scholarly journals Investigation of the presence of G354A (Cys87Tyr) mutation in osteoprotegerin gene in women with osteoporosis in Chaharmahal and Bakhtiari province

2019 ◽  
Vol 21 (6) ◽  
pp. 250-253
Author(s):  
Seyedeh Zahra Mousavi ◽  
Morteza Dehghan ◽  
Razieh Pourahmad
Keyword(s):  
Gel Electrophoresis ◽  
Analytical Study ◽  
Competitive Inhibitor ◽  
Osteoclast Formation ◽  
Bone Diseases ◽  
Mineral Density ◽  
Sequencing Method ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Osteoprotegerin Gene

Background and aims: Osteoprotegerin (OPG) is a competitive inhibitor of the differentiation and activity of osteoclasts, which inhibits the final stages of osteoclast formation and induces its apoptosis. In addition, OPG is considered as one of the most important candidate genes in the pathogenesis of bone diseases such as osteoporosis and idiopathic hyperphosphatasia. The G354A (Cys87Tyr) mutation in the OPG gene leads to idiopathic hyperphosphatasia. This mutation is probably related to osteoporosis. The purpose of this study was to investigate the presence of G354A (Cys87Tyr) in women with osteoporosis in Chaharmahal and Bakhtiari province. Methods: In this descriptive-analytical study, the bone mineral density (BMD) of the femoral neck and lumbar spine of women referring to Shahrekord bone densitometry centers was measured by the X-ray absorptiometry technique in 2013-2014. Based on T-scores, people with osteoporosis were identified and 70 patients were enrolled in the study after receiving their consent. Finally, DNA was extracted from blood samples, amplified by polymerase chain reaction (PCR) technique, and sequenced by DNA sequencing method. Results: After DNA extraction from the blood, the quality and quantity were determined by gel electrophoresis and spectrophotometry, respectively. Then, the gene was amplified by the PCR method and the product was detected by gel electrophoresis, followed by sequencing the samples to investigate the presence of the mutation. Eventually, genotypes associated with Cys87Tyr mutation were not observed in the studied population. Conclusion: In the present study, the G354A (Cys87Tyr) mutation associated with idiopathic hyperphosphatasia was not found in women with osteoporosis.

scholarly journals Kalkitoxin Reduces Osteoclast Formation and Resorption and Protects against Inflammatory Bone Loss

2021 ◽  
Vol 22 (5) ◽  
pp. 2303
Author(s):  
Liang Li ◽  
Ming Yang ◽  
Saroj Kumar Shrestha ◽  
Hyoungsu Kim ◽  
William H. Gerwick ◽  
...  
Keyword(s):  
Bone Loss ◽  
Transmembrane Protein ◽  
Osteoclast Differentiation ◽  
Mapk Signaling ◽  
Osteoclast Formation ◽  
Bone Diseases ◽  
Tartrate Resistant Acid Phosphatase ◽  
Nuclear Factor ◽  
Mineral Density ◽  
Multinucleated Cells

Osteoclasts, bone-specified multinucleated cells produced by monocyte/macrophage, are involved in numerous bone destructive diseases such as arthritis, osteoporosis, and inflammation-induced bone loss. The osteoclast differentiation mechanism suggests a possible strategy to treat bone diseases. In this regard, we recently examined the in vivo impact of kalkitoxin (KT), a marine product obtained from the marine cyanobacterium Moorena producens (previously Lyngbya majuscula), on the macrophage colony-stimulating factor (M-CSF) and on the receptor activator of nuclear factor κB ligand (RANKL)-stimulated in vitro osteoclastogenesis and inflammation-mediated bone loss. We have now examined the molecular mechanism of KT in greater detail. KT decreased RANKL-induced bone marrow-derived macrophages (BMMs) tartrate-resistant acid phosphatase (TRAP)-multinucleated cells at a late stage. Likewise, KT suppressed RANKL-induced pit area and actin ring formation in BMM cells. Additionally, KT inhibited several RANKL-induced genes such as cathepsin K, matrix metalloproteinase (MMP-9), TRAP, and dendritic cell-specific transmembrane protein (DC-STAMP). In line with these results, RANKL stimulated both genes and protein expression of c-Fos and nuclear factor of activated T cells (NFATc1), and this was also suppressed by KT. Moreover, KT markedly decreased RANKL-induced p-ERK1/2 and p-JNK pathways at different time points. As a result, KT prevented inflammatory bone loss in mice, such as bone mineral density (BMD) and osteoclast differentiation markers. These experiments demonstrated that KT markedly inhibited osteoclast formation and inflammatory bone loss through NFATc1 and mitogen-activated protein kinase (MAPK) signaling pathways. Therefore, KT may have potential as a treatment for destructive bone diseases.

scholarly journals A revisit to a low-cost method for the isolation of microsatellite markers: the case of the endangered Malayan tapir (Tapirus indicus)

2018 ◽  
Author(s):  
Qi Luan Lim ◽  
Nurul Adilah Ismail ◽  
Ramitha Arumugam ◽  
Wei Lun Ng ◽  
Christina Seok Yien Yong ◽  
...  
Keyword(s):  
Microsatellite Markers ◽  
Microsatellite Loci ◽  
Low Cost ◽  
Pcr Cloning ◽  
Chain Reaction ◽  
Sequencing Method ◽  
Rapid Polymerase Chain Reaction ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tapirus Indicus

AbstractThere are many approaches to develop microsatellite markers. We revisited an easy and rapid Polymerase Chain Reaction (PCR)-cloning-sequencing method to design microsatellite markers for Tapirus indicus. Using six random amplified microsatellite (RAM) markers, this study had rapidly generated 45 unique genomic sequences containing microsatellites. After screening 15 terminal and seven intermediate microsatellite loci, we shortlisted five and seven which were amplified either by single- or multiplex PCR using the economical three-primer PCR method. Genotyping attempts were made with ten Tapirus indicus individuals using three of the terminal microsatellite loci and all seven intermediate loci. However, none of the terminal microsatellite loci were considered useful for population genotyping studies, while the seven intermediate loci showed good amplification but were monomorphic in the ten samples. Despite successful detection of amplified loci, we would like to highlight that, researchers who are interested in this alternative method for isolation of microsatellite loci to be cautious and be aware of the limitations and downfalls reported herein that could render these loci unsuitable for population genotyping.

scholarly journals Antifungal strains and gene mapping of secondary metabolites in mangrove sediments from Semarang city and Karimunjawa islands, Indonesia

2021 ◽  
Vol 7 (4) ◽  
pp. 499-512
Author(s):  
Delianis Pringgenies ◽  
◽  
Wilis Ari Setyati
Keyword(s):  
Secondary Metabolites ◽  
Gene Mapping ◽  
Pathogenic Bacteria ◽  
Fungal Species ◽  
Mangrove Forests ◽  
Detection Approach ◽  
Sequencing Method ◽  
Genes Encoding ◽  
Pcr Method ◽  
Polymerase Chain

<abstract> <p>Infection caused by pathogenic fungal species is one of the most challenging disease to be tackled today. The antifungal bacteria candidate can be found in terrestrial as well as aquatic ecosystems, with mangrove forests being one of them. The purpose of this study is to obtain candidate isolates of antifungal strains with a detection approach and gene mapping simulation of bioactive compounds producers and screening to determine qualitative antifungal activity. The research will be carried out by collecting sediment samples from the mangrove ecosystems of Karimunjawa and Mangkang sub-district of Semarang city, isolating and purifying bacteria with Humic Acid Vitamin Agar (HVA), International Streptomyces Project 1 (ISP 1) and Zobell (Marine Agar). added with antibiotics, qualitative antifungal ability screening of each isolate obtained, detection of the presence of PKS gene and NRPS using special primers using the Polymerase Chain Reaction (PCR) method, and molecular identification of each isolate by 16s rRNA sequencing method. Of the total 59 isolates produced from the sample isolation process, 31 isolates from Karimunjawa sediments and 8 isolates from Semarang sediments showed activity against test pathogenic bacteria, namely <italic>Candida albicans</italic>, <italic>Trichoderma</italic> sp., and <italic>Aspergillus niger</italic>. Detection of Biosynthethic Gene Cluster (BGC) showed that the genes encoding secondary metabolites (NRPS, PKS 1 and PKS 2) were detected in KI 2-2 isolates from Karimunjawa. NRPS were detected only in isolates SP 3-9, SH 3-4, KI 1-6, KI 2-2, KI 2-4. The secondary metabolite-encoding gene, PKS1, was detected in isolates SP 3-5, SP 3-8, KI 2-2. PKS II genes were detected only on isolates SP 2-4, SH 3-8, KI 1-6, KI 2-2, and KI 2-4. Isolate SP 3-5 was revealed as <italic>Pseudomonas aeruginosa</italic> (93.14%), isolate SP 2-4 was <italic>Zhouia amylolytica</italic> strain HN-181 (100%) and isolate SP 3-8 was <italic>P. aeruginosa</italic> strain QK -2 (100%).</p> </abstract>

Investigation of duck plague virus in hoar areas of Bangladesh

2020 ◽  
Vol 26 (1-2) ◽  
pp. 73-78
Author(s):  
A Hossen ◽  
MH Rahman ◽  
MZ Ali ◽  
MA Yousuf ◽  
MZ Hassan ◽  
...  
Keyword(s):  
Cytopathic Effect ◽  
Clinical Sample ◽  
Chorioallantoic Membrane ◽  
Clinical Samples ◽  
Isolation And Identification ◽  
Duck Plague Virus ◽  
Pcr Method ◽  
Polymerase Chain ◽  
The Individual ◽  
Free Ranging

Duck plague (DP) is the most important infectious disease of geese, ducks and free-ranging water birds. The present study was conducted to determine the prevalence of duck plague virus followed by isolation and identification. For these purposes, a total of 155 cloacal swabs samples were collected randomly from duck of different haor areas of Bangladesh including 45 (41 surveillance and 4 clinical) samples from Netrokona; 42 (40 surveillance and 2 clinical) samples from Kishoregonj; 30 samples from Brahmanbaria and 38 samples from Sunamganj. The samples were processed and pooled (1:5 ratio) for initial screening of target polymerase gene of duck plague virus by polymerase chain reaction (PCR) method. All the samples of a positive pool were then tested individually for identifying the individual positive samples. The result showed that out of 155 samples, 41 (26.45%) were found positive in which 17 were from Netrokona, where 15 (36.58%) were from surveillance samples and 2 (50%) were from clinical sample; 16 were from Kishoregonj, where 14 (35%) were from surveillance samples and 2 (100%) were from clinical sample; 2 (6.6%) were from Brahmanbaria and 5 (13.15%) were from Sunamganj. These positive samples were inoculated into 9-10 days embryonated duck eggs (EDE) through chorioallantoic membrane (CAM) route for the isolation of virus. The EDE died earlier was also chilled, and in a similar way, the CAMs were collected and again performed PCR for id entification of virus. Out of 41 PCR positive samples, 26 samples were isolated and reconfirmed by PCR. Subsequently, DPV was isolated in primary duck embryo fibroblasts cell culture and confirmed by observing cytopathic effect (CPE). Bang. J. Livs. Res. Vol. 26 (1&2), 2019: P. 73-78

Prevalence of SHV-extended spectrum β-lactamase producing carbapenem –resistantKlebsiellapneumoniae among patients with lower respiratory tractinfections in Babylon Province-Iraq.

2018 ◽  
Vol 9 (SPL1) ◽  
Author(s):  
Fatima Moeen Abbas
Keyword(s):  
Resistance Rate ◽  
Combination Method ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Carbapenem Resistant ◽  
Extended Spectrum ◽  
Pcr Method ◽  
Polymerase Chain ◽  
Tract Infections ◽  
Positive Results

This study was carried out to screen the prevalence of Klebsiella pneumoniae isolated from patients with lower respiratory tract infections in Babylon province.From December,2015 to the end of March,2016,a total of 100 sputum samples were collected from patients visited or hospitalized Merjan Teaching Hospital and Al- Hashimya General Hospital. Fifteenth (65%) isolates were identified as Klebsiellapneumoniae. All bacterial isolates were evaluated for extended spectrum β-lactamase (ESBL) production phenotypically using disk combination method. Eleven (73.3%) isolates were detected as ESBL-producers. Kirby-Bauer disk diffusion method was employed to determine resistance profile of ESBLs-positive isolates. Higher rates of resistance were observed for ampicillin and piperacillin antibiotics with (81.8%) and (72.7%) resistance rate, respectively, while the lowest rate was noticed for imipenem antibiotic (14.28%). Carbapenem-resistant isolates were investigated for blaSHV gene by Polymerase Chain Reaction (PCR) method, 2 (100%) isolates gave positive results.

Calcium and Vitamin D Supplementation. Myths and Realities with Regard to Cardiovascular Risk

2019 ◽  
Vol 17 (6) ◽  
pp. 610-617 ◽  
Author(s):  
Giovanna Muscogiuri ◽  
Luigi Barrea ◽  
Barbara Altieri ◽  
Carolina Di Somma ◽  
Harjit pal Bhattoa ◽  
...  
Keyword(s):  
Vitamin D ◽  
Side Effects ◽  
Secondary Hyperparathyroidism ◽  
Calcium Supplementation ◽  
Physiological Role ◽  
Bone Diseases ◽  
Current Evidence ◽  
Bone Homeostasis ◽  
Mineral Density ◽  
Muscle Health

Vitamin D and calcium are considered crucial for the treatment of bone diseases. Both vitamin D and calcium contribute to bone homeostasis but also preserve muscle health by reducing the risk of falls and fractures. Low vitamin D concentrations result in secondary hyperparathyroidism and contribute to bone loss, although the development of secondary hyperparathyroidism varies, even in patients with severe vitamin D deficiency. Findings from observational studies have shown controversial results regarding the association between bone mineral density and vitamin D/calcium status, thus sparking a debate regarding optimum concentrations of 25-hydroxyvitamin D and calcium for the best possible skeletal health. Although most of the intervention studies reported a positive effect of supplementation with calcium and vitamin D on bone in patients with osteoporosis, this therapeutic approach has been a matter of debate regarding potential side effects on the cardiovascular (CV) system. Thus, the aim of this review is to consider the current evidence on the physiological role of vitamin D and calcium on bone and muscle health. Moreover, we provide an overview on observational and interventional studies that investigate the effect of vitamin D and calcium supplementation on bone health, also taking into account the possible CV side-effects. We also provide molecular insights on the effect of calcium plus vitamin D on the CV system.

scholarly journals Intra-Bone Marrow Administration of Mesenchymal Stem/Stromal Cells Is a Promising Approach for Treating Osteoporosis

2019 ◽  
Vol 2019 ◽  
pp. 1-10
Author(s):  
Hideki Agata ◽  
Yoshinori Sumita ◽  
Tatsuro Hidaka ◽  
Mayumi Iwatake ◽  
Hideaki Kagami ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Bone Mineral Content ◽  
Bone Mineral ◽  
Intravenous Administration ◽  
Stromal Cells ◽  
Mineral Content ◽  
Bone Diseases ◽  
Research Progress ◽  
Bone Thickness ◽  
Mineral Density

Mesenchymal stem/stromal cells (MSCs) are known to be useful for treating local bone diseases. However, it is not known if MSCs are effective for treating systemic bone diseases, as the risk for mortality following intravenous MSC administration has hindered research progress. In this study, we compared the safety and efficacy of intra-bone marrow and intravenous administration of MSCs for the treatment of ovariectomy- (OVX-) induced osteoporosis. Cells capable of forming bone were isolated from the murine compact bones and expanded in culture. Relatively pure MSCs possessing increased potential for cell proliferation, osteogenic differentiation, and inhibition of osteoclastogenesis were obtained by magnetic-activated cell sorting with the anti-Sca-1 antibody. Sca-1-sorted MSCs were administered to OVX mice, which were sacrificed 1 month later. We observed that 22% of the mice died after intravenous administration, whereas none of the mice died after intra-bone marrow administration. With respect to efficacy, intravenous administration improved bone mineral density (BMD) by increasing bone mineral content without affecting bone thickness, whereas intra-bone marrow administration improved BMD by increasing both bone mineral content and bone thickness. These results indicate that intra-bone marrow administration of pure MSCs is a safer and more effective approach for treating osteoporosis.

scholarly journals Clinical utility of the HCV drug resistance assay using the direct sequencing method and the cycleave PCR method in the dual therapy with daclatasvir and asunaprevir

Kanzo ◽  
2015 ◽  
Vol 56 (10) ◽  
pp. 533-535 ◽  
Author(s):  
Hideyuki Kudoh ◽  
Yoko Nagasawa ◽  
Michiru Ito ◽  
Nobuko Watanabe ◽  
Isao Naruse ◽  
...  
Keyword(s):  
Drug Resistance ◽  
Clinical Utility ◽  
Direct Sequencing ◽  
Dual Therapy ◽  
Sequencing Method ◽  
Direct Sequencing Method ◽  
Pcr Method
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