Estudio comparativo entre un test de antígeno y la prueba de la reacción en cadena de la polimerasa en tiempo real (RT-PCR) para el diagnóstico de infección por COVID-19 en pediatría / Comparative study between an antigen test and the reverse transcription-polymerase chain reaction (RT-PCR) test for the diagnosis of COVID-19 in pediatrics

Introducción. La pandemia por COVID-19 ha puesto de manifiesto la necesidad de pruebas diagnósticas rápidas. La prueba de referencia es la reacción en cadena de la polimerasa en tiempo real (RT-PCR). Requiere un equipo y personal capacitado, y su resultado puede llevar un tiempo de espera prolongado. El sistema BD Veritor® es el método rápido cromatográfico utilizado para la detección del antígeno del coronavirus de tipo 2 del síndrome respiratorio agudo grave, en individuos sintomáticos. El objetivo primario del siguiente trabajo es evaluar sensibilidad y especificidad del test de antígeno (TA) comparadas con la RT-PCR en población pediátrica. Población y métodos. Estudio prospectivo, de prueba diagnóstica. Se incluyó a todo menor de 17 años en los primeros 5 días de inicio de síntomas, que consultó desde julio de 2021 hasta febrero de 2022. Se calculó un mínimo de 300 muestras para lograr una precisión de ± 8,76 % y de ± 3,68 % para sensibilidad y especificidad respectivamente. Se analizaron en paralelo las muestras por ambas metodologías. Resultados. De 316 muestras pareadas, 33 fueron positivas por ambos métodos; 6 fueron positivas solo por RT-PCR. La especificidad del TA fue del 100 %; la sensibilidad, del 84,6 %, con un valor predictivo positivo y negativo del 100 % y del 98 % respectivamente. Conclusiones. El TA demostró ser útil en el diagnóstico de pacientes pediátricos con COVID-19 en los primeros 5 días de inicio de síntomas, aunque aquellos con TA negativo y alta sospecha clínica deberían confirmar su resultado con la RT-PCR.

Introduction. The COVID-19 pandemic has brought to light the need for rapid diagnostic tests. The gold standard test is reverse transcription-polymerase chain reaction (RT-PCR). RT-PCR requires equipment and trained personnel, and results may take a long waiting time. The BD Veritor® System is a rapid chromatographic method used for the detection of severe acute respiratory syndrome coronavirus 2 antigen in symptomatic individuals. The primary objective of this study is to assess the sensitivity and specificity of the antigen test (AT) compared to the RT-PCR in the pediatric population. Population and methods. Prospective study with a diagnostic test. All children younger than 17 years in the first 5 days of symptom onset, who consulted between July 2021 and February 2022, were included. A minimum of 300 specimens was estimated to achieve an accuracy of ±8.76% and ±3.68% for sensitivity and specificity, respectively. Specimens were analyzed in parallel using both methodologies. Results. Of 316 paired samples, 33 were positive by both methods; 6 were positive only by RT-PCR. The specificity of the AT was 100%; sensitivity was 84.6%, with a positive and negative predictive value of 100% and 98%, respectively. Conclusions. The AT proved to be useful in the diagnosis of pediatric patients with COVID-19 in the first 5 days of symptom onset, although those with a negative AT and high clinical suspicion should confirm their result with a RT-PCR.

Study of long non-coding RNA Tug 1 expression in Egyptian colorectal adenocarcinoma patients

Purpose: Colorectal cancer (CRC) is one of the most fatal tumors worldwide. In Egypt, most CRC cases occur in individuals > 40 years old. TUG1 has been proved to be disrupted in different malignancies and may have a critical role in tumor progression, invasion, and metastasis. However, its role in CRC has not been adequately studied. Materials / Methods: Quantitative real-time polymerase chain reaction (PCR) was used to evaluate the expression levels of long non-coding RNA (LncRNA) taurine upregulated gene 1 (TUG1), in nonmetastatic and metastatic CRC tissues and adjacent noncancerous tissues as control. Results: LncRNA TUG1 expression was significantly upregulated in both nonmetastatic and metastatic CRC tissues, in comparison with the adjacent noncancerous tissue. It was found that TUG1 could have a possible prognostic role in CRC, by comparing the sensitivity and specificity of TUG1 with those of CEA and CA19-9. Conclusion: The results of the current study suggest that the LncRNA TUG1 participates in the malignant behaviors of CRC cells. (AU)

Risque de transmission de certains arbovirus à travers le don de sang à Bamako / Risk of transmission of certain arbovirus through blood donation in the city of Bamako

Introduction. Au Mali, le dépistage de certains virus tels que la dengue, Zika et la fièvre de la vallée du Rift n'est pas systématique au centre national de transfusion sanguine (CNTS). Le risque peut être considérable en raison de leurs courtes périodes de virémie asymptomatique dans la population dont l'incidence est variable et parfois extrêmement élevée. Cette étude avait pour objectif d'explorer la possibilité de transmission de certains arbovirus à travers le don de sang au CNTS de Bamako. Méthodes. Il s'agissait d'une étude transversale, de juillet 2019 à juin 2020 à Bamako. Au total deux cents (200) donneurs de sang du CNTS ont été inclus. Les examens ont été réalisés au Centre d'Infectiologie Charles Mérieux (CICM) de Bamako avec le dépistage du génome des virus responsables de la Dengue, de la fièvre de la Vallée du Rift, et du Zika à l'aide de la technique de la RT-PCR en temps réel. Le Test de Dépistage Rapide (TDR) a été utilisé pour la détection des anticorps IgG et IgM spécifiques de la Dengue. Résultats. Le sexe masculin représente 84% (168/200). Le TDR a détecté 4,5% (9/200) de Dengue IgG positifs et aucun cas de Dengue IgM positif. La technique de RT-PCR n'a détecté aucun des trois virus. Conclusion. Cette étude prouve que le risque de transmission de certains arbovirus à travers le don de sang existe, mais il semble être minime au CNTS de Bamako

Background. In Mali, screening for certain viruses such as dengue, Zika, and Rift Valley fever is not systematic at the national blood transfusion center (CNTS). The risk can be considerable due to their short periods of asymptomatic viremia in the population with variable and sometimes extremely high incidence. The objective of this study was to explore the possibility of transmission of certain arboviruses through blood donation at the CNTS of Bamako. Methods. This was a cross-sectional study, from July 2019 to June 2020 in Bamako. A total of two hundred (200) blood donors from the CNTS were included. The examinations were performed at the Centre d'Infectiologie Charles Mérieux (CICM) in Bamako with the screening of the genome of viruses responsible for Dengue, Rift Valley fever, and Zika using the real-time RT-PCR technique. The Rapid Screening Test (RST) was used for the detection of Dengue-specific IgG and IgM antibodies. Results. Male sex represented 84% (168/200). The RDT detected 4.5% (9/200) of IgG positive Dengue and no IgM positive Dengue cases. The RT-PCR technique did not detect any of the three viruses. Conclusion. This study proves that the risk of transmission of certain arboviruses through blood donation exists, but it seems to be minimal at the CNTS of Bamako.

SARS-CoV-2 RT-PCR Ct Value and Laboratory Tests: Clinicopathologic characteristics among adult Filipino Inpatients diagnosed with COVID-19 in a tertiary medical center

INTRODUCTION@# The role of the laboratory during the COVID-19 pandemic is not limited to just diagnosis of the disease, but also in clinical decision-making, by providing information on relevant laboratory biomarkers. Clinicians also use Ct value to guide patient management. There are limited studies available locally regarding the significance of Ct value and pertinent laboratory biomarkers in COVID-19 patients. This study aimed to assess the aforementioned laboratory data, along with the clinicopathologic characteristics of affected patients, and determined if this information may be useful for robust clinical decision-makin@*METHODOLOGY@#In this retrospective analytic study, we identified 325 out of 1,049 adult Filipino inpatients diagnosed with COVID-19 and analyzed their Ct values and pertinent laboratory biomarkers such as neutrophil and lymphocyte count, platelet count, LDH, ferritin, procalcitonin, CRP, AST/SGOT, ALT/SGPT, PT/ INR, and D-dimer, and correlated them with the severity of the disease.@*RESULTS@#Two hundred twenty (67.7%) patients had non-severe disease, while 105 (32.3%) had severe disease. Lower Ct values of ORF1ab (median = 26.4) and N (median = 24.8) genes were seen in the severe group compared to the non-severe group and were found to be significant (p<0.001). Laboratory markers (neutrophil, platelet counts, LDH, ferritin, procalcitonin, CRP, AST, PT/INR, and D-dimer) were associated with severe COVID-19. On the other hand, ALT was not associated with severe disease.@*CONCLUSION@#The laboratory biomarkers together with Ct value and overall clinical picture may provide valuable information to physicians for more robust clinical decision-making.

Molecular detection and prevalence of Rotavirus with acute gastroenteritis among the children of rural and urban areas / Detecção molecular e prevalência de rotavírus com gastroenterite aguda entre as crianças de áreas rurais e urbanas

Rotavirus is the main infective agent of acute gastroenteritis (AGE) in children under the age of five years and causing significant morbidity as well as mortality throughout the world. The study was carried out to detect the prevalence rate, genotypes strain and risk factors of Rotavirus among the children of rural and urban areas of district Bannu Khyber Pakhtunkhwa Pakistan. A total of 180 stool samples were collected from children under the age of 5 years from two major hospitals of Bannu from January to December (2015). The samples were analyzed by Reverse-transcriptase Polymerase Chain Reaction (RT-PCR) for the detection of Rotavirus, positive samples were further processed for genotyping (G and P type) through specific PCR. Of the total, 41 (23%) samples were positive for Rotavirus. The most prevalent G genotypes found were: G3, G8, G9 (each 29%), followed by G10 (15%), and G11 (10%). Whereas the prevalent P genotypes were: P-8 (25%), P-4 and P-10 (each 20%), P-9 (15%), followed by P-6 and P-11 (each 10%). Moreover, Rotavirus infection was more prevalent in summer (23.73%) and winter (22.7%) than spring (20%) and autumn (21.4%). Rotavirus infection exhibited high frequency in June (14%), October (8%) and November (6%). It is concluded that Rotavirus is more prevalent in children and various genotypes (G and P) of Rotavirus are present in the study area. Lack of studies, awareness and rarer testing of Rotavirus are the principal reasons of virus prevalence in district Bannu, Pakistan.

O rotavírus é o principal agente infeccioso da gastroenterite aguda (AGE) em crianças menores de 5 anos e causa de morbidade e mortalidade significativas em todo o mundo. O estudo foi realizado para detectar a taxa de prevalência, cepa de genótipos e fatores de risco de rotavírus entre as crianças de áreas rurais e urbanas do distrito de Bannu Khyber Pakhtunkhwa, Paquistão. Um total de 180 amostras de fezes foi coletada de crianças menores de 5 anos de dois grandes hospitais de Bannu de janeiro a dezembro (2015). As amostras foram analisadas por reação em cadeia da polimerase transcriptase reversa (RT-PCR) para detecção de rotavírus; as amostras positivas foram posteriormente processadas para genotipagem (tipo G e P) através de PCR específica. Do total, 41 (23%) amostras foram positivas para rotavírus. Os genótipos G mais prevalentes encontrados foram: G3, G8, G9 (cada 29%), seguidos de G10 (15%) e G11 (10%). Considerando que os genótipos P prevalentes foram: P-8 (25%), P-4 e P-10 (cada 20%), P-9 (15%), seguido por P-6 e P-11 (cada 10%). Além disso, a infecção por rotavírus foi mais prevalente no verão (23,73%) e inverno (22,7%) do que na primavera (20%) e no outono (21,4%). A infecção por rotavírus apresentou alta frequência em junho (14%), outubro (8%) e novembro (6%). Conclui-se que o rotavírus é mais prevalente em crianças e vários genótipos (G e P) do rotavírus estão presentes na área de estudo. A falta de estudos, conhecimento e testes mais raros de rotavírus são as principais razões da prevalência do vírus no distrito de Bannu, Paquistão.

Rapid detection and genotyping of SARS-CoV-2 Omicron BA.4/5 variants using a RT-PCR and CRISPR-Cas12a-based assay / 南方医科大学学报

OBJECTIVE@#To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology.@*METHODS@#We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated.@*RESULTS@#This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively.@*CONCLUSION@#By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.

RGD peptide inhibits tumor growth by affecting angiogenesis related signaling pathway of laryngeal cancer stem cells / El péptido RGD inhibe el crecimiento tumoral al afectar la vía de señalización relacionada con la angiogénesis de las células madre del cáncer de laringe

SUMMARY: This study is to investigate the role and mechanism of RGD peptide in laryngeal cancer stem cells (CSCs). Laryngeal cancer CD133+Hep-2 CSCs were sorted by flow cytometry. RGD peptide was co-cultured with sorted laryngeal CSCs. Cell proliferation was detected with CCK-8 assay. The mRNA levels of VEGF/VEGFR2/STAT 3/HIF-1α were detected with RT-PCR. The proteins of VEGF/ VEGFR2/STAT 3/HIF-1α were detected with Western blot. The sorted CSCs were inoculated into nude mice. Tumor volume was measured. Integrin αvβ3 expression in tumor tissues was analyzed with immunohistochemistry. The results showed that the ratio of CD133+ CSCs to the total number of cells was 1.34±0.87 %, while CD133-non-tumor stem cells accounted for 95.0±5.76 %. The sorted cancer stem cells grew well. The RGD peptide significantly inhibited the proliferation of CD133+Hep-2 laryngeal CSCs in a dose-dependent manner. The RGD peptide significantly inhibited the mRNA of VEGFR2, STAT3 and HIF-1α in laryngeal CSCs in a concentration-dependent manner. Consistently, the RGD peptide significantly inhibited the protein expression of VEGFR2, STAT3 and HIF-1α in laryngeal CSCs in a dose-dependent manner. At the same time, in vivo tumor experiments showed that the RGD peptide significantly inhibited tumor volume but not the body weight. Furthermore, RGD peptide significantly inhibited the expression of tumor angiogenesis-related protein integrin αvβ3. Our findings demonstrate that RGD peptide inhibits tumor cell proliferation and tumor growth. The underlying mechanism may that RGD inhibits tumor angiogenesis-related signaling pathways, thus affecting the tumor angiogenesis, and decreasing the progression of human laryngeal CSCs.

Este estudio se realizó para investigar el papel y el mecanismo del péptido RGD en las células madre del cáncer de laringe (CSC). Las CSC CD133+Hep-2 de cáncer de laringe se clasificaron mediante citometría de flujo. El péptido RGD se cocultivó con CSC laríngeas clasificadas. La proliferación celular se detectó con el ensayo CCK-8. Los niveles de ARNm de VEGF/VEGFR2/ STAT 3/HIF-1α se detectaron con RT-PCR. Las proteínas de VEGF/ VEGFR2/STAT 3/HIF-1α se detectaron con Western blot. Las CSC clasificadas se inocularon en ratones nudos. Se midió el volumen del tumor. La expresión de integrina αvβ3 en tejidos tumorales se analizó con inmunohistoquímica. Los resultados mostraron que la proporción de CSC CD133+ con respecto al número total de células fue de 1,34 ± 0,87 %, mientras que las células madre no tumorales CD133 representaron el 95,0 ± 5,76 %. Las células madre cancerosas clasificadas crecieron bien. El péptido RGD inhibió significativamente la proliferación de CSC laríngeas CD133+Hep-2 de una manera dependiente de la dosis. El péptido RGD inhibió significativamente el ARNm de VEGFR2, STAT3 y HIF-1α en CSC laríngeas de manera dependiente de la concentración. De manera consistente, el péptido RGD inhibió significativamente la expresión proteica de VEGFR2, STAT3 y HIF-1α en CSC laríngeas, de manera dependiente de la dosis. Al mismo tiempo, los experimentos con tumores in vivo mostraron que el péptido RGD inhibía significativamente el volumen del tumor pero no el peso corporal. Además, el péptido RGD inhibió significativamente la expresión de la proteína integrina αvβ3 relacionada con la angiogénesis tumoral. Nuestros hallazgos demuestran que el péptido RGD inhibe la proliferación de células tumorales y el crecimiento tumoral. El mecanismo subyacente puede ser que RGD inhiba las vías de señalización relacionadas con la angiogénesis tumoral, afectando así la angiogénesis tumoral y disminuyendo la progresión de las CSC laríngeas humanas.

Expression Levels of the CA9, WT1, and PRAME genes and genotyping-associated antigens for the diagnosis and prognosis of colorectal cancer

Background: Colorectal cancer (CRC) is the third most prevalent type of cancer worldwide, and is one of the major health problems in Asia, Africa, Europe, and America. The tumor antigens recently are of interesting indicators as diagnostic and prognostic tools, The aim of the present study is to detect the expression levels of carbonic anhydrase IX (CA9), the Wilms tumor gene (WT1), and the preferentially expressed antigen in melanoma (PRAME) in the peripheral blood of CRC patients in comparison with healthy controls. Methods: A prospective case-control study of CRC patients was conducted. We included 25 newly-diagnosed CRC eligible patients and obtained peripheral blood samples of them as well as 10 blood samples from the control group. All samples were then submitted to deoxyribonucleic acid (DNA) extraction and a molecular study through real-time polymerase chain reaction (PCR). Results: The CRC group consisted of 15 (60%) female and 10 (40%) male patients with a mean age of 50.52 ± 9.8 years, while the control group included 4 (40%) female and 6 (60%) male patients with a mean age of 47.7 ± 7.9 years. The CRC group, 24 (96%) of patient samples were CA9-positive with strong statistically significant differences (p < 0.00001; sensitivity: 96%; specificity: 90%). Regarding the WT1 gene, there were 11 (44%) positive samples in the CRC group, with no statistically significant differences (p = 0.055; sensitivity: 44%; specificity: 90%). The PRAME gene was positive in 9 (36%) samples in the CRC group, with no statistically significant differences (p = 0.357; sensitivity: 36%; specificity: 80%. Among CA9 (24 patients; 96%) of patients with CRC expressed positive results, in WT1 11(91.6%) CRC patients expressed gene, and in PRAME gene, 9 patients with CRC (81.8%) expressed positive results. Conclusion: Overexpression of the CA9 gene in CRC of high sensitivity and specificity to be used as a tool to discriminate CRC from benign associate with high accuracy compare to WT1 and PRAME genes. (AU)

Regulation of Notch1 and Foxp1 by MiR-34a in psoriasis vulgaris / Regulación de Notch1 y Foxp1 por MiR-34ª en psoriasis vulgaris

SUMMARY: This study is to investigate the regulation of Notch1 and Foxp1 by miR-34a in the development of psoriasis vulgaris. RT-PCR was used to compare the levels of miR-34a in the skin lesions of 20 patients with psoriasis vulgaris and 20 normal skin tissues. Immunohistochemistry was used to detect the expression of Notch1 and Foxp1 in 51 patients with psoriasis vulgaris, which were further compared with that in 29 normal control tissues. In addition, in HaCaT cells, we used miR-34a mimics and inhibitors to overexpress and inhibit miR-34a, respectively, and detected the mRNA and protein levels of miR-34a, Notch1, and Foxp1. The level of miR-34a in the skin lesions of patients with psoriasis vulgaris was significantly higher than that in normal skin tissues (t=2.192, P<0.05). The positive rate of Notch1 in the skin lesions of patients with psoriasis vulgaris was 76.47 %, which was significantly higher than that in normal skin tissues (13.79 %) (t=29.215, P<0.01). The positive rate of FOXP1 in the psoriasis vulgaris group was 92.16 %, which was also significantly higher than that in the normal skin group (65.52 %) (t=9.087, P<0.01). In addition, overexpression of miR-34a significantly promoted the expression of Notch1 and Foxp1. However, inhibition of miR-34a significantly reduced Notch1 and Foxp1 levels. miR- 34a is highly expressed in the skin tissues of patients with psoriasis vulgaris, and may participate in the development of psoriasis vulgaris by regulating Notch1 and Foxp1.

RESUMEN: El objetivo de este estudio fue investigar la regulación de Notch1 y Foxp1 por miR-34a en el desarrollo de la psoriasis vulgar. Se utilizó RT-PCR con el fin de comparar los niveles de miR-34a en las lesiones cutáneas de 20 pacientes con psoriasis vulgar y 20 tejidos de piel normales. Se utilizó inmunohistoquímica para detectar la expresión de Notch1 y Foxp1 en 51 pacientes con psoriasis vulgar, que se compararon además con la de 29 tejidos normales control. Además, en las células HaCaT, usamos miméticos e inhibidores de miR-34a para sobreexpresar e inhibir miR-34a, respectivamente, y detectamos los niveles de ARNm y proteína de miR-34a, Notch1 y Foxp1. El nivel de miR- 34a en las lesiones cutáneas de pacientes con psoriasis vulgar fue significativamente mayor que en los tejidos normales de la piel (t=2,192, P<0,05). La tasa de positividad de Notch1 en las lesiones cutáneas de pacientes con psoriasis vulgar fue del 76,47 %, que fue significativamente mayor que la de los tejidos normales de la piel (13,79 %) (t=29,215, P<0,01). La tasa positiva de FOXP1 en el grupo de psoriasis vulgar fue del 92,16 %, que también fue significativamente mayor que la del grupo de piel normal (65,52 %) (t=9,087, P<0,01). Además, la sobreexpresión de miR-34a promovió significativamente la expresión de Notch1 y Foxp1. Sin embargo, la inhibición de miR-34a redujo de manera importante los niveles de Notch1 y Foxp1. miR-34a se expresa en gran medida en los tejidos de la piel en pacientes con psoriasis vulgar y puede participar en el desarrollo de la psoriasis vulgar mediante la regulación de Notch1 y Foxp1.

Resultados del embarazo en mujeres con y sin COVID-19 en un hospital nacional de nivel III de Perú / Pregnancy outcomes among women with and without COVID-19 in a national level III hospital in Peru

OBJETIVO: Comparar los resultados del embarazo entre mujeres con y sin COVID-19 atendidas en un hospital nacional de nivel III de Perú. MÉTODO: Estudio observacional, retrospectivo y comparativo. Participaron gestantes con RT-PCR positiva y negativa en razón 1:1, y 1:2 con gestantes del año 2019. Se recogió información materna y perinatal. Se usó la prueba exacta de Fisher con significancia de 0,05 y razones de prevalencia (RP) con intervalo de confianza del 95% (IC95%). RESULTADOS: Participaron 51 gestantes con RT-PCR positiva, 51 gestantes con RT-PCR negativa y 102 gestantes del año 2019. Se observó asociación entre los resultados de la RT-PCR y el parto pretérmino (p < 0,05). La RP de parto pretérmino en las gestantes con RT-PCR positiva fue de 3,14 (IC95%: 1,29-7,64) veces en comparación con las gestantes de 2019 y de 4,0 (IC95%: 1,13-14,17) veces en comparación con las gestantes con RT-PCR negativa. CONCLUSIONES: Los hallazgos sugieren que puede existir asociación entre COVID-19 y parto pretérmino. Sin embargo, se requieren estudios más amplios para analizar el papel de otros factores maternos en esta asociación.

OBJECTIVE: To compare pregnancy outcomes among women with and without COVID-19 infection attended in a national level III hospital in Peru. METHOD: Observational, retrospective and comparative study. RT-PCR positive pregnant women participated in a 1:1 ratio with negative RT-PCR, and 1:2 with pregnant women of 2019. Maternal and perinatal information was collected. Fishers exact test was used with a significance level of 0.05 and prevalence ratios (PR) with their confidence interval of 95% (CI95%). RESULTS: 51 pregnant women with positive RT-PCR, 51 with negative RT-PCR and 102 pregnant women in 2019 participated. RT-PCR test were associated to preterm delivery (p < 0.05). The PR for preterm delivery in women with positive RT-PCR compared to pregnant women in 2019 was 3.14 (CI95%: 1.29-7.64); and compared to women with negative RT-PCR was 4.0 (CI95%: 1.13-14.17). CONCLUSIONS: The studys findings suggest the existence of an association between maternal COVID-19 and preterm birth. However, more studies are required to analyze the role of maternal factors.

4-hydroxy-2-nonenal as a marker of the oxidative stress in brains of dogs with canine distemper / 4-hidroxi-2-nonenal como marcador de estresse oxidativo no cérebro de cães com cinomose

Canine Distemper is a disease caused by Canine morbillivirus (CM), a pantropic virus that can affect the central nervous system (CNS), causing demyelination. However, the pathogenesis of this lesion remains to be clarified. Brain samples of 14 naturally infected dogs by CM were analyzed to evaluate the presence of oxidative stress and demyelination. RT-PCR assay was performed to confirm a diagnosis of canine distemper in the brain, immunohistochemistry anti-CM was used to localize the viral proteins in the tissue, and anti-4-hydroxy-2-nonenal (4-HNE) was a marker of a product of lipid peroxidation. The results showed the presence of viral proteins in the demyelinated area with the presence of 4-HNE. Our results suggest that the CM virus infection causes oxidative stress leading to lipid peroxidation, which causes tissue damage and demyelination. In conclusion, oxidative stress plays a significant role in canine distemper pathogenesis in the CNS.(AU)

A cinomose canina é uma doença causada pelo Morbilivírus canino (CM), um vírus pantrópico que pode afetar o sistema nervoso central (SNC), causando desmielinização. No entanto, a patogênese dessa lesão não está totalmente esclarecida. RT-PCR e imuno-histoquímica foram realizadas para confirmação do diagnóstico de cinomose em amostras de encéfalo de 14 cães naturalmente infectados. Após confirmação, foi realizada uma avaliação do estresse oxidativo por imuno-histoquímica com uso de anti-4-hidroxi-nonenal (4HNE) como marcador de produtos resultantes da peroxidação lipídica. Os resultados sugerem que a infecção pelo CM causa estresse oxidativo no tecido, levando a peroxidação lipídica, a qual causa danos ao tecido, culminando com desmielinização. Conclui-se que o estresse oxidativo tem papel importante na patogênese da cinomose canina no sistema nervoso central.(AU)

Pattern of chest computerized tomography scan findings in symptomatic RT-PCR positive COVID-19 patients at the Korle Bu Teaching Hospital, Ghana

Background: Chest Computerized Tomography (CT) features of Corona Virus Disease 2019 (COVID-19) pneumonia are nonspecific, variable and sensitive in detecting early lung disease. Hence its usefulness in triaging in resource-limited regions. Objectives: To assess the pattern of chest CT scan findings of symptomatic COVID-19 patients confirmed by a positive RTPCR in Ghana. Methods: This study retrospectively reviewed chest CT images of 145 symptomatic RT-PCR positive COVID-19 patients examined at the Radiology Department of the Korle Bu Teaching Hospital (KBTH) from 8th April to 30th November 2020. Chi-Squared test was used to determine associations among variables. Statistical significance was specified at p≤0.05. Results: Males represent 73(50.3%). The mean age was 54.15±18.09 years. The age range was 5 months-90 years. Consolidation 88(60.7%), ground glass opacities (GGO) 78(53.8%) and crazy paving 43(29.7%) were the most predominant features. These features were most frequent in the elderly (≥65years). Posterobasal, peripheral and multilobe disease were found bilaterally. The most common comorbidities were hypertension 72(49.7%) and diabetes mellitus 42(29.2%) which had significant association with lobar involvement above 50%. Conclusion: The most predominant Chest CT scan features of COVID-19 pneumonia were GGO, consolidation with air bronchograms, crazy paving, and bilateral multilobe lung disease in peripheral and posterior basal distribution

Profil épidémiologique des patients consultants et des cas positifs Covid-19 de l'unité de triage EHU Oran (mars 2020 - août 2020) / Epidemiological profile of consulting patients and Covid-19 positive cases in the triage unit - EHU Oran (March 2020 - August 2020)

Introduction. En décembre 2019, la population de Wuhan, une grande ville chinoise de 11 millions d'habitants, est atteinte par une pneumonie virale extrêmement contagieuse due au coronavirus SARSCoV-2. En Algérie, le premier cas a été identifié le 25 février 2020, un plan de riposte national élaboré par le ministère de la santé a été mis en place pour affronter l'épidémie. Dès le début de la pandémie, l'établissement hospitalier et universitaire d'Oran (EHUO), centre de soins de première ligne, d'une capacité 780 lits desservant environ 2 millions d'habitants s'est complètement réorganisé, en aménageant principalement des circuits pour le tri des patients « suspects Covid 19 ¼. L'objectif de notre travail consiste à décrire le profil épidémiologique des patients consultants et des cas incidents Covid-19 au niveau de l'unité de triage EHUO depuis le début de l'épidémie au 07 Aout 2020. Méthodes. Les données sont collectées à partir des enquêtes épidémiologiques auprès des patients consultant au niveau de l'unité de triage Covid-19 de l'EHUO. Résultats. À la date du 7 aout 2020, un total de 7110 cas suspects de Covid-19 ont été enregistrés. Du total des cas suspects, 3036 (42,7 %) étaient positifs par RT-PCR. Il s'agissait de 1421 de sexe masculin (46,8 %) et de 1615 de sexe féminin (53,2 %), soit une sex-ratio de 0,88. La répartition globale des cas Covid19 par tranches d'âge était la suivante : 21,9 % des patients avaient un âge entre 30 et 39 ans, 17,3 % entre 40 et 49 ans. Le tableau clinique était dominé par la toux (26,5%), suivi de céphalées (19 %), de fièvre (16,2 %) et d'anosmie (15,9 %). 234 patients (7,7 %) avaient une SpO2 < 95 %. Conclusion. L'épidémiologie de l'infection Covid 19 reste encore peu connue à l'heure actuelle dans notre wilaya. Un dépistage de masse permettrait d'avoir une vision plus claire sur le nombre réel de patients atteints. La Covid 19 est une infection avec un large panel de symptômes autres que respiratoires, souvent non spécifiques, faisant d'elle une pathologie difficile à diagnostiquer cliniquement, d'où la nécessité de la confirmation biologique (PCR).

Application of two RT-PCR methods for detection of norovirus in market-sold oysters and norovirus genetic characteristic analysis, a survey conducted in Beijing / 中华流行病学杂志

Objective: To evaluate the application of real-time RT-PCR and semi-nested RT-PCR in the detection of norovirus in oysters and analyzing the genetic characteristics of the isolates. Methods: Real-time fluorescent RT-PCR and semi-nested RT-PCR were used to detect norovirus GⅠ/GⅡ in fresh oysters collected from the markets in Beijing from November 2014 to October 2015. The detection rate of the parallel test was also analyzed. In addition, the reliability of semi-nested RT-PCR was evaluated by agreement rate and consistency test (Kappa value). The positive products of norovirus GⅠ/GⅡ capsid protein region gene by semi-nested RT-PCR were sequenced. Software BioEdit 7.0.9.0 was used for sequence alignment, and software Mega 6.0 was used to construct the evolutionary tree. Results: In 72 samples, the detection rate of norovirus was 31.94% (23/72) by real-time RT-PCR, 38.89% (28/72) by semi-nested RT-PCR and 48.61% (35/72) by parallel test. The coincidence rate of the two methods was 73.61%, a moderate degree (Kappa value =0.43). A total of 13 norovirus strains were successfully sequenced, and 11 strains (7 GⅡ.17 strains, 2 GⅡ. 4 Sydney_ 2012 strains, 1 GⅡ. 1 strain and 1 GⅡ. 21 strain) were obtained from norovirus positive samples by two RT-PCR methods, two strains (1 GⅡ. 17 strain and 1 GⅡ. 3 strain) were obtained from real-time RT-PCR negative samples which were positive for norovirus by semi-nested RT-PCR. The similarity between these strains and reference strains from diarrhea patients, environmental sewage, and shellfish products were 84.4% - 100.0%. Conclusions: The parallel test of norovirus in oysters by two RT-PCR methods can improve the detection rate and detect more genotypes. Norovirus strains in oysters were highly homologous with reference strains from diarrheal patients, environmental sewage, and shellfish products. Therefore, surveillance, prevention and control for norovirus should be carried out in people who have frequent contacts with oysters and related environments.

Hisopado endocervical en mujeres con síntomas de COVID-19 provenientes de departamentos del Paraguay / Endocervical swabbing in women with COVID-19 symptoms from departments of Paraguay

INTRODUCCIÓN: El COVID-19 es eminentemente una infección de transmisión e inicio respiratorio, se discute la existencia de otras fuentes de contagio. El receptor viral ACE2 también ha sido detectado en el útero y en la vagina; de allí se ha planteado el compromiso del virus SARS-CoV-2 sobre el sistema genitourinario y sus posibles repercusiones en el embarazo. OBJETIVO: Determinar la presencia de SARS-CoV-2 en muestras endocervicales de mujeres con COVID-19 en departamentos del Paraguay. PACIENTES Y MÉTODOS: Diseño observacional prospectivo, de corte transverso. Se reclutaron 200 mujeres desde agosto 2020 hasta febrero 2021, con no más de 48/72 h de un resultado previo positivo de hisopado nasofaríngeo para SARS-CoV-2 por retrotranscriptasa reversa-reacción en cadena de la polimerasa (en inglés rt-RT-PCR) y que aceptaron ingresar al estudio. Se llenó un cuestionario clínico epidemiológico. Las tomas de muestras se realizaron en servicios de salud del Ministerio de Salud Pública y Bienestar Social (MSP y BS), domicilios y albergues de los distintos departamentos de Paraguay. Cada paciente fue sometida a un hisopado con hisopos de dacron o citobrush endocervical para la detección de SARS-CoV-2 por rt RT-PCR. Resultados: Las mujeres estudiadas tenían una edad media de 46,5 años (IC 95% 31,5-62,5). Refirieron contagio comunitario con SARS-CoV-2 en 75,5%, 13,5% en el hogar, 8,5% en el lugar de trabajo y 1,5% en el extranjero. Las manifestaciones clínicas fueron: 30%, síndrome gripal, fiebre 22,5%, tos 20%, anosmia 15,5%, trastornos digestivos 15,5%, y otros se presentaron con menor frecuencia. Las muestras de hisopados o citobrush endocervical sometidas a rt-RT-PCR para la deteccción de SARS Cov-2, resultaron negativas en las 200 mujeres de estudio. Discusión: Cabe destacar que las muestras vaginales fueron tomadas dentro de las 24-72 h de haber obtenido un resultado positivo para SARS-CoV-2 en el hisopado nasofaríngeo y que 62,5% de las mujeres se encontraban internadas en módulos respiratorios. Se discute la razón de la negatividad de los exámenes y su trascendencia. CONCLUSIÓN: No se detectó infección con SARS-CoV-2 en la región endocervical de 200 mujeres con manifestaciones clínicas de COVID 19 y evaluadas dentro de las 48/72 h de un resultado positivo nasofaríngeo para SARS Cov-2. Los resultados en la población de estudio concuerdan con otros estudios reportados en la literatura científica.

BACKGROUND: COVID-19 is an eminently respiratory transmissible infection of respiratory initiation, the existence of other sources of contagion is discussed. The ACE2 viral receptor has also been detected in the uterus and vagina; Hence, the involvement of the SARS-CoV-2 virus on the genitourinary system and its possible repercussions on pregnancy has been raised. AIM: To determine the presence of SARS-CoV-2 in endocervical samples of women with COVID-19 in the departments of Paraguay. METHODS: Designed as a prospective observational of transverse cohort. Two hundred women were recruited from August 2020 to February 2021, with no more than 48/72 hours of a previous positive nasopharyngeal swab result for SARS-CoV-2 by reverse transcriptase-polymerase chain reaction (rt-RT-PCR) and who agreed to participate in the study. A clinical epidemiological questionnaire was completed. The samples were taken in health services of the MSPYBS (Public Ministry of Health and Social Welfare), homes and shelters in the different departments of Paraguay. Each patient underwent a swab (dacron swabs) or endocervical cytobrush for the detection of SARS-CoV-2 by rt-RT-PCR. RESULTS: Women recruited had a mean age of 46.5 years (95% CI 31,562.5). They reported contagion with SARS-CoV-2: 75.5% in the community, 13.5% at home, 8.5% in the place of work and 1.5% abroad. The clinical manifestations were: 30% flu syndrome, 22.5% fever, 20% cough, 15.5% anosmia, 15.5% digestive disorders, among other symptoms. The swabs or endocervical cytobrush samples subjected to rt-RT-PCR for the detection of SARS-CoV-2 were negative in the 200 study women. Discussion: It should be noted that the vaginal samples were taken within 24-72 hours after obtaining a positive result for SARS-CoV-2 in the nasopharyngeal swab and that 62.5% of the women were hospitalized in respiratory modules. The reason for the negativity of the exams and their significance are discussed. CONCLUSION: No SARS Cov-2 infection was detected in the endocervical region of 200 women with clinical manifestations of COVID 19 and evaluated within 48/72 hours of a positive nasopharyngeal result for SARS Cov-2. The results in the study population agree with the findings of other studies reported in the literature.

Effect of Huangkui on mouse podocytes under high glucose environment / Efecto de Huangkui en podocitos de ratón en un entorno de glucosa alta

To explore a new underlying molecular mechanism of Huangkui Extract Powder (HKEP) in the alleviation of diabetic nephropathy (DN). Murine immortalized podocytes were divided into (i) normal glucose (NG, 5.6 mM), (ii) NG + HKEP (0.45 g/L), (iii) HG, and (iv) HG + HKEP (0.45 g/L) groups. MTT assay and flow cytometry were used to detect the podocyte proliferation, apoptosis and cell cycle. Cell viability was inhibited, and apoptosis increased in(iii) HG group compared with (i) NG group (p<0.05). mRNA and protein expression of nephrin and podocin significantly decreased in (iii) HG group compared with (i) NG group (p<0.05). When compared with (iii) HG group, (iv) HG + HKEP group had higher cell viability, lower apoptotic rate and higher mRNA and protein expression of nephrin and podocin (p<0.05). HKEP can attenuate HG-induced podocyte damage, which may be one of the mechanisms of HKEP for attenuating DN.

Explorar un nuevo mecanismo molecular subyacente del extracto del polvo de Huangkui (HKEP) en el alivio de la nefropatía diabética (ND). Los podocitos murinos inmortalizados se dividieron en (i) grupos de glucosa normal (NG, 5,6 mM), (ii) NG + HKEP (0,45 g/L), (iii) HG y (iv) HG + HKEP (0,45 g/L). Se utilizaron el ensayo MTT y la citometría de flujo para detectar la proliferación de podocitos, la apoptosis y el ciclo celular. La viabilidad celular se inhibió y la apoptosis aumentó en el grupo (iii) HG en comparación con el grupo (i) NG (p<0,05). La expresión de ARNm y proteínas de nefrina y podocina disminuyó significativamente en el grupo (iii) HG en comparación con el grupo (i) NG (p<0,05). En comparación con el grupo (iii) HG, el grupo (iv) HG + HKEP tuvo una mayor viabilidad celular, una tasa de apoptosis más baja y una expresión de ARNm y proteínas más altas de nefrina y podocina (p<0,05). HKEP puede atenuar el daño de los podocitos inducido por HG, que puede ser uno de los mecanismos de HKEP para atenuar la DN.

Gene expression and apoptosis response in hepatocellular carcinoma cells induced by biocompatible polymer/magnetic nanoparticles containing 5-fluorouracil

BACKGROUND: Super-paramagnetic iron oxide nanoparticles (SPION) contain a chemotherapeutic drug and are regarded as a promising technique for improving targeted delivery into cancer cells. RESULTS: In this study, the fabrication of 5-fluorouracil (5-FU) was investigated with loaded Dextran (DEXSPION) using the co-precipitation technique and conjugated by folate (FA). These nanoparticles (NPs) were employed as carriers and anticancer compounds against liver cancer cells in vitro. Structural, magnetic, morphological characterization, size, and drug loading activities of the obtained FA-DEX-5-FUSPION NPs were checked using FTIR, VSM, FESEM, TEM, DLS, and zeta potential techniques. The cellular toxicity effect of FA-DEX-5-FU-SPION NPs was evaluated using the MTT test on liver cancer (SNU-423) and healthy cells (LO2). Furthermore, the apoptosis measurement and the expression levels of NF-1, Her-2/neu, c-Raf-1, and Wnt-1 genes were evaluated post-treatment using flow cytometry and RT-PCR, respectively. The obtained NPs were spherical with a suitable dispersity without noticeable aggregation. The size of the NPs, polydispersity, and zeta were 74 ± 13 nm, 0.080 and 45 mV, respectively. The results of the encapsulation efficiency of the nano-compound showed highly colloidal stability and proper drug maintenance. The results indicated that FA-DEX-5-FU-SPION demonstrated a sustained release profile of 5-FU in both phosphate and citrate buffer solutions separately, with higher cytotoxicity against SNU-423 cells than against other cells types. These findings suggest that FA-DEX-SPION NPs exert synergistic effects for targeting intracellular delivery of 5-FU, apoptosis induction, and gene expression stimulation. CONCLUSIONS: The findings proved that FA-DEX-5-FU-SPION presented remarkable antitumor properties; no adverse subsequences were revealed against normal cells.

Impact of rt-pcr test false-negative results for SARS-CoV-2 surveillance in Mexico

ABSTRACT Background: Underestimation of the number of cases during the coronavirus disease 2019 (COVID-19) pandemic has been a constant concern worldwide. Detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) RNA using real-time reverse-transcription polymerase chain reaction (RT-PCR) is the most common method to confirm a case. However, these tests have suboptimal sensitivity. Objective: The objective of the study was to estimate the number of COVID-19 confirmed cases, intensive care unit (ICU) admissions and deaths in Mexico, accounting for the probabilities of false-negative tests. Methods: We used publicly available, national databases of all SARS-CoV-2 tests performed at public laboratories in Mexico between February 27 and October 31, 2020. We used the estimated probabilities of false-negative tests based on the day of clinical sample collection after symptom initiation calculated previously. With the resulting model, we estimated the corrected daily number of cases, ICU admissions, and deaths. Results: Among 2,024,822 people tested in Mexico between February 27 and October 31 with an available result, we estimated 1,248,583 (95% confidence interval 1,094,850-1,572,818) cases, compared to 902,343 cases reported with positive tests. ICU admissions and deaths were 15% and 8% higher than reported, respectively. Conclusion: Accounting for SARS-CoV-2 RT-PCR-based diagnostic testsҠprecision is a simple way to improve estimations for the true number of COVID-19 cases among tested persons.

Genotypic evaluation of antimicrobial resistance in Staphylococcus spp. isolated from bovine clinical mastitis / [Avaliação genotípica da resistência antimicrobiana em Staphylococcus spp. isolados de mastite clínica bovina]

Bovine clinical mastitis caused by Staphylococcus spp. is a serious and widespread disease in the world of dairy farming. Antimicrobial therapy is of fundamental importance in the prevention and treatment of infectious mastitis, but the indiscriminate use of antimicrobials acts as a determining factor for the spread of the disease. The present study evaluated the resistance profiles of 57 Staphylococcus spp. isolated from bovine clinical mastitis to beta-lactams and gentamicin, relating characteristics of phenotype (in vitro susceptibility tests) and genotype (detection and expression of genes encoding resistance - mecA, mecALGA251, blaZ, femA, femB, and aacA-aphD - using PCR and RT-PCR, respectively). One or more genes coding for resistance to different antimicrobials were detected in 50 Staphylococcus spp. isolates. The femA and femB genes were the most frequent (75.4% for both). The observed expression of the genes was as follows: blaZ (60%), femA (39.5%), aacA-aphD (50%), femB (32.6%), mecA (8.3%), and mecALGA251 (0%). Considering the relevance of the genus Staphylococcus to bovine mastitis, this study aimed to elucidate aspects regarding the genotypic and phenotypic profiles of these microorganisms so as to contribute to the development of effective strategies for mastitis control.(AU)

A mastite clínica bovina causada por Staphylococcus spp. é uma doença grave e generalizada no mundo da pecuária leiteira. A terapia antimicrobiana é de fundamental importância na prevenção e no tratamento da mastite infecciosa, mas o uso indiscriminado de antimicrobianos atua como fator determinante para a disseminação da doença. O presente estudo avaliou os perfis de resistência de 57 Staphylococcus spp. isolados de mastite clínica bovina em relação ao uso de betalactâmicos e gentamicina, relacionando características do fenótipo (testes de suscetibilidade in vitro) e genótipo (detecção e expressão de genes que codificam resistência - mecA, mecALGA251, blaZ, femA, femB, e aacA-aphD - usando PCR e RT-PCR, respectivamente). Um ou mais genes que codificam resistência a diferentes antimicrobianos foram detectados em 50 Staphylococcus spp. isolados. Os genes femA e femB foram os mais frequentes (75,4% para ambos). A expressão observada dos genes foi a seguinte: blaZ (60%), femA (39,5%), aacA-aphD (50%), femB (32,6%), mecA (8,3%) e mecALGA251 (0%). Considerando-se a relevância do gênero Staphylococcus para a mastite bovina, este estudo teve como objetivo elucidar aspectos referentes aos perfis genotípico e fenotípico desses microrganismos, a fim de contribuir para o desenvolvimento de estratégias eficazes para o controle da mastite.(AU)

Sensibilidad y especificidad de pruebas moleculares en odontología / Sensitivity and specificity of molecular tests in dentistry

La biología molecular tiene mayor afinidad en las áreas de la salud, en odontología su principal aplicación ha sido en la identificación de microorganismos orales patógenos mediante el uso de secuencias genéticas específicas (ácido desoxirribonucleico [DNA], ácido ribonucleico [RNA] y proteínas). Las pruebas a nivel molecular se caracterizan por su rapidez, reproductibilidad, sensibilidad y especificidad de los microorganismos diana. El presente artículo de revisión bibliográfica servirá como herramienta para comprender los principios de las técnicas más destacadas como son: PCR estándar y RT-PCR en tiempo real, PCR con transcriptasa inversa, microarreglos y ensayo por inmunoabsorción ligado a enzimas (ELISA), además de sus ventajas y desventajas respecto a las pruebas convencionales (AU)

Molecular biology has a greater affinity in the areas of health. In dentistry, its main application has been the identification of pathogenic oral microorganisms, through the use of specific genetic sequences (deoxyribonucleic acid [DNA], ribonucleic acid [RNA] and proteins). Molecular tests are characterized by their rapidity, reproducibility, sensitivity and specificity of target microorganisms. This literature review article will serve as a tool to understand the principles of the most prominent techniques such as: Standard PCR, Real-time RT-PCR, Reverse transcriptase PCR, microarrays and Enzyme-linked immunosorbent assay (ELISA), in addition to their advantages and disadvantages with respect to conventional tests (AU)