Genomic Epidemiology of Treponema pallidum and Circulation of Strains With Diminished tprK Antigen Variation Capability in Seattle, 2021-2022.

BACKGROUND: The incidence of syphilis continues to increase in the United States, yet little is known about Treponema pallidum genomic epidemiology within American metropolitan areas. METHODS: We performed whole-genome sequencing and tprK deep sequencing of 28 T. pallidum-containing specimens, collected mostly from remnant Aptima swab specimens from 24 individuals from Seattle Sexual Health Clinic during 2021-2022. RESULTS: All 12 individuals infected with Nichols-lineage strains were men who have sex with men, while a specific SS14 cluster (mean, 0.33 single-nucleotide variant) included 1 man who has sex with women and 5 women. All T. pallidum strains sequenced were azithromycin resistant via 23S ribosomal RNA A2058G mutation. Identical T. pallidum genomic sequences were found in pharyngeal and rectal swab specimens taken concurrently from the same individuals. The tprK sequences were less variable between patient-matched specimens and between epidemiologically linked clusters. We detected a 528-base pair deletion in the tprK donor site locus, eliminating 9 donor sites, in T. pallidum genomes of 3 individuals with secondary syphilis, associated with diminution of TprK diversity. CONCLUSIONS: We developed an end-to-end workflow for public health genomic surveillance of T. pallidum from remnant Aptima swab specimens. tprK sequencing may assist in linking cases beyond routine T. pallidum genome sequencing. T. pallidum strains with deletions in tprK donor sites currently circulate and are associated with diminished TprK antigenic diversity.

Extension of shelf life of Nile tilapia (Oreochromis niloticus) fillets using seaweed extracts during refrigerated storage.

The effects of seaweed (Padina tetrastromatica, Sargassum natans, and Sargassum fluitans) ethanolic extracts on the quality and shelf life extension of Nile tilapia (Oreochromis niloticus) fillets were investigated during refrigerated storage for 20 days. Each of the seaweed ethanolic extracts solution (2%, w/v) was used for dipping the fish fillets for 10 min at 4°C. The control and seaweed extract-treated fillets were stored at 4 ± 1°C in air-tight polyethylene bags, and chemical, bacteriological, and sensory evaluation were performed at every 4 days' intervals. During the storage period, P. tetrastromatica extract significantly (p < .05) reduced the increment of pH, peroxide value, thiobarbituric acid reactive substances, and total volatile basic nitrogen values in Nile tilapia fillets compared to other seaweed extracts-treated and untreated fillets. The maximal total viable count of control, P. tetrastromatica, S. natans, and S. fluitans extracts-treated fillets was 6.53, 7.11, 6.75, and 7.10 log CFU/g at the 8th, 20th, 12th, and 16th days of storage, respectively. The total psychrotrophic count of control and seaweed extracts-treated fillets was also significantly increased (p < .05) throughout the storage period. The P. tetrastromatica extracts-treated fillets showed better sensory characteristics than other seaweed extracts-treated and control fillets. Results of this study suggest that ethanolic extracts (2%, w/v) of P. tetrastromatica extend the shelf life for 12 days longer than the control fillets in refrigerated conditions.

Genomic Characterization of Respiratory Syncytial Virus during 2022-23 Outbreak, Washington, USA.

We sequenced 54 respiratory syncytial virus (RSV) genomes collected during 2021-22 and 2022-23 outbreaks in Washington, USA, to determine the origin of increased RSV cases. Detected RSV strains have been spreading for >10 years, suggesting a role for diminished population immunity from low RSV exposure during the COVID-19 pandemic.

Antioxidant and antimicrobial potential of water lily extracts and their effects on the quality of frozen Nile tilapia (Oreochromis niloticus) fillets.

This study was investigated to evaluate the antioxidant and antimicrobial potential of water lily extracts and their effects on the quality of Nile tilapia (Oreochromis niloticus) fillets during frozen storage (-18 ± 1°C). Antioxidant and antimicrobial activities of water lily extracts, and chemical, microbiological, and sensory qualities of fish fillets were assessed. Results showed that the highest total phenolic content (34.07 mg GAE/g) and total flavonoid content (32.67 mg QE/g extract) were found in the ethanolic extract and the lowest in water extract of water lily. The ethanolic extracts of water lily also exhibited the highest antioxidant capacities and antimicrobial activities than other hydroethanolic and water extracts. The water lily extracts-treated fish fillets showed the highest potentiality in lowering the pH, total volatile basic nitrogen, and thiobarbituric acid reactive substances than the untreated fillets throughout the storage period. Moreover, ethanolic extracts of water lily exhibited comparatively higher efficacy in inhibiting bacterial growth in fish fillets than other extracts-treated fillets. The ethanolic extracts-treated fillets also showed better sensory attributes than hydroethanolic and control fillets. Therefore, ethanolic extract of water lily can be used as a natural preservative in enhancing the quality and prolonging the shelf life of Nile tilapia fillets during frozen storage.

ORF-Interrupting Mutations in Monkeypox Virus Genomes from Washington and Ohio, 2022.

Monkeypox virus, the causative agent of the 2022 monkeypox outbreak, is a double-stranded DNA virus in the Orthopoxvirus genus of the Poxviridae family. Genes in terminal regions of Orthopoxvirus genomes mostly code for host-pathogen interaction proteins and are prone to selective pressure and modification events. Using viral whole genome sequencing, we identified twenty-five total clinical samples with ORF-disrupting mutations, including twenty samples encoding nonsense mutations in MPXVgp001/191 (OPG001), MPXVgp004/188 (OPG015), MPXVgp010 (OPG023), MPXVgp030 (OPG042), MPXVgp159 (OPG0178), or MPXVgp161 (OPG181). Additional mutations include a frameshift leading to an alternative C-terminus in MPXVgp010 (OPG023) and an insertion in an adenine homopolymer at the beginning of the annotated ORF for MPXVgp153 (OPG151), encoding a subunit of the RNA polymerase, suggesting the virus may instead use the start codon that encodes Met9 as annotated. Finally, we detected three samples with large (>900 bp) deletions. These included a 913 bp deletion that truncates the C-terminus of MPXVgp010 (OPG023); a 4205 bp deletion that eliminates MPXVgp012 (OPG025), MPXVgp013 (OPG027), and MPXVgp014 (OPG029) and truncates MPXVgp011 (OPG024; D8L) and MPXVgp015 (OPG030); and a 6881 bp deletion that truncates MPXVgp182 (OPG210) and eliminates putative ORFs MPXVgp184, MPXVgp185 (OPG005), and MPXVgp186, as well as MPXVgp187 (OPG016), and MPXVgp188 (OPG015) from the 3' ITR only. MPXVgp182 encodes the monkeypox-specific, highly immunogenic surface glycoprotein B21R which has been proposed as a serological target. Overall, we find greater than one-tenth of our sequenced MPXV isolates have at least one gene inactivating mutation and these genes together comprised greater than one-tenth of annotated MPXV genes. Our findings highlight non-essential genes in monkeypox virus that may be evolving as a result of selective pressure in humans, as well as the limitations of targeting them for therapeutics and diagnostic testing.

De novo emergence of a remdesivir resistance mutation during treatment of persistent SARS-CoV-2 infection in an immunocompromised patient: a case report.

SARS-CoV-2 remdesivir resistance mutations have been generated in vitro but have not been reported in patients receiving treatment with the antiviral agent. We present a case of an immunocompromised patient with acquired B-cell deficiency who developed an indolent, protracted course of SARS-CoV-2 infection. Remdesivir therapy alleviated symptoms and produced a transient virologic response, but her course was complicated by recrudescence of high-grade viral shedding. Whole genome sequencing identified a mutation, E802D, in the nsp12 RNA-dependent RNA polymerase, which was not present in pre-treatment specimens. In vitro experiments demonstrated that the mutation conferred a ~6-fold increase in remdesivir IC50 but resulted in a fitness cost in the absence of remdesivir. Sustained clinical and virologic response was achieved after treatment with casirivimab-imdevimab. Although the fitness cost observed in vitro may limit the risk posed by E802D, this case illustrates the importance of monitoring for remdesivir resistance and the potential benefit of combinatorial therapies in immunocompromised patients with SARS-CoV-2 infection.

Effect of different wavelengths of LED light on the growth, chlorophyll, ß-carotene content and proximate composition of Chlorella ellipsoidea.

Chlorella ellipsoidea is a freshwater green microalga that has great prospect for the sustainable development of aquaculture industry. Microalgae require optimal lighting conditions for efficient photosynthesis. The key to cost-effective algal biomass production is to optimize algae growth conditions. This study aimed to investigate the effects of various wavelengths viz. white (380-750 nm), green (510 nm), blue (475 nm), and red (650 nm) light-emitting diodes (LEDs) on the growth, pigment content (chlorophyll-a, chlorophyll-b, and ß-carotene), and proximate composition of C. ellipsoidea with a photoperiod of 12 h:12 h light: dark cycle under indoor environmental conditions. C. ellipsoidea was cultured in Bold's Basal Medium for 18 days. The cell density (125.36×105 cells ml-1), cell dry weight (58.9 ± 4.57 mg L-1), optical density (1.66 ± 0.08 g L-1), chlorophyll-a (7.31 ± 0.04 µg ml-1), chlorophyll-b (2.73 ± 0.13 µg ml-1), and ß-carotene (0.39 ± 0.04 µg ml-1) content of C. ellipsoidea were significantly (P < 0.05) higher at 15th-day when cultured under blue LED light exposure. Significantly lower growth and nutritional values were obtained under red LED light exposure compared to the control and other LEDs spectra. In Pearson correlation analysis, the cell density and cell dry weight values showed a strong positive correlation with the values of pigment contents of C. ellipsoidea in all the treatments. The LEDs light spectra showed significant effects on proximate composition of C. ellipsoidea. Protein and lipid contents of C. ellipsoidea were significantly higher in blue LED growth conditions compared to white, green, and red LEDs. C. ellipsoidea cells were 3-7.04 µm in size and the maximum area of the cell was 38.94 µm2 in blue LED treatment. Results of this study demonstrated that blue LED light spectra was the most suitable condition to induce nutritionally rich biomass production of C. ellipsoidea, which can be used as a potential source of fish feed towards sustainable aquaculture.

Bilateral idiopathic temporal bone meningoencephaloceles - An unusual presentation.

Meningoencephaloceles (MECs) occur due to herniation of brain tissue through a bony defect in the skull base. They can be spontaneous or acquired. These are secondary to trauma, infection or neoplasia. Adult-onset spontaneous or idiopathic MECs are rare. Temporal bone MECs can present with watery discharge from the ear, conductive hearing loss or symptoms of meningitis like headache, fever, vomiting or seizures. These symptoms mimic chronic suppurative or serous otitis media. Computed tomography scan and magnetic resonance imaging differentiate between them. Awareness of this disease entity is necessary for early detection to avoid complications. We report a case of bilateral idiopathic temporal bone MECs with a rare presentation of autophony as the chief complaint. The differentiating features on computed tomography scan and magnetic resonance imaging and the surgical management are discussed.

Structure-function relationships and modifications of cardiac sarcoplasmic reticulum Ca2+-transport.

Sarcoplasmic reticulum (SR) is a specialized tubular network, which not only maintains the intracellular concentration of Ca2+ at a low level but is also known to release and accumulate Ca2+ for the occurrence of cardiac contraction and relaxation, respectively. This subcellular organelle is composed of several phospholipids and different Ca2+-cycling, Ca2+-binding and regulatory proteins, which work in a coordinated manner to determine its function in cardiomyocytes. Some of the major proteins in the cardiac SR membrane include Ca2+-pump ATPase (SERCA2), Ca2+-release protein (ryanodine receptor), calsequestrin (Ca2+-binding protein) and phospholamban (regulatory protein). The phosphorylation of SR Ca2+-cycling proteins by protein kinase A or Ca2+-calmodulin kinase (directly or indirectly) has been demonstrated to augment SR Ca2+-release and Ca2+-uptake activities and promote cardiac contraction and relaxation functions. The activation of phospholipases and proteases as well as changes in different gene expressions under different pathological conditions have been shown to alter the SR composition and produce Ca2+-handling abnormalities in cardiomyocytes for the development of cardiac dysfunction. The post-translational modifications of SR Ca2+ cycling proteins by processes such as oxidation, nitrosylation, glycosylation, lipidation, acetylation, sumoylation, and O GlcNacylation have also been reported to affect the SR Ca2+ release and uptake activities as well as cardiac contractile activity. The SR function in the heart is also influenced in association with changes in cardiac performance by several hormones including thyroid hormones and adiponectin as well as by exercise-training. On the basis of such observations, it is suggested that both Ca2+-cycling and regulatory proteins in the SR membranes are intimately involved in determining the status of cardiac function and are thus excellent targets for drug development for the treatment of heart disease.

Production of a Health-Beneficial Food Emulsifier by Enzymatic Partial Hydrolysis of Phospholipids Obtained from the Head of Autumn Chum Salmon.

Phospholipids and their partial hydrolysates, namely lysophospholipids (LPLs), have been widely used in food, pharmaceutical, and cosmetic products as highly efficient emulsifiers. This study was conducted to produce docosahexaenoic acid (DHA)-esterified LPLs by enzymatic modification of phospholipids obtained from the head of autumn chum salmon (Oncorhynchus keta). The emulsifying properties of the obtained LPLs were also evaluated. Two different types of substrates of salmon head phospholipids were prepared via silica gel and cold acetone precipitation. Enzymatic partial hydrolysis was carried out using immobilized phospholipase A1 (PLA1) and Lipozyme RM IM. Results showed that the increase in DHA in the LPLs was much higher in the silica-separated phospholipids than in the acetone-precipitated phospholipids. When silica-separated phospholipids were used as the substrate, the DHA content of the LPLs increased from 23.1% to 40.6% and 42.6% after 8 h of partial hydrolysis with Lipozyme RM IM and immobilized PLA1, respectively. The yield of the LPLs was comparatively higher in the Lipozyme RM IM than in the immobilized PLA1 hydrolysis reaction. The critical micelle concentration values of the LPLs and purified lysophosphatidylcholine (LPC) were 100 mg/L and 5 mg/L, respectively. The surface tension values of the LPLs and LPC were reduced to 30.0 mN/m and 30.5 mN/m, respectively. The hydrophilic-lipophilic balance of the LPLs and LPC were 6.0 and 9.4, respectively. Based on the emulsifying properties observed, we conclude that LPLs derived from the phospholipids of salmon head lipids could be used as a health-beneficial emulsifier in the food industry.

Intracranial EEG analysis in tumor-related epilepsy: Evidence of distant epileptic abnormalities.

OBJECTIVE: In patients with tumor-related epilepsy (TRE), surgery traditionally focuses on tumor resection; but identification and removal of associated epileptogenic zone may improve seizure outcome. Here, we study spatial relationship of tumor and seizure onset and early spread zone (SOSz). We also perform quantitative analysis of interictal epileptiform activities in patients with both TRE and non-lesional epilepsy in order to better understand the electrophysiological basis of epileptogenesis. METHODS: Twenty-five patients (11 with TRE and 14 with non-lesional epilepsy) underwent staged surgery using intracranial electrodes. Tumors were outlined on MRI and images were coregistered with post-implantation CT images. For each electrode, distance to the nearest tumor margin was measured. Electrodes were categorized based on distance from tumor and involvement in seizure. Quantitative EEG analysis studying frequency, amplitude, power, duration and slope of interictal spikes was performed. RESULTS: At least part of the SOSz was located beyond 1.5 cm from the tumor margin in 10/11 patients. Interictally, spike frequency and power were higher in the SOSz and spikes near tumor were smaller and less sharp. Interestingly, peritumoral electrodes had the highest spike frequencies and sharpest spikes, indicating greatest degree of epileptic synchrony. A complete resection of the SOSz resulted in excellent seizure outcome. CONCLUSIONS: Seizure onset and early spread often involves brain areas distant from the tumor. SIGNIFICANCE: Utilization of epilepsy surgery approach for TRE may provide better seizure outcome and study of the intracranial EEG may provide insight into pathophysiology of TRE.

False-negative rate of intraoperative frozen section margin analysis for complex head and neck nonmelanoma skin cancer excisions.

INTRODUCTION: Intraoperative frozen section analysis (IFSA) is traditionally performed for complex and high-risk non-melanoma skin cancer (NMSC) resections, particularly when surgery under a general anaesthetic and a complex reconstruction is required, and where Mohs micrographic surgery (MMS) is not available. METHODS: A retrospective audit of 253 cases between 1999 and 2009 was undertaken, investigating the accuracy and efficacy of IFSA for the treatment of NMSC in our tertiary skin tumour unit based in a university hospital setting. RESULTS: The combined incomplete and very narrow (<1 mm) excision margin rates were 28.7% and 27.5% for basal cell and squamous cell carcinoma, respectively. Unrepresentative sampling of the excision margins intraoperatively was the overwhelming cause of error (94%). CONCLUSION: After a thorough audit of our data, IFSA has been abandoned for the treatment of NMSC in our unit. MMS is practised intraoperatively, even in advanced cases. We believe that IFSA no longer has any role in our complex, multidisciplinary skin cancer practice.

Common site of urinary calculi in kidney, ureter and bladder region.

Urolithiasis is an ancient disease with global distribution. It refers to stones originating anywhere in the urinary system. Urinary calculi or stones are the most common cause of acute urinary system obstruction. The study was aimed with finding out the common site of urinary calculus in kidney ureter bladder (KUB) region. This was a prospective cross-sectional study conducted from June 2012 to September 2012 at Tribhuvan University, Teaching Hospital, Maharajgunj. A total 240 urolitiasis patients were enrolled for plain KUB examination. Site of urinary calculus was identified by observing KUB film of the subjects under the supervision of radiologist. The data were analyzed prospectively with outcome measures of gender & stone location. Out of 240 patients, 138 were male and 102 were female with male to female ratio of 1.35:1. The age ranged from 9 to 83 years. Out of total 240 patients, 71.9% (187) patients belonged to productive age group (20-60 years). Total number of urinary calculi was 345 in which 208 were found in male patients and 137 were found in female patients. Of total 345 calculi, 237 were renal stones, 47 were ureteric stones, 22 of the stones were found in pelviureteric junction (PUJ), 33 of stones were found in vesicoureteric junction (VUJ), and 6 were in bladder. In conclusion, urinary stone disease is a major public health problem in a developing country like Nepal with male and productive age group predominance. Kidney stones are most common. Distal ureter is the most common site of ureteric stone.

Safety profile of statins alone or combined with ezetimibe: a pooled analysis of 27 studies including over 22,000 patients treated for 6-24 weeks.

Aims: The aim of this analysis was to assess the overall safety and tolerability profiles of various statins + ezetimibe vs. statin monotherapy and to explore tolerability in sub-populations grouped by age, race, and sex. Methods: Study-level data were combined from 27 double-blind, placebo-controlled or active-comparator trials that randomized adult hypercholesterolemic patients to statin or statin + ezetimibe for 6-24 weeks. In the full cohort, % patients with AEs within treatment groups (statin: N = 10,517; statin + ezetimibe: N = 11,714) was assessed by logistic regression with terms for first-/second-line therapy (first line = drug-naïve or rendered drug-naïve by washout at study entry; second line = ongoing statin at study entry or statin run-in), trial within first-/second-line therapy, and treatment. The same model was fitted for age (< 65, ≥ 65 years), sex, race (white, black, other) and first-/second-line subgroups with additional terms for subgroup and subgroup-by-treatment interaction. Results: In the full cohort, the only significant difference between treatments was consecutive AST or ALT elevations ≥ 3 × upper limit of normal (ULN) (statin: 0.35%, statin + ezetimibe: 0.56%; p = 0.017). Significantly more subjects reported ≥ 1 AE; drug-related, hepatitis-related and gastrointestinal-related AEs; and CK elevations ≥ 10 × ULN (all p ≤ 0.008) in first-line vs. second-line therapy studies with both treatments. AEs were generally similar between treatments in subgroups, and similar rates of AEs were reported within age and race subgroups; however, women reported generally higher AE rates. Conclusions: In conclusion, in second-line studies, ongoing statin treatment at study entry likely screened out participants for previous statin-related AEs and tolerability issues. These results describe the safety profiles of widely used lipid-lowering therapies and encourage their appropriate and judicious use in certain subpopulations.

Morphology-based automatic seizure detector for intracerebral EEG recordings.

In this paper, a new seizure detection system aimed at assisting in a rapid review of prolonged intracerebral EEG recordings is described. It is based on quantifying the sharpness of the waveform, one of the most important electrographic EEG features utilized by experts for an accurate and reliable identification of a seizure. The waveform morphology is characterized by a measure of sharpness as defined by the slope of the half-waves. A train of abnormally sharp waves resulting from subsequent filtering are used to identify seizures. The method was optimized using 145 h of single-channel depth EEG from seven patients, and tested on another 158 h of single-channel depth EEG from another seven patients. Additionally, 725 h of depth EEG from 21 patients was utilized to assess the system performance in a multichannel configuration. Single-channel test data resulted in a sensitivity of 87% and a specificity of 71%. The multichannel test data reported a sensitivity of 81% and a specificity of 58.9%. The new system detected a wide range of seizure patterns that included rhythmic and nonrhythmic seizures of varying length, including those missed by the experts. We also compare the proposed system with a popular commercial system.

The congenital cleft nipple and its surgical treatment.

Congenital variations of the shape and structure of the nipple lacks coverage in the medical literature. We present four cases of a specific and unique congenital nipple anomaly, the cleft nipple. Each woman had a normal contralateral breast and nipple-areola complex. Three of the four cases were corrected surgically with no complications experienced. Included in the discussion is a description of the techniques used in the corrective surgery and a review of the current literature on associated variations of normality.