Reengineering the specificity of the highly selective Clostridium botulinum protease via directed evolution.

The botulinum neurotoxin serotype A (BoNT/A) cuts a single peptide bond in SNAP25, an activity used to treat a wide range of diseases. Reengineering the substrate specificity of BoNT/A's protease domain (LC/A) could expand its therapeutic applications; however, LC/A's extended substrate recognition (≈ 60 residues) challenges conventional approaches. We report a directed evolution method for retargeting LC/A and retaining its exquisite specificity. The resultant eight-mutation LC/A (omLC/A) has improved cleavage specificity and catalytic efficiency (1300- and 120-fold, respectively) for SNAP23 versus SNAP25 compared to a previously reported LC/A variant. Importantly, the BoNT/A holotoxin equipped with omLC/A retains its ability to form full-length holotoxin, infiltrate neurons, and cleave SNAP23. The identification of substrate control loops outside BoNT/A's active site could guide the design of improved BoNT proteases and inhibitors.

Future Challenges of Covid-19 and Oral Manifestations in Daily Dental Practice: A Literature Review.

OBJECTIVE: Some patients reportedly present with oral manifestations of coronavirus disease 2019 (Covid-19). It is unknown if this is due to the virus itself or a side effect of treatment; however, severe acute respiratory syndrome coronavirus 2 has been shown to have a predilection for angiotensin-converting enzyme receptors that are present in the respiratory tract, oral mucosa, tongue, and salivary glands, causing alterations in taste and smell. Therefore, the objective of this review was to present the future challenges of Covid-19 and oral manifestations in daily dental practice through a literature review. MATERIALS AND METHODS: Scientific evidence in the databases Scopus and PubMed was searched using the Boolean operators. Articles published in Spanish and English between January and December 2020 were included. RESULTS: Of the 89 articles found in Scopus, 69 were of the open access type and 20 were in the "others" category; only 22 scientific articles were found in PubMed. CONCLUSION: It is important to take into account these clinical manifestations such as loss of taste and smell in order to detect the disease. Good oral hygiene is also recommended as a preventive measure to reduce viral load, which considerably reduces the probability of infecting other people who are in contact with the infected patient.

Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score.

Effective methods for predicting COVID-19 disease trajectories are urgently needed. Here, enzyme-linked immunosorbent assay (ELISA) and coronavirus antigen microarray (COVAM) analysis mapped antibody epitopes in the plasma of COVID-19 patients (n = 86) experiencing a wide range of disease states. The experiments identified antibodies to a 21-residue epitope from nucleocapsid (termed Ep9) associated with severe disease, including admission to the intensive care unit (ICU), requirement for ventilators, or death. Importantly, anti-Ep9 antibodies can be detected within 6 days post-symptom onset and sometimes within 1 day. Furthermore, anti-Ep9 antibodies correlate with various comorbidities and hallmarks of immune hyperactivity. We introduce a simple-to-calculate, disease risk factor score to quantitate each patient's comorbidities and age. For patients with anti-Ep9 antibodies, scores above 3.0 predict more severe disease outcomes with a 13.42 likelihood ratio (96.7% specificity). The results lay the groundwork for a new type of COVID-19 prognostic to allow early identification and triage of high-risk patients. Such information could guide more effective therapeutic intervention.IMPORTANCE The COVID-19 pandemic has resulted in over two million deaths worldwide. Despite efforts to fight the virus, the disease continues to overwhelm hospitals with severely ill patients. Diagnosis of COVID-19 is readily accomplished through a multitude of reliable testing platforms; however, prognostic prediction remains elusive. To this end, we identified a short epitope from the SARS-CoV-2 nucleocapsid protein and also a disease risk factor score based upon comorbidities and age. The presence of antibodies specifically binding to this epitope plus a score cutoff can predict severe COVID-19 outcomes with 96.7% specificity.

Predicting COVID-19 Severity with a Specific Nucleocapsid Antibody plus Disease Risk Factor Score.

Effective methods for predicting COVID-19 disease trajectories are urgently needed. Here, ELISA and coronavirus antigen microarray (COVAM) analysis mapped antibody epitopes in the plasma of COVID-19 patients (n = 86) experiencing a wide-range of disease states. The experiments identified antibodies to a 21-residue epitope from nucleocapsid (termed Ep9) associated with severe disease, including admission to the ICU, requirement for ventilators, or death. Importantly, anti-Ep9 antibodies can be detected within six days post-symptom onset and sometimes within one day. Furthermore, anti-Ep9 antibodies correlate with various comorbidities and hallmarks of immune hyperactivity. We introduce a simple-to-calculate, disease risk factor score to quantitate each patients comorbidities and age. For patients with anti-Ep9 antibodies, scores above 3.0 predict more severe disease outcomes with a 13.42 Likelihood Ratio (96.7% specificity). The results lay the groundwork for a new type of COVID-19 prognostic to allow early identification and triage of high-risk patients. Such information could guide more effective therapeutic intervention.

Optimization of a laccase-mediator system with natural redox-mediating compounds for pesticide removal.

This study proposed the optimization of a laccase-mediator system to reduce pesticide levels (bentazone, carbofuran, diuron, clomazone, tebuconazole, and pyraclostrobin) on aqueous medium. Firstly, the mediator concentration of 1 mM was established (average removal of 36%). After that, seven redox-mediating compounds, namely, 2,20-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, caffeic acid, chlorogenic acid, p-coumaric acid, ferulic acid, gallic acid, protocatechuic acid, and vanillin, were compared regarding their removal efficiency. The highest removal (77%) was achieved with the laccase-vanillin system. After this screening, the optimization was carried out by a 22 full factorial design. Variables under study were the enzyme (laccase) activity and vanillin concentration. Maximum removal (53-85%) was achieved with 0.95 U/mL laccase and 1.8 mM vanillin. Pesticide removal in reaction media was fitted to the first-order kinetics equation with an average half-time life of 2.2 h. This is the first study of the use of this natural compound as a mediator in the degradation of the pesticides under investigation. The results of this study contribute, with alternative methods, to decrease pesticide levels since they are highly persistent in aqueous samples and, as a result, mitigate the environmental impact.

Factores de riesgo del estado nutricional al nacer / Risk factors of nutritional status at birth

Objetivo: Identificar los factores maternos que se asocian al estado nutricional del recién nacido según edad materna. Método: Estudio de casos y controles, con revisión de 3091 historias clínicas en el Nuevo Hospital El Milagro de Salta Capital, año 2007. Se conformó un grupo de casos integrado por neonatos con inadecuado estado nutricional y otro de control con neonatos con estado nutricional normal. Se utilizaron medidas descriptivas y el test de Chi cuadrado. Resultados: El grupo más numeroso de los neonatos fueron nacidos a término, con estado nutricional normal y peso adecuado para la edad gestacional. El mayor porcentaje fueron hijos de madres adultas (20-34 años), el 10 por ciento de añosas y el 14 por ciento de adolescentes. El índice de masa corporal pregestacional se relacionó significativamente con la ganancia ponderal. Estas dos variables junto al número de controles prenatales presentaron un marcado efecto sobre el estado nutricional del neonato (p <0,05). Las mujeres adultas tuvieron un mayor porcentaje de neonatos con estado nutricional excesivo, las añosas en déficit y las adolescentes adecuado. Sin embargo, no hubo relación significativa entre estas variables.Conclusión: La edad materna no condicionó el estado nutricional al nacer. El índice de masa corporal se relacionó significativamente con la ganancia ponderal y, a su vez, estas dos condicionaron el estado nutricional neonatal, junto al número de controles prenatales.

Objective: To identify factors that influences the nutritional status of the newborn by maternal age. Method: Case-control study, with review of 3091 medical records in the Nuevo Hospital El Milagro of Salta Capital city, 2007. A group of cases was formed comprised infants with inadequate nutrition and a control group of neonates with normal nutritional status. We used descriptive measures and Chi square test to analyze the information. Results: The largest groups of infants were born at term with normal nutritional status and weight for gestational age. The highest percentages were children of adult mothers (20-34 years), 10 per cent of older mothers and 14 per cent of adolescents. The pre-pregnancy body mass index was associated significantly and compensating with weight gain (p <0.05). And these two variables with the number of prenatal visits had a marked effect on the nutritional status of the newborn (p <0.05). Adult women had a higher percentage of infants with excessive nutritional status, the deficit older mothers and suitable, the adolescents. However, there was no significant relationship between these variables. Conclusion: Maternal age did not affect the nutritional status at birth. The pre-pregnancy body mass index was significantly associated with weight gain and in turn, these two conditioned neonatal nutritional status, with the number of prenatal visits.

Factores de riesgo del estado nutricional al nacer / Risk factors of nutritional status at birth

Objetivo: Identificar los factores maternos que se asocian al estado nutricional del recién nacido según edad materna. Método: Estudio de casos y controles, con revisión de 3091 historias clínicas en el Nuevo Hospital El Milagro de Salta Capital, año 2007. Se conformó un grupo de casos integrado por neonatos con inadecuado estado nutricional y otro de control con neonatos con estado nutricional normal. Se utilizaron medidas descriptivas y el test de Chi cuadrado. Resultados: El grupo más numeroso de los neonatos fueron nacidos a término, con estado nutricional normal y peso adecuado para la edad gestacional. El mayor porcentaje fueron hijos de madres adultas (20-34 años), el 10 por ciento de añosas y el 14 por ciento de adolescentes. El índice de masa corporal pregestacional se relacionó significativamente con la ganancia ponderal. Estas dos variables junto al número de controles prenatales presentaron un marcado efecto sobre el estado nutricional del neonato (p <0,05). Las mujeres adultas tuvieron un mayor porcentaje de neonatos con estado nutricional excesivo, las añosas en déficit y las adolescentes adecuado. Sin embargo, no hubo relación significativa entre estas variables.Conclusión: La edad materna no condicionó el estado nutricional al nacer. El índice de masa corporal se relacionó significativamente con la ganancia ponderal y, a su vez, estas dos condicionaron el estado nutricional neonatal, junto al número de controles prenatales.(AU)

Objective: To identify factors that influences the nutritional status of the newborn by maternal age. Method: Case-control study, with review of 3091 medical records in the Nuevo Hospital El Milagro of Salta Capital city, 2007. A group of cases was formed comprised infants with inadequate nutrition and a control group of neonates with normal nutritional status. We used descriptive measures and Chi square test to analyze the information. Results: The largest groups of infants were born at term with normal nutritional status and weight for gestational age. The highest percentages were children of adult mothers (20-34 years), 10 per cent of older mothers and 14 per cent of adolescents. The pre-pregnancy body mass index was associated significantly and compensating with weight gain (p <0.05). And these two variables with the number of prenatal visits had a marked effect on the nutritional status of the newborn (p <0.05). Adult women had a higher percentage of infants with excessive nutritional status, the deficit older mothers and suitable, the adolescents. However, there was no significant relationship between these variables. Conclusion: Maternal age did not affect the nutritional status at birth. The pre-pregnancy body mass index was significantly associated with weight gain and in turn, these two conditioned neonatal nutritional status, with the number of prenatal visits.(AU)

Factores de riesgo del estado nutricional al nacer / Risk factors of nutritional status at birth

Objetivo: Identificar los factores maternos que se asocian al estado nutricional del recién nacido según edad materna. Método: Estudio de casos y controles, con revisión de 3091 historias clínicas en el Nuevo Hospital El Milagro de Salta Capital, año 2007. Se conformó un grupo de casos integrado por neonatos con inadecuado estado nutricional y otro de control con neonatos con estado nutricional normal. Se utilizaron medidas descriptivas y el test de Chi cuadrado. Resultados: El grupo más numeroso de los neonatos fueron nacidos a término, con estado nutricional normal y peso adecuado para la edad gestacional. El mayor porcentaje fueron hijos de madres adultas (20-34 años), el 10 por ciento de añosas y el 14 por ciento de adolescentes. El índice de masa corporal pregestacional se relacionó significativamente con la ganancia ponderal. Estas dos variables junto al número de controles prenatales presentaron un marcado efecto sobre el estado nutricional del neonato (p <0,05). Las mujeres adultas tuvieron un mayor porcentaje de neonatos con estado nutricional excesivo, las añosas en déficit y las adolescentes adecuado. Sin embargo, no hubo relación significativa entre estas variables.Conclusión: La edad materna no condicionó el estado nutricional al nacer. El índice de masa corporal se relacionó significativamente con la ganancia ponderal y, a su vez, estas dos condicionaron el estado nutricional neonatal, junto al número de controles prenatales.(AU)

Objective: To identify factors that influences the nutritional status of the newborn by maternal age. Method: Case-control study, with review of 3091 medical records in the Nuevo Hospital El Milagro of Salta Capital city, 2007. A group of cases was formed comprised infants with inadequate nutrition and a control group of neonates with normal nutritional status. We used descriptive measures and Chi square test to analyze the information. Results: The largest groups of infants were born at term with normal nutritional status and weight for gestational age. The highest percentages were children of adult mothers (20-34 years), 10 per cent of older mothers and 14 per cent of adolescents. The pre-pregnancy body mass index was associated significantly and compensating with weight gain (p <0.05). And these two variables with the number of prenatal visits had a marked effect on the nutritional status of the newborn (p <0.05). Adult women had a higher percentage of infants with excessive nutritional status, the deficit older mothers and suitable, the adolescents. However, there was no significant relationship between these variables. Conclusion: Maternal age did not affect the nutritional status at birth. The pre-pregnancy body mass index was significantly associated with weight gain and in turn, these two conditioned neonatal nutritional status, with the number of prenatal visits.(AU)

Infarto testicular en la pubertad secundario a hemangioma intratesticular / Testicular infarction in puberty secondary to intratesticular hemangioma

Se presenta un caso de hemangioma cavernoso intratesticular en un varón de 12 años que se manifestó como aumento de volumen escrotal y dolor de 10 días de evolución. Estuvo asociado a hemorragia peritumoral e infarto hemorrágico del tumor y el testículo, lo que provocó el cuadro agudo. Se revisan seis casos adicionales de la literatura médica, tres de ellos en niños. Se concluye que el hemangioma testicular es una causa excepcional de escroto agudo en individuos de cualquier edad