Racial and ethnic disparities in the association between financial hardship and self-reported weight change during the first year of the pandemic in the U.S.

Studies have shown that financial hardship can impact weight change; however, it is unclear what the economic impact of the COVID-19 pandemic has had on weight change in U.S. adults, or whether racial-ethnic groups were impacted differentially. We estimated the association between financial hardship and self-reported weight change using data from the cross-sectional COVID-19's Unequal Racial Burden (CURB) survey, a nationally representative online survey of 5,500 American Indian/Alaska Native, Asian, Black/African American, Latino (English- and Spanish-speaking), Native Hawaiian/Pacific Islander, White, and multiracial adults conducted from 12/2020 to 2/2021. Financial hardship was measured over six domains (lost income, debt, unmet general expenses, unmet healthcare expenses, housing insecurity, and food insecurity). The association between each financial hardship domain and self-reported 3-level weight change variable were estimated using multinomial logistic regression, adjusting for sociodemographic and self-reported health. After adjustment, food insecurity was strongly associated with weight loss among American Indian/Alaska Native (aOR = 2.18, 95% CI = 1.05-4.77), Black/African American (aOR = 1.77, 95% CI = 1.02-3.11), and Spanish-speaking Latino adults (aOR = 2.32, 95% CI = 1.01-5.35). Unmet healthcare expenses were also strongly associated with weight loss among Black/African American, English-speaking Latino, Spanish-speaking Latino, and Native Hawaiian/Pacific Islander adults (aORs = 2.00-2.14). Other domains were associated with weight loss and/or weight gain, but associations were not as strong and less consistent across race-ethnicity. In conclusion, food insecurity and unmet healthcare expenses during the pandemic were strongly associated with weight loss among racial-ethnic minority groups. Using multi-dimensional measures of financial hardship provides a comprehensive assessment of the effects of specific financial hardship domains on weight change among diverse racial-ethnic groups.

Fight COVID Milwaukee protective behaviors and risk communications associated with the COVID-19 pandemic.

The COVID-19 pandemic has had a major impact on society, causing significant disruptions to everyday life. Risk communication strategies can play an important role in risk management as they allow individuals to prepare for and respond to public health emergencies appropriately. The aim of this study is to investigate public risk behaviors, perceptions of risk and risk communication, and experiences with COVID-19 to better understand the impact of COVID-19 on our community and to better inform public health decisions about communicating and reducing personal risk. Nine virtual focus groups were conducted with 79 residents of Milwaukee County. Audio transcripts of focus group recordings were qualitatively analyzed using MAXQDA. Predominant themes identified include public risk protective behaviors, the emotional toll associated with lockdown measures, and risk communication. Our findings provide a better understanding of how adults, African American and Hispanic groups in particular, viewed the risk communications and protective behaviors associated with COVID-19, how their lives were impacted by the pandemic, and how to effectively communicate public information about personal risk. These findings can help guide risk communication efforts and public health policy interventions for potential infection outbreaks in the future.

Complementary mechanisms of modulation of spontaneous phasic contractions by the gaseous signalling molecules NO, H2S, HNO and the polysulfide Na2S3 in the rat colon.

OBJECTIVES: Reactive oxygen and nitrogen species may be produced during inflammation leading to the formation of NO, H2S or HNO. Enzymes such as iNOS, CSE and CBS might also be responsible for polysulfide production. Since these signalling molecules might have an impact on colonic motility, the aim of this study was to compare their effect on rat colonic slow phasic contractions (SPC). METHODS: Organ bath measurements with strips obtained from rat proximal colon were performed using the polysulfide Na2S3, sodium nitroprusside (NaNP), sodium hydrogen sulfide (NaHS), Angeli's salt as NO, H2S, and HNO donors, respectively. TTX (1 µM) was used to block neuronal activity. RESULTS: All four molecules, concentration-dependently, inhibited the amplitude and frequency of SPC both in the circular and longitudinal muscle layer. The relative potency was NaNP>Angeli's salt>NaHS>Na2S3. The inhibitory response induced by NaNP (1 µM) and Angeli's salt (50 µM) was reversed by ODQ (10 µM) whereas the inhibitory effect of NaHS (1 mM) was reversed by apamin (1 µM) and glibenclamide (10 µM). Na2S3 (1 mM) response was partially reversed by apamin (1 µM) and glibenclamide (10 µM). High concentrations of Na2S3 caused an increase in tone. Low concentrations of NaHS or Na2S3 did not potentiate NaNP responses. CONCLUSIONS: All signalling molecules inhibit SPC in both muscle layers. The effect is independent of neural activity and involves guanylyl cyclase (NO and HNO) and SKCa and KATP channels (NaHS or Na2S3). Other pathways might also be involved in Na2S3 responses. Accordingly, complementary mechanisms of inhibition might be attributable to these signalling molecules.

Oxidative Stress-Induced Male Infertility: Role of Antioxidants in Cellular Defense Mechanisms.

Male infertility is linked to several environmental and mutagenic factors. Most of these factors, i.e., lifestyle, radiations, and chemical contaminations, work on the fundamental principles of physics, chemistry, and biology. Principally, it may induce oxidative stress (OS) and produce free radicals within the cells. The negative effect of OS may enhance the reactive oxygen species (ROS) levels in male reproductive organs and impair basic functions in a couple's fertility. Evidence suggests that infertile men have significantly increased ROS levels and a reduced antioxidant capacity compared with fertile men. Although, basic spermatic function and fertilizing capacity depend on a delicate balance between physiological activity of ROS and antioxidants to protect from cellular oxidative injury in sperm, that is essential to achieve pregnancy. The ideal oxidation-reduction (REDOX) equilibrium requires a maintenance of a range of ROS concentrations and modulation of antioxidants. For this reason, the chapter focuses on the effects of ROS in sperm functions and the current concepts regarding the benefits of medical management in men with diminished fertility and amelioration of the effect to improve sperm function. Also, this evidence-based study suggests an increasing rate of infertility that poses a global challenge for human health, urging the need of health care professionals to offer a correct diagnosis, comprehension of the process, and an individualized management of the patients.

CHARACTERIZATION OF SPERMATOZOON ULTRASTRUCTURE IN TETRAGONOCEPHALUM SP. (CESTODA: LECANICEPHALIDEA: TETRAGONOCEPHALIDAE) FROM THE WHIPRAY, UROGYMNUS ASPERRIMUS 1 (DASYATIDAE: UROGYMNINAE).

The Lecanicephalidea represents one of 11 currently recognized elasmobranch-hosted tapeworm lineages. It is quite speciose, with nearly 100 validly recognized species to date. Yet spermatozoon features have been fully characterized for only a single species; spermatozoon characters for a second species were previously included in a data matrix used to infer phylogenetic relationships among cestode orders, but data are limited and no images were provided. Specimens of Tetragonocephalum sp. were collected from the whipray, Urogymnus asperrimus 1, from the Solomon Sea off the Solomon Islands. The mature spermatozoa of Tetragonocephalum sp. are distinctly different from the other lecanicephalidean species for which spermatozoon ultrastructure has been thoroughly investigated. Tetragonocephalum sp. spermatozoa represent the Type IV (sensu Levron, 2010) morphology possessing a single axoneme, crested bodies, cortical microtubules running parallel to the axoneme, and nucleus, which is helical to the axoneme. Although spermatozoa for both lecanicephalideans examined to date are of the Type IV spermatozoon morphology, they differ in the number of crested bodies and cortical microtubules, the relative size of the spermatozoon-specifically as it relates to the nuclear region-and the presence or absence of the anterior spiral structure. Significant spermatozoon character variation is present across these 2 representatives of 2 lecanicephalidean families, indicating that additional studies of representatives of the other families in the order are needed to better understand the extent of character variation present in the Lecanicephalidea. Overall, spermatozoon ultrastructure is understudied in elasmobranch-hosted tapeworm lineages compared to other vertebrate-hosted tapeworm orders at both the species and family levels.

Investigation of the Optimal Prime Boost Spacing Regimen for a Cancer Therapeutic Vaccine Targeting Human Papillomavirus.

Therapeutic vaccine studies should be designed to elicit durable, high magnitude, and efficacious T cell responses, all of which can be impacted by the choice of the vaccination schedule. Here, we compare different prime-boost intervals (PBI) in a human papillomavirus (HPV) model using a HPV16E7E6 Venezuelan equine encephalitis virus replicon particle (VRP) vaccination to address the optimal boosting schedule, quality of immune response, and overall in vivo efficacy. Six different vaccine regimens were tested with each group receiving booster vaccinations at different time intervals. Analysis of T-cell responses demonstrated a significant HPV16 E7 specific CD8+ T cell response with at minimum a one-week PBI between antigen re-exposure. Significant E7-specific in vivo cytotoxicity was also observed with longer PBIs. Additionally, longer PBIs led to an enhanced memory recall response to tumor challenge, which correlated with differential expansion of T cell memory subsets. Our findings imply that when using alphavirus vector platforms as a vaccination strategy, a one-week PBI is sufficient to induce high magnitude effector T cells with potent anti-tumor activity. However, longer PBIs lead to enhanced long-term protective anti-tumor immunity. These findings have implications for therapeutic vaccine clinical trials in which shorter intervals of prime-boost regimens may lead to suboptimal durable immune responses.

Involving society in science: Reflections on meaningful and impactful stakeholder engagement in fundamental research.

Open Science calls for transparent science and involvement of various stakeholders. Here are examples of and advice for meaningful stakeholder engagement.

Biology and Life Stages of Pine Spittle Bug Ocoaxo assimilis Walker (Hemiptera: Cercopidae).

The first records of outbreaks of the Pine Spittle bug Ocoaxo assimilis Walker were recently identified from Puebla, Mexico, which promoted more than 2600 ha of forest foliar fall. Beyond the taxonomic and distribution information of this species, the basic traits of its biology remain unknown. This study aims to describe some biological aspects of O. assimilis, in a natural pine forest at Nicolás Bravo, Puebla (NB). Using morphological characteristics and a phylogenetic analysis of a fragment of cytochrome oxidase subunit I mtDNA (COI), immature instars with adults were studied; the instar number was determined by means of a multivariate analysis of 19 morphological characteristics of 121 specimens. The systematic sampling to evaluate the occurrence of nymphal specimens during a year, plus host selection experiments, allowed for determination of the abundance over time, voltism, and host preferences. Phylogenetic analysis of the COI supported that both nymphs and adults collected in NB correspond to O. assimilis. Principal coordinate analysis supported the existence of five nymphal stages. Field sampling and host selection experiments indicated that this species displays a single generation per year, is associated with the rainy season, and that specimens from the three first nymphal stages feed on roots of eight host species (one grass, four herbaceous species, one bush, and two trees). From the fourth instar, the insects feed on pine roots to complete their development, and when they are adults, they migrate to needles of young or mature pine stands of Pinus pseudostrobus to feed and reproduce.

El uso del portafolio para el pregrado en Ciencias Médicas / The Usage of the Portfolio for the Undergraduate Level in Medical Sciences

En los últimos años ha surgido la utilización del portafolio o carpeta como herramienta en los procesos formativos, al adoptarse una nueva y renovada visión sobre la enseñanza, el aprendizaje desarrollador y la evaluación educativa. El objetivo de este artículo es evaluar la competencia del uso del portafolio en el pregrado de Ciencias Médicas. Se demostró que en las universidades donde se utilizaba el portafolio, este favorecía el desarrollo profesional de los estudiantes, lo que propiciaba el dominio de competencias clínicas, y el desarrollo de actitudes como la responsabilidad y la sistematicidad; implicaba la documentación del aprendizaje y la recopilación de evidencias; y permitía la retroalimentación del desempeño del alumno. Aunque en los escenarios cubanos no se encontraron evidencias del empleo del portafolio, ya existen experiencias en varias latitudes que dan cuenta de los beneficios potenciales que esta herramienta está generando en el ámbito universitario, por lo que puede ser una buena opción o alternativa de evaluación(AU)

In recent years, the usage of the portfolio or folder as a tool in training processes has emerged as a new and renewed vision has been adopted regarding teaching, developing learning and educational evaluation. The objective of this article is to assess the competence associated to the usage of the portfolio in the undergraduate level of medical sciences. It was shown that, in the universities where the portfolio was used, it favored the students' professional development, which led to the mastery of clinical skills and the development of attitudes such as responsibility and systematicity; it involved the documentation of learning and the collection of evidence; and allowed feedback on student performance. Although there was no evidence of the usage of the portfolio in Cuban settings, there are already experiences in several sites that account for the potential benefits that this tool is generating in the university context, so it can be a good option or alternative for evaluation(AU)

Adherencia a la terapia inhalada en el ámbito ambulatorio / Adherence to inhaled therapy in the outpatient setting

Objetivo: Conocer el grado de adherencia a la terapia inhalada en pacientes con enfermedad pulmonar obstructiva crónica (EPOC) y asma en una consulta de Neumología. Identificar el tipo de incumplimiento y otros factores que influyen en el mismo. Método: Estudio descriptivo transversal en el que se incluyó consecutivamente a 124 pacientes. La adherencia se evaluó a través del Cuestionario Test de Adhesión a los Inhaladores (TAI). Resultados: Se reclutó a 62 mujeres y 62 hombres, y la mediana de edad fue 63 años. El 51,6% fueron pacientes con EPOC y el 48,4% con asma. El 38,7% presentó buena adherencia, el 37,9% intermedia y el 23,4% mala. En cuanto al incumplimiento, un 13,7% fue errático, un 4,8% deliberado y un 28,2% inconsciente. Los pacientes con estudios básicos tuvieron mejor cumplimiento que los demás (p=0,05) y los pacientes con EPOC presentan mejor adherencia que los asmáticos (p=0,02). La edad, el género y el formador no mostraron diferencias estadísticas significativas. Conclusiones: El 38,7% de los pacientes presentan una buena adherencia a la terapia inhalada, predominando el incumplimiento inconsciente. Pacientes con EPOC y con estudios básicos presentan mayores niveles de adhesión. La formación de neumólogo y enfermera de neumología parece mejorar la adherencia, aunque no se encontraron diferencias significativas

Objective: To find out the degree of adherence to inhaled therapy in patients with Chronic Obstructive Pulmonary Disease (COPD) and asthma in a pulmonology clinic. To identify the type of non-compliance and the different factors affecting it. Method: A descriptive, cross-sectional study in which 124 patients were included. Adherence was assessed using the 'Test of Adherence to Inhalers' (TAI) questionnaire. Results: A total of 62 women and 62 men were recruited and the average age was 63. Fifty-one point 6percent of the patients had COPD, and 48.4% were asthmatics. Thirty-eight point 7percent of the them showed high adherence, 37.9% showed intermediate adherence and 23.4%, low adherence. In terms of non-compliance, 13.7% was erratic, 4.8% was deliberate, and 28.2%, unwitting. Patients with a basic level of education had better compliance than the others (P=.05), and patients with COPD showed higher adherence than the asthmatics (P=.02). Age, gender and the trainer did not indicate significant statistical differences. Conclusion: 38.7% of patients show high adherence to inhaled therapy, with a predominance of unwitting non-compliance. Patients with COPD and with a basic level of education showed higher levels of adherence. The training given by the pulmonologist and the respiratory nurse seemed to improve adherence to these treatments, although no significant differences were found

Adherence to inhaled therapy in the outpatient setting. / Adherencia a la terapia inhalada en el ámbito ambulatorio.

OBJECTIVE: To find out the degree of adherence to inhaled therapy in patients with Chronic Obstructive Pulmonary Disease (COPD) and asthma in a pulmonology clinic. To identify the type of non-compliance and the different factors affecting it. METHOD: A descriptive, cross-sectional study in which 124 patients were included. Adherence was assessed using the 'Test of Adherence to Inhalers' (TAI) questionnaire. RESULTS: A total of 62 women and 62 men were recruited and the average age was 63. Fifty-one point 6percent of the patients had COPD, and 48.4% were asthmatics. Thirty-eight point 7percent of the them showed high adherence, 37.9% showed intermediate adherence and 23.4%, low adherence. In terms of non-compliance, 13.7% was erratic, 4.8% was deliberate, and 28.2%, unwitting. Patients with a basic level of education had better compliance than the others (P=.05), and patients with COPD showed higher adherence than the asthmatics (P=.02). Age, gender and the trainer did not indicate significant statistical differences. CONCLUSION: 38.7% of patients show high adherence to inhaled therapy, with a predominance of unwitting non-compliance. Patients with COPD and with a basic level of education showed higher levels of adherence. The training given by the pulmonologist and the respiratory nurse seemed to improve adherence to these treatments, although no significant differences were found.

Leche humana y su efecto sobre la mineralización del esmalte: revisión de literatura

La leche materna es el alimento que provee al niño desde su nacimiento de todos los nutrientes que necesita para su crecimiento y desarrollo físico y psicológico. Su efecto sobre el tejido dental ha sido objeto de controversias, ya que se ha visto que actúa sobre la mineralización del esmalte y se ha asociado a caries de la infancia temprana. La leche humana tiene diversos componentes protectores para el esmalte dental pero al mismo tiempo contiene factores que pueden generar la pérdida de minerales. El propósito de esta revisión es exponer los elementos que producen la desmineralización y la remineralización del esmalte y proponer algunas medidas que apoyen la integridad de este tejido, así como recomendar la lactancia materna como la alimentación principal que el infante debe recibir durante el primer año de vida. La leche humana tiene diversas proteínas y elementos que ayudan en el proceso de desmineralización y remineralización del esmalte dental de la primera dentición. Se puede concluir que, la leche materna es el alimento ideal para los niños en los primeros años de vida. Por lo que hay que promover la lactancia materna como el método preferido para alimentar a los bebés. Se debe implementar la higiene desde que el primer diente erupciona en boca. Esta medida evitará que el posible efecto desmineralizante de la leche ejerza un efecto irreversible sobre el esmalte dental. Es conveniente orientar a las madres sobre el almacenamiento de la leche materna de tal manera que al ser consumida por el niño, no se encuentre acidificada, y no produzca un efecto dañino sobre el esmalte.

Breast milk provides the child of all the nutrients he needs for his growth and physical and psychological development beginning at birth. Its effect on dental tissue has been the subject of controversy, since it has been seen to act on the mineralization of enamel and has been associated with early childhood caries. Human milk has various protective components for tooth enamel but at the same time it contains factors that can cause the loss of minerals. The purpose of this review is to review the elements that produce the demineralization and remineralization of enamel and propose some measures that support the integrity of this tissue, as well as to recommend breastfeeding as the main feeding source that the infant should receive during the first year of life. Human milk has various proteins and elements that help in the process of demineralization and remineralization of the tooth enamel of first dentition. It can be concluded that, breast milk is the ideal food for children during the first years of life. Therefore, breastfeeding should be promoted as the preferred method of feeding babies. Oral hygiene should be implemented as soon as the first tooth erupts. This measure will prevent the possible demineralizing effect of milk from exerting an irreversible effect on tooth enamel. It is advisable to guide mothers on the storage of breast milk so that when consumed by the child, it is not acidified, and does not produce harmful effects on the enamel.

Suicidality in primary care patients who present with sadness and anhedonia: a prospective European study.

BACKGROUND: Sadness and anhedonia (loss of interest in activities) are central symptoms of major depression. However, not all people with these symptoms meet diagnostic criteria for major depression. We aimed to assess the importance of suicidality in the outcomes for primary care patients who present with sadness and anhedonia. METHOD: Cohort study of 2,599 unselected primary care attenders in six European countries followed up at 6 and 12 months. RESULTS: 1) In patients with sadness and/or anhedonia who were not depressed at entry to the study, suicide plans (OR = 3.05; 95 % CI = 1.50-6.24; p = 0.0022) and suicide attempts (OR = 9.08; 95 % CI = 2.57-32.03; p = 0.0006) were significant predictors of developing new onset depression at 6 or 12 months. 2) In patients with sadness and/or anhedonia who met CIDI criteria for major depression at entry, suicidal ideation (OR = 2.93; 95 % CI = 1.70-5.07; p = 0.0001), suicide plans (OR = 3.70; 95 % CI = 2.08-6.57; p < 0.0001), and suicide attempts (OR = 3.33; 95 % CI = 1.47-7.54; p = 0.0040) were significant predictors of persistent depression at 6 or 12 months. CONCLUSIONS: Three questions on suicidality could help primary care professionals to assess such patients more closely without necessarily establishing whether they meet criteria for major depression.

Neuron specific Rab4 effector GRASP-1 coordinates membrane specialization and maturation of recycling endosomes.

The endosomal pathway in neuronal dendrites is essential for membrane receptor trafficking and proper synaptic function and plasticity. However, the molecular mechanisms that organize specific endocytic trafficking routes are poorly understood. Here, we identify GRIP-associated protein-1 (GRASP-1) as a neuron-specific effector of Rab4 and key component of the molecular machinery that coordinates recycling endosome maturation in dendrites. We show that GRASP-1 is necessary for AMPA receptor recycling, maintenance of spine morphology, and synaptic plasticity. At the molecular level, GRASP-1 segregates Rab4 from EEA1/Neep21/Rab5-positive early endosomal membranes and coordinates the coupling to Rab11-labelled recycling endosomes by interacting with the endosomal SNARE syntaxin 13. We propose that GRASP-1 connects early and late recycling endosomal compartments by forming a molecular bridge between Rab-specific membrane domains and the endosomal SNARE machinery. The data uncover a new mechanism to achieve specificity and directionality in neuronal membrane receptor trafficking.

Contribución al conocimiento etnomicológico de los hongos comestibles silvestres de mercados regionales y comunidades de la Sierra Nevada (México) / Contribution to ethnomycological knowledge of wild edible mushrooms from regional markets and communities of Sierra Nevada (Mexico)

En el presente trabajo se estudiaron 67 especies de hongos comestibles silvestres, 65 de ellas comercializadas en cuatro mercados regionales y varias en comunidades rurales, todas en la región de la Sierra Nevada, ubicada ésta en parte de las entidades del Estado de México, Puebla y Tlaxcala, al E y NE de la Ciudad de México. Se determinó el valor diferencial de importancia etnomicológica de las especies identificadas, por medio de la medición del valor de venta, así como de la presencia y la temporalidad de los hongos en los mercados, a lo largo de un año, en 1998. Se registró la preferencia de los hongos comestibles en una encuesta en 10 comunidades de la región, durante 1999. De los hongos considerados, Hebeloma fastibile fue la especie con mayor valor de importancia etnomicológica en los mercados y Lyophyllum decastes en las comunidades.

In this study, 67 species of wild edible mushrooms were studied, 65 of them were sold in four regional markets and some of them in communities, all on the Sierra Nevada found in the states of Mexico, Puebla, and Tlaxcala, at E and NE of Mexico City. The differential value of ethnomycological importance of the identified species was determined in terms of selling price as well as the presence and seasonality of the mushrooms, throughout one year, in 1998. Also, preference for the edible mushrooms mentioned was recorded through a survey of 10 communities in this region, during 1999. Of the mushrooms identified, Hebeloma fastibile was the species of highest value in terms of ethnomycological importance in the markets and Lyophyllum decastes was in the communities.

Contribuição ao conhecimento etnomicológico dos cogumelos comestiveis silvestres de mercados regionais e comunidades da Serra Nevada (México).

No presente trabalho se estudaram 67 espécies de cogumelos comestíveis silvestres, 65 delas comercializadas em quatro mercados regionais e varias em comunidades rurais, todas na região da Serra Nevada, situada esta em parte das entidades do Estado de México, Puebla e Tlaxcala, al E e NE da Cidade do México. Determinou-se o valor diferencial de importância etnomicológica das espécies identificadas, por meio da medição do valor de venda, assim como da presença e a temporalidade dos cogumelos nos mercados, ao longo de um ano, em 1998. Registrou-se a preferência dos cogumelos comestíveis em uma encosta em 10 comunidades da região, durante 1999. Dos cogumelos considerados, Hebeloma fastibile foi a espécie com maior valor de importância etnomicológica nos mercados e Lyophyllum decastes nas comunidades.

Pharmacologic characterization of intrinsic mechanisms controlling tone and relaxation of porcine lower esophageal sphincter.

The neurotransmitters mediating relaxation of lower esophageal sphincter (LES) were studied using circular LES strips from adult pigs in organ baths. LES relaxation by sodium nitroprusside (1 nM-3 microM), vasoactive intestinal peptide (VIP) and pituitary adenylate cyclase-activating peptide (PACAP; 1 nM-1 microM), ATP (10 microM-30 mM), and tricarbonyldichlororuthenum dimer (1 microM-1 mM) was unaffected by tetrodotoxin (1 microM) or l-N(G)-nitroarginine methyl ester (l-NAME; 100 microM). Calcitonin gene-related peptide (CGRP; 1 nM-1 microM) did not affect LES tone. ATP relaxation was blocked by 1 microM apamin and the P2Y(1) antagonist MRS 2179 (N6-methyl 2'-deoxyadenosine 3',5'-bisphosphate; 10 microM). Apamin inhibited PACAP relaxation. VIP and PACAP relaxation was blocked by 10 U/ml alpha-chymotrypsin. L-NAME (-62.52 +/- 13.13%) and 1H-[1,2,4]oxadiazole-[4,3-alpha]quinoxalin-1-one (ODQ; 10 microM, -67.67 +/- 6.80%) similarly inhibited electrical LES relaxation, and apamin blocked non-nitrergic relaxation. Nicotine relaxation (100 microM) was inhibited by L-NAME (-60.37 +/- 10.8%) and ODQ (-41.90 +/- 7.89%), and apamin also blocked non-nitrergic relaxation. Non-nitrergic and apamin-sensitive LES relaxation by electrical stimulation or nicotine was strongly inhibited by MRS 2179, slightly inhibited by alpha-chymotrypsin and the P2X(1,2,3) receptor antagonist NF 279 (8,8 cent-[carbonylbis(imino-4,1-phenylenecarbonylimino-4,1-phenylenecarbonylimino)]bis-1,3,5-naphthalenetrisulfonic acid hexasodium salt; 10 microM), and unaffected by tin protoporphyrin IX (100 microM). Porcine LES relaxation after stimulation of intrinsic inhibitory motor neurons is mediated by two main neuromuscular pathways: nitric oxide through guanylate cyclase signaling and apamin-insensitive mechanisms and by non-nitrergic apamin-sensitive neurotransmission mainly mediated by ATP, ADP, or a related purine acting on P2Y1 receptors and a minor contribution of purinergic P2X1,2,3 receptors and PACAP. Nitrergic and purinergic co-transmitters show parallel effects of similar magnitude without major interplay. Our study shows no role for CGRP and only a minor one for VIP and carbon monoxide in porcine LES relaxation.

Comparison of two methods of PCR followed by enzymatic restriction digestion for detection and typing of herpes simplex viruses isolated from patients with mucocutaneous or cutaneous lesions.

A multitude of different polymerase chain reactions (PCRs) have been described for detection and typing of Herpes simplex virus (HSV). This paper compares two PCRs coupled to enzymatic restriction (PCR/RFLP) to detect and type HSV. A primers set was designed to amplify a HSV DNA fragment from UL30 and UL 15 genes. Typing was done by restriction of the UL30 and UL15 amplicons with Ava II and Hpa II enzymes, respectively. This strategy was tested with two reference strains (HSV-1 McIntyre, and HSV-2 G), and 47 clinical HSV isolates. Both PCRs produced the expected amplicons (a 492 bp UL30, and 305 bp UL15). The restriction of both amplicons clearly differentiated HSV- from HSV-2, and produced equal results. Thirty one (66%) of the isolates were identified as HSV-1, and the other 16 (34%), as HSV-2. Most of the HSV-1 isolates (27/31) were from orofacial and thoracic lesions; and also, one half of the HSV-2 isolates (8/16) were from the same anatomical regions. Our results showed that either of the two PCR/RFLP could be used to detect and type HSV. Furthermore, our results of the anatomical site of HSV-1 and HSV-2 infections are consistent with previous reports which have shown changes in the classical anatomical localization of herpesvirus infections.