Role of DPY-30 Proteins in Flagella

Cilia and flagella are similar organelles found in eukaryotic cells. A microtubule-based axonemal cytoskeleton supports these organelles that project into the extracellular environment to detect various stimuli and propel fluid. These functions enable cells to sense and respond to the changing environment. Because of their importance, the fundamental mechanisms have been conserved throughout evolution. However, it remains largely unknown how axonemes are assembled and how the motility is regulated. This thesis investigated two molecules, RSP2 and RSP23, positioned in the axonemes within a T-shaped complex, the Radial Spoke (RS). Among several axonemal complexes, flagella lacking the evolutionarily conserved RS are paralyzed suggesting that this complex is crucial for motility. It was proposed that the RS transduces mechanical and chemical signals to regulate flagellar beating. These two molecules, RSP2 and RSP23, in Chlamydomonas bind the prototype calcium sensor, calmodulin. They also share a highly conserved DPY-30 domain that resembles the RIIa domain known for dimerization and targeting of cAMP-dependent protein kinase A to specific intracellular locations. Moreover, RSP2 links the head and stalk modules in the RS while RSP23 contains a Nucleoside Diphosphate Kinase (NDK) domain presumably for maintaining the equilibrium of nucleotide species. To reveal the mechanisms mediated by these molecules, mutants defective in these domains were generated. The calmodulin-binding region in RSP2, that is absent in mammalian homologs, is not required for the assembly of RS or the oscillatory beating. However, the motile mutants cannot maintain the typical helical trajectory when cells are exposed to bright light and glass barrier simultaneously. In contrast, mutants lacking the DPY-30 domain in RSP2 are paralyzed, despite the presence of all RSPs. Mutants expressing a fraction of RSP23 with inactive NDK activity generate shorter flagella with reduced amounts of RS. Together these results suggest that DPY-30 domains targets to key locations conserved molecular modules critical for flagellar elongation and motility and the diverged calmodulin-binding regions for steering the biflagellate. Models are proposed to illustrate the various roles of these molecular domains. These discoveries provide new insight on the extraordinary mechanisms in cilia and flagella

DPY-30 Domain and its Flanking Sequence Mediate the Assembly Modulation of Flagellar Radial Spoke Complexes

RIIa is known as the dimerization and docking (D/D) domain of the cyclic AMP (cAMP)-dependent protein kinase. However, numerous molecules, including radial spoke protein 2 (RSP2) in Chlamydomonas flagella, also contain an RIIa or a similar DPY-30 domain. To elucidate new roles of D/D domain-containing proteins, we investigated a panel of RSP2 mutants. An RSP2 mutant had paralyzed flagella defective in RSP2 and multiple subunits near the spokehead. New transgenic strains lacking only the DPY-30 domain in RSP2 were also paralyzed. In contrast, motility was restored in strains that lacked only RSP2’s calmodulin- binding C-terminal region. These cells swam normally in dim light but could not maintain typical swimming trajectories under bright illumination. In both deletion transgenic strains, the subunits near the spokehead were restored, but their firm attachment to the spokestalk required the DPY-30 domain. We postulate that the DPY-30–helix dimer is a conserved two-prong linker, required for normal motility, organizing duplicated subunits in the radial spoke stalk and formation of a symmetrical spokehead. Further, the dispensable calmodulin-binding region appears to fine-tune the spokehead for regulation of “steering” motility in the green algae. Thus, in general, D/D domains may function to localize molecular modules for both the assembly and modulation of macromolecular complexes

The Efficacy of Aerobic Exercise and Relaxation Technique in Fatigue among Children with Acute Lymphoblastic Leukemia: A Comparative study

INTRODUCTION : Childhood cancer is one of the leading cause of death by disease among which Acute Lymphoblastic Leukemia (ALL) accounts for 30% of all the malignancies. ALL is the most common form of pediatric leukemia which is also known as Acute Lymphocytic or Acute Lymphoid Leukemia. Incidence is approximately 3-4 cases per 100,000 children below 15 years with peak incidence below 2-5 years of age. In most of the countries, the incidence rate in children was approximately four times than that of the adults. Studies have shown a 98% cure rate in ALL. But even with these survival rates in pediatric population, only half of the ALL patients survived 5 years. Exact etiology of ALL is still unknown but common etiological factors include ionizing radiations, congenital anomalies, chemical agents like Benzene and Immunodeficiency syndromes. Many studies have reported the risk factors like Down syndrome, Erbstien Barr Virus (EBV) infections, parental history of fetal loss, miscarriage, perinatal exposure to pesticides and still birth to be reasons for developing leukemia. The pathogenesis of ALL is assumed to be the neoplasm arising from the clonal proliferation of immature cells of hemopoetic system which are characterized by the aberrant or arrested differentiation. The associated complications of ALL include musculoskeletal, Central Nervous System (CNS) and other systemic complications. Recent study have shown that 2% of the boys diagnosed with ALL shows testicular enlargement. Neural complications like pain, paresthesia and reduced deep tendon reflexes have been reported in various studies. Musculoskeletal complications like impaired gross motor and fine motor performances, increased fatigue, compromised ROM, strength and endurance have also been identified. Unfortunately, fatigue has become the most debilitating symptom in patients with cancer. It was reported to be an intensive disability among cancer patients as it induces inactivity which on prolonged rest lead to muscle wasting and loss of endurance. Fatigue affects quality of life to about 60% in cancer patients. But limited focus was given to those groups recognized with fatigue complaining unique experiences. AIM OF THE STUDY : This study was aimed to find out the effect of aerobic training and relaxation technique on fatigue in children with ALL. OBJECTIVES : 1. To find out the effectiveness of CSF in aerobic training among children with ALL. 2. To find out the effectiveness of PSF in aerobic training among children with ALL. 3. To find out the effectiveness of CSF in relaxation technique among children with ALL. 4. To find out the effectiveness of PSF in relaxation technique among children with ALL. 5. To compare the outcomes of CSF between the aerobic training and relaxation technique among children with ALL. 6. To compare the outcomes of PSF between the aerobic training and relaxation technique among children with ALL. METHODOLOGY : Study Design : A comparative study. Study Setting : Pediatric inpatient oncology ward, Sri Ramakrishna Institute of Oncology and Research, Coimbatore. Sampling Technique : A convenience sampling. Sample Size : A total of 30 acute lymphoblastic leukemia patients was selected on the basis of inclusion criteria and conveniently allocated into two groups of 15 each. Both the groups were designed for experimentation. Study Duration : The study was conducted over a period of 10 months. Treatment Duration : The treatment duration was 4 weeks. Inclusion Criteria : • Children undergoing chemotherapy. • Aged between 5-12. • Both gender. • Children with preserved cardiac function. • Children able to accept and follow verbal instructions. • Children complaining of fatigue. Exclusion Criteria : • Children with unstable cardiac function. • Children with platelet count less than 3000/uL. • Children with recent surgeries(less than 3 months). • Children with congenital musculoskeletal anomalies. • Children with visual and hearing deficits. • Children with systemic complications. RESULTS : The mean values of aerobic training group (27.7) is greater than the relaxation training group (23.3). The calculated ‘t’ value is 2.848 which is greater than the table value (2.048). The study showed that there is significant difference in child fatigue scale level of aerobic training group and relaxation training group at the level of p<0.05. This shows that aerobic training is effective than the relaxation training in improvement of fatigue among ALL children. CONCLUSION : The study confirmed the effects of both Aerobic exercise and Relaxation technique which could reduce the level of fatigue among children with ALL. The aerobic training yield better results compared to relaxation training in reducing fatigue level among ALL children

A DYNAMIC PROPOSAL FOR PROTECTING MULTIMEDIA DATA IN CLOUD SYSTEM

Distribution of copyrighted multimedia objects by way of uploading these to online hosting sites can effect in main lack of revenues for content designers. Systems needed to locate illegal copies of multimedia objects take time and effort and important. We recommend a manuscript the perception of important multimedia content protection systems.  We concentrate on the method for safeguarding multimedia content that is content-based copy recognition by which signatures are removed from original objects. Our bodies for multimedia content protection finds out unlawfully made copies of multimedia objects over the Internet. Our design attains fast employment of content protection systems, as it is according to cloud infrastructures that provide computing hardware in addition to software sources. Its two new components like a approach to generate signatures of three-dimensional and distributed matching engine for multimedia objects

Seroprevalence of transfusion transmissible infections among healthy blood donors at KIMS blood bank

Background: Safe blood is a critical component in improving health care and in preventing the spread of infectious diseases. Aims & Objectives: Blood transfusion can cause the transmission of infectious to recipients. This is an important mode of infection. The aim of the study was to assess the prevalence of such type of infection among blood donors and to compare the seroprevalence of transfusion transmitted diseases in blood donors. Retrospective study was conducted for 5 years from January-2009 to December-2013 at KIMS Blood Bank, Secunderabad. Materials and methods: All donors reporting to the blood bank during the period January-2009 to December-2013 were screened for human immunodeficiency virus (HIV) 1 & 2, hepatitis C viruses, malaria and syphilis. Screening of HIV, hepatitis B and hepatitis C viruses were done by chemillumiencies and syphilis was screened by RPR method. Results: A total of 39780 voluntary blood donors were screened, of which 38697 were males and 1083 were females. Seropositivity of HIV, hepatitis B, hepatitis C viruses & syphilis were 0.26%, 1.28%, 0.51% and 0.03% respectively. No blood donors test showed positivity for malaria parasite. Conclusion: With the implementation of strict donor selection criteria and use of sensitive screening test, it may be possible to reduce the incidence of TTIs

Analysis of HLA-A, HLA-B and HLA-DRB1 allelic frequencies in tertiary care from Telangana and Andhra Pradesh

Introduction: The high polymorphism of the HLA system and its typification is used as valuable tool in establishing association to various illnesses, immune and genetic profiles. It also provides a guide to identify compatibility among donor and recipient of organ transplants. Objective: To analyse HLA-A, HLA-B and HLA-DRB1 allelic frequencies among renal transplant recipients and their prospective related and unrelated donors attending Nephrology department at KIMS Hospital, Secunderabad. Methods: 146 kidney recipients and 154 related or unrelated donors coming from different regions of Telangana and Andhra Pradesh for kidney transplantation from September 2007 to December 2013 were included in this study. All were screened for HLA-A, B and DRB, via SSP PCR based methodology and allelic frequency was established. Result: 1738 and 17 different alleles were identified for HLA-A, B and DRB1 respectively. Alleles found more frequently are A*24, A*02, A*01, B*35, B*61, B*44 and DRB1*15, DRB1*04, DRB1*07. Conclusion: The results obtained are useful as a reference for organ transplantations and also for allowing compatible donors within specific area for patients requiring renal transplant

General and Specific Promotion of Flagellar Assembly by a Flagellar Nucleoside Diphosphate Kinase

Nucleoside diphosphate kinases (NDKs) play a central role in diverse cellular processes using the canonical NDK activity or alternative mechanisms that remain poorly defined. Our study of dimeric NDK5 in a flagellar motility control complex, the radial spoke (RS), has revealed new modalities. The flagella in Chlamydomonas ndk5 mutant were paralyzed, albeit only deficient in three RS subunits. RS morphology appeared severely changed in averaged cryo-electron tomograms, suggesting that NDK5 is crucial for the intact spokehead formation as well as RS structural stability. Intriguingly, ndk5’s flagella were also short, resembling those of an allelic spoke-less mutant. All ndk5’s phenotypes were rescued by expressions of NDK5 or a mutated NDK5 lacking the canonical kinase activity. Importantly, the mutated NDK5 that appeared fully functional in ndk5 cells elicited a dominant-negative effect in wild-type cells, causing paralyzed short flagella with hypophosphorylated, less abundant, but intact RSs, and accumulated hypophosphorylated NDK5 in the cell body. We propose that NDK5 dimer is an RS structural subunit with an additional mechanism that uses cross-talk between the two NDK monomers to accelerate phosphorylation-related assembly of RSs and entire flagella

Seroprevalance of HBsAg, anti-HIV 1&2 and anti-HCV by chemiluminescence method - A hospital based study from 2007 to 2014

Introduction: Hepatitis B virus (HBV), human immunodeficiency virus 1&2 (HIV1&2) and hepatitis C virus (HCV) are major public health problem. Detection of hepatitis B surface antigen (HBsAg), antibodies to human immunodeficiency virus 1&2 and hepatitis C virus in patients (both inpatients and outpatients) can help in patient management. Screening of patient samples for HBsAg, anti-HIV 1&2 and anti-HCV is important in managing surgical and medical care. The objective is to know prevalence of hepatitis B surface antigen (HBsAg), antibodies to HIV 1&2 and HCV inpatient and outpatient, their gender, age wise distribution and department wise distribution. Material & methods: The present study was conducted in the Department of Microbiology, Krishna Institute of Medical Sciences (KIMS), Secunderabad, Telangana, during a study period of 8 years from January 2007 to December 2014. It is a cross sectional study. It is a laboratory based study, no clinical correlation has been done. The positive/reactive samples were studied for their gender, age group and department. Analysis was done on a yearly basis. A cumulative data of 8 years is being presented. The samples were screened by enhanced chemiluminescence (ECLIA) technology by using an instrument, Vitros ECiQ(Ortho Clinical Diagnostic). Results: 1,00,590 patients were screened 1,805 (1.79 %) were positive for HBsAg. 92,819 patients screened 501(0.53%) were reactive for HIV 1&2 antibodies. 61,072 screened 744(1.21%) were reactive for HCV infection. The seroprevalence of all viral markers is more in males than females. In the present study seropositivity of HBsAg was higher in 41-50 year & 51-60 years followed by 31-40 years & 61-70 years. Highest prevalence of HIV was observed in 31-40 years followed by 41-50 years. Seropositivity of HCV was higher in 51-60 years followed by 41-50 years & 61-70 years. Conclusion: Emphasizes the need of screening of viral markers for inpatients and out patients. Adherence to universal precautions

Effect of Hydrocortisone on Mortality and Organ Support in Patients With Severe COVID-19: The REMAP-CAP COVID-19 Corticosteroid Domain Randomized Clinical Trial.

Importance: Evidence regarding corticosteroid use for severe coronavirus disease 2019 (COVID-19) is limited. Objective: To determine whether hydrocortisone improves outcome for patients with severe COVID-19. Design, Setting, and Participants: An ongoing adaptive platform trial testing multiple interventions within multiple therapeutic domains, for example, antiviral agents, corticosteroids, or immunoglobulin. Between March 9 and June 17, 2020, 614 adult patients with suspected or confirmed COVID-19 were enrolled and randomized within at least 1 domain following admission to an intensive care unit (ICU) for respiratory or cardiovascular organ support at 121 sites in 8 countries. Of these, 403 were randomized to open-label interventions within the corticosteroid domain. The domain was halted after results from another trial were released. Follow-up ended August 12, 2020. Interventions: The corticosteroid domain randomized participants to a fixed 7-day course of intravenous hydrocortisone (50 mg or 100 mg every 6 hours) (n = 143), a shock-dependent course (50 mg every 6 hours when shock was clinically evident) (n = 152), or no hydrocortisone (n = 108). Main Outcomes and Measures: The primary end point was organ support-free days (days alive and free of ICU-based respiratory or cardiovascular support) within 21 days, where patients who died were assigned -1 day. The primary analysis was a bayesian cumulative logistic model that included all patients enrolled with severe COVID-19, adjusting for age, sex, site, region, time, assignment to interventions within other domains, and domain and intervention eligibility. Superiority was defined as the posterior probability of an odds ratio greater than 1 (threshold for trial conclusion of superiority >99%). Results: After excluding 19 participants who withdrew consent, there were 384 patients (mean age, 60 years; 29% female) randomized to the fixed-dose (n = 137), shock-dependent (n = 146), and no (n = 101) hydrocortisone groups; 379 (99%) completed the study and were included in the analysis. The mean age for the 3 groups ranged between 59.5 and 60.4 years; most patients were male (range, 70.6%-71.5%); mean body mass index ranged between 29.7 and 30.9; and patients receiving mechanical ventilation ranged between 50.0% and 63.5%. For the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively, the median organ support-free days were 0 (IQR, -1 to 15), 0 (IQR, -1 to 13), and 0 (-1 to 11) days (composed of 30%, 26%, and 33% mortality rates and 11.5, 9.5, and 6 median organ support-free days among survivors). The median adjusted odds ratio and bayesian probability of superiority were 1.43 (95% credible interval, 0.91-2.27) and 93% for fixed-dose hydrocortisone, respectively, and were 1.22 (95% credible interval, 0.76-1.94) and 80% for shock-dependent hydrocortisone compared with no hydrocortisone. Serious adverse events were reported in 4 (3%), 5 (3%), and 1 (1%) patients in the fixed-dose, shock-dependent, and no hydrocortisone groups, respectively. Conclusions and Relevance: Among patients with severe COVID-19, treatment with a 7-day fixed-dose course of hydrocortisone or shock-dependent dosing of hydrocortisone, compared with no hydrocortisone, resulted in 93% and 80% probabilities of superiority with regard to the odds of improvement in organ support-free days within 21 days. However, the trial was stopped early and no treatment strategy met prespecified criteria for statistical superiority, precluding definitive conclusions. Trial Registration: ClinicalTrials.gov Identifier: NCT02735707