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.

Alzheimer's Disease Prediction Using Fly-Optimized Densely Connected Convolution Neural Networks Based on MRI Images.

Alzheimer's is a degenerative brain cell disease that affects around 5.8 million people globally. The progressive neurodegenerative disease known as Alzheimer's Disease (AD), affects the frontal cortex, the part of the brain in charge of memory, language, and cognition. As a result, researchers are utilizing a variety of machine-learning techniques to create an automated method for AD detection. The massive data collected during ROI and biomarker identification takes longer to handle using current methods. This study uses metaheuristic-tuned deep learning to detect the AD-affected region. The research utilizes advanced deep learning and image processing techniques to enhance early and accurate diagnosis of Alzheimer's disease, potentially enhancing patient outcomes and prompt therapy. The capacity of deep neural networks to extract complex patterns from magnetic resonance imaging (MRI) scans makes them indispensable in the diagnosis of AD since they allow the detection of minor aberrations and complex alterations in brain structure and composition. An adaptive histogram approach processes the collected photographs, and a weighted median filter is used in place of the noisy pixels. The next step is to identify the issue region using a deep convolution network-based clustering segmentation process. A correlated information theory approach is used to extract various textural and statistical features from the separated regions. Lastly, the selected features are probed by the fly-optimized densely linked convolution neural networks. The method surpasses state-of-the-art techniques in sensitivity (15.52%), specificity (15.62%), accuracy (9.01%), error rate (11.29%), and F-measure (10.52%) for recognizing AD-impacted regions in MRI scans using the Kaggle dataset.

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.

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.

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.

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.

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.

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.

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.

RNAMotif, an RNA secondary structure definition and search algorithm.

RNA molecules fold into characteristic secondary and tertiary structures that account for their diverse functional activities. Many of these RNA structures are assembled from a collection of RNA structural motifs. These basic building blocks are used repeatedly, and in various combinations, to form different RNA types and define their unique structural and functional properties. Identification of recurring RNA structural motifs will therefore enhance our understanding of RNA structure and help associate elements of RNA structure with functional and regulatory elements. Our goal was to develop a computer program that can describe an RNA structural element of any complexity and then search any nucleotide sequence database, including the complete prokaryotic and eukaryotic genomes, for these structural elements. Here we describe in detail a new computational motif search algorithm, RNAMotif, and demonstrate its utility with some motif search examples. RNAMotif differs from other motif search tools in two important aspects: first, the structure definition language is more flexible and can specify any type of base-base interaction; second, RNAMotif provides a user controlled scoring section that can be used to add capabilities that patterns alone cannot provide.

Prediction of rho-independent transcriptional terminators in Escherichia coli.

A new algorithm called RNAMotif containing RNA structure and sequence constraints and a thermodynamic scoring system was used to search for intrinsic rho-independent terminators in the Escherichia coli K-12 genome. We identified all 135 reported terminators and 940 putative terminator sequences beginning no more than 60 nt away from the 3'-end of the annotated transcription units (TU). Putative and reported terminators with the scores above our chosen threshold were found for 37 of the 53 non-coding RNA TU and for almost 50% of the 2592 annotated protein-encoding TU, which correlates well with the number of TU expected to contain rho-independent terminators. We also identified 439 terminators that could function in a bi-directional fashion, servicing one gene on the positive strand and a different gene on the negative strand. Approximately 700 additional termination signals in non-coding regions (NCR) far away from the nearest annotated gene were predicted. This number correlates well with the excess number of predicted 'orphan' promoters in the NCR, and these promoters and terminators may be associated with as yet unidentified TU. The significant number of high scoring hits that occurred within the reading frame of annotated genes suggests that either an additional component of rho-independent terminators exists or that a suppressive mechanism to prevent unwanted termination remains to be discovered.

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.

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.

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.

Management of ascending aortic aneurysm complicating coarctation of the aorta.

Four patients with coarctation of the aorta complicated by an aneurysm of the ascending aorta are described. One patient, treated only medically, died suddenly. For the 3 patients who underwent operation, management was similar. Pharmacological control of blood pressure and repair of the aortic coarctation to achieve anatomical reduction in afterload (stage I) were followed after an interval of one to five weeks by surgical repair of the ascending aortic aneurysm (stage II). Initial repair of the coarctation relieves proximal hypertension, thereby decreasing the chance of progressive dissection or rupture of the aneurysm. It also permits safe arterial cannulation for perfusion during correction of the aneurysm in the second stage. In patients not requiring valve replacement in stage II, continued long-term follow-up for progression of aortic valvular disease appears to be necessary.

Use of Proplast II as a subperiosteal implant for the correction of anophthalmic enophthalmos.

BACKGROUND: A variety of autogenous and alloplastic materials have been used as subperiosteal implants to correct anophthalmic enophthalmos. Proplast II is a synthetic porous composite of Teflon polymer and alumina. Proplast II offers a number of advantages over other commonly used alloplastic materials such as silicone and polymethyl methacrylate. It is light, porous, resilient, malleable, and easy to shape. It can be readily sterilised after shaping. It has been found to integrate with the surrounding tissues, thereby minimising the risk of subsequent implant migration and extrusion. METHODS: Proplast II was used as a subperiosteal implant in a total of 15 anophthalmic patients during the period June 1990 to March 1994. The indication for this procedure in all patients was poor orbital volume replacement despite the prior insertion of an adequately sized spherical socket implant. RESULTS: The results were excellent with a good correction of preoperative upper eyelid sulcus deformity. There were no operative complications nor any serious postoperative complications. The implants were well tolerated. CONCLUSION: Proplast II can be highly recommended for use as a subperiosteal implant.

Erroneous automated refraction in a case of asteroid hyalosis.

To our knowledge, this is the first report of asteroid hyalosis giving rise to spurious automated refraction readings. A patient with asteroid hyalosis had uneventful phacoemulsification with intraocular lens implantation. Using a Nikon autorefractor, the postoperative refraction was +13.0 diopters (D); retinoscopy revealed that it should be +1.0 D. Asteroid hyalosis changes the optical quality of the vitreous and can cause an incorrect reading from automated equipment such as autorefractors and A-scan ultrasound scanners. In patients with asteroid hyalosis, results from autorefractors should be viewed in conjunction with clinical findings.

Deltamethrin treated bednets for control of malaria transmitted by Anopheles culicifacies (Diptera: Culicidae) in India.

In a malaria endemic area in Orissa state in eastern India baseline (November 1989 to October 1990) malaria incidence ranged front 215 to 328 cases/1,000 population/yr in different groups of villages. In November 1990, nylon bednets treated with deltamethrin at 25 mg/m2 were given out in two villages (population 1062), untreated bednets were given out in five villages (population 1,226) and in one village (population 786) nets were not given. Nets were retreated in October 1991 and June 1992 in treated-net villages. The trial continued until October 1992. The treated nets caused significant reduction in indoor resting density, biting (landing), light trap catches, human engorgement rate, and parous rate of malaria vector Anopheles culicifacies Giles as compared with untreated nets or no nets. Untreated nets also caused reductions in biting and indoor density. Treated nets retained insecticidal action well over 6 mo. In the final year, malaria incidence was reduced 8.9% in the no-net village, 34.9% in the villages with untreated nets, and 59.1% in villages with treated nets. The relative risk of malaria and parasite rates declined significantly in villages with treated nets. Pediatric splenomegaly rate did not change in the no-net village, increased significantly in villages with untreated nets, but decreased significantly in those with treated nets. Treated nets also reduced pediatric anemia rates, but Hb concentration increased in all villages. Considering the benefits of treated bednets and development of resistance among vectors to DDT and malathion, bednets treated with deltamethrin could be an effective alternative strategy to control malaria in forested areas in India.