scholarly journals Sperm dynein ATPase and ATPases associated with various cellular activities (AAA+): regulation in oligo-astheno-teratozoospermia and necrozoospermia as extreme sperm motility disorders

2019 ◽  
Vol 92 (2) ◽  
Author(s):  
Silvia W. Lestari ◽  
Fitrah Firdaus ◽  
Dessy Noor Miati ◽  
Asmarinah
Keyword(s):  
Atpase Activity ◽  
Sperm Motility ◽  
Atp Hydrolysis ◽  
Sperm Concentration ◽  
Binding Pocket ◽  
World Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cellular Mechanisms ◽  
Motility Disorders ◽  
Health Organization

Asthenozoospermia is the most frequent sperm motility disorder, but there are other more extreme sperm motility disorders, namely oligo-astheno-teratozoospermia (OAT) and necrozoospermia. There are several cellular mechanisms known for OAT and necrozoospermia, but there are limited data on dynein ATPase and ATPases associated with various cellular activities (AAA+). AAA1 is involved in ATP hydrolysis, while AAA2 is entangled in ATP-binding pocket. This study was conducted to investigate the role of dynein ATPase activity and quantification of AAA dynein. Spermatozoa from 14 men with OAT, 11 men with necrozoospermic and 17 men with normoozspermic samples were used in this study. Makler chamber was used to determine sperm concentration and motility, while Papanicolaou stained semen smears using World Health Organization-fifth edition criteria was performed to determine sperm morphology, and dynein ATPase was quantified by calculation of released inorganic phosphate. AAA was quantified by enzyme-linked immunosorbent assay, whereas the distribution was determined by immunocytochemistry. This study showed that the dynein ATPase activity in OAT and necrozoospermia was lower than in the normozoospermic group (2.68±0.76, 1.01±0.31, 7.22±1.08 μmol Pi/mg protein/h, respectively, P<0.05), as well as the amounts of AAA1 and AAA2. In addition, staining for AAA in the sperm tail paralleled the dynein ATPase activity and quantity of AAA, being the highest in sperm from normozoospermic samples, lower in sperm from OAT samples, and almost undetectable in sperm from necrozoospermic samples. The structure and function of damaged sperm dynein may alter dynein ATPase activity and levels of AAA1 and AAA2.

scholarly journals Sperm Dynein AAA1 and AAA2 Expression in Human Sperm : A Regulation in Sperm Preparation

2018 ◽  
Vol 11 (1) ◽  
pp. 77-84 ◽  
Author(s):  
Silvia W. Lestari ◽  
Manggiasih D. Larasati ◽  
Indra G. Mansur ◽  
Muhammad F. Soelaeman ◽  
Favian A. Rahmat ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Atp Hydrolysis ◽  
Intrauterine Insemination ◽  
Semen Analysis ◽  
Gradient Centrifugation ◽  
Motor Protein ◽  
Human Sperm ◽  
World Health ◽  
Sperm Preparation ◽  
Health Organization

Axoneme structures in sperm tail, is a supramolecular protein complex with motor protein and regulatory which playing a crucial role in determining sperm motility. Dynein, one of the three members of cytoskeletal motor protein, has a ring of six AAA+ which linked together into one large polypeptide that contribute to the formation of sperm flagella bending. The previously research reported that the first two AAA motor domains, AAA1 and AAA2, were a major site in ATP hydrolysis associated with motility in the flagellum. Intrauterine Insemination (IUI) as a management of infertility requires sperm preparation procedures, by Swim-up (SU) and Density Gradient Centrifugation (DGC), in order to enhance the quality regarding concentration and motility of the initial sperm. This study aimed to evaluate the efficiency of the DGC and SU methods in selecting sperm, based the expression of sperm dynein AAA1 and AAA2. Semen samples were obtained from men underwent sperm preparation for IUI and divided into two groups, normozoospermia and asthenozoospermia, according to World Health Organization 2010 guideline. Semen analysis was performed to measure the sperm motility and velocity, before and after sperm preparation. The axoneme was isolated from the obtained samples from SU and DGC methods, while the level of AAA1 and AAA2 was measured by ELISA. This study showed that the percentage of motile sperm and velocity of prepared sperm in both groups in prepared sperm (post-SU and post-DGC) was higher compared to whole semen. The expression of sperm dynein AAA1 of prepared sperm in normozoospermia group showed higher, while in asthenozoospermia group showed lower activities compared to whole semen. The expression of sperm dynein AAA2 of prepared sperm in both groups showed lower activities compared to whole semen. The sperm preparation enhanced the quality of sperm and may increase the expression of sperm dynein AAA1 compared to the whole semen, without the involvement of sperm dynein AAA2.

scholarly journals Effects of Varicocelectomy on Abnormal Semen Parameters in Patients with Clinically Palpable Varicocele

2018 ◽  
Vol 43 (2) ◽  
pp. 94-100
Author(s):  
Md. Selim Morshed ◽  
AKM Khurshidul Alam ◽  
AKM Anwarul Islam ◽  
Sojib Bin Zaman ◽  
Mohammad Saruar Alam ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Surgical Correction ◽  
World Health ◽  
Semen Parameters ◽  
Semen Parameter ◽  
Quasi Experimental ◽  
Health Organization ◽  
The Impact

Prevalence of varicocele is approximately 15.1% of the general population. Studies showed that surgical correction of clinically palpable varicocele could improve the semen parameters. However, there is scarce of study to demonstrate post-operative outcome of varicocelectomy among the patients with abnormal semen parameter. Therefore, this study tried to compare the microscopic changes in sperm concentration, sperm motility, and sperm morphology before and after varicocelectomy. This quasi-experimental study was done in Urology Department of Bangabandhu Sheikh Mujib Medical University (BSMMU), Dhaka, Bangladesh from November 2014 to April 2016. Forty five patients with clinically palpable varicocele were finally includedin the study. Semen parameters of these patients were assessed by following the World Health Organization′s 5th manual of semen parameter, 2010. Subinguinal varicocelectomy was performed in every case. Follow up was done three months and six months after surgery. Data were analysed with statistical software SPSS 20 and level of significance was assessed by paired t-test.Almost 60.0% of patients were from 25 to 35 years age group. Of them, 87.1% patients were presented with left-sided varicocele, and 80% were grade III. All the patients showed oligospermia on semen analysis, whereas the number of abnormal sperm motility and abnormal morphology was 39, and 09, respectively. After surgical correction of varicocele, mean improvement of sperm concentration, motility and morphology found in 35 patients (77.7%), 31 patients (79.4%) and six patients (66.6%) respectively. The improvement was statistically significant (p<0.05). This study showed that surgical treatment could lead to the betterment of semen parameters in patients with clinically palpable varicocele, which will guide in deciding the management of varicocele patients presented with abnormal semen parameter. However, to recommend varicocelectomy as a most useful procedure in treating infertility cases; further longitudinal studies are recommended to establish the impact of varicocelectomy on

T26248G-transversion mutation in exon7 of the putative methyltransferase Nsun7 gene causes a change in protein folding associated with reduced sperm motility in asthenospermic men

2015 ◽  
Vol 27 (3) ◽  
pp. 471 ◽  
Author(s):  
Nahid Khosronezhad ◽  
Abasalt Hosseinzadeh Colagar ◽  
Syed Golam Ali Jorsarayi
Keyword(s):  
Protein Folding ◽  
Sperm Motility ◽  
Protein Function ◽  
Direct Sequencing ◽  
Protein Structures ◽  
Single Strand Conformation Polymorphism ◽  
Extraction Methods ◽  
World Health ◽  
Transversion Mutation ◽  
Health Organization

The NOP2/Sun domain family, member 7 (Nsun7) gene, which encodes putative methyltransferase Nsun7, has a role in sperm motility in mice. In humans, this gene is located on chromosome 4 with 12 exons. The aim of the present study was to investigate mutations of exon 7 in the normospermic and asthenospermic men. Semen samples were collected from the Fatemezahra IVF centre (Babol, Iran) and analysed on the basis of World Health Organization (WHO) guidelines using general phenol–chloroform DNA extraction methods. Exon 7 was amplified using Sun7-F and Sun7-R primers. Bands on samples from asthenospermic men that exhibited different patterns of movement on single-strand conformation polymorphism gels compared with normal samples were identified and subjected to sequencing for further identification of possible mutations. Direct sequencing of polymerase chain reaction (PCR) products, along with their analysis, confirmed C26232T-transition and T26248G-transversion mutations in asthenospermic men. Comparison of normal and mutant protein structures of Nsun7 indicated that the amino acid serine was converted to alanine, the structure of the helix, coil and strand was changed, and the protein folding and ligand binding sites were changed in samples from asthenospermic men with a transversion mutation in exon 7, indicating impairment of protein function. Because Nsun7 gene products have a role in sperm motility, if an impairment occurs in exon 7 of this gene, it may lead to infertility. The transversion mutation in exon 7 of the Nsun7 gene can be used as an infertility marker in asthenospermic men.

scholarly journals Advances in Diagnostic Methods for Zika Virus Infection

2018 ◽  
Vol 12 (4) ◽  
Author(s):  
Carlos A. Herrada ◽  
Md. Alamgir Kabir ◽  
Rommel Altamirano ◽  
Waseem Asghar
Keyword(s):  
Reverse Transcription ◽  
Zika Virus ◽  
Vaccine Development ◽  
Genome Structure ◽  
Diagnostic Methods ◽  
World Health ◽  
Health Concern ◽  
Localized Surface Plasmon ◽  
Enzyme Linked Immunosorbent Assay ◽  
Health Organization

The Zika virus (ZIKV) is one of the most infamous mosquito-borne flavivirus on recent memory due to its potential association with high mortality rates in fetuses, microcephaly and neurological impairments in neonates, and autoimmune disorders. The severity of the disease, as well as its fast spread over several continents, has urged the World Health Organization (WHO) to declare ZIKV a global health concern. In consequence, over the past couple of years, there has been a significant effort for the development of ZIKV diagnostic methods, vaccine development, and prevention strategies. This review focuses on the most recent aspects of ZIKV research which includes the outbreaks, genome structure, multiplication and propagation of the virus, and more importantly, the development of serological and molecular detection tools such as Zika IgM antibody capture enzyme-linked immunosorbent assay (Zika MAC-ELISA), plaque reduction neutralization test (PRNT), reverse transcription quantitative real-time polymerase chain reaction (qRT-PCR), reverse transcription-loop mediated isothermal amplification (RT-LAMP), localized surface plasmon resonance (LSPR) biosensors, nucleic acid sequence-based amplification (NASBA), and recombinase polymerase amplification (RPA). Additionally, we discuss the limitations of currently available diagnostic methods, the potential of newly developed sensing technologies, and also provide insight into future areas of research.

scholarly journals Development and Analytical Validation of an Immunoassay for Quantifying Serum Anti-Pertussis Toxin Antibodies Resulting from Bordetella pertussis Infection

2009 ◽  
Vol 16 (12) ◽  
pp. 1781-1788 ◽  
Author(s):  
Sandra L. Menzies ◽  
Vijay Kadwad ◽  
Lucia C. Pawloski ◽  
Tsai-Lien Lin ◽  
Andrew L. Baughman ◽  
...  
Keyword(s):  
Pertussis Toxin ◽  
Bordetella Pertussis ◽  
Collaborative Study ◽  
World Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Validation ◽  
Igg Antibody ◽  
Pertussis Infection ◽  
Validation Parameters ◽  
Health Organization

ABSTRACT Adequately sensitive and specific methods to diagnose pertussis in adolescents and adults are not widely available. Currently, no Food and Drug Administration-approved diagnostic assays are available for the serodiagnosis of Bordetella pertussis. Since concentrations of B. pertussis-specific antibodies tend to be high during the later phases of disease, a simple, rapid, easily transferable serodiagnostic test was developed. This article describes test development, initial evaluation of a prototype kit enzyme-linked immunosorbent assay (ELISA) in an interlaboratory collaborative study, and analytical validation. The data presented here demonstrate that the kit met all prespecified criteria for precision, linearity, and accuracy for samples with anti-pertussis toxin (PT) immunoglobulin G (IgG) antibody concentrations in the range of 50 to 150 ELISA units (EU)/ml, the range believed to be most relevant for serodiagnosis. The assay met the precision and linearity criteria for a wider range, namely, from 50 to 200 EU/ml; however, the accuracy criterion was not met at 200 EU/ml. When the newly adopted World Health Organization International Standard for pertussis antiserum (human) reference reagent was used to evaluate accuracy, the accuracy criteria were met from 50 to 200 international units/ml. In conclusion, the IgG anti-PT ELISA met all assay validation parameters within the range considered most relevant for serodiagnosis. This ELISA was developed and analytically validated as a user-friendly kit that can be used in both qualitative and quantitative formats. The technology for producing the kit is transferable to public health laboratories.

scholarly journals Comparison of Western Immunobloting to an Enzyme-Linked Immunosorbent Assay for the Determination of Anti-Bordetella pertussis Antibodies

2011 ◽  
Vol 18 (4) ◽  
pp. 615-620 ◽  
Author(s):  
Stephen D. Merrigan ◽  
Ryan J. Welch ◽  
Christine M. Litwin
Keyword(s):  
Immunosorbent Assay ◽  
Bordetella Pertussis ◽  
World Health ◽  
Enzyme Linked Immunosorbent Assay ◽  
Recent Infection ◽  
Content Type ◽  
Igm Elisa ◽  
Iga Antibodies ◽  
Health Organization

ABSTRACTDuringBordetella pertussisinfection, it has been established that an increase of anti-pertussis toxin (PT) and anti-filamentous hemagglutinin (FHA) antibodies occurs. Immunoblots from two manufacturers using FHA and PT antigens were compared with an enzyme-linked immunosorbent assay (ELISA) that used both FHA and PT. One manufacturer used two concentrations of PT bands for the IgG immunoblot, calibrated to the World Health Organization standard for PT in international units (IU/ml), 100 IU/ml (PT-100) and 8 IU/ml (PT). The second immunoblot kit measured antibodies to a single calibrated PT band. Both kits measured IgA antibodies, and one additionally measured IgM antibodies. Two of 41 (5%) ELISA IgM positives were confirmed positive by IgM immunoblotting, suggesting poor specificity of the IgM ELISA. The agreements of the IgG and IgA immunoblots with the ELISA ranged from 72.5% to 85.3%, with only 38 to 51% of IgA positives confirmed by immunoblotting and only 61 to 68% of IgG positives confirmed by immunoblotting. The two immunoblots correlated well with each other, with 91.7% and 94.3% agreement for IgG and IgA, respectively. When the FHA band was used with the PT band as the criterion for positivity, significant differences existed in specificity compared to the ELISA (IgG, 84.1% versus 33.3%; IgA, 82.4% versus 71.0%). When the positive IgA immunoblots (evidence of natural recent infection) were compared to the positive PT-100 IgG immunoblots (evidence of recent infection or vaccination), the PT-100 blot showed a 71% sensitivity in detecting natural recent infection.B. pertussisimmunoblots, alone or in combination with ELISAs, can aid in the diagnosis ofB. pertussisinfection.

Effect of Panax notoginseng Extracts on Inferior Sperm Motility in vitro

1999 ◽  
Vol 27 (01) ◽  
pp. 123-128 ◽  
Author(s):  
Jung-Chou Chen ◽  
Ming-Xiong Xu ◽  
Leih-Der Chen ◽  
Yan-Nian Chen ◽  
Tsan Hung Chiu
Keyword(s):  
Sperm Motility ◽  
Sperm Count ◽  
Semen Analysis ◽  
Panax Notoginseng ◽  
World Health ◽  
Computer Assisted ◽  
Butanol Extract ◽  
The World ◽  
Health Organization

The purpose of this study was to investigate the effects of Panax notoginseng extracts on inferior sperm motility in vitro. Semen samples were collected from 23 patients with sperm motility between 20% and 40%. The sperm count was over 20 × 106/ml in accordance with the World Health Organization standard. 1.0 mg/ml and 2.0 mg/ml of Panax notoginseng extracts including aqueous extract, n-butanol extract, and polysaccharide fraction on sperm motility and progression were evaluated by computer assisted semen analysis. The results demonstrated that sperm motility as well as progression on inferior sperm motility were enhanced at 1 hour and 2 hours after incubation with all three types of extracts.

scholarly journals The untapped cell biology of neglected tropical diseases

2016 ◽  
Vol 27 (5) ◽  
pp. 739-743 ◽  
Author(s):  
William Sullivan
Keyword(s):  
World Health Organization ◽  
Cell Biology ◽  
High Probability ◽  
Neglected Tropical Diseases ◽  
World Health ◽  
Tropical Diseases ◽  
Cellular Mechanisms ◽  
The World ◽  
Health Organization

The World Health Organization lists a constellation of 17 tropical diseases that afflict approximately one in six individuals on the planet and, until recently, few resources have been devoted to the treatment and eradication of those diseases. They are often referred to as the diseases of the “bottom billion,” because they are most prevalent among the poorest individuals in impoverished tropical nations. However, the few studies that have been performed reveal an extraordinary world of molecular and cellular adaptations that facilitate the pathogens’ survival in hosts ranging from insects to humans. A compelling case can be made that even a modest investment toward understanding the basic molecular and cell biology of these neglected pathogens has a high probability of yielding exciting new cellular mechanisms and insights into novel ways of combating these diseases.

scholarly journals Adherence to WHO 2010 Recommendations in Relation to Semen Analysis Reports in the Laboratories of West Bengal

2019 ◽  
Vol 01 (04) ◽  
pp. 187-192
Author(s):  
Sujoy Dasgupta
Keyword(s):  
Observational Study ◽  
Reference Values ◽  
West Bengal ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Reproductive Technology ◽  
World Health ◽  
The World ◽  
Health Organization ◽  
Better Than

Background: In 2010, The World Health Organization (WHO) suggested the standards of reporting of semen analysis and the reference values. We tried to determine the adherence to the WHO 2010 standard regarding semen analysis among the laboratories of West Bengal. Methods: An observational study was carried out by collecting the semen analysis reports from different laboratories. Compliance with the WHO 2010 recommendations regarding the reporting of semen analysis and references mentioned was subsequently analyzed. Results: A total of 211 laboratory reports were collected; of which 15 were ART (Assisted Reproductive Technology)-laboratories (7%) and 196 were non-ART-laboratories (93%). More than half of the laboratories did not mention any reference values. Only 7.5% used the phrase “WHO 2010” as the reference. Only 3% of the laboratories reported all the six “important” parameters (volume, pH, sperm concentration, motility, morphology and vitality) and used the WHO 2010 references for all of them. The ART laboratories performed significantly better than their non-ART counterparts in reporting and quoting the WHO 2010 reference values. Conclusion: Even nine years after its introduction, the compliance with the WHO 2010 recommendations on semen analysis was still low among our laboratories. There is need for increased awareness for the laboratory persons in this regard.

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