Hospital presentations for self-poisoning during COVID-19 in Sri Lanka: an interrupted time-series analysis.

BACKGROUND: There is widespread concern over the impact of public health measures, such as lockdowns, associated with COVID-19 on mental health, including suicide. High-quality evidence from low-income and middle-income countries, where the burden of suicide and self-harm is greatest, is scarce. We aimed to determine the effect of the pandemic on hospital presentations for self-poisoning. METHODS: In this interrupted time-series analysis, we established a new self-poisoning register at the tertiary care Teaching Hospital Peradeniya in Sri Lanka, a lower-middle-income country. Using a standard extraction sheet, data were gathered for all patients admitted to the Toxicology Unit with self-poisoning between Jan 1, 2019, and Aug 31, 2020. Only patients classified by the treating clinician as having intentionally self-poisoned were included. Data on date of admission, age or date of birth, sex, and poisoning method were collected. No data on ethnicity were available. We used interrupted time-series analysis to calculate weekly hospital admissions for self-poisoning before (Jan 1, 2019-March 19, 2020) and during (March 20-Aug 31, 2020) the pandemic, overall and by age (age <25 years vs ≥25 years) and sex. Individuals with missing date of admission were excluded from the main analysis. FINDINGS: Between Jan 1, 2019, and Aug 31, 2020, 1401 individuals (584 [41·7%] males, 761 [54·3%] females, and 56 [4·0%] of unknown sex) presented to the hospital with self-poisoning and had date of admission data. A 32% (95% CI 12-48) reduction in hospital presentations for self-poisoning in the pandemic period compared with pre-pandemic trends was observed (rate ratio 0·68, 95% CI 0·52-0·88; p=0·0032). We found no evidence that the impact of the pandemic differed by sex (rate ratio 0·64, 95% CI 0·44-0·94, for females vs 0·85, 0·57-1·26, for males; pinteraction=0·43) or age (0·64, 0·44-0·93, for patients aged <25 years vs 0·81, 0·57-1·16, for patients aged ≥25 years; pinteraction=0·077). INTERPRETATION: This is the first study from a lower-middle-income country to estimate the impact of the pandemic on self-harm (non-fatal) accounting for underlying trends. If the fall in hospital presentations during the pandemic reflects a reduction in the medical treatment of people who have self-poisoned, rather than a true fall in incidence, then public health messages should emphasise the importance of seeking help early. FUNDING: Elizabeth Blackwell Institute University of Bristol, Wellcome Trust, and Centre for Pesticide Suicide Prevention. TRANSLATIONS: For the Sinhalese and Tamil translations of the abstract see Supplementary Materials section.

Biopharmaceutical Industry Practices for Sequence Variant Analyses of Recombinant Protein Therapeutics.

The application of advanced methodologies such as next-generation sequencing (NGS) and mass spectrometry (MS) to the characterization of cell lines and recombinant proteins has enabled the highly sensitive detection of sequence variants (SVs). However, although these approaches can be leveraged to provide deep insight into product microheterogeneity caused by SVs, they are not used in a standardized manner across the industry. Currently, there is little clarity and consensus on the utilization, timing, and significance of SV findings. This white paper addresses the current practices, logistics, and strategies for the analysis of SVs using a benchmarking survey coordinated by the International Consortium for Innovation & Quality in Pharmaceutical Development (IQ) as well as a series of deliberations among a panel of experts assembled from across the biopharmaceutical industry. Discussion includes current industry experiences including approaches for detection and quantitation of SVs during cell-line and process development, risk assessments, and regulatory feedback. Although SVs are a potential issue for all recombinant protein therapeutics, the scope of this discussion will be limited to SVs produced in mammalian cells. Ultimately, it is our hope that the findings from the survey and deliberations of the committee are useful to decision makers in industry and positions them to respond to findings of SVs in recombinant proteins that are destined for clinical or commercial use in a strategic manner.LAY ABSTRACT: This white paper addresses the current practices, logistics, and strategies for the analysis of amino acid sequence variants using a benchmarking survey coordinated by the International Consortium for Innovation & Quality in Pharmaceutical Development (IQ) as well as a series of deliberations among a panel of experts assembled from across the biopharmaceutical industry. Discussion includes current industry experiences regarding detection and quantitation of SVs during cell-line and process development, risk assessments, and regulatory feedback.

Earlier detection of Alzheimer disease using N-fold cross validation approach.

According to the recent study, world-wide 40 million patients are affected by Alzheimer disease (AD) because it is one of the dangerous neurodegenerative disorders. This AD disease has less symptoms such as short term memory loss, mood swings, problem with language understanding and behavioral issues. Due to these low symptoms, AD disease is difficult to recognize in the early stage. So, the automated computer aided system need to be developed for recognizing the AD disease for minimizing the mortality rate. Initially, brain MRI image is collected from patients which are processed by applying different processing steps such as noise removal, segmentation, feature extraction, feature selection and classification. The captured MRI image has noise that is eliminated by applying the Lucy-Richardson approach which examines the each pixel in the image and removes the Gaussian noise which also eliminates the blur image. After eliminating the noise pixel from the image, affected region is segmented by Prolong adaptive exclusive analytical Atlas approach. From the segmented region, different GLCM statistical features are extracted and optimal features subset is selected by applying the hybrid wrapper filtering approach. This selected features are analyzed by N-fold cross validation approach which recognizes the AD related features successfully. Then the efficiency of the system is evaluated with the help of MATLAB based experimental results, in which Alzheimer's Disease Neuroimaging Initiative (ADNI) dataset images are utilized for examining the efficiency in terms of sensitivity, specificity, ROC curve and accuracy.

Implementation of Plate Imaging for Demonstration of Monoclonality in Biologics Manufacturing Development.

Monoclonality of mammalian cell lines used for production of biologics is a regulatory expectation and one of the attributes assessed as part of a larger process to ensure consistent quality of the biologic. Historically, monoclonality has been demonstrated through statistics generated from limiting dilution cloning or through verified flow cytometry methods. A variety of new technologies are now on the market with the potential to offer more efficient and robust approaches to generating and documenting a clonal cell line.Here we present an industry perspective on approaches for the application of imaging and integration of that information into a regulatory submission to support a monoclonality claim. These approaches represent the views of a consortium of companies within the BioPhorum Development Group and include case studies utilising imaging technology that apply scientifically sound approaches and efforts in demonstrating monoclonality. By highlighting both the utility of these alternative approaches and the advantages they bring over the traditional methods, as well as their adoption by industry leaders, we hope to encourage acceptance of their use within the biologics cell line development space and provide guidance for regulatory submission using these alternative approaches.LAY ABSTRACT: In the manufacture of biologics produced in mammalian cells, one recommendation by regulatory agencies to help ensure product consistency, safety, and efficacy is to produce the material from a monoclonal cell line derived from a single, progenitor cell. The process by which monoclonality is assured can be supplemented with single-well plate images of the progenitor cell. Here we highlight the utility of that imaging technology, describe approaches to verify the validity of those images, and discuss how to analyze that information to support a biologic filing application. This approach serves as an industry perspective to increased regulatory interest within the scope of monoclonality for mammalian cell culture-derived biologics.

The detection of microbial DNA but not cultured bacteria is associated with increased mortality in patients with suspected sepsis-a prospective multi-centre European observational study.

OBJECTIVES: Blood culture results inadequately stratify the mortality risk in critically ill patients with sepsis. We sought to establish the prognostic significance of the presence of microbial DNA in the bloodstream of patients hospitalized with suspected sepsis. METHODS: We analysed the data collected during the Rapid Diagnosis of Infections in the Critically Ill (RADICAL) study, which compared a novel culture-independent PCR/electrospray ionization-mass spectrometry (ESI-MS) assay with standard microbiological testing. Patients were eligible for the study if they had suspected sepsis and were either hospitalized or were referred to one of nine intensive care units from six European countries. The blood specimen for PCR/ESI-MS assay was taken along with initial blood culture taken for clinical indications. RESULTS: Of the 616 patients recruited to the RADICAL study, 439 patients had data on outcome, results of the blood culture and PCR/ESI-MS assay available for analysis. Positive blood culture and PCR/ESI-MSI result was found in 13% (56/439) and 40% (177/439) of patients, respectively. Either a positive blood culture (p 0.01) or a positive PCR/ESI-MS (p 0.005) was associated with higher SOFA scores on enrolment to the study. There was no difference in 28-day mortality observed in patients who had either positive or negative blood cultures (35% versus 32%, p 0.74). However, in patients with a positive PCR/ESI-MS assay, mortality was significantly higher in comparison to those with a negative result (42% versus 26%, p 0.001). CONCLUSIONS: Presence of microbial DNA in patients with suspected sepsis might define a patient group at higher risk of death.

Repeatability of Spectral Domain Optical Coherence Tomography Measurements in High Myopia.

PURPOSE: The purpose of this study was to compare the repeatability of spectral domain optical coherence tomography (SDOCT) parameters in high-myopic and emmetropic healthy subjects, and to evaluate the influence of axial length on the repeatability of SDOCT parameters in high myopia. METHODS: In a prospective study, 93 eyes of 63 high-myopic subjects (spherical refractive error, -6 to -12 D; median age, 25 y) and 28 eyes of 14 emmetropic (spherical refractive error, 0 D; median age, 30 y) subjects underwent optic nerve head, retinal nerve fiber layer (RNFL), and ganglion cell complex imaging with SDOCT. For the repeatability analysis, 31 eyes of 31 high-myopic subjects and 14 eyes of 14 emmetropic subjects underwent 3 repeated scans in the same session. RESULTS: Among the optic nerve head parameters, within-subject coefficient of variation (CVw) measurements of the disc area (0.6% vs. 0.2%), rim area (8.7 vs. 2.8), and rim volume (16.7 vs. 8.9) were significantly larger (worse) in high-myopic compared with the emmetropic subjects. CVw measurements of all RNFL (range, 1.7 to 22.4) and ganglion cell complex (range, 1.8 to 2.5) parameters in high-myopic subjects were comparable to that in emmetropic subjects (2.4 to 24.0 and 1.7 to 2.0, respectively). Axial length significantly affected the CVw of nasal (coefficient, 0.01; P=0.04) and average RNFL (coefficient, 0.004; P=0.001) parameters but not that of the other SDOCT parameters. CONCLUSIONS: Repeatabilities of most of the SDOCT parameters in high-myopic subjects were good and comparable to that of emmetropic subjects. This suggests that SDOCT can be useful for following up high-myopic glaucoma patients to detect progression.

Tenofovir alafenamide fumarate for the treatment of HIV infection.

Tenofovir alafenamide fumarate is a recently developed prodrug of tenofovir, a nucleotide analogue reverse transcriptase inhibitor with potent inhibitory activity against HIV. The utility of a previously developed tenofovir prodrug, tenofovir disoproxil fumarate, had been hampered by renal and bone mineral adverse events. Tenofovir alafenamide fumarate overcomes the shortcomings of tenofovir disoproxil fumarate by delivering high intracellular concentrations of the parent drug, tenofovir, while substantially reducing systemic exposure. Tenofovir alafenamide fumarate is currently available as a component of three fixed-dose products: i) coformulation with emtricitabine; ii) coformulation with elvitegravir, cobicistat and emtricitabine; and iii) coformulation with rilpivirine and emtricitabine.

Industrial production of clotting factors: Challenges of expression, and choice of host cells.

The development of recombinant forms of blood coagulation factors as safer alternatives to plasma derived factors marked a major advance in the treatment of common coagulation disorders. These are complex proteins, mostly enzymes or co-enzymes, involving multiple post-translational modifications, and therefore are difficult to express. This article reviews the nature of the expression challenges for the industrial production of these factors, vis-à-vis the translational and post-translational bottlenecks, as well as the choice of host cell lines for high-fidelity production. For achieving high productivities of vitamin K dependent proteins, which include factors II (prothrombin), VII, IX and X, and protein C, host cell limitation of γ-glutamyl carboxylation is a major bottleneck. Despite progress in addressing this, involvement of yet unidentified protein(s) impedes a complete cell engineering solution. Human factor VIII expresses at very low levels due to limitations at several steps in the protein secretion pathway. Protein and cell engineering, vector improvement and alternate host cells promise improvement in the productivity. Production of Von Willebrand factor is constrained by its large size, complex structure, and the need for extensive glycosylation and disulfide-bonded oligomerization. All the licensed therapeutic factors are produced in CHO, BHK or HEK293 cells. While HEK293 is a recent adoption, BHK cells appear to be disfavored.

Identification and typing of Francisella tularensis with a highly automated genotyping assay.

A PCR assay was developed to genotypically characterize Francisella tularensis and F. novicida. An integrated and partially redundant set of markers was selected to provide positive identification of these species, identify subspecies of F. tularensis and genotype 14 variable number tandem repeat (VNTR) markers. Assay performance was evaluated with 117 Francisella samples. Sample DNA was amplified, and the masses of the PCR products were determined with electrospray ionization/time of flight mass spectrometry (ESI-MS). The base compositions of the PCR amplicons were derived from these high-accuracy mass measurements and contrasted with databased information associated with each of the 25 assay markers. Species and subspecies determinations for all samples were fully concordant with results from established typing methods, and VNTR markers provided additional discrimination among samples. Sequence variants were observed with a number of assay markers, but these did not interfere with sample characterization, and served to increase the genetic diversity detected by the assay.

5 years review of periocular basal cell carcinoma and proposed follow-up protocol.

AIM: (1) To investigate the recurrence of periocular basal cell carcinoma (BCC) reported as completely excised on histology. (2) To identify risks associated with recurrence. (3) To recommend a rational follow-up protocol. METHODS: This is a cohort study by case note review of consecutive patients undergoing excision of periocular BCC between 2000 and 2006 at University Hospitals of Leicester. All lesions were excised with 3 mm clinical margin and the defect reconstructed only after the excision margin was declared clear. RESULTS: A total of 413 episodes of surgical excision were recorded for 270 patients over the 7-year period of 2000-2006. All of them have 5 years follow-up. Mean age 73.7 (±12.5). In all, 67% were nodular BCC and 45.4% located in the lower eyelid. The main outcome measure was the recurrence rate. None of the patients with primary nodular BCC suffered recurrence. The recurrence rate for primary morphoeaform BCC following complete excision is 3.8%. In total, 8.1% of patients had several lesions simultaneously whereas 7.8% patients had BCC in multiple locations subsequently (metachronous). Three patients who had previously recurrent BCC (rBCC) treated elsewhere or not using this method had orbital/lacrimal drainage system involvement requiring exenteration. CONCLUSION: We recommend that patients with a single, completely excised primary solid or nodular BCC can be discharged after one 6-monthly review, although they should be instructed to monitor for the development of further lesions. The incidence of recurrence for primary morphoeaform BCC is 3.8% and for rBCC is 3.6% over 5 years and these patients should stay under review for this period.

Isolated ocular cutaneous involvement in antiphospholipid syndrome.

BACKGROUND: Antiphospholipid syndrome (APS) is defined as the presence of venous or arterial thrombosis, and/or recurrent miscarriage with evidence of antiphospholipid antibodies (aPL). In both primary and secondary APS, ocular and neurophthalmic manifestations such as retinal arteritis, retinal venous occlusion, ischemic optic neuropathy, transient loss of vision - amaurosis fugax, diplopia have been reported. MATERIALS AND METHODS: We present an unusual case of APS in a healthy 24-year old male who had isolated ocular presentation with recurrent right periocular oedema and non-healing ulceration of the biopsy site without systemic involvement. Ocular examinations and investigations including inflammatory markers were normal. CONCLUSION: Atypical presentations of APS may result in initial difficulty in making diagnosis.

Multidisciplinary management of periocular necrotising fasciitis: a series of 11 patients.

BACKGROUND: Necrotising fasciitis is an uncommon but life-threatening soft tissue infection characterised by rapidly spreading inflammation and necrosis of skin, subcutaneous fat and fascia. Left untreated, the mortality can be more than 70%. Early surgical intervention can reduce morbidity and mortality. PATIENTS AND METHODS: This is a series of 11 patients who presented to our oculoplastic and orbit unit with periocular necrotising fasciitis over a period of five years. We present the modes of presentation, predisposing factors, diagnosis, and the multidisciplinary team management of these patients. RESULTS: Of the 11 patients, 1 patient died and 2 patients required intensive care management. Of the 10 surviving patients, 8 patients needed further surgical interventions for correction of complications, like eyelid malposition, ptosis and protective or corrective surgery in the form of ectropion correction, skin grafting and other rehabilitative procedures. CONCLUSION: To the best of our knowledge, this is the largest series of periocular necrotising fasciitis in the literature. Necrotising fasciitis is a potentially fatal condition, resulting in a high rate of mortality and morbidity. Early surgical intervention reduces the mortality. A high index of suspicion is needed to make a prompt diagnosis. These patients need expeditious intervention and may require a long follow-up and subsequent surgery for complications related to scarring and other sequelae.

Adenoid reservoir for pathogenic biofilm bacteria.

Biofilms of pathogenic bacteria are present on the middle ear mucosa of children with chronic otitis media (COM) and may contribute to the persistence of pathogens and the recalcitrance of COM to antibiotic treatment. Controlled studies indicate that adenoidectomy is effective in the treatment of COM, suggesting that the adenoids may act as a reservoir for COM pathogens. To investigate the bacterial community in the adenoid, samples were obtained from 35 children undergoing adenoidectomy for chronic OM or obstructive sleep apnea. We used a novel, culture-independent molecular diagnostic methodology, followed by confocal microscopy, to investigate the in situ distribution and organization of pathogens in the adenoids to determine whether pathogenic bacteria exhibited criteria characteristic of biofilms. The Ibis T5000 Universal Biosensor System was used to interrogate the extent of the microbial diversity within adenoid biopsy specimens. Using a suite of 16 broad-range bacterial primers, we demonstrated that adenoids from both diagnostic groups were colonized with polymicrobial biofilms. Haemophilus influenzae was present in more adenoids from the COM group (P = 0.005), but there was no significant difference between the two patient groups for Streptococcus pneumoniae or Staphylococcus aureus. Fluorescence in situ hybridization, lectin binding, and the use of antibodies specific for host epithelial cells demonstrated that pathogens were aggregated, surrounded by a carbohydrate matrix, and localized on and within the epithelial cell surface, which is consistent with criteria for bacterial biofilms.

Modified visual field test for ptosis surgery (Leicester Peripheral Field Test).

INTRODUCTION: There is lack of consensus among Primary Health Care Trusts (PCTs) and health insurers on how to reimburse ptosis surgery and upper lid blepharoplasty, as these procedures can be regarded as cosmetic. Standardised photographs are expensive and difficult to achieve, whilst the routine 24-2 visual field lacks the range to detect visually significant superior field defects.AimTo introduce a modified visual field designed to assess the functional disability associated with ptosis and dermatochalasis and to demonstrate the effectiveness of surgery in improving the visual field. METHODS: Patients who had surgery for ptosis or dermatochalasis between January 2006 and December 2009 were prospectively invited to perform a modified visual field test pre- and post-operatively. RESULTS: In total, 97 patients amounting to 194 eyes were included in the study. Ninety five eyes had aponeurotic repair with or without blepharoplasty and 77 eyes had blepharoplasty alone. This modified test has a sensitivity of 98.8% of detecting ptosis. For patients who underwent ptosis surgery with or without blepharoplasty, 84.2% recorded an improvement in points seen with the test and 81% recorded an improvement in visual field height. For those who had blepharoplasty alone, 90.9% recorded an improvement in points seen in the modified visual field test and 80.6% had improvement in visual field height. CONCLUSION: Our modified visual field assessment is a quick and easy way to assess patient disability associated with ptosis and dermatochalasis. Surgery improves the demonstrated defect, confirming that ptosis and dermatochalasis can be considered a functional rather than cosmetic issue.

Lacrimal fossa lesions: pictorial review of CT and MRI features.

A wide range of disease process involve the lacrimal gland/fossa. In this pictorial review, we use histology-proven cases to illustrate conditions that affect the lacrimal gland/fossa. CT and MRI features of neoplastic, inflammatory, infiltrative, and developmental conditions are discussed.

Identification of bacterial plant pathogens using multilocus polymerase chain reaction/electrospray ionization-mass spectrometry.

Polymerase chain reaction/electrospray ionization-mass spectrometry (PCR/ESI-MS, previously known as "TIGER") utilizes PCR with broad-range primers to amplify products from a wide array of organisms within a taxonomic group, followed by analysis of PCR amplicons using mass spectrometry. Computer analysis of precise masses allows for calculations of base compositions for the broad-range PCR products, which can then be compared to a database for identification. PCR/ESI-MS has the benefits of PCR in sensitivity and high-throughput capacity, but also has the distinct advantage of being able to detect and identify organisms with no prior characterization or sequence data. Existing broad range PCR primers, designed with an emphasis on human pathogens, were tested for their ability to amplify DNA of well characterized phytobacterial strains, as well as to populate the existing PCR/ESI-MS bacterial database with base counts. In a blinded panel study, PCR/ESI-MS successfully identified 93% of unknown bacterial DNAs to the genus level and 73% to the species/subspecies level. Additionally, PCR/ESI-MS was capable of detecting and identifying multiple bacteria within the same sample. The sensitivity of PCR/ESI-MS was consistent with other PCR based assays, and the specificity varied depending on the bacterial species. Preliminary tests with real life samples demonstrate a high potential for using PCR/ESI-MS systems for agricultural diagnostic applications.

Current trends in the diagnosis and management of renal nutcracker syndrome: a review.

Nutcracker syndrome is caused by compression of the left renal vein between the aorta and the superior mesenteric artery where it passes in the fork formed at the bifurcation of these arteries. The phenomenon results in left renal venous hypertension. The syndrome is manifested by left flank and abdominal pain, with or without unilateral haematuria. Other common presentation is as "pelvic congestion syndrome" characterized by symptoms of dysmenorrhea, dyspareunia, post-coital ache, lower abdominal pain, dysuria, pelvic, vulvar, gluteal or thigh varices and emotional disturbances. Likewise compression of the left renal vein can cause left renal-to-gonadal vein reflux resulting in lower limb varices and varicoceles in males. Its diagnosis is based on history and physical examination, basic lab tests to exclude other causes of haematuria, cystoscopy and ureteroscopy to confirm unilateral haematuria and exclude other causes of this sinister symptom. Sequence of imaging has more or less been rationalised to USS with Doppler studies, CT or MR angiography and finally phlebography with renal vein and IVC manometery to confirm the diagnosis.