Capture, Release, and Identification of Newly Synthesized Proteins for Improved Profiling of Functional Translatomes.

New protein synthesis is regulated both at the level of mRNA transcription and translation. RNA-Seq is effective at measuring levels of mRNA expression, but techniques to monitor mRNA translation are much more limited. Previously, we reported results from O-propargyl-puromycin (OPP) labeling of proteins undergoing active translation in a 2-h time frame, followed by biotinylation using click chemistry, affinity purification, and on-bead digestion to identify nascent proteins by mass spectrometry (OPP-ID). As with any on-bead digestion protocol, the problem of nonspecific binders complicated the rigorous categorization of nascent proteins by OPP-ID. Here, we incorporate a chemically cleavable linker, Dde biotin-azide, into the protocol (OPP-IDCL) to provide specific release of modified proteins from the streptavidin beads. Following capture, the Dde moiety is readily cleaved with 2% hydrazine, releasing nascent polypeptides bearing OPP plus a residual C3H8N4 tag. When results are compared side by side with the original OPP-ID method, change to a cleavable linker led to a dramatic reduction in the number of background proteins detected in controls and a concomitant increase in the number of proteins that could be characterized as newly synthesized. We evaluated the method's ability to detect nascent proteins at various submilligram protein input levels and showed that, when starting with only 100 µg of protein, ∼1500 nascent proteins could be identified with low background. Upon treatment of K562 cells with MLN128, a potent inhibitor of the mammalian target of rapamycin, prior to OPP treatment, we identified 1915 nascent proteins, the majority of which were downregulated upon inhibitor treatment. Repressed proteins with log2 FC <-1 revealed a complex network of functionally interacting proteins, with the largest cluster associated with translational initiation. Overall, incorporation of the Dde biotin-azide cleavable linker into our protocol has increased the depth and accuracy of profiling of nascent protein networks.

Health systems readiness and quality of inpatient malaria case-management in Kano State, Nigeria.

BACKGROUND: Nigeria was among the first African countries to adopt and implement change of treatment policy for severe malaria from quinine to artesunate. Seven years after the policy change health systems readiness and quality of inpatient malaria case-management practices were evaluated in Kano State of Nigeria. METHODS: A cross-sectional survey was undertaken in May 2019 at all public hospitals. Data collection comprised hospital assessments, interviews with inpatient health workers and data extraction from medical files for all suspected malaria patients admitted to the paediatric and medical wards in April 2019. Descriptive analyses included 22 hospitals, 154 health workers and 1,807 suspected malaria admissions analysed from malaria test and treat case-management perspective. RESULTS: 73% of hospitals provided malaria microscopy, 27% had rapid diagnostic tests and 23% were unable to perform any parasitological malaria diagnosis. Artemisinin-based combination therapy (ACT) was available at 96% of hospitals, artemether vials at 68% while injectable quinine and artesunate were equally stocked at 59% of hospitals. 32%, 21% and 15% of health workers had been exposed to relevant trainings, guidelines and supervision respectively. 47% of suspected malaria patients were tested while repeat testing was rare (7%). 60% of confirmed severe malaria patients were prescribed artesunate. Only 4% of admitted non-severe test positive cases were treated with ACT, while 76% of test negative patients were prescribed an anti-malarial. Artemether was the most common anti-malarial treatment for non-severe test positive (55%), test negative (43%) and patients not tested for malaria (45%). In all categories of the patients, except for confirmed severe cases, artemether was more commonly prescribed for adults compared to children. 44% of artesunate-treated patients were prescribed ACT follow-on treatment. Overall compliance with test and treat policy for malaria was 13%. CONCLUSIONS: Translation of new treatment policy for severe malaria into inpatient practice is compromised by lack of malaria diagnostics, stock-outs of artesunate and suboptimal health workers' practices. Establishment of the effective supply chain and on-going supportive interventions for health workers accompanied with regular monitoring of the systems readiness and clinical practices are urgently needed.

Transmembrane Polar Relay Drives the Allosteric Regulation for ABCG5/G8 Sterol Transporter.

The heterodimeric ATP-binding cassette (ABC) sterol transporter, ABCG5/G8, is responsible for the biliary and transintestinal secretion of cholesterol and dietary plant sterols. Missense mutations of ABCG5/G8 can cause sitosterolemia, a loss-of-function disorder characterized by plant sterol accumulation and premature atherosclerosis. A new molecular framework was recently established by a crystal structure of human ABCG5/G8 and reveals a network of polar and charged amino acids in the core of the transmembrane domains, namely, a polar relay. In this study, we utilize genetic variants to dissect the mechanistic role of this transmembrane polar relay in controlling ABCG5/G8 function. We demonstrated a sterol-coupled ATPase activity of ABCG5/G8 by cholesteryl hemisuccinate (CHS), a relatively water-soluble cholesterol memetic, and characterized CHS-coupled ATPase activity of three loss-of-function missense variants, R543S, E146Q, and A540F, which are respectively within, in contact with, and distant from the polar relay. The results established an in vitro phenotype of the loss-of-function and missense mutations of ABCG5/G8, showing significantly impaired ATPase activity and loss of energy sufficient to weaken the signal transmission from the transmembrane domains. Our data provide a biochemical evidence underlying the importance of the polar relay and its network in regulating the catalytic activity of ABCG5/G8 sterol transporter.

Structural snapshot of the cholesterol-transport ATP-binding cassette proteins 1.

The ATP-binding cassette (ABC) proteins play critical roles in maintaining lipid and sterol homeostasis in higher eukaryotes. In humans, several subfamily-A and -G members function as cholesterol transporters across the cellular membranes. Deficiencies of these ABC proteins can cause dyslipidemia that is associated with health conditions such as atherosclerosis, diabetes, fatty liver disease, and neurodegeneration. The physiological roles of ABC cholesterol transporters have been implicated in mediating cholesterol efflux for reverse cholesterol transport and in maintaining membrane integrity for cell survival. The precise role of these ABC transporters in cells remains elusive, and little is known about the sterol-transport mechanism. The membrane constituents of ABC transporters have been postulated to play a key role in determining the transport substrates and the translocation mechanisms via the transmembrane domains. Recent breakthroughs in determining high-resolution structures of the human sterol transporter ABCG5/G8 and its functional homologs have shed light on new structural features of ABC transporters, providing a more relevant framework for mechanistic analysis of cholesterol-transport ABC proteins. This minireview outlines what is known about ABCG cholesterol transporters, addresses key structural features in the putative sterol translocation pathway on the transmembrane domains, and concludes by proposing a mechanistic model of ABC cholesterol transporters.

Assessing a thermal spike model of swift heavy ion-matter interactions via Pd1-xNix/Si interface mixing.

The thermal spike model (TSM), a widely accepted mechanism of swift heavy ion (SHI)-matter interactions, provides explanation for various SHI induced effects, including mixing across interfaces. We assess the validity of the model via tuning the electron-phonon coupling strength (G) by taking a series Pd1-xNix of a completely solid soluble binary, and then observing Pd1-xNix/Si interface mixing induced by a combination of 100 MeV Au ion irradiation and 4 keV Ar ion sputtering. If the TSM truly describes the SHI-matter interaction mechanism, any non-linearity in x-variation of G must also result in a similar non-linearity in the x-dependence of mixing. Experimentally, the extent of mixing has been parametrized by the irradiation induced change Δσ2 in variances of Pd and Ni depth profiles derived from XPS. Computationally, G determined using density functional theory has been used to solve the equations appropriate to the TSM, and then an equivalent quantity L2, proportional to Δσ2, has been calculated. Both Δσ2(x) and L2(x) show non-linearities, albeit in slightly dissimilar ways, leading to a conjecture that the present work at least does not invalidate the TSM.

Structural stability of purified human CFTR is systematically improved by mutations in nucleotide binding domain 1.

The Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) is an ABC transporter containing two transmembrane domains forming a chloride ion channel, and two nucleotide binding domains (NBD1 and NBD2). CFTR has presented a formidable challenge to obtain monodisperse, biophysically stable protein. Here we report a comprehensive study comparing effects of single and multiple NBD1 mutations on stability of both the NBD1 domain alone and on purified full length human CFTR. Single mutations S492P, A534P, I539T acted additively, and when combined with M470V, S495P, and R555K cumulatively yielded an NBD1 with highly improved structural stability. Strategic combinations of these mutations strongly stabilized the domain to attain a calorimetric Tm > 70 °C. Replica exchange molecular dynamics simulations on the most stable 6SS-NBD1 variant implicated fluctuations, electrostatic interactions and side chain packing as potential contributors to improved stability. Progressive stabilization of NBD1 directly correlated with enhanced structural stability of full-length CFTR protein. Thermal unfolding of the stabilized CFTR mutants, monitored by changes in intrinsic fluorescence, demonstrated that Tm could be shifted as high as 67.4 °C in 6SS-CFTR, more than 20 °C higher than wild-type. H1402S, an NBD2 mutation, conferred CFTR with additional thermal stability, possibly by stabilizing an NBD-dimerized conformation. CFTR variants with NBD1-stabilizing mutations were expressed at the cell surface in mammalian cells, exhibited ATPase and channel activity, and retained these functions to higher temperatures. The capability to produce enzymatically active CFTR with improved structural stability amenable to biophysical and structural studies will advance mechanistic investigations and future cystic fibrosis drug development.

Provider and patient perceptions of malaria rapid diagnostic test use in Nigeria: a cross-sectional evaluation.

BACKGROUND: Nigeria commenced a phased programmatic deployment of rapid diagnostic tests (RDT) at the primary health care (PHC) facility levels since 2011. Despite various efforts, the national testing rate for malaria is still very low. The uptake of RDT has been variable. This study was undertaken to determine the provider and patient perceptions to RDT use at the PHC level in Nigeria with their implications for improving uptake and compliance. METHODS: A cross-sectional survey was conducted in 120 randomly selected PHCs across six states, across the six-geopolitical zones of Nigeria in January 2013. Health facility staff interviews were conducted to assess health workers (HW) perception, prescription practices and determinants of RDT use. Patient exit interviews were conducted to assess patient perception of RDT from ten patients/caregivers who met the eligibility criterion and were consecutively selected in each PHC, and to determine HW's compliance with RDT test results indirectly. Community members, each selected by their ward development committees in each Local Government Area were recruited for focus group discussion on their perceptions to RDT use. RESULTS: Health workers would use RDT results because of confidence in RDT results (95.4%) and its reduction in irrational use of artemisinin-based combination therapy (ACT) (87.2%). However, in Enugu state, RDT was not used by health workers because of the pervasive notion RDT that results were inaccurate. Among the 1207 exit interviews conducted, 549 (45.5%) had received RDT test. Compliance rate (administering ACT to positive patients and withholding ACT from negative patients) from patient exit interviews was 90.2%. Among caregivers/patients who had RDT done, over 95% knew that RDT tested for malaria, felt it was necessary and liked the test. Age of patients less than 5 years (p = 0.04) and "high" educational status (p = 0.0006) were factors influencing HW's prescription of ACT to RDT negative patients. CONCLUSION: The study demonstrated positive perception to RDT use by HW and among community members with good compliance rate among health workers at the PHC level. This positive perception should be explored in improving the current low level of malaria testing in Nigeria while addressing the influence of age on HW administration of ACT to RDT negative cases.

Challenges encountered and lessons learnt from venous injuries at Sri Lankan combat theatres.

PURPOSE: The objective of this study was to characterise the spectrum of peripheral venous injury in the Sri Lankan war theatres, including categorisation of anatomic patterns, mechanism and management of casualties, including short-term results of surgical repair of traumatic venous injuries versus ligation. In addition, the effects and outcome of combined arterial and venous injuries versus arterial injury alone are compared. METHODS: All adults with extremity vascular injuries admitted to a military base hospital during an 8-month period were prospectively recorded and those with a venous injury were analysed. RESULT: A total of 123 vascular injuries were recorded in the study period, of which 70 had a venous injury, combined with an arterial injury in 58 (83%) and in isolation in 12 (17%). There were 43 transections, 26 lateral tears and a single through and through penetrating injury. Twenty-five (36%) vein injures were repaired and 45 ligated. Only six popliteal veins were repaired in 21 lower limbs that underwent arterial revascularisation. In the combined arterial/venous injuries group 13 primary amputations were performed and five delayed amputations were necessary. There were no amputations in the isolated venous injury group. There were three deaths (4.3%), 18 infections with four cultures positive for pseudomonas species, five arterial graft thromboses were recorded. There were significantly more blood transfusions and concomitant skeletal injuries, resulting in more amputations, in combined arterial and venous injuries in comparison with arterial injury alone (all p values<0.05). CONCLUSIONS: In an ideal setting, venous injuries should be repaired when possible and tolerated by the patient in order to ameliorate the risk of thrombotic and infectious complications. An aggressive use of shunting, fasciotomies and venous repair in wartime limb injuries at echelon structured care may prevent preventable limb loss in these challenging case scenarios.

Prognostic factors in stage III-IV adrenocortical carcinomas (ACC): an European Network for the Study of Adrenal Tumor (ENSAT) study.

BACKGROUND: The clinical course of advanced adrenocortical carcinoma (ACC) is heterogeneous. Our study aimed primarily to refine and make headway in the prognostic stratification of advanced ACC. PATIENTS AND METHODS: Patients with advanced ENSAT ACC (stage III or stage IV) at diagnosis registered between 2000 and 2009 in the ENSAT database were enrolled. The primary end point was overall survival (OS). Parameters of potential prognostic relevance were selected. Univariate and multivariate analyses were carried out: model 1 'before surgery'; model 2 'post-surgery'. RESULTS: Four hundred and forty-four patients with advanced ENSAT ACC (stage III: 210; stage IV: 234) were analyzed. After a median follow-up of 55.2 months, the median OS was 24 months. A modified ENSAT (mENSAT) classification was validated: stage III (invasion of surrounding tissues/organs or the vena renalis/cava) and stage IVa, IVb, IVc (2, 3 or >3 metastatic organs, including N, respectively). Two- or 5-year OS was 73%, 46%, 26% and 15% or 50%, 15%, 14% and 2% for stages III, IVa, IVb and IVc, respectively. In the multivariate analysis, mENSAT stages (stages IVa, IVb, or IVc, respectively) were significantly correlated with OS (P < 0.0001), as well as additional parameters: age ≥ 50 years (P < 0.0001), tumor- or hormone-related symptoms (P = 0.01 and 0.03, respectively) in model 1 but also the R status (P = 0.001) and Grade (Weiss >6 and/or Ki67 ≥ 20%, P = 0.06) in model 2. CONCLUSION: The mENSAT classification and GRAS parameters (Grade, R status, Age and Symptoms) were found to best stratify the prognosis of patients with advanced ACC.

[Obesity and heart failure]. / Adipositas und Herzinsuffizienz.

BACKGROUND: Obesity is an important risk factor for the development of heart failure. DIAGNOSTICS: In normotensive obese patients, a reduced peripheral resistance is typically observed and is accompanied by an increased fluid volume and an increase in cardiac work, resulting in hypertrophy and diastolic heart failure, which can be visualized with echocardiography. However, in the presence of arterial hypertension cardiac geometry is not different to hypertensive heart disease without obesity. Furthermore, the typical changes found with obesity, such as reduced peripheral resistance and increased blood volume, are no longer present. Obstructive sleep apnea (OSA) is very common in obesity and warrants screening but levels of the heart failure marker N-terminal pro-brain natriuretic peptide (NT-ProBNP) might be misleading as the values are lower in obesity than in normal weight controls. THERAPY: Body weight reduction is advisable but difficult to achieve and much more difficult to maintain. Furthermore, diet and exercise has not been proven to enhance life expectancy in obesity. However, with bariatric surgery, long-term weight reduction can be achieved and mortality can be reduced. CONCLUSIONS: With effective weight loss and improved clinical outcome after bariatric surgery, treatment of obesity has shifted much more into focus. Regardless of technical challenges in the work-up of obese patients, clinical symptoms suggestive of cardiac disorders warrant prompt investigation with standard techniques following recommendations as established for normal weight patients.

Quantum error channels in high energetic photonic systems.

In medical applications-such as positron emission tomography (PET)-511 keV photons that experience Compton scattering are studied. We present a consistent framework based on quantum error-correction channels-intensively studied in quantum computing-to fully describe the quantum information-theoretic content of high energetic photons undergoing Compton scattering, characterized by the Klein-Nishina formula in unoriented matter. In this way, we can predict the expected spatial distribution of two or more, pure or mixed, polarization entangled or separable photons. This framework allows us to characterize the accessible and inaccessible information for different parameter ranges. It also answers the question of how to describe successive multi-photon scattering. In addition our formalism provides a complete framework for dealing with single and all multi-partite errors that can occur in the propagation, providing the basis for modeling future dedicated experiments that will then have applications in medicine, such as reducing errors in PET imaging or exploring possibilities for quantum-based diagnostic indicators.

Cervical cancer awareness, perception, and attitude among tertiary health institution students in northeastern Nigeria.

Background: The devastating scourge of cervical cancer in Africa is largely due to the absence of preventive interventions, driven by low awareness and poor perception of the disease in the continent. This work is a preliminary effort toward understanding key social drivers promoting this disease in our immediate environment with a view to mitigating it. Method: Female students of two tertiary health institutions in Azare, northeastern Nigeria, were approached to participate in this cross-sectional descriptive study. A structured self-administered questionnaire was administered to consenting participants and covered questions on their socio-demographics, awareness, perception, and attitude about/toward cervical cancer and its prevention. The responses were scrutinized for coherency and categorized into themes using summary statistics, while a chi-square test was used to determine the association between awareness of cervical cancer and participant age, marital status, religion, screening uptake, and willingness to undergo screen. Results: Awareness of cervical cancer was recorded among 174/230 (75.7%) respondents who enrolled in this study; 117 (67.2%) knew that it was preventable, but only three (1.3%) respondents had undergone screening. Among the aware participants, 91 (52.3%) and 131 (75.3%) knew that sexual intercourse and multiple sexual partners are risk factors for the disease, respectively. In contrast, knowledge of the etiology was poor; 82 (47.1%) respondents who knew it was preventable had heard about human papillomavirus (HPV), while 72 (41.4%) knew that HPV causes cervical cancer. Most (78%) of the participants expressed willingness to take a human papillomavirus vaccine or undergo screening (84.6%) if made available to them. Awareness was significantly associated with participants' age (p = 0.022) and willingness to undergo screening (p = 0.016). Conclusion: This study revealed discordance between awareness and knowledge about cervical cancer. Educational initiatives reflective of population perception/knowledge of cervical cancer are needed to mitigate the rising incidence of this disease, especially among female healthcare providers.

Outreach Training and Supportive Supervision for Quality Malaria Service Delivery: A Qualitative Evaluation in 11 Sub-Saharan African Countries.

Quality improvement of malaria services aims to ensure that more patients receive accurate diagnosis, appropriate treatment, and referral. The Outreach Training and Supportive Supervision Plus (OTSS+) approach seeks to improve health facility readiness and provider competency through onsite supportive supervision, troubleshooting, and on-the-job training. As part of a multicomponent evaluation, qualitative research was conducted to understand the value of the OTSS+ approach for malaria quality improvement. Semistructured key informant interviews, focus group discussions, and structured health facility-based interviews were used to gather stakeholder perspectives at subnational, national, and global levels. Data were collected globally and in 11 countries implementing OTSS+; in-depth data collection was done in four: Cameroon, Ghana, Niger, and Zambia. Study sites and participants were selected purposively. Verbatim transcripts were analyzed thematically, following the Framework approach. A total of 262 participants were included in the analysis; 98 (37.4%) were supervisees, 99 (37.8%) were supervisors, and 65 (24.8%) were other stakeholders. The OTSS+ approach was perceived to improve provider knowledge and skills in malaria service delivery and to improve data and supply management indirectly. Improvements were attributed to a combination of factors. Participants valued the relevance, adaptation, and digitization of supervision checklists; the quality and amount of contact with problem-solving supervisors; and the joint identification of problems and solutions, and development of action plans. Opportunities for improvement were digitized checklist refinement, assurance of a sufficient pool of supervisors, prioritization of health facilities, action plan dissemination and follow-up, and data review and use. The OTSS+ approach was perceived to be a useful quality improvement approach for malaria services.

Case fatality and factors associated with Maternal mortality in patients with eclampsia: A retrospective review of 761 cases in a tertiary institution in Nigeria.

To determine the case fatality and factors associated with maternal mortality in patients with eclampsia in UMTH, we conducted a retrospective review of 761 consecutive cases of eclampsia managed UMTH, Nigeria, from 2005 to 2020. Case fatality was 7.6%, most being unbooked primigravidae with no formal education. Common causes of death identified were aspiration, cerebrovascular accident, and haemorrhage. Admission delayed >10 hours was found to be independently associated with maternal death, and Caesarean delivery.

Efficacy and safety of transcranial magnetic stimulation for treating major depressive disorder: An umbrella review and re-analysis of published meta-analyses of randomised controlled trials.

OBJECTIVES: We re-analysed data from published meta-analyses testing the effects of Transcranial Magnetic Stimulation (TMS) on Major Depressive Disorder (MDD) in adults. We applied up-to-date meta-analytic techniques for handling heterogeneity including the random-effects Hartung-Knapp-Sidik-Jonkman method and estimated 95% prediction intervals. Heterogeneity practices in published meta-analyses were assessed as a secondary aim. STUDY DESIGN AND SETTING: We performed systematic searches of systematic reviews with meta-analyses that included randomised controlled trials assessing the efficacy, tolerability, and side effects of TMS on MDD. We performed risk of bias assessment using A MeaSurement Tool to Assess Reviews (AMSTAR) 2 and re-analysed meta-analyses involving 10 or more primary studies. RESULTS: We included 29 systematic reviews and re-analysed 15 meta-analyses. Authors of all meta-analyses interpreted findings to suggest TMS is safe and effective for MDD. Our re-analysis showed that in 14 out of 15 meta-analyses, the 95% prediction intervals included the null and captured values in the opposite effect direction. We also detected presence of small-study effects in some meta-analyses and 24 out of 25 systematic reviews received an AMSTAR 2 rating classed as critically low. CONCLUSION: Authors of all included meta-analyses interpreted findings to suggest TMS is safe and effective for MDD despite lack of comprehensive investigation of heterogeneity. Our re-analysis revealed the direction and magnitude of treatment effects vary widely across different settings. We also found high risk of bias in the majority of included systematic reviews and presence of small-study effects in some meta-analyses. Because of these reasons, we argue TMS for MDD may not be as effective and potentially less tolerated in some populations than current evidence suggests.

Effectiveness of buffered and non-buffered local anaesthetic in inferior alveolar nerve block: a randomised study.

The objective of this study was to compare the effectiveness of buffered and non-buffered local anaesthetic (LA) following inferior alveolar nerve block (IANB). This study was conducted at Usmanu Danfodiyo University Teaching Hospital Sokoto from June 2020 to January 2021. Subjects were randomised into Groups A and B. Group A received 2 ml of freshly prepared 2% lignocaine with 1:100,000 adrenaline buffered with 0.18 ml of 8.4% sodium bicarbonate solution while those in Group B received non-buffered 2% lignocaine with 1:100,000 adrenaline LA solutions. The onset of action of the LA was assessed by subjective and objective methods, while pain at the injection site was assessed using a numerical rating scale. Data obtained were analysed using statistical package for social sciences (IBM SPSS) version 21. The mean (SD) ages for Groups A and B were 37.4 (14.9) and 40.1 (14.4) years, respectively. The mean (SD) onset times of the LA by subjective testing were 126 (31.7) and 201 (66.8) seconds for Groups A and B, respectively. Similarly, the mean (SD) onset times of the local anaesthesia by objective testing for Groups A and B were 186 (41.0) and 287 (85.0) seconds, respectively, and both were significant (p<0.001). Pain at the injection site assessed objectively and subjectively were also statistically different (p<0.001). The findings of this study suggest that buffered LA is more effective than non-buffered LA of the same composition when used for IANB particularly concerning a significantly faster rate of onset and less pain at the injection site.

Non-convulsive status epilepticus as the initial manifestation in a family with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL).

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is an autosomal dominant small-vessel disease caused by mutations of the NOTCH3 gene. It typically presents with migraine, recurrent brain ischaemia, and cognitive disorders. Seizures rarely present as the initial manifestation, with non-convulsive status epilepticus being even less frequent. We present a series of 3 related patients with this arteriopathy, 2 of whom presented status epilepticus as a manifestation of the disease.

Evaluation of an opa gene-based nucleic acid amplification test for detection of Neisseria gonorrhoeae in urogenital samples in North India.

Due to the poor positive predictive value of nucleic acid amplification tests (NAATs) for gonorrhoea when applied to a low-prevalence setting, current guidelines recommend the use of supplementary polymerase chain reaction (PCR) targeting a different gene for confirmation of true positives in urogenital specimens. This study sought to standardize and evaluate performance of an in-house opa gene-based PCR assay for gonorrhoea compared to assays targeting the porA pseudogene and 16S rRNA gene. Four hundred samples (300 endocervical, 100 urethral swabs) from patients attending STD clinics in New Delhi, India were used. The sensitivity, specificity, positive predictive value and negative predictive value of the opa-based PCR were 100%, 97·9%, 89·5% and 100%, respectively. In females, the use of NAATs provided enhanced diagnosis of gonorrhoea.

Risk factors and microbial isolates of puerperal sepsis at the University of Maiduguri Teaching Hospital, Maiduguri, North-eastern Nigeria.

PURPOSE: To determine the risk factors and microbial isolates of puerperal sepsis. METHODS: A 12 year retrospective review of patients with puerperal sepsis from January 1999 to December 2010 at the University of Maiduguri Teaching Hospital (UMTH) was conducted. Information on socio-demographic characteristics, clinical presentations, place and mode of delivery and microorganism isolated from the endocervical swabs was collected and analysed. Comparison was also made with the other women that delivered during the period of study. RESULTS: The incidence of puerperal sepsis was 0.78%. Majority (88.0%) of the patients were unbooked, 59.1% delivered at home and 23.1% delivered in other peripheral hospitals. The major risk factors for developing puerperal sepsis were unbooked status, home delivery, perineal trauma, caesarean section (C/S) and maternal age <24 years with OR of 56.60, 39.25, 8.52, 4.99 and 1.32, respectively. The commonest microorganism isolated were Staphylococcus aureus and Escherichia coli seen in 35.4 and 20.9%, respectively. Streptococcus species was found in 6.9%, while 20.3% had polymicrobials isolated. CONCLUSION: Puerperal sepsis continues to present a significant risk of maternal mortality in developing countries. The risk factors are unbooked women, home delivery, perineal trauma, caesarean section and maternal age <24 years. S. aureus and E. coli are the commonest isolated organisms. There is a need to enlighten the populace on the need for booking, skilled attendant at delivery and hospital delivery under aseptic conditions.

The benefit of dynamic neutrophil-lymphocyte ratio and systemic immune-inflammation index in predicting survival in patients undergoing hemiarthroplasty.

OBJECTIVE: We aimed at investigating the relationship between the values of neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), systemic immune-inflammation index (SII) in the preoperative and postoperative periods in patients who underwent hemiarthroplasty. PATIENTS AND METHODS: The medical records of patients aged >65 years who underwent hemiarthroplasty due to hip fracture in our institution between January 2017 and December 2020 were examined. Dynamic Neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (dPLR), and systemic immune-inflammation index (dSII) were calculated using biomarker data (neutrophil, lymphocyte, and platelet counts) obtained at admission and on postoperative day 2 and 5. RESULTS: A total of 176 individuals (119 females, 57 males) were included in the study. The PLR on postoperative day 2, and NLR and SII measurement on postoperative day 2 and 5 in patients were found to have significant predictive value in 1-year mortality rates. A 1-year increase in age was associated with a 4.9% increased risk of death at 1-year follow-up (p=0.006). A 1-unit increase in NLR on postoperative day 5 was associated with a 4.3% increased risk of death at 1-year follow-up (p=0.004). CONCLUSIONS: The present study shows that our model for predicting the absolute mortality risk in patients can be improved by evaluating dSII, dPLR, and dNLR in the preoperative period and postoperative day 2 and 5. However, it cannot be said that the model has a discriminatory ability to exactly predict postoperative mortality. Nevertheless, these parameters may aid in the development of early therapeutic intervention to improve patient outcomes.