Does Blacklight Illumination Improve Speed and Accuracy of Foot Pedal Activation in the Low-Light Operating Room?

Background: Urologists frequently activate foot pedals in a low-light operating room (OR). Pedal activation in low-light conditions poses the potential for incorrect pedal activation, potentially leading to increased radiation exposure, patient burns, or OR fires. This study compares speed, accuracy, dark adaptation, and surgeon preference for pedal activation in 4 lighting conditions. Materials and Methods: During a simulated percutaneous nephrolithotomy (PCNL), pedals for C-arm, laser, and ultrasonic lithotripter (USL) were randomized to 3 different positions. Urology attendings, residents, and medical students activated pedals in a randomized order in 4 settings: a dark OR with no illumination, an OR with overhead illumination, a dark OR with glowstick illumination, and a dark OR with blacklight illumination. Endpoints included pedal activation time; number of attempted, incomplete, and incorrect activations; dark adaptation; and subjective pedal preference. ANOVA was used for analysis with p < 0.05 considered significant. Results: In our study with 20 participants, the mean pedal activation times were significantly faster when using glowstick illumination (6.77 seconds) and blacklight illumination (5.34 seconds) compared with the no illumination arm (8.47 seconds, p < 0.001). Additionally, individual pedal activations for the C-arm, laser, and USL were significantly faster with glowstick and blacklight illumination compared with a dark OR (p < 0.001 for all). The blacklight illumination arm demonstrated decreased attempted (0.30 vs. 3.45, p < 0.001), incomplete (1.25 vs. 7.75, p < 0.001), and incorrect activations (0.35 vs. 1.25, p < 0.001) compared with the dark setting, while demonstrating no difference compared with having room lights on. Dark adaptation was significantly improved with blacklight illumination compared with having the room lights on (134.5 vs. 140.5 luminance, p < 0.001). All participants (100%) preferred illuminated pedals compared with the dark OR, with 90% favoring the blacklight illumination. Conclusions: During a simulated PCNL, blacklight illumination significantly improved accuracy and efficiency of pedal activation compared with the conventional dark OR, while maintaining the surgeon's dark adaptation.

Relationship between perceived physical literacy and obesity-related outcomes in adolescents: the EHDLA study.

Purpose: The aim of the present study was to examine the relationship between perceived physical literacy and obesity-related outcomes among adolescents from Spain. Methods: This is a secondary cross-sectional analysis including a total sample of 845 Spanish adolescents (55.3% girls) aged 12-17 years from the Valle de Ricote (Region of Murcia) from the Eating Healthy and Daily Life Activities (EHDLA) project. Physical literacy was evaluated using the Spanish Perceived Physical Literacy Instrument for adolescents (S-PPLI). Body mass index was computed by taking the participants' body weight in kilograms and dividing it by the square of their height in meters, and body mass index (z score) and overweight/obesity and obesity were computed by the World Health Organization age- and sex-specific thresholds. Waist circumference was measured using a constant tension tape. Moreover, the waist-to-height ratio was calculated, and therefore, abdominal obesity was determined. Skinfold measurements were taken at the triceps and medial calf using calibrated steel calipers. Results: In general, the overall trend was downward (i.e., the higher the PPLI score the lower the obesity-related indicators), with the approximate significance of smooth terms being statistically significant for all models examined (p < 0.001). Adolescents with lower perceived physical literacy (PPL) showed the highest estimated marginal means of body mass index, body mass index z score, waist circumference, waist-to-height ratio, and skinfold (triceps and calf) and predictive probabilities of having excess weight, obesity, and abdominal obesity, while their counterparts with high PPL had the lowest. In addition, significant differences were observed for all the obesity-related indications between adolescents with low PPL and those with medium PPL (p-adjusted < 0.05 for all indicators), as well as with those with high PPL (p-adjusted < 0.05 for all indicators). Moreover, these significant differences were also shown for most indicators between adolescents with medium PPL and those with high PPL (except for obesity). Conclusion: Physical literacy could play a crucial role in maintaining more desirable obesity-related outcomes in adolescents. Adolescents with high perceived physical literacy showed lower obesity-related indicators (i.e., body mass index, body mass index z score, waist circumference, waist-to-height ratio, skinfolds), as well as a lower probability of having excess weight, obesity, and abdominal obesity.

Total mercury and selenium levels in commercial shrimp along the Pacific coast of Mexico.

The present study analyzed the content of total mercury (THg) and selenium (Se) in the muscle of shrimp collected from local markets in the 11 Pacific coastal states of Mexico. Methylmercury (MeHg) concentration, Se:Hg ratio, health benefits value from selenium consumption (HBVSe) and the permissible weekly consumption were estimated to assess the health risk to consumers. All THg and Se concentrations were below the maximum permissible limits. All hazard quotient (HQ) values were <1, however in Hermosillo, Culiacán and Guadalajara, the Se:Hg ratio and HBVSe were <1 and negative, due to the low concentrations of Se. As a general conclusion, there is no risk nor benefit from the consumption of shrimp from the Pacific coast of Mexico due to its Hg and Se content.

Exploring the link between perceived physical literacy and academic performance outcomes: insights from the EHDLA study.

Objective: The aim of this study was to verify the relationship between perceived physical literacy (PPL) and academic performance outcomes among Spanish adolescents aged 12-17 years. Methods: This cross-sectional study is a secondary examination utilizing data derived from the Eating Healthy and Daily Life Activities (EHDLA) project. The Spanish Perceived Physical Literacy Instrument for Adolescents (S-PPLI) was used to evaluate the PPL of the participants. To determine academic performance, the numerical grades for each subject were added together and then divided by the total number of subjects. Results: Overall, a positive trend in the association between S-PPLI scores and academic performance was observed. We identified two significant ranges within the S-PPLI scores. First, there was a significant range between 9 and 33 points on the S-PPLI, indicating an inverse association with academic performance. Second, another significant range was observed at scores of 34 points or higher on the S-PPLI, suggesting a positive association with academic performance. Participants with low PPL had the lowest academic performance (mean = 6.4; bias-corrected and accelerated (BCa) bootstrapped 95% confidence interval (CI): 6.2-6.6). Conversely, those adolescents with high PPL had the highest academic performance (mean = 6.9; BCa bootstrapped 95% CI: 6.6-7.1). Significant differences were found between low PPL and medium PPL (p-adjusted = 0.031) and between low PPL and high PPL (p-adjusted = 0.031). Conclusions: Increased physical literacy could be a relevant factor for achieving greater academic performance in adolescents. This study has potential implications for physical education instructors, school leaders, and healthcare practitioners.

Evaluation of usability, adherence, and clinical efficacy of therapeutic footwear in persons with diabetes at moderate to high risk of diabetic foot ulcers: A multicenter prospective study.

OBJECTIVE: To evaluate therapeutic footwear expectations and usability of individuals with diabetes and foot complications. DESIGN: A prospective multicenter study was conducted on participants with a high risk of developing a diabetic foot ulcer. SETTING: Participants were enrolled in 11 different specialized diabetic foot units in Spain between March 2022 and June 2023. SUBJECTS: Patients with diabetes at moderate to high risk of foot ulceration receiving first therapeutic footwear prescription. INTERVENTIONS: All the patients included in the research were prescribed with their first pair of therapeutic footwear. MAIN MEASURES: Primary outcome measures were MOS-pre and MOS-post questionnaires evaluating use and usability of prescribed therapeutic footwear. Secondary outcome measures aimed to evaluate footwear clinical efficacy as ulceration rate and self-reported perceived walking distance per day. RESULTS: The use of therapeutic footwear exceeded the patient's pre-provision prediction of their anticipated use in 94% of people (n = 126). Based on the visual analogic satisfaction scale, the median satisfaction of daily wearing their therapeutic footwear was 7 points, Interquartile Range (IQR) [5-8.25]. During the follow-up period, 39 participants (29.1%) experienced diabetic foot ulcer. Perceived walking distance participants reported an improvement in their perceived walking ability during various daily life activities. CONCLUSIONS: Diabetes patients at moderate to high risk of diabetic foot ulcer improved their perception of walking ability after therapeutic footwear prescription. Adherence to the therapeutic footwear prescription resulted in less ulcerations.

What factors influence the rediscovery of lost tetrapod species?

We created a database of lost and rediscovered tetrapod species, identified patterns in their distribution and factors influencing rediscovery. Tetrapod species are being lost at a faster rate than they are being rediscovered, due to slowing rates of rediscovery for amphibians, birds and mammals, and rapid rates of loss for reptiles. Finding lost species and preventing future losses should therefore be a conservation priority. By comparing the taxonomic and spatial distribution of lost and rediscovered tetrapod species, we have identified regions and taxa with many lost species in comparison to those that have been rediscovered-our results may help to prioritise search effort to find them. By identifying factors that influence rediscovery, we have improved our ability to broadly distinguish the types of species that are likely to be found from those that are not (because they are likely to be extinct). Some lost species, particularly those that are small and perceived to be uncharismatic, may have been neglected in terms of conservation effort, and other lost species may be hard to find due to their intrinsic characteristics and the characteristics of the environments they occupy (e.g. nocturnal species, fossorial species and species occupying habitats that are more difficult to survey such as wetlands). These lost species may genuinely await rediscovery. However, other lost species that possess characteristics associated with rediscovery (e.g. large species) and that are also associated with factors that negatively influence rediscovery (e.g. those occupying small islands) are more likely to be extinct. Our results may foster pragmatic search protocols that prioritise lost species likely to still exist.

Retinal shadows produced by myopia control spectacles.

PURPOSE: To analyse ocular coherence tomography (OCT) images of the retinal shadows caused by defocus and diffusion optics spectacles. METHODS: One eye was fitted successively with the Hoya Defocus Incorporated Multiple Segments (DIMS) spectacle lens, two variations of the +3.50 D peripheral add spectacle (DEFOCUS) and the low-contrast dot lens (Diffusion Optics Multiple Segments, DOMS); each at a vertex distance of 12 mm. Simultaneously, a retinal image of the macular region with central fixation was obtained using infrared OCT. The corneal power and intraocular distances were determined using an optical biometer. RESULTS: The retinal images for the DIMS and DOMS lenses showed patterns of obvious retinal shadows in the periphery, while the central 10-11° remained clear. The DEFOCUS lens produced a darkened peripheral area. Dividing the size of the retinal pattern, measured with the calliper of the OCT software, by the actual size on the spectacle lens gave a magnification of -0.57 times. This is consistent with the incoming OCT beam being imaged to a position approximately 31 mm beyond the front of the eye. [Correction added on 26 October 2023 after first online publication: The preceding paragraph was corrected.] CONCLUSION: With device-specific correction, retinal OCT images can help visualise the regions affected by the defocus or lowered contrast induced by myopia control spectacles. This is of potential value for improving myopia therapies.

Cross-Sectional Association between Perceived Physical Literacy and Mediterranean Dietary Patterns in Adolescents: The EHDLA Study.

PURPOSE: The objective of the current study was to analyze the relationship between perceived physical literacy (PPL) and adherence to the Mediterranean Diet (MedDiet) and its specific components in a sample of Spanish adolescents. METHODS: In this cross-sectional study, we examined a sample of 845 adolescents (58.1% boys) aged 12-17 years from the Valle de Ricote (Region of Murcia, Spain). PPL was assessed by the Spanish Perceived Physical Literacy Instrument for adolescents (S-PPLI). To assess adherence to the MedDiet, the Mediterranean Diet Quality Index in children and adolescents (KIDMED) was applied. RESULTS: In unadjusted analyses, adolescents with medium or high PPL were more likely to have optimal adherence to the MedDiet (medium PPL: OR = 1.69, 95% CI 1.20-2.40, p-adjusted = 0.003; high PPL: OR = 2.90, 95% CI 2.03-4.17, p-adjusted < 0.001). These findings remained significant after adjusting for sex, age, socioeconomic status, body mass index, energy intake, overall sleep duration, physical activity, and sedentary behavior (medium PPL: OR = 1.51, 95% CI 1.05-2.19, p-adjusted = 0.003; high PPL: OR = 2.27, 95% CI 1.53-3.39, p-adjusted < 0.001). CONCLUSIONS: PPL could play a relevant role in the adoption of healthy eating habits among adolescents. Adolescents with low or medium PPL were less likely to report optimal adherence to the MedDiet. Adolescents with high PPL seem to consume more fruits, vegetables, fish, pulses, nuts, and dairies (for breakfast). Conversely, these adolescents tend to skip breakfast less, as well as to consume less commercially baked goods or pastries at this meal.

Natural life cycle of Versteria cuja (Taeniidae) in Argentina and histopathology of metacestodiasis in intermediate hosts.

Using morphological and molecular studies, the life cycle of Versteria cuja (Cestoda: Taeniidae) was elucidated, involving subterranean rodents (Ctenomyidae) as intermediate hosts, and the lesser grison, Galictis cuja (Mustelidae), as definitive host. Metacestodes (cysticerci and polycephalic larvae) were found mainly in the liver but also in spleen, pancreas, lungs and small intestine of 2 species of tuco-tucos (Ctenomys spp.) from Chubut, Argentina. Identity of the metacestodes with the adult was based primarily on the number, size and shape of rostellar hooks: 40­48 hooks in 2 rows, particularly small (10­16 µm total length by 6­10 µm wide), composed of handle, blade and guard with characteristic shapes. Genetic analysis (cox1 gen mtDNA) performed on metacestodes from both intermediate hosts corroborated their conspecificity with adults of V. cuja from lesser grisons in the same locality. Histopathological study showed the hepatic parenchyma altered by the presence of cysts containing larvae, each surrounded by a capsule of connective tissue with inflammatory infiltrate, atrophied hepatocytes and an increase of bile ducts. In the lung, in addition to the cysts, dilated alveoli, oedema and hyperaemic blood vessels were observed. This is the first report of a natural life cycle of a Versteria species from South America. It shows strong similarities with that described for a North American zoonotic lineage of Versteria, confirming a close relationship between V. cuja and this North American lineage, as previously demonstrated by molecular studies. Consequently, the zoonotic potential of V. cuja should not be disregarded.

Total Mercury and Selenium in wild Shrimp from Coastal Lagoons of Northwest Mexico: Human Health risk Assessment.

This study analyzed total mercury (THg), and selenium (Se) in edible tissues of white shrimp (Litopenaeus vannamei), blue shrimp (L. stylirostris) and brown shrimp (F. californiensis), from three states of the Northwest of Mexico in September and October 2017. Concentrations of THg and Se in the muscle were between 0.026 and 0.829 and 0.126-1.741 µg/g dry weight (dw), respectively. Significant differences were observed among Hg concentration of Sonora and Nayarit and among Se concentration of Sinaloa and Nayarit. In addition, the health risk assessment (HQ) in the three species of shrimp was between 0.550 and 0.607. All Se:Hg molar ratios were > 1 and positive HBVSe values that showed that shrimp from Northwest of Mexico does not represent a risk to human health.

One-year breakthrough SARS-CoV-2 infection and correlates of protection in fully vaccinated hematological patients.

The long-term clinical efficacy of SARS-CoV-2 vaccines according to antibody response in immunosuppressed patients such as hematological patients has been little explored. A prospective multicenter registry-based cohort study conducted from December 2020 to July 2022 by the Spanish Transplant and Cell Therapy group, was used to analyze the relationship of antibody response over time after full vaccination (at 3-6 weeks, 3, 6 and 12 months) (2 doses) and of booster doses with breakthrough SARS-CoV-2 infection in 1551 patients with hematological disorders. At a median follow-up of 388 days after complete immunization, 266 out of 1551 (17%) developed breakthrough SARS-CoV-2 infection at median of 86 days (range 7-391) after full vaccination. The cumulative incidence was 18% [95% confidence interval (C.I.), 16-20%]. Multivariate analysis identified higher incidence in chronic lymphocytic leukemia patients (29%) and with the use of corticosteroids (24.5%), whereas female sex (15.5%) and more than 1 year after last therapy (14%) were associated with a lower incidence (p < 0.05 for all comparisons). Median antibody titers at different time points were significantly lower in breakthrough cases than in non-cases. A serological titer cut-off of 250 BAU/mL was predictive of breakthrough infection and its severity. SARS-CoV-2 infection-related mortality was encouragingly low (1.9%) in our series. Our study describes the incidence of and risk factors for COVID-19 breakthrough infections during the initial vaccination and booster doses in the 2021 to mid-2022 period. The level of antibody titers at any time after 2-dose vaccination is strongly linked with protection against both breakthrough infection and severe disease, even with the Omicron SARS-CoV-2 variant.

Biomonitoring of potentially toxic elements through oysters (Saccostrea palmula and Crassostrea corteziensis) from coastal lagoons of Southeast Gulf of California, Mexico: health risk assessment.

The coastal lagoons of the Gulf of California support important traditional fisheries and mollusc cultures (generally oysters) and receive important volumes of agricultural, industrial and urban effluents, consumption of the oysters could pose risk to human health. The concentrations of arsenic (As), cadmium (Cd), copper (Cu), iron (Fe), lead (Pb), and zinc (Zn) in the oysters Saccostrea palmula and Crassostrea corteziensis, from four coastal lagoons (Altata, AL; Macapule, ML; Navachiste, NL; El Colorado, ECL) in the Southeast Gulf of California, were seasonally evaluated (summer 2019-spring 2020). The order of magnitude of potentially toxic elements concentrations in the soft tissue in both oyster species and at all sites was Zn > Fe > Cu > As > Cd > Pb. Cadmium, Cu, Pb, and Zn exceeded the maximum permissible limits in more than one sampling site. The highest concentrations (mg kg-1, wet weight) of As (4.2 ± 1.1, spring) and Cd (3.3 ± 0.7, autumn) were registered in S. palmula et al. and NL sampling sites, respectively. Crassostrea corteziensis presented higher levels of Cu (40.5 ± 6.7, spring), Pb (2.0 ± 0.4, spring), and Zn (96.9 ± 20.4, spring) in ECL and Fe (62.2 ± 25.4, autumn) in ML. The hazard quotient (HQ) values exceeded the safe level of 1 for Cd in S. palmula and C. corteziensis in NL for children (~ 16 kg weight). In addition, in children, the hazard index (HI) values in both species of oysters ranged from 0.7 to 2.1 and 0.6 to 1.9, respectively. On the other hand, the intake of the studied elements through the consumption of oysters would not induce adverse effects to human health (men and women weighing 70 and 60 kg, respectively); HQ and HI values were < 1.

Vestibular Drop Attack: An Analysis of the Therapeutic Response.

The present study evaluates the response to betahistine in patients who presented vestibular drops attacks in the context of Ménière's disease (MD) and the factors that can predict an unfavorable response to it. A total of 43 patients were analyzed, out of which 33 were diagnosed with MD. This is a descriptive, cross-sectional study with retrospective data collection. Data as regards age, accompanying symptoms, etiological diagnosis and response to MD treatment were collected. A statistical analysis was carried out, and we found that the disease evolution time and specific alterations in the vestibulospinal and oculomotor physical examination present an unfavorable response to betahistine. Failures for betahistine were treated with intratympanic gentamicin, with which symptomatic control was achieved in all cases.

Trace Metals in Two Geoduck Clams (Panopea generosa and P. Globosa) Exploited for the Regional Market from Two Areas of Northwest Mexico.

Biological and fishery features of Panopea sp. clams have been studied in northwest Mexico because of their importance for human consumption. However, the content of pollutants in their tissues, along with their implications have not been addressed yet. The concentrations of cadmium (Cd), copper (Cu), mercury (Hg), and zinc (Zn) in soft tissues of Panopea generosa and P. globosa clams were examined in this region by atomic absorption spectroscopy. The metal concentrations (µg/g wet weight) ranged from 6.5 to 14.2, 0.97-8.09, 0.60-1.18, and 0.01-0.07, for Zn, Cu, Cd, and Hg, respectively. This study proposes that metal presence is related to weathering, upwelling, and drainage from adjacent agricultural lands to the coast. According to the Official Mexican Standard (NOM-242-SSA1-2009) and the World Health Organization (WHO, 2022), the metal content in siphon tissue indicates safe levels for human consumption.

Cadmium and copper mixture effects on immunological response and susceptibility to Vibrio harveyi in white shrimp Litopenaeus vannamei.

Cadmium (Cd2+) and copper (Cu2+) are considered immunotoxic metals and their presence in combination in the aquatic environment may cause effects on shrimp species as Litopenaeus vannamei. Thus, this research evaluates the combined effects of Cd2+ and Cu2+ on shrimp inoculated with Vibrio harveyi bacteria. The experiments were performed at 96-h of exposure to sublethal concentrations of both metals. No mortality was observed in organisms exposed to the sum of Criterion of Continuous Concentration (ΣCCC) in Cd + Cu mixture and those inoculated with V. harveyi. Higher clotting times were recorded in Cd + Cu + V. harveyi treatment at higher metal concentrations. No significant differences (P > 0.05) were recorded in hemocyanin content between shrimp exposed to metals and those experimentally infected. Significantly higher (P < 0.05) total hemocyte count (THC) was recorded at 96 h exposure in the ΣCCC and 10% treatments of Cd + Cu + V. harveyi experiment. Regarding Cd + Cu + V. harveyi bioassay, the highest phenoloxidase (PO) activity was recorded in shrimp inoculated with V. harveyi (0.326 ± 0.031 PO units/mg protein) at 96-h exposure. The lowest PO activity was observed in organisms exposed to Cd + Cu + V. harveyi. Regarding superoxide dismutase (SOD) activity, shrimp exposed to higher metal concentrations at 96 h showed the lowest hemolymph activity (6.03 ± 0.62 SOD units/mL). Protein decrease was observed in organisms exposed to metal mixture. The results showed that L. vannamei could be more susceptible to V. harveyi when exposed to Cd + Cu.

A new species of Versteria (Cestoda: Taeniidae) parasitizing Galictis cuja (Carnivora: Mustelidae) from Patagonia, Argentina: Morphological and molecular characterization.

Via morphological and molecular analysis, we describe a new species of taeniid from Patagonia (Argentina): Versteria cuja n. sp., parasitizing the Lesser grison Galictis cuja (Molina) (Carnivora: Mustelidae). This is the first report of a species of Versteria in Argentina and for a native mustelid. The new species (the third in the genus Versteria) is proposed using an integrative taxonomic approach, based on traditional morphology (distinctive morphological and morphometric diagnostic characters), genetic distances and phylogeny based on molecular data, the distinct geographical distribution, and the different definitive host species. Versteria cuja n. sp. mainly differs from Versteria mustelae (Gmelin, 1790) (from Europe) in the number of testes (54-85 vs. 83-127 in V. mustelae), the rostellum size (39-75 vs. 85-180 µm in V. mustelae), the genital atrium size (170-420 vs. 68-91 µm in V. mustelae) and in the hooks' shape. It also differs from the African species Versteria brachyacantha (Baer and Fain, 1951) by having smaller measurements regarding the main diagnostic characters, i.e. size of scolex, rostellum and suckers, number, size and shape of rostellar hooks, number of testes, and by having smooth cirrus (vs. cirrus covered with hair-like bristles in V. brachyacantha). Phylogenetical analysis using cox1 showed our specimens clustering with North American isolates of Versteria sp. in a well-supported American clade (mean genetic divergence 0.024), separated from another clade composed of different isolates of V. mustelae (0.093). The close relationship between the new species and the North American species, known as "zoonotic" Versteria sp., and responsible for fatal infections by metacestodes in free-ranging wildlife (rodents), captive primates and immunosuppressed people, lead us to think that the zoonotic potential of Versteria cuja n. sp. should not be discarded.

SARS-CoV-2 vaccine response and rate of breakthrough infection in patients with hematological disorders.

BACKGROUND: The clinical efficacy of SARS-CoV-2 vaccines according to antibody response in immunosuppressed patients such as hematological patients has not yet been established. PATIENTS AND METHODS: A prospective multicenter registry-based cohort study conducted from December 2020 to December 2021 by the Spanish transplant and cell therapy group was used to analyze the relationship of antibody response at 3-6 weeks after full vaccination (2 doses) with breakthrough SARS-CoV-2 infection in 1394 patients with hematological disorders. RESULTS: At a median follow-up of 165 days after complete immunization, 37 out of 1394 (2.6%) developed breakthrough SARS-CoV-2 infection at median of 77 days (range 7-195) after full vaccination. The incidence rate was 6.39 per 100 persons-year. Most patients were asymptomatic (19/37, 51.4%), whereas only 19% developed pneumonia. The mortality rate was 8%. Lack of detectable antibodies at 3-6 weeks after full vaccination was the only variable associated with breakthrough infection in multivariate logistic regression analysis (Odds Ratio 2.35, 95% confidence interval 1.2-4.6, p = 0.012). Median antibody titers were lower in cases than in non-cases [1.83 binding antibody units (BAU)/mL (range 0-4854.93) vs 730.81 BAU/mL (range 0-56,800), respectively (p = 0.007)]. We identified 250 BAU/mL as a cutoff above which incidence and severity of the infection were significantly lower. CONCLUSIONS: Our study highlights the benefit of developing an antibody response in these highly immunosuppressed patients. Level of antibody titers at 3 to 6 weeks after 2-dose vaccination links with protection against both breakthrough infection and severe disease for non-Omicron SARS-CoV-2 variants.

Enhanced Detection of Viral RNA Species Using FokI-Assisted Digestion of DNA Duplexes and DNA/RNA Hybrids.

The accurate detection of nucleic acids from certain biological pathogens is critical for the diagnosis of human diseases. However, amplified detection of RNA molecules from a complex sample by direct detection of RNA/DNA hybrids remains a challenge. Here, we show that type IIS endonuclease FokI is able to digest DNA duplexes and DNA/RNA hybrids when assisted by a dumbbell-like fluorescent sensing oligonucleotide. As proof of concept, we designed a battery of sensing oligonucleotides against specific regions of the SARS-CoV-2 genome and interrogated the role of FokI relaxation as a potential nicking enzyme for fluorescence signal amplification. FokI-assisted digestion of SARS-CoV-2 probes increases the detection signal of ssDNA and RNA molecules and decreases the limit of detection more than 3.5-fold as compared to conventional molecular beacon approaches. This cleavage reaction is highly specific to its target molecules, and no detection of other highly related B-coronaviruses was observed in the presence of complex RNA mixtures. In addition, the FokI-assisted reaction has a high multiplexing potential, as the combined detection of different viral RNAs, including different SARS-CoV-2 variants, was achieved in the presence of multiple combinations of fluorophores and sensing oligonucleotides. When combined with isothermal rolling circle amplification technologies, FokI-assisted digestion reduced the detection time of SARS-CoV-2 in COVID-19-positive human samples with adequate sensitivity and specificity compared to conventional reverse transcription polymerase chain reaction approaches, highlighting the potential of FokI-assisted signal amplification as a valuable sensing mechanism for the detection of human pathogens.

Comparison of in-house SARS-CoV-2 genome extraction procedures. A need for COVID-19 pandemic.

Among the methods used to diagnose COVID-19, those based on genomic detection by q(RT)-PCR are the most sensitive. To perform these assays, a previous genome extraction of the sample is required. The dramatic increase in the number of SARS-CoV-2 detection assays has increased the demand for extraction reagents hindering the supply of commercial reagents. Homemade reagents and procedures could be an alternative. Nasopharyngeal samples were extracted by seven different methods as well as the automatic method MagNaPure96, to detect SARS-CoV-2. All protocols show sensitivity higher than 87 %, in comparison with reference method, for detecting SARS-CoV-2 as well as human ß- globin. Our results support that these procedures, using common and cheap reagents, are effective to extract RNA (from SARS-CoV-2) or DNA (from human ß-globin) genome from nasopharyngeal swabs. Furthermore, these procedures could be easily adopted by routine diagnostic laboratories to implement detection methods to help to fight against COVID-19 pandemic.