Nodule-specific Cu+ -chaperone NCC1 is required for symbiotic nitrogen fixation in Medicago truncatula root nodules.

Cu+ -chaperones are a diverse group of proteins that allocate Cu+ ions to specific copper proteins, creating different copper pools targeted to specific physiological processes. Symbiotic nitrogen fixation carried out in legume root nodules indirectly requires relatively large amounts of copper, for example for energy delivery via respiration, for which targeted copper deliver systems would be required. MtNCC1 is a nodule-specific Cu+ -chaperone encoded in the Medicago truncatula genome, with a N-terminus Atx1-like domain that can bind Cu+ with picomolar affinities. MtNCC1 is able to interact with nodule-specific Cu+ -importer MtCOPT1. MtNCC1 is expressed primarily from the late infection zone to the early fixation zone and is located in the cytosol, associated with plasma and symbiosome membranes, and within nuclei. Consistent with its key role in nitrogen fixation, ncc1 mutants have a severe reduction in nitrogenase activity and a 50% reduction in copper-dependent cytochrome c oxidase activity. A subset of the copper proteome is also affected in the ncc1 mutant nodules. Many of these proteins can be pulled down when using a Cu+ -loaded N-terminal MtNCC1 moiety as a bait, indicating a role in nodule copper homeostasis and in copper-dependent physiological processes. Overall, these data suggest a pleiotropic role of MtNCC1 in copper delivery for symbiotic nitrogen fixation.

Suberin deficiency and its effect on the transport physiology of young poplar roots.

The precise functions of suberized apoplastic barriers in root water and nutrient transport physiology have not fully been elucidated. While lots of research has been performed with mutants of Arabidopsis, little to no data are available for mutants of agricultural crop or tree species. By employing a combined set of physiological, histochemical, analytical, and transport physiological methods as well as RNA-sequencing, this study investigated the implications of remarkable CRISPR/Cas9-induced suberization defects in young roots of the economically important gray poplar. While barely affecting overall plant development, contrary to literature-based expectations significant root suberin reductions of up to 80-95% in four independent mutants were shown to not evidently affect the root hydraulic conductivity during non-stress conditions. In addition, subliminal iron deficiency symptoms and increased translocation of a photosynthesis inhibitor as well as NaCl highlight the involvement of suberin in nutrient transport physiology. The multifaceted nature of the root hydraulic conductivity does not allow drawing simplified conclusions such as that the suberin amount must always be correlated with the water transport properties of roots. However, the decreased masking of plasma membrane surface area could facilitate the uptake but also leakage of beneficial and harmful solutes.

Symplasmic and transmembrane zinc transport is modulated by cadmium in the Cd/Zn hyperaccumulator Sedum alfredii.

This study investigated the influence of Cd (25 µM) on Zn accumulation in a hyperaccumulating (HE) and a non-hyperaccumulating (NHE) ecotype of Sedum alfredii Hance at short-term supply of replete (Zn5, 5 µM) and excess (Zn400, 400 µM) Zn. Cd inhibited Zn accumulation in both ecotypes, especially under Zn400, in organs with active metal sequestration, i.e. roots of NHE and shoots of HE. Direct biochemical Cd/Zn competition at the metal-protein interaction and changes in transporter gene expression contributed to the observed accumulation patterns in the roots. Specifically, in HE, Cd stimulated SaZIP4 and SaPCR2 under Zn5, but downregulated SaIRT1 and SaZIP4 under Zn400. However, Cd downregulated related transporter genes, except for SaNRAMP1, in NHE, irrespective of Zn. Cadmium stimulated casparian strip (CSs) development in NHE, as part of the defense response, while it had a subtle effect on the (CS) in HE. Moreover, Cd delayed the initiation of the suberin lamellae (SL) in HE, but stimulated SL deposition in NHE under both Zn5 or Zn400. Changes in suberization were mainly ascribed to suberin-biosynthesis-related genes and hormonal signaling. Altogether, Cd regulated Zn accumulation mainly via symplasmic and transmembrane transport in HE, while Cd inhibited both symplasmic and apoplasmic Zn transport in NHE.

Does obesity predict morbidity and mortality amongst patients undergoing transfemoral amputations?

BACKGROUND: We investigated the role of obesity on morbidity and mortality in patients undergoing above knee amputation. METHODS: Data of 4225 patients undergoing AKAs was extracted from NIS Database (2016-2019) for a retrospectively matched case-control study and were grouped into; Non-obese (N-Ob-BMI <29.9 kg/m2; n = 1413), class I/II obese (Ob-I/II-BMI: 30-39.9 kg/m2; n = 1413), and class III obese groups (Ob-IIIBMI > 40; n = 1399). Morbidity, mortality, length of stay, and hospital charges were analyzed. RESULTS: Blood loss anemia (OR = 1.42; 95% CI = 1.19-1.64), superficial SSI (OR = 5.10; 95% CI = 1.4717.63) and acute kidney injury (AKI- OR = 1.42; 95% CI = 1.21-1.67) were higher in Ob-III patients. Mortality was 5.8%, 4.5%, and 6.4% in N-Ob, Ob-I/II and Ob-III patients (p < 0.001; Ob-I/II vs. Ob-III), respectively. Hospital LOS was 3 days higher in Ob-III (16.1 ± 18.0), comparatively resulting in $25,481 higher inpatient-hospital charge. CONCLUSION: Patients in Ob-III group were noted to have increased morbidity, higher LOS, and inpatient-hospital cost.

Prevalence and treatment of HIV/AIDS in Pakistani population: A retrospective study.

HIV/AIDS epidemic poses an emerging threat around the world. An estimated 0.1% of people aged 15 to 49 years have been infected with HIV/AIDS in Pakistan during the last few years. This research study aims to analyse the prevalence and treatment/management of HIV/AIDS among the Pakistani population aged 15 to 45 years from 1990 to 2020 and this research data was collected during the year 2021. The analysis was done by using E-views software. Descriptive statistical analysis, correlation coefficient, unit root analysis, and linear regression analysis were used. Results indicated that there is low awareness regarding AIDS in the population and there is a significant link between prevalence and population increase, i.e. with the increase in population the prevalence of AIDS also increases. Findings are discussed in terms of implications for prevention and education of HIV/AIDS in Pakistan's population.

A comparative study of early functional outcomes in undisplaced neck of femur fracture treated with partially threaded and fully threaded cannulated screw fixation in patients above 60 years age.

Objective: To compare the partially threaded versus fully threaded cannulated screw fixation methods in stable neck of femur fracture in terms of early functional outcomes in patients of age 60 and above. Methods: A randomized controlled trial was conducted at Orthopedic Unit-II, Mayo Hospital Lahore, Pakistan from July 2021 to July 2022. A total of 82 (41 in each group) patients of both genders, aged 60 or above were included. All patients had garden Type-I or II and were mobile before they fractured the femoral neck were included. In Group-A, cannulated screw fixation (cancellous screws 6 mm) was done using partially threaded screws while fully threaded screws (cancellous screws 6 mm) were used in Group-B. Patients were followed up at six weeks, three months and six months interval for "Radiographic Union Scale for Hip (RUSH)" and "Harris Hip score". Results: In a total of 82 (41 in each group) patients, 66 (80.5%) we male. At 6th week (p=0540) and 3rd month (p=0.653) postoperatively, no significant differences were seen between groups for Hip Harris score. However, at 6th month, functional outcome of Group-B patients was significantly better as compared to Group-A (p=0.038). Mean RUSH score in Group-A and in Group-B at 6th month postoperative was 25.45±2.73 and 30.52±2.39 (p<0.001). Conclusion: Fully threaded cannulated screw fixation is better in treating undisplaced neck of femur fracture as compared to partially threaded cannulated screw fixation in terms of early functional outcomes among the age group of 60 years and above.RCT Registration number: NCT05587660.

An epidemiological, strategic and response analysis of the COVID-19 pandemic in South Asia: a population-based observational study.

INTRODUCTION: South Asia has had a dynamic response to the ongoing COVID-19 pandemic. The overall burden and response have remained comparable across highly-burdened countries within the South Asian Region. METHODOLOGY: Using a population-based observational design, all eight South Asian countries were analyzed using a step-wise approach. Data were obtained from government websites and publicly-available repositories for population dynamics and key variables. RESULTS: South Asian countries have a younger average age of their population. Inequitable distribution of resources centered in urban metropolitan cities within South Asia is present. Certain densely populated regions in these countries have better testing and healthcare facilities that correlate with lower COVID-19 incidence per million populations. Trends of urban-rural disparities are unclear given the lack of clear reporting of the gaps within these regions. COVID-19 vaccination lag has become apparent in South Asian countries, with the expected time to complete the campaign being unfeasible as the COVID-19 pandemic progresses. CONCLUSION: With a redesigning of governance policies on preventing the rise of COVID-19 promptly, the relief on the healthcare system and healthcare workers will allow for adequate time to roll out vaccination campaigns with equitable distribution. Capacity expansion of public health within the Region is required to ensure a robust healthcare response to the ongoing pandemic and future infectious disease outbreaks.

Pharmacognostic and phytochemical study of the flowers of Cordia sebestena L.

The present study shows the pharmacognostic and phytochemical studies on the flowers of Cordia sebestena L. belongs to the family Boraginaceae. C. sebestena L. is found primarily in tropical and subtropical regions of the American, Asian and African continents. Though it is an important plant, until date no pharmacognostic work is found on its parts such as flowers. Various organoleptic characters were recorded by macroscopic study. Microscopic study of the flowers were also conducted which shows the presence of fibers, calcium oxalate crystals and multiple types of trichomes, along with fluorescence analysis. The present study also deals with the Fourier transform infrared spectroscopic analysis of C. sebestena L. FT-IR spectra revealed the presence of C-H, C=C, C-N, C-O and aromatic groups. Chemical composition of the hexane extract of the flowers of C. sebestena L. was detected through GC-MS and spectrum achieved through GC-MS were correlated with the database of National Institute of Standards and Technology (NIST) which comprise of beyond 62000 outlines of the mass spectrum. GC-MS analysis of n-hexane extract shown the existence of Retinoic acid, lupeol, ß-sitosterol, stigmasterol, hexadecanoic acid along with fatty acids, esters, alkaloids and alcohols. These pharmacognostic and phytochemical studies can be valuable towards giving reliable proof of the quality of the plant which can benefit health professionals and herbal medicine manufacturers.

Alkaline lipase production by novel meso-tolerant psychrophilic Exiguobacterium sp. strain (AMBL-20) isolated from glacier of northeastern Pakistan.

Lipase is an important commercial enzyme with unique and versatile biotechnological applications. This study was conducted to biosynthesize and characterizes alkaliphilic lipase by Exiguobacterium sp. strain AMBL-20T isolated from the glacial water samples of the northeastern (Gilgit-Baltistan) region of Pakistan. The isolated bacterium was identified as Exiguobaterium sp. strain AMBL-20T on the basis of morphological, biochemical, and phylogenetic analysis of 16S rRNA sequences with GenBank accession number MW229267. The bacterial strain was further screened for its lipolytic activity, biosynthesis, and characterization by different parameters with the aim of maximizing lipase activity. Results showed that 2% Olive oil, 0.2% peptone at 25 °C, pH 8, and 24 h of incubation time found optimal for maximum lipase production. The lipase enzyme was partially purified by ammonium sulphate precipitation and its activity was standardized at pH 8 under 30 °C temperature. The enzyme showed functional stability over a range of temperature and pH. Hence, extracellular alkaliphilic lipase from Exiguobacterium sp. is a potential candidate with extraordinary industrial applications, particularly in bio-detergent formulations.

Clinical efficacy of scaling and root planing with and without metronidazole on glycemic control: three-arm randomized controlled trial.

BACKGROUND: Treating periodontitis through non-surgical periodontal therapy (NSPT) may improve glycemic control in type-2 Diabetes Mellitus (T2DM) patients. However, the evidence to maintain this improvement beyond four months is insufficient. Hence, this trial was conducted to assess clinical efficacy of NSPT on glycemic control in T2DM patients. METHODS: This three-arm randomized controlled trial recruited 150 known T2DM participants (35-65 years), suffering from moderate to severe periodontitis, having HbA1c level ≥ 6.5% at baseline. Participants were followed up at 3 and 6 months. Intervention for test group-1 included scaling and root planing (SRP) with metronidazole (MET) and oral hygiene instructions (OHI). Test group-2 was intervened with SRP + OHI and control group with OHI only. Stata v. 14 was used to observe inter and intragroup mean changes in glycemic [glycated hemoglobin (HbA1c), fasting blood glucose (FBG)] and periodontal variables [bleeding on probing (BOP), periodontal pocket depth (PPD), clinical attachment loss (CAL)] using ANOVA and RMANOVA. Proportion of change in outcome variable (HbA1c) was assessed between treatment groups using chi-square test. Change was considered significant at p-value ≤ 0.05. RESULTS: A significant reduction was observed in BOP, PPD, CAL, HbA1c and FBG over time [p < 0.05]. Significant reductions were observed in same variables in both test groups in comparison to control arm [p < 0.05]. No change between the two test groups was observed [p > 0.05]. CONCLUSION: Scaling and root planing improves glycemic control of T2DM patients independently of the use of MET. Therefore, SRP after every 6 months may be suggested and included as a part of overall diabetes management for patients suffering from T2DM. Clinical trial registration NCT 03,343,366 [Date of Registration: 17/11/2017].

Dual mobility versus unipolar total hip arthroplasty for neck of femur fractures : a single centre study.

Current national guidelines (NICE) recommends that all medically fit, independently-mobile patients without cognitive impairment receive a total hip arTHAoplasty(THA) for displaced intracapsular neck of femur (NOF) fractures. Dislocation is a concern(2-10%). Dual mobility cups have been suggested to address this complication. Our study sets out to compare dislocation rates between dual mobility cups versus unipolar cups. We performed a retrospective single centre multiple surgeon study of all THAs performed for NOFs between January 2012 and May 2018. A total of 322 total hip replacements (127 dual mobility and 195 unipolar ; Age range of patients, 29 to 91, mean 70 years) were identified for analysis using a database. Data was obtained from electronic patient records and radiographs. 12 patients sustained a dislocation of their THA out of our 322 patients. Of these, 10 dislocations occurred in the unipolar group (5.13%). From the dual mobility cups, 2 had dislocations(1.57%), both with a 28mm head. Both of these dislocations were in alcohol dependent patients with increased susceptibility to falls. Statistical analysis of our data was performed using chi-squared test (p value = 0.0723) In 'Getting It Right First Time' (GIRFT), the authors recommend that all patients that sustain a NOF fracture meeting the criteria of a THA to be offered a dual mobility acetabular cup to reduce the risk of dislocation. The cost of the dual mobility acetabular cup is offset from the cost of overall revision surgery. Limitations of our study are its retrospective nature and selection bias.

Chronic exposure of soybean plants to nanomolar cadmium reveals specific additional high-affinity targets of cadmium toxicity.

Solving the global environmental and agricultural problem of chronic low-level cadmium (Cd) exposure requires better mechanistic understanding. Here, soybean (Glycine max) plants were exposed to Cd concentrations ranging from 0.5 nM (background concentration, control) to 3 µM. Plants were cultivated hydroponically under non-nodulating conditions for 10 weeks. Toxicity symptoms, net photosynthetic oxygen production and photosynthesis biophysics (chlorophyll fluorescence: Kautsky and OJIP) were measured in young mature leaves. Cd binding to proteins [metalloproteomics by HPLC-inductively coupled plasma (ICP)-MS] and Cd ligands in light-harvesting complex II (LHCII) [X-ray absorption near edge structure (XANES)], and accumulation of elements, chloropyll, and metabolites were determined in leaves after harvest. A distinct threshold concentration of toxicity onset (140 nM) was apparent in strongly decreased growth, the switch-like pattern for nutrient uptake and metal accumulation, and photosynthetic fluorescence parameters such as Φ RE10 (OJIP) and saturation of the net photosynthetic oxygen release rate. XANES analyses of isolated LHCII revealed that Cd was bound to nitrogen or oxygen (and not sulfur) atoms. Nutrient deficiencies caused by inhibited uptake could be due to transporter blockage by Cd ions. The changes in specific fluorescence kinetic parameters indicate electrons not being transferred from PSII to PSI. Inhibition of photosynthesis combined with inhibition of root function could explain why amino acid and carbohydrate metabolism decreased in favour of molecules involved in Cd stress tolerance (e.g. antioxidative system and detoxifying ligands).

Evaluation of HbA1c in type-2 diabetes mellitus patients with periodontitis: preliminary findings of three-arm clinical trial.

OBJECTIVE: To assess a relationship and predict changes in glycaemic control due to chronic periodontitis in type-2 diabetic patients. METHODS: Chronic periodontitis (CP) of a cross-section of 118 patients (age ?32 years) was determined using bleeding on probing (BOP), periodontal pocket depth (PPD) and clinical attachment loss (CAL). Their HbA1c, fasting blood glucose (FBG), fasting serum insulin (FSI) and homeostasis assessment model of insulin resistance (HOMA-IR) levels were also tested. Stata 11.0 was used to assess correlation between periodontal and glycaemic measures. Univariate analysis followed by multiple regression analysis through stepwise forward selection process was used to explore significant variables that may predict change in HbA1c. Multi-collinearity and heteroskedasticity were assessed at p-value > 0.05. RESULTS: Results of participants (n=95) showed significantly positive correlations (r) between HbA1c and BOP [r = 0.34, p-value = 0.002], PPD [r = 0.36, p-value = 0.003] and CAL [r=0.42, p-value = 0.002]. However, FBG and HOMA-IR were not found correlated with any of the periodontal measures; except CAL with FBG [r=0.27, p-value=0.02]. In addition to healthy controlled diet, cultural background, education and FBG, CAL was found significant [coef. = 0.746, p-value = 0.001, CI = 0.339, 1.153] to predict change in HbA1c in the final model [R2 = 0.549, p-value < 0.001] with no multicollinearity [mean VIF =1.30] and heteroskedasticity [chi 2 =0.02, p-value= 0.878] in fi n al model. CONCLUSIONS: Increased CAL is estimated to increase HbA1c level by 0.75% in T2DM patients after controlling other factors. It is suggested that periodontal prophylactic management may be planned with an aim to reduce HbA1c. CLINICAL TRIAL REGISTRATION NO: NCT03343366.

Ultratrace Metal Speciation Analysis by Coupling of Sector-Field ICP-MS to High-Resolution Size Exclusion and Reversed-Phase Liquid Chromatography.

Techniques for metal speciation analysis with subnanomolar (ppt) detection limits in complex matrices, with simultaneous quantification of matrix elements, have become a necessity for investigating targets of trace metal binding to macromolecules and pigments at environmentally relevant concentrations. In this work we optimized the analysis of such metal binding in a custom-built HPLC-ICP-MS system. Key elements of the optimization were the choice of components for the metal-free HPLC-DAD system and sector-field ICP-MS detection (ICF-sfMS) with desolvating injection and optimization of sample handling. Protein analysis was done using ammonium bicarbonate buffer and size exclusion chromatography (SEC-ICP-sfMS), with possible addition of anion exchange chromatography. Detection of metal exchange in pigments (chlorophylls and bacteriochlorophylls) was based on reversed-phase chromatography with a methanol-acetone gradient and coupling to the ICP-sfMS via a dedicated organic matrix interface (RPC-ICP-sfMS). The resulting HPLC-DAD-ICP-sfMS system has detection limits in the picomolar range in protein buffer, limited by the maximal achievable purity of buffers/solvents and not by system sensitivity. Tests for method optimization showed that sonication, meant to increase protein solubilization, leads to artifacts of metal loss from metalloproteins. Examples for Cd binding to soybean proteins and chlorophyll, Cr binding to Arabidopsis thaliana proteins, La binding to Desmodesmus quadricauda proteins, and Cu binding to Rhodospirillum rubrum proteins and pigments are shown. These application examples demonstrate that the system is sensitive enough to detect binding of metals to proteins and pigments at background concentration levels of typical nutrient solutions made from analytical grade chemicals, equivalent to ultratrace metal concentrations in nonpolluted environments.

Evaluation of Turmeric Nanoparticles as Anti-Gout Agent: Modernization of a Traditional Drug.

Background and objectives: Turmeric has assisted in the control of inflammation and pain for decades and has been used in combination with other nutraceuticals to treat acute and chronic osteoarthritis pain. Recently, the effect of turmeric, turmeric extract, or curcuminoids on musculoskeletal pain, either by themselves or in conjunction with other substances, has been reported. The aim of this study was to develop and characterize turmeric nanoparticles (T-NPs) for various parameters, both in vitro and in vivo. Materials and Methods: The T-NPs were successfully synthesized and characterized using particle size analysis, solubility improvement, SEM, EDX, X-ray diffraction, and in vivo antigout activity in mice model. Results: The T-NPs were of about 46 nm in size with a positive zeta potential +29.55 ± 3.44 and low polydispersity index (PDI) (0.264). Furthermore, the diseased mice, with induced gout via monosodium urate crystals, were treated with 5, 10, and 20 ppm T-NPs, administered orally, and the anti-gout potential was observed through measurement of joint diameter and changes in biochemical parameters, including lipid profile, renal function test, and liver function tests which significantly reduced the levels of these biochemical parameters. Conclusions: Uric acid levels were significantly reduced after the treatment with T-NPs. indicating that T-NPs show superior potential against gout management. Thus, T-NPs can be developed as an efficient antigout agent with minimum toxicities.

Dental graduates perspective of professionalism competences from a developing country.

OBJECTIVE: To evaluate the perception of fresh dental graduates regarding professional competences essential for a graduating dentist. METHODS: This quantitative, descriptive, cross-sectional study was conducted at four public and four private sector dental colleges in Punjab, Pakistan, from June to December 2014, and comprised dental graduates of the years 2013 and 2014. A comprehensive document containing 32 professionalism competences, divided into 3 sub-domains, was developed based on the competence documents of local and foreign accrediting bodies. Responses were recorded on a five-point Likert scale. The competences were categorised as 'essential' and 'good to have' based on the agreement cut-off of 75%. SPSS 21 was used for data analysis. RESULTS: Of the 386 participants, 152(39.4%) were from public-sector colleges and 234(60.6%) from private ones. The overall mean age was 23.69±1.06 years. Of the 32 competences, the participants marked 18(56.25%) as 'essential' and 15(46.9%) as 'good to have'. In the latter category, 3(20%) competences were from the domain of professional attitude and behaviour, 7(46.6%) from ethics and jurisprudence and 4(26.7%) from the domain of communication and interpersonal skills. CONCLUSIONS: Pakistani dental graduates had an overall positive attitude regarding professionalism competences.

If You Twinkle When You Tinkle, Stones Are Found on Ultrasound.

Urolithiasis in children is an underrecognized cause of pediatric abdominal pain. Our case describes a child who presented to the emergency department with right lower quadrant pain, in whom a point-of-care ultrasound detected an ureterovesicular stone, which obviated the need for any further radiographic studies. We review the current literature on pediatric urolithiasis with a focus on sonographic diagnosis.

Understanding and practice of evidence based search strategy among postgraduate dental students: a preliminary study.

BACKGROUND: One of the core attributes of competent practice is the ability to locate and analyze high-quality evidence. Research on information seeking knowledge of dental graduates is scarce. OBJECTIVE: The objective was to evaluate pertinent knowledge and skills on formulation of answerable questions and searching skills through the understanding and application of EBP. MATERIAL AND METHODS: Participants' understanding and abilities to develop answerable questions in application of EBP were assessed through a quasi-experimental study design among freshly inducted postgraduate dental students at Dow University of Health Sciences. Pre and post workshop activity sessions were conducted and assessment was made through a tool that was an adaptation of the Fresno Test. The assessments performed were calculated for mean scores and standard deviations to draw descriptive results for the participants' understanding of the search items and clinical question formulation skills. Wilcoxon-signed rank test was performed to compare the pre and post workshop mean scores. RESULTS: Forty two participants (females = 20, males = 22) attended the workshop. Pre-workshop knowledge and understanding about the terminologies used in EBP was low among participants. Post-workshop performance of the participants to formulate a question, find evidence and search on PubMed was improved to a "Limited" level. The improvements were statistically significant but could not attain "Strong" and "Excellent" grades as far as the EBP skills were concerned. CONCLUSION: This study observed a potential to improve search skills of dental graduates after their training and exposure to search strategies available on databases.