluation of the therapeutic efficacy of this combination which confirmed this synergistic effect. The in vitro and in vivo techniques showed synergistic interaction between tested drugs with fractional inhibitory concentration indices (FICIs) of ≤0.5. Conventional PCR and quantitative real-time PCR techniques were used in molecular detection of bla IMP and aac(6′)-Ib as 35.5% and 42.2% of P. aeruginosa harbored bla IMP and aac(6′)-Ib respectively. Drug combination viewed statistically significant reduction in bacterial counts (p value < 0.5). The lowest bla IMP and aac(6′)-Ib expression was observed after treatment with 0.25 MIC of imipenem + 0.5 MIC of amikacin. Morphological changes in P. aeruginosa isolates were detected by scanning electron microscope (SEM) showing cell shrinkage and disruption in the outer membrane of P. aeruginosa that were more prominent with combination therapy than with monotherapy.

Differences and similarities of biophysical and biological characteristics between GBM and astrocyte cells

The current cancer studies focus on the molecular-targeting diagnostics and their interaction with surrounding microenvironment, however, there are some missing points on the characterization of the cells with their topological differences and elemental composition. Glioblastoma multiforme (GBM) which is an astrocytic aggressive brain tumour with short survival time. GBM and astrocyte cells may differ at molecular level and the elemental and topological evaluation of these cells are vital for a definition of new potential targets for cancer research. Here, we report the topology and chemistry of cancer (GBM) and healthy (astrocyte) cells by atomic force microscopy (AFM), scanning electron microscopy (SEM) supported with energy dispersive X-ray spectroscopy (EDS) and X-ray photoelectron spectroscopy (XPS), for the first time. Additionally, F-actin Immunofluorescence staining and Real-Time Polymerase chain reaction (RT-PCR) techniques were performed. Actin related genes were similar in level of gene expression; however, F-actin protein intensities were different. The gene expressions related to the invasion were elevated in GBM cells. Morphologically, GBM cells were found to be longer and narrower while astrocytes were shorter and more disseminated based on AFM. Furthermore, roughness values of these cells were relatively close to each other. SEM-EDS analysis demonstrated that elongated GBM cells exhibited several filopodial protrusions whereas the astrocyte surfaces were rougher in lamellipodial area. Our investigation provides considerable further insight into rapid cancer cell characterization field in terms of its combinatorial spectroscopic and microscopic approach.

An investigation of the genotoxic and cytotoxic effects of myclobutanil fungicide on plants

Myclobutanil is a chemical pesticide commonly used in the production of some vegetables and fruits like greenhouse peppers, grapes, and apples. The aim of this study was to investigate the genotoxic and cytotoxic effects of myclobutanil fungicide on the Allium cepa plant, the model organism. Randomly amplified polymorphic DNA and inter simple sequence repeat-PCR techniques were performed on the DNA of A. cepa exposed to the different myclobutanil doses and time periods. The nucleus anomalies and abnormal anaphases were investigated using a light microscope. PCR analyses showed that myclobutanil causes some DNA sequence changes on the onion genome depending on the increase in the fungicide dose and exposure time. It was determined that myclobutanil has a serious genotoxic effect, even in low doses like 25–50 ppm.

Teknik Diagnostik Konvensional dan Lanjutan Untuk Pemeriksaan Mikrobiologi pada Infeksi Nosokomial di Indonesia

Introduction: Nosocomial infections are infections caused by microbial such as bacteria, viruses, and fungi.that are acquired during the process of receiving health care. Diagnostic techniques for the examination of nosocomial infections play an important role in determining the accuracy of the infection of microorganisms causing infectious agents, so that the treatment given can be appropriate and minimize drug resistance. Purpose: This literature review is structured to provide an overview of diagnostic techniques fornosokomialinfection using conventional and advanced methods. Methods: The preparation of this review is based on the development of diagnostic techniques in the medical laboratory. Results: Conventional diagnostic techniques are generally carried out bymeans of culture on artificial media, macroscopic observations and biochemical tests. Further tests that can be applied are serological tests, antigen tests, and molecular tests such as PCR techniques. Conclusion: Conventional diagnostic techniques for microbiological examination of nosococomial infections require further tests to help establish a rapid and accurate diagnosis.