'Nothing About Us Without Us': exploring benefits and challenges of peer support for people with disability in peer support organisations - protocol paper for a qualitative coproduction project.

INTRODUCTION: One in six people live with disability in Australia with higher levels of disability of people from diverse communities, such as those with culturally and linguistically diverse (CALD) backgrounds. In Australia, CALD refers to people from diverse ethnicity and cultures, nationalities, societal structures and religions that may or may not speak a language other than English. This study employs researchers with lived experience of disability and peer support to study the impact of peer support for people with disability, including people from CALD backgrounds, in two peer-led organisations in New South Wales (NSW) Australia. METHODS AND ANALYSIS: This study uses participatory action research and inclusive research design with researchers with lived experience, having lived experience of disability and a peer in the disability community, leading the research.Over three years, three different groups will be recruited through Community Disability Alliance Hunter (CDAH) and Diversity and Disability Alliance (DDAlliance): (1) peers with disability, (2) peer leaders with disability and (3) researchers with lived experience of disability and peer support. Data collection and creation methods include semistructured interviews, surveys and focus groups. Qualitative data will be analysed using thematic analysis through the lens of the researchers with lived experience. ETHICS AND DISSEMINATION: Ethical approval was granted by the University of Newcastle Human Research Ethics Committee (Approval No: H-2021-0088). Dissemination includes peer-reviewed publications, presentations at local, national and international conferences and written reports for user-led organisations, disability service providers, disability agencies and people with disability.

Effect of long chain fatty acids on the memory switching behavior of tetraindolyl derivatives.

Non-volatile memory devices using organic materials have attracted much attention due to their excellent scalability, fast switching speed, low power consumption, low cost etc. Here, we report both volatile as well as non-volatile resistive switching behavior of p-di[3,3'-bis(2-methylindolyl)methane]benzene (Indole2) and its mixture with stearic acid (SA). Previously, we have reported the bipolar resistive switching (BRS) behavior using 1,4-bis(di(1H-indol-3-yl)methyl)benzene (Indole1) molecules under ambient conditions [Langmuir 37 (2021) 4449-4459] and complementary resistive switching (CRS) behavior when the device was exposed to 353 K or higher temperature [Langmuir 38 (2022) 9229-9238]. However, the present study revealed that when the H of -NH group of Indole1 is replaced by -CH3, the resultant Indole2 molecule-based device showed volatile threshold switching behaviour. On the other hand, when Indole2 is mixed with SA at a particular mole fraction, dynamic evolution of an Au/Indole2-SA/ITO device from volatile to non-volatile switching occurred with very good device stability (>285 days), memory window (6.69 × 102), endurance (210 times), data retention (6.8 × 104 s) and device yield of the order of 78.5%. Trap controlled SCLC as well as electric field driven conduction was the key behind the observed switching behaviour of the devices. In the active layer, trap centers due to the SA network may be responsible for non-volatile characteristics of the device. Observed non-volatile switching may be a potential candidate for write once read many (WORM) memory applications in future.

Ophthalmic genetic counselling: emerging trends in practice perspectives in Asia.

Genetic counselling (GC) provides information to the patient and the family to make informed choices. Among the advanced Western countries and a few Asian countries, there are certified or trained professionals who perform GC. The Human Genome Project and next-generation sequencing diagnostics have provided an opportunity for increased genetic testing in the field of ophthalmology. The recent interventional therapeutic research strategies have also generated additional interest to seek GC globally, including in Asia. However, GC has several barriers to practise in the developing countries in Asia, namely, (a) shortage of qualified or trained genetic counsellors, (b) poor knowledge and reluctance in clinical adoption of genomics among the physicians in clinical practice, (c) overstretched public health services, and (d) negligible ophthalmic GC-related research and publications. The GC inadequacy in Asia is glaring in the most populous countries like China and India. Cultural differences, religious beliefs, misogyny, genetic discrimination, and a multitude of languages in Asia create unique challenges that counsellors in the West may only encounter with the immigrant minorities. Since there are currently 500 or more specific Mendelian genetic eye disorders, it is important for genetic counsellors to translate the genetic results at a level that the patient and family understand. There is therefore a need for governmental and healthcare organisations to train genetic counsellors in Asia and especially this practice must be included in the routine comprehensive ophthalmic care practice.

Impact of the National Amyotrophic Lateral Sclerosis Registry: Analysis of Registry-funded Research.

OBJECTIVE: This research aims to examine the impact of the National Amyotrophic Lateral Sclerosis (ALS) Registry-funded research activities. METHODS: Registry-funded research and related publications were identified through the National ALS Registry website, the National Institutes of Health (NIH) Reporter website, and verified by Principal Investigators. Key study characteristics (e.g., study population, sample size) and key impact features (e.g., risk factors) were abstracted and recorded on study abstraction forms. Descriptive statistics were used to analyze the volume, productivity, and findings of the Registry-funded research. RESULTS: Since 2012, the National ALS Registry funded 21 research projects. Of these, 14 were through extramural research grants and included in the analysis. These studies are often related to environmental, medical conditions, and genetic risk factors. On average, the funded grants produced 1 to 2 publications which were cited 114 times by other researchers. The relative citation ratio averaged 1.81 with a weighted relative citation ratio of 16.28. These studies supported the identification and confirmation of candidate risk factors. Environmental and occupational risk factors typically related to heavy metal exposure (e.g., lead, mercury) and agricultural chemicals (e.g., pesticides, herbicides), and the occupations associated with exposure to these substances were most frequently explored. INTERPRETATION: The National ALS Registry is a multifaceted research platform, one component of which is funded research. This Registry-funded research fills an essential gap in the overall ALS scientific community as it is difficult to prevent and treat a disease without a deeper understanding of its causes.

Differential dysregulation of granule subsets in WASH-deficient neutrophil leukocytes resulting in inflammation.

Dysregulated secretion in neutrophil leukocytes associates with human inflammatory disease. The exocytosis response to triggering stimuli is sequential; gelatinase granules modulate the initiation of the innate immune response, followed by the release of pro-inflammatory azurophilic granules, requiring stronger stimulation. Exocytosis requires actin depolymerization which is actively counteracted under non-stimulatory conditions. Here we show that the actin nucleator, WASH, is necessary to maintain azurophilic granules in their refractory state by granule actin entrapment and interference with the Rab27a-JFC1 exocytic machinery. On the contrary, gelatinase granules of WASH-deficient neutrophil leukocytes are characterized by decreased Rac1, shortened granule-associated actin comets and impaired exocytosis. Rac1 activation restores exocytosis of these granules. In vivo, WASH deficiency induces exacerbated azurophilic granule exocytosis, inflammation, and decreased survival. WASH deficiency thus differentially impacts neutrophil granule subtypes, impairing exocytosis of granules that mediate the initiation of the neutrophil innate response while exacerbating pro-inflammatory granule secretion.

Complementary Resistive Switching Behavior in Tetraindolyl Derivative-Based Memory Devices.

Complementary resistive switching (CRS) devices are more advantageous compared to bipolar resistive switching (BRS) devices for memory applications as they can minimize the sneak path problem observed in the case of BRS having a crossbar array structure. Here, we report the CRS behavior of 1,4-bis(di(1H-indol-3-yl)methyl)benzene (Indole1) molecules. Our earlier study revealed that Au/Indole1/Indium tin oxide (ITO) devices showed BRS under ambient conditions. However, the present investigations revealed that when the device is exposed to 353 K or higher temperatures, dynamic evolution of the Au/Indole1/ITO device from BRS to CRS occurred with a very good memory window (∼103), data retention (5.1 × 103 s), stability (50 days), and device yield (∼ 60%). This work explores the application possibility of indole derivatives toward future ultradense resistive random access memory.

An Evaluation of Jaw Tracking Movements in Patients with Total Joint Replacements versus a Control Group.

Background and Objectives: One form of treatment for degenerative temporomandibular joint diseases such as osteoarthritis, rheumatic arthritis, TMJ ankylosis, and condylar resorption is total joint replacement. The aim of this study was to examine the function of the temporomandibular joint after prosthetic joint replacement. Materials and methods: Fifteen patients with unilateral or bilateral TMJ total joint replacements and 15 healthy controls were evaluated via a SICAT JMT+ device. This non-invasive system measures 3D position and linear movements in all degrees of freedom and allows undisturbed functional mandibular movements to provide a quantitative evaluation. In addition, a TMJ questionnaire consisting of the subjective symptoms was also obtained. To date, no similar studies have been cited in the literature. Results: Mandibular movements after prosthetic joint replacement were recorded during opening, closing, protrusion, and lateral excursive movements and were all significantly decreased compared to those of controls. In the treatment group, the maximum incisal opening was 33.46 ± 5.47 mm, left lateral movement was 1.91 ± 2.7 mm, right lateral movement was 1.74 ± 1.74 mm, and protrusive movement was 2.83 ± 2.05 mm. The p-value comparison study and control group indicated significant difference (p < 0.0001) between the two groups. The study group stated a high level of satisfaction with the total joint replacement. Conclusion: Within the limitations of the study, the following conclusions can be drawn: (1) TMJ replacement patients showed significantly limited jaw movements compared to the control group; (2) a small percentage of TMJ replacement patients still present low levels of pain but improved chewing ability and quality of life.

Prevalence of chronic obstructive pulmonary disease (COPD) among rural population: A national survey in Bangladesh.

Background: In Bangladesh, there is a scarcity of nationally representative data on the burden of chronic obstructive pulmonary disease (COPD). Methods: To estimate the COPD prevalence in rural settings, this cross-sectional, population-based study was conducted in all eight administrative divisions of Bangladesh, and involved adults aged 40 years and above. By using multi-stage random sampling, 2,458 individuals were enrolled. The Global Initiative for Chronic Obstructive Lung Disease (GOLD) guidelines were used to diagnose COPD based on post-bronchodilator lung function, while additional participant data was gathered through computer-assisted personal interviews. Results: A 2% COPD prevalence (95% CI: 1.45, 2.55) was found in the study sample with a statistically significant difference between males (2.7%; 95% CI: 1.8, 3.6) and females (1.2%; 95% CI: 0.59, 1.81). Increasing age significantly inflated the odds of having COPD irrespective of sex (OR: 1.03; 95% CI: 1.00, 1.05; P value < 0.05). Furthermore, prevalence of COPD was higher among manual workers, cigarette smokers, and those that used the indoor kitchen and did not have a primary education. Sex-based analysis showed that smokeless tobacco consumption was significantly associated with COPD occurrence among males (OR: 2.14; 95% CI: 1.05, 4.37; P value < 0.05), but not females. Further, using an indoor kitchen increased the odds of developing COPD by 400% among female participants (OR: 4.39; 95% CI: 1.37, 14.10; P value < 0.05). Conclusion: This study provides a comprehensive sex-based estimation of COPD prevalence among rural population and imparts significant contribution to the growing database on COPD prevalence in Bangladesh.

DYNC1LI2 regulates localization of the chaperone-mediated autophagy receptor LAMP2A and improves cellular homeostasis in cystinosis.

The dynein motor protein complex is required for retrograde transport but the functions of the intermediate-light chains that form the cargo-binding complex are not elucidated and the importance of individual subunits in maintaining cellular homeostasis is unknown. Here, using mRNA arrays and protein analysis, we show that the dynein subunit, DYNC1LI2 (dynein, cytoplasmic 1 light intermediate chain 2) is downregulated in cystinosis, a lysosomal storage disorder caused by genetic defects in CTNS (cystinosin, lysosomal cystine transporter). Reconstitution of DYNC1LI2 expression in ctns-/- cells reestablished endolysosomal dynamics. Defective vesicular trafficking in cystinotic cells was rescued by DYNC1LI2 expression which correlated with decreased endoplasmic reticulum stress manifested as decreased expression levels of the chaperone HSPA5/GRP78, and the transcription factors ATF4 and DDIT3/CHOP. Mitochondrial fragmentation, membrane potential and endolysosomal-mitochondrial association in cystinotic cells were rescued by DYNC1LI2. Survival of cystinotic cells to oxidative stress was increased by DYNC1LI2 reconstitution but not by its paralog DYNC1LI1, which also failed to decrease ER stress and mitochondrial fragmentation. DYNC1LI2 expression rescued the localization of the chaperone-mediated autophagy (CMA) receptor LAMP2A, CMA activity, cellular homeostasis and LRP2/megalin expression in cystinotic proximal tubule cells, the primary cell type affected in cystinosis. DYNC1LI2 failed to rescue phenotypes in cystinotic cells when LAMP2A was downregulated or when co-expressed with dominant negative (DN) RAB7 or DN-RAB11, which impaired LAMP2A trafficking. DYNC1LI2 emerges as a regulator of cellular homeostasis and potential target to repair underlying trafficking and CMA in cystinosis, a mechanism that is not restored by lysosomal cystine depletion therapies.Abbreviations: ACTB: actin, beta; ATF4: activating transcription factor 4; CMA: chaperone-mediated autophagy; DYNC1LI1: dynein cytoplasmic 1 light intermediate chain 1; DYNC1LI2: dynein cytoplasmic 1 light intermediate chain 2; ER: endoplasmic reticulum; LAMP1: lysosomal associated membrane protein 1; LAMP2A: lysosomal associated membrane protein 2A; LIC: light-intermediate chains; LRP2/Megalin: LDL receptor related protein 2; PTCs: proximal tubule cells; RAB: RAB, member RAS oncogene family; RAB11FIP3: RAB11 family interacting protein 3; RILP: Rab interacting lysosomal protein.

Heavy Metals in the Fish Tenualosa ilisha Hamilton, 1822 in the Padma-Meghna River Confluence: Potential Risks to Public Health.

Hilsa shad (Tenulosa ilisha) is Bangladesh's most important single-species fishery that contributes to 11% of total catch and employment for millions of people. However, heavy metals (HMs) toxicity in the edible organs of T. ilisha and their plausible public health threats have received weak attention. To provide insights on this issue, we determined, using ICP-MS, the concentration of Zn, Cu, Cr (VI), Pb, and Cd in the edible organs of five different sizes of T. ilisha and the surface water collected from the Padma-Meghna River confluence, Chandpur (Bangladesh). Multivariate analysis indicated that T. ilisha gills and liver contained higher HMs than muscle, and the surface water was below the safety limits. The study revealed that only Cr crossed the safety limits and bioaccumulated in the smaller-sized gills and liver. To assess the public health risks, target hazard quotient (THQ), total THQ (TTHQ) and carcinogenic (CR) risks were calculated. Only Cr imposed non-carcinogenic risks to consumers, while TTHQ showed higher chronic health risks. There was no CR risk measured for consumers, except for the largest-sized gills for children. Randomly positive relations between HMs and sizes were found; whereas, consistently positive relations were found among the tissue types. The outcomes of our study may aid policymakers in managing pollutants, especially the Cr sources in the greater Chandpur regions.

Impact of Literacy on Hypertension Knowledge and Control of Blood Pressure in a Southern Indian Tertiary Hospital.

BACKGROUND: Hypertension is a global public health concern. Awareness and knowledge about the disease in a community collectively would allow adequate prevention, promote self-care practices, adherence to medication and ultimately effective management of hypertension. AIMS: To ascertain the level of education associated with the knowledge of hypertension and control of blood pressure. METHODS: A cross-sectional questionnaire survey consisting of item questions about awareness and knowledge of hypertension. Hypertensive patients (n = 424) of both genders and more than 20 years of age were included in the study. Hypertensive patients were divided into two groups (school group and school pass-out group) to assess the level of knowledge. Chi-square test was performed to determine the assessment, and p-value < 0.05 was considered significant. RESULTS: Out of 424 participants, 71.2% were school group and 28.7% school pass-out group. School pass-out group had significant knowledge about dangerous natural course of hypertension (p = 0.00069), hypertension can lead to death if untreated (p = 0.015), benefits of cessation of smoking (p = 0.03), advantage of limiting alcohol (p = 0.019) and performing regular exercise (p = 0.013) reduces blood pressure. School pass-out group had significant (p = 0.04) hypertension control compared to the school group. CONCLUSION: Educational status plays a vital role in increasing knowledge and improving the management of hypertension through better self-care practices and strict adherence to medication. Community- based health education interventional programs targeting the lower socioeconomic group of a population would help to reduce the gap in awareness and effective control of hypertension.

Current concepts and molecular mechanisms in pharmacogenetics of essential hypertension.

Hypertension is a leading age-related disease in our society and if left untreated, leads to fatal cardiovascular complications. The prevalence of hypertension has increased and becomes a significant global health economic burden, particularly in lower-income societies. Many loci associated with blood pressure and hypertension have been reported by genome-wide association studies that provided potential targets for pharmacotherapy. Pharmacogenetic research had shown interindividual variations in drug efficacy, safety, and tolerability. This could be due to genetic polymorphisms in the pharmacokinetics (genes involved in a transporter, plasma protein binding, and metabolism) or pharmacodynamic pathway (receptors, ion channels, enzymes). Pharmacogenetics promises great hope toward targeted therapy, but challenges remain in implementing pharmacogenetic aided antihypertensive therapy in clinical practice. Using various databases, we analyzed the underlying mechanisms between the candidate gene polymorphisms and antihypertensive drug interactions and the challenges of implementing precision medicine. We review the emergence of pharmacogenetics and its relevance to clinical pharmacological practice.

The atypical small GTPase GEM/Kir is a negative regulator of the NADPH oxidase and NETs production through macroautophagy.

Despite the important function of neutrophils in the eradication of infections and induction of inflammation, the molecular mechanisms regulating the activation and termination of the neutrophil immune response is not well understood. Here, the function of the small GTPase from the RGK family, Gem, is characterized as a negative regulator of the NADPH oxidase through autophagy regulation. Gem knockout (Gem KO) neutrophils show increased NADPH oxidase activation and increased production of extracellular and intracellular reactive oxygen species (ROS). Enhanced ROS production in Gem KO neutrophils was associated with increased NADPH oxidase complex-assembly as determined by quantitative super-resolution microscopy, but normal exocytosis of gelatinase and azurophilic granules. Gem-deficiency was associated with increased basal autophagosomes and autolysosome numbers but decreased autophagic flux under phorbol ester-induced conditions. Neutrophil stimulation triggered the localization of the NADPH oxidase subunits p22phox and p47phox at LC3-positive structures suggesting that the assembled NADPH oxidase complex is recruited to autophagosomes, which was significantly increased in Gem KO neutrophils. Prevention of new autophagosome formation by treatment with SAR405 increased ROS production while induction of autophagy by Torin-1 decreased ROS production in Gem KO neutrophils, and also in wild-type neutrophils, suggesting that macroautophagy contributes to the termination of NADPH oxidase activity. Autophagy inhibition decreased NETs formation independently of enhanced ROS production. NETs production, which was significantly increased in Gem-deficient neutrophils, was decreased by inhibition of both autophagy and calmodulin, a known GEM interactor. Intracellular ROS production was increased in Gem KO neutrophils challenged with live Gram-negative bacteria Pseudomonas aeruginosa or Salmonella Typhimurium, but phagocytosis was not affected in Gem-deficient cells. In vivo analysis in a model of Salmonella Typhimurium infection indicates that Gem-deficiency provides a genetic advantage manifested as a moderate increased in survival to infections. Altogether, the data suggest that Gem-deficiency leads to the enhancement of the neutrophil innate immune response by increasing NADPH oxidase assembly and NETs production and that macroautophagy differentially regulates ROS and NETs in neutrophils.

Ex vivo model for studying endothelial tip cells: Revisiting the classical aortic-ring assay.

A drug undergoes several in silico, in vitro, ex vivo and in vivo assays before entering into the clinical trials. In 2014, it was reported that only 32% of drugs are likely to make it to Phase-3 trials, and overall, only one in 10 drugs makes it to the market. Therefore, enhancing the precision of pre-clinical trial models could reduce the number of failed clinical trials and eventually time and financial burden in health sciences. In order to attempt the above, in the present study, we have shown that aortic ex-plants isolated from different stages of chick embryo and different regions of the aorta (pulmonary and systemic) have differential sprouting potential and response to angiogenesis modulatory drugs. Aorta isolated from HH37 staged chick embryo showed 16% (p < 0.001) and 11% (p < 0.001) increase in the number of tip cells at 72 h of culture compared to that of HH35 and HH29 respectively. The ascending order of the number of tip cells was found as central (Gen II), proximal (Gen I) and distal (Gen III) in a virtual zonal segmentation of endothelial sprouting. The HH37 staged aortas displayed differential responses to pro- and anti-angiogenic drugs like Vascular endothelial growth factor (VEGF), nitric oxide donor (spNO), and bevacizumab (avastin), thalidomide respectively. The human placenta tissue-culture however evinced endothelial sprouting only on day 12, with a gradual decrease in the number of tip cells until 21 days. In summary, this study provides an avant-garde angiogenic model emphasized on tip cells that would enhance the precision to test next-generation angiogenic drugs.

A comprehensive treatment approach for idiopathic condylar resorption and anterior open bite with 3D virtual surgical planning and self-ligated customized lingual appliance.

INTRODUCTION: We report the successful treatment of a 38-year-old woman with bilateral idiopathic condylar resorption and anterior open bite. She had incompetent lips, a gummy smile, increased lower facial height, high mandibular plane angle, skeletal and dental Class II malocclusion with mild mandibular crowding, increased overjet, and mandibular midline deviation to the right. METHODS: The treatment plan included: (1) presurgical alignment and leveling of the teeth in both arches; (2) jaw motion tracking (JMT) to detect mandibular movement; (3) 3-piece maxillary osteotomies with mandibular reconstruction and bilateral coronoidectomies; and (4) postsurgical correction of the malocclusion. The orthodontic treatment was performed with the use of custom lingual braces and clear brackets and the orthognathic surgery was planned with the use of virtual surgical planning. RESULTS: The idiopathic condylar resorption and anterior open bite were treated, crowding was eliminated in the lower anterior segment, correction of skeletal and dental Class II malocclusion was obtained, mandibular plane angle was reduced, and facial profile improved. CONCLUSIONS: The results suggest that esthetic and functional results can be achieved with the cooperation of 2 specialties and with the use of state-of-the-art technology.

Cross-regulation of defective endolysosome trafficking and enhanced autophagy through TFEB in UNC13D deficiency.

Several lines of evidence support the occurrence of cross-regulation between the endocytic pathway and autophagy, but the molecular mechanisms regulating this process are not well-understood. Here, we show that the calcium sensor UNC13D regulates the molecular mechanism of late endosomal trafficking and endosomal maturation, and defects in UNC13D lead to macroautophagy upregulation. unc13d-null cells showed impaired endosomal trafficking and defective endocytic flux. The defective phenotypes were rescued by the expression of UNC13D but not by its STX7-binding-deficient mutant. This defective endosomal function in UNC13D-deficient cells resulted in increased autophagic flux, increased long-lived protein degradation, decreased SQSTM1/p62 protein levels and increased autolysosome formation as determined by biochemical, microscopy and structural methods. The autophagic phenotype was not associated with increased recruitment of the UNC13D-binding proteins and autophagy regulators, RAB11 or VAMP8, but was caused, at least in part, by TFEB-mediated upregulation of a subset of autophagic and lysosomal genes, including Atg9b. Downregulation of TFEB decreased Atg9b levels and decreased macroautophagy in unc13d-null cells. UNC13D upregulation corrected the defects in endolysosomal trafficking and decreased the number of accumulated autophagosomes in a cellular model of the lysosomal-storage disorder cystinosis, under both fed and starvation conditions, identifying UNC13D as an important new regulatory molecule of autophagy regulation in cells with lysosomal disorders. Abbreviations ACTB: actin, beta; CTSB: cathepsin B; EEA1: early endosome antigen 1; ESCRT: endosomal sorting complex required for transport; FHL3: familial hemophagocytic; lymphohistiocytosis type 3; HEX: hexosaminidase; HLH: hemophagocytic lymphohistiocytosis; LSD: lysosomal storage disorder; MEF: mouse embryonic fibroblast; SEM: standard errors of the mean; SNARE: soluble n-ethylmaleimide-sensitive-factor attachment receptor; STX: syntaxin; SYT7: synaptotagmin VII; TFE3: transcription factor E3; TFEB: transcription factor EB; TIRF: total internal reflection fluorescence ULK1: unc-51 like kinase 1; UNC13D: unc-13 homolog d; VAMP: vesicle-associate membrane protein; WT: wild-type.

Chaperone-Mediated Autophagy Upregulation Rescues Megalin Expression and Localization in Cystinotic Proximal Tubule Cells.

Cystinosis is a lysosomal storage disorder caused by defects in CTNS, the gene that encodes the lysosomal cystine transporter cystinosin. Patients with nephropathic cystinosis are characterized by endocrine defects, defective proximal tubule cell (PTC) function, the development of Fanconi syndrome and, eventually, end-stage renal disease. Kidney disease is developed despite the use of cysteamine, a drug that decreases lysosomal cystine overload but fails to correct overload-independent defects. Chaperone-mediated autophagy (CMA), a selective form of autophagy, is defective in cystinotic mouse fibroblasts, and treatment with cysteamine is unable to correct CMA defects in vivo, but whether the vesicular trafficking mechanisms that lead to defective CMA in cystinosis are manifested in human PTCs is not currently known and whether PTC-specific mechanisms are corrected upon CMA upregulation remains to be elucidated. Here, using CRISPR-Cas9 technology, we develop a new human PTC line with defective cystinosin expression (CTNS-KO PTCs). We show that the expression and localization of the CMA receptor, LAMP2A, is defective in CTNS-KO PTCs. The expression of the lipidated form of LC3B, a marker for another form of autophagy (macroautophagy), is decreased in CTNS-KO PTCs indicating decreased autophagosome numbers under basal conditions. However, the autophagic flux is functional, as measured by induction by starvation or by blockage using the v-ATPase inhibitor bafilomycin A, and by degradation of the macroautophagy substrate SQSTM1 under starvation and proteasome-inhibited conditions. Previous studies showed that LAMP2A accumulates in Rab11-positive vesicles in cystinotic cells. Here, we show defective Rab11 expression, localization and trafficking in CTNS-KO PTCs as determined by confocal microscopy, immunoblotting and TIRFM. We also show that both Rab11 expression and trafficking in cystinotic PTCs are rescued by the upregulation of CMA using small-molecule CMA activators. Cystinotic PTCs are characterized by PTC de-differentiation accompanied by loss of the endocytic receptor megalin, and megalin recycling is regulated by Rab11. Here we show that megalin plasma membrane localization is defective in CTNS-KO PTCs and its expression is rescued by treatment with CMA activators. Altogether, our data support that CMA upregulation has the potential to improve PTC function in cystinosis.

Effect of age on bite force.

This study compared bite force in adults older than 60 years with that of young adults. The participants were 20 healthy adults (9 men) older than 60 years (median age, 66 years) and 44 healthy young adults (22 men; age range, 18-25 years; median age, 22 years) at the International Medical University, Malaysia. All participants had at least 20 teeth, and bite force was measured and evaluated using the Dental Prescale system. Average (SD) bite force was 420.5 (242.0) N for the older adults and 541.4 (296.3) N for the young adults. Although mean bite force was higher for the young adults, the difference was not significant. These findings suggest that bite force is unaffected by age in adults with adequate dentition. (J Oral Sci 58, 361-363, 2016).

Plasmodium falciparum Bloom homologue, a nucleocytoplasmic protein, translocates in 3' to 5' direction and is essential for parasite growth.

Malaria caused by Plasmodium, particularly Plasmodium falciparum, is the most serious and widespread parasitic disease of humans. RecQ helicase family members are essential in homologous recombination-based error-free DNA repair processes in all domains of life. RecQ helicases present in each organism differ and several homologues have been identified in various multicellular organisms. These proteins are involved in various pathways of DNA metabolism by providing duplex unwinding function. Five members of RecQ family are present in Homo sapiens but P. falciparum contains only two members of this family. Here we report the detailed biochemical and functional characterization of the Bloom (Blm) homologue (PfBlm) from P. falciparum 3D7 strain. Purified PfBlm exhibits ATPase and 3' to 5' direction specific DNA helicase activity. The calculated average reaction rate of ATPase was ~13 pmol of ATP hydrolyzed/min/pmol of enzyme. The immunofluorescence assay results show that PfBlm is expressed in all the stages of intraerythrocytic development of the P. falciparum 3D7 strain. In some stages of development in addition to nucleus PfBlm also localizes in the cytoplasm. The gene disruption studies of PfBlm by dsRNA showed that it is required for the ex-vivo intraerythrocytic development of the parasite P. falciparum 3D7 strain. The dsRNA mediated inhibition of parasite growth suggests that a variety of pathways are affected resulting in curtailing of the parasite growth. This study will be helpful in unravelling the basic mechanism of DNA transaction in the malaria parasite and additionally it may provide leads to understand the parasite specific characteristics of this protein.

Plasmodium falciparum Werner homologue is a nuclear protein and its biochemical activities reside in the N-terminal region.

RecQ helicases, also addressed as a gatekeeper of genome, are an inevitable family of genome scrutiny proteins conserved from prokaryotes to eukaryotes and play a vital role in DNA metabolism. The deficiencies of three RecQ proteins out of five are involved in genetic abnormalities like Bloom syndrome (BS), Werner syndrome (WS), and Rothmund-Thomson syndrome (RTS). It is noteworthy that Plasmodium falciparum contains only two members of the RecQ family as opposed to five members present in the host Homo sapiens. In the present study, we report the biochemical characterization of the homologue of Werner (Wrn) helicase from P. falciparum 3D7 strain. Although there are significant sequence conservations between Wrn helicases of both H. sapiens and P. falciparum as well as among all the other Plasmodium species, they contain some peculiar differences also. In silico studies reveal that PfWrn is evolutionarily close to the bacterial RecQ protein. The N-terminal fragment (PfWrnN) contains all the helicase motifs along with all the functional domains and the predicted structure resembles with the human RecQ1 protein, whereas the C-terminal fragment (PfWrnC) contains no significant domain. Biochemical characterization further revealed that purified recombinant PfWrnN shows ATPase and DNA helicase activity in 3' to 5' direction, but PfWrnC lacks the ATPase and helicase activities. Immunofluorescence study shows that PfWrn is expressed in all the stages of intraerythrocytic development of the P. falciparum 3D7 strain and localizes distinctly in the nucleus. This study can be used for further characterization of RecQ helicases that will aid in understanding the physiological significance of these helicases in the malaria parasite.