Expression of genes associated with fertility in the uterus and oviduct of heifers challenged with lipopolysaccharide.

Lipopolysaccharide (LPS) endotoxemia has been negatively associated with fertility. This study aimed to investigate the effect of LPS-induced inflammation on gene expression associated with bovine fertility in the uterus and oviduct. Sixteen healthy heifers were divided into two groups. The LPS group (n = 8) received two intravenous (i.v.) injections of 0.5 µg/kg of body weight of LPS with a 24-h interval, and the control group (n = 8) received two i.v. injections of saline solution with the same interval of time. All the animals had the follicular wave synchronized. Three days after the second injection of LPS, all animals were slaughtered and uterine and oviduct samples were collected. Gene expression associated with inflammatory response, thermal and oxidative stresses, oviduct environment quality, and uterine environment quality was evaluated. Body temperature and leucogram demonstrated that LPS induced an acute systemic inflammatory response. In the uterus, the expression of PTGS2 and NANOG genes was downregulated by the LPS challenge. However, no change in expression was observed in the other evaluated genes in the uterus, nor those evaluated in the oviduct. In conclusion, the inflammatory process triggered by LPS did not persist in the uterus and oviduct 3 days after challenge with LPS. Nonetheless, reduction in PTGS2 and NANOG expression in the uterus suggested that, indirectly, LPS may have a prolonged effect, which may affect corpus luteum and endometrial functions.

Ventromedial Prefrontal Cortex Tracks Multiple Environmental Variables during Search.

To make efficient foraging decisions, we must combine information about the values of available options with nonvalue information. Some accounts of ventromedial PFC (vmPFC) suggest that it has a narrow role limited to evaluating immediately available options. We examined responses of neurons in area 14 (a putative macaque homolog of human vmPFC) as 2 male macaques performed a novel foraging search task. Although many neurons encoded the values of immediately available offers, they also independently encoded several other variables that influence choice, but that are conceptually distinct from offer value. These variables include average reward rate, number of offers viewed per trial, previous offer values, previous outcome sizes, and the locations of the currently attended offer. We conclude that, rather than serving as specialized economic value center, vmPFC plays a broad role in integrating relevant environmental information to drive foraging decisions.SIGNIFICANCE STATEMENT Decision makers must often choose whether to take an immediately available option or continue to search for a better one. We hypothesized that this process, which is integral to foraging theory, leaves neural signatures in the brain region ventromedial PFC. Subjects performed a novel foraging task in which they searched through differently valued options and attempted to balance their reward threshold with various time costs. We found that neurons not only encode the values of immediately available offers, but multiplexed these with environmental variables, including reward rate, number of offers viewed, previous offer values, and spatial information. We conclude that vmPFC plays a rich role in encoding and integrating multiple foraging-related variables during economic decisions.

Minimally invasive fetal surgery for myelomeningocele: preliminary report from a single center.

OBJECTIVE: Recent trials have shown the safety and benefits of fetoscopic treatment of myelomeningocele (MMC). The authors' aim was to report their preliminary results of prenatal fetoscopic treatment of MMC using a biocellulose patch, focusing on neurological outcomes, fetal and maternal complications, neonatal CSF leakage, postnatal hydrocephalus, and radiological outcomes. METHODS: Preoperative assessment included clinical examination, ultrasound imaging, and MRI of the fetus. Patients underwent purely fetoscopic in utero MMC repair, followed by postoperative in utero and postnatal MRI. All participants received multidisciplinary follow-up. RESULTS: Five pregnant women carrying fetuses affected by MMC signed informed consent for the fetoscopic treatment of the defect. The mean MMC size was 30.4 mm (range 19-49 mm). Defect locations were L1 (2 cases), L5 (2 cases), and L4 (1 case). Hindbrain herniation and ventriculomegaly were documented in all cases. The mean gestational age at surgery was 28.2 weeks (range 27.8-28.8 weeks). Fetoscopic repair was performed in all cases. The mean gestational age at delivery was 33.9 weeks (range 29.3-37.4 weeks). After surgery, reversal of hindbrain herniation was documented in all cases. Three newborns developed signs of hydrocephalus requiring CSF diversion. Neurological outcomes in terms of motor level were favorable in all cases, but a premature newborn died due to CSF infection and sepsis. CONCLUSIONS: The authors' preliminary results suggest that fetoscopic treatment of MMC is feasible, reproducible, and safe for mothers and their babies. Neurological outcomes were favorable and similar to those in the available literature. As known, prematurity was the greatest complication.

Neuronal responses support a role for orbitofrontal cortex in cognitive set reconfiguration.

We are often faced with the need to abandon no-longer beneficial rules and adopt new ones. This process, known as cognitive set reconfiguration, is a hallmark of executive control. Although cognitive functions like reconfiguration are most often associated with dorsal prefrontal structures, recent evidence suggests that the orbitofrontal cortex (OFC) may play an important role as well. We recorded the activity of OFC neurons while rhesus macaques performed an analogue of the Wisconsin Card Sorting Task that involved a trial and error stage. The OFC neurons demonstrated two types of switch-related activity, an early (switch-away) signal and a late (switch-to) signal, when the new task set was established. We also found a pattern of match modulation: a significant change in activity for the stimulus that matched the current perceptual rule (and would therefore be selected). These results extend our understanding of the executive functions of the OFC. They also allow us to directly compare the OFC with the complementary datasets we previously collected in the ventral (VS) and dorsal (DS) striatum. Although both effects are observed in all three areas, the timing of responses aligns the OFC more closely with DS than with VS.

MicroRNA Profiling Using a PCR-Based Method.

MicroRNAs (miRNAs) are small non-coding RNA molecules involved in the post-transcriptional regulation of specific mRNA targets, thus possibly controlling many biological processes. The miRNA profiling analysis can contribute to understanding several signaling pathways, as biomarkers for molecular diagnostic, as well as potential to be used as therapeutic targets. The miRNAs expression can be analyzed by quantitative reverse transcription PCR (RT-qPCR), microarrays, and RNA sequencing. The RT-qPCR method is sensitive and specific and has a lower cost when compared to other techniques as microarrays and RNA sequencing. Therefore, the protocol presented in this chapter describes step by step all the details to perform miRNA analysis using primer-based RT-qPCR.

[Acute myocardial infarction in a young woman with systemic lupus erythematosus]. / Infarto agudo de miocardio en mujer joven con diagnóstico de lupus eritematoso sistémico.

We repor a case of acute ST elevation myocardial infarction in a 22-year-old patient with SLE, hypertension and nephropathy who underwent successful coronary angioplasty to a middle third of the left anterior descending artery. She evolved without signs of heart failure however, due to the delay in diagnosis, she presented severe deterioration of ventricular function. ST segment elevation myocardial infarction is a very rare event in young premenopausal women, but compared to the general population, patients with lupus have at least a 50% higher risk of suffering it regardless their age. In this population, the most frequent causes are vasculitis, early atherosclerosis and secondary thrombosis to antiphospholipid syndrome. In the context of lupus, conditions such as the presence of nephritis have been described as favoring the appearance of myocardial infarction, constituting subgroups of higher risk. The increased risk of AMI in patients with SLE must be taken into account and must be suspected as a differential diagnosis of precordial pain in young women, even those under 25 years of age, a population categorized as having low CV risk according to traditional scores. This would avoid delays in diagnosis and treatment with adverse consequences such as extensive myocardial necrosis and its impact on ventricular systolic function, as occurred in this patient.

Se presenta un caso de infarto agudo de miocardio con elevación del segmento ST en una paciente de 22 años de edad, con LES, HTA y nefropatía a la cual se le realizó angioplastia coronaria exitosa a tercio medio de arteria descendente anterior. Evolucionó sin signos de insuficiencia cardiaca, sin embargo, debido al retraso en el diagnóstico presentó deterioro grave de la función ventricular. El infarto agudo de miocardio con elevación del ST es un evento muy poco frecuente en mujeres pre menopáusicas pero, en comparación con la población general las pacientes con lupus eritematoso sistémico presentan al menos un 50% más de riesgo de padecerlo independientemente de la edad. En esta población, las etiologías más frecuentes son la vasculitis, la aterosclerosis precoz y la trombosis secundaria a síndrome antifosfolipídico. A su vez, en contexto de lupus, se han descripto condiciones como la presencia de nefritis lúpica, que favorecen aún más a la aparición del infarto de miocardio, constituyendo subgrupos de mayor riesgo. El incremento del riesgo de IAM en los pacientes con LES debe tenerse en cuenta y hay que sospechar como diagnóstico diferencial del dolor precordial aún en mujeres jóvenes, incluso menores de 25 años, población categorizada como de bajo riesgo cardiovascular según los scores y criterios tradicionales. Esto evitaría las demoras en el diagnóstico y tratamiento con consecuencias pronósticas adversas como la necrosis miocárdica extensa y su impacto negativo sobre la función sistólica ventricular como ocurrió en esta paciente.

Development of low-cost cage-like particles to formulate veterinary vaccines.

Low-cost adjuvants are urgently needed for the development of veterinary vaccines able to trigger strong immune responses. In this work, we describe a method to obtain a low-cost cage-like particles (ISCOMATRIX-like) adjuvant useful to formulate veterinary vaccines candidates. The main components to form the particles are lipids and saponins, which were obtained from egg yolk by ethanolic extraction and by dialyzing a non-refined saponins extract, respectively. Lipids were fully characterized by thin layer chromatography (TLC) and gas-chromatography (GC) and enzymatic methods, and saponins were characterized by TLC, HPLC and MALDI-TOF. Cage-like particles were prepared with these components or with commercial inputs. Both particles and the traditional Alum used in veterinary vaccines were compared by immunizing mice with Ovalbumin (OVA) formulated with these adjuvants and assessing IgG1, IgG2a anti OVA antibodies and specific Delayed-type Hypersensitivity (DTH). In the yolk extract, a mixture of phospholipids, cholesterol and minor components of the extract (e.g. lyso-phospholipids) with suitable proportions to generate cage-like particles was obtained. Also, semi-purified saponins with similar features to those of the QuilA® were obtained. Cage-like particles prepared with these components have 40-50 nm and triggers similar levels of Anti-OVA IgG1 and DTH than with commercial inputs but higher specific-IgG2a. Both adjuvants largely increased the levels of IgG1, IgG2a and DTH in relation to the formulation with Alum. The methods described to extract lipids from egg yolk and saponins from non-refined extract allowed us to obtain an inexpensive and highly effective adjuvant.

Long-term effect of oxycodone/naloxone on the management of postoperative pain after hysterectomy: a randomized prospective study.

BACKGROUND: The analgesic efficacy of oxycodone prolonged-release (PR) combined with naloxone PR (OXN) in postoperative pain management is recognized, however, few studies have examined the efficacy of OXN on pain relief and bowel function following hysterectomy. This study compared the effect of OXN vs. standard treatment for post-operative pain management and bowel function following hysterectomy. METHODS: This randomized prospective study included 83 women who underwent laparoscopic/laparotomic hysterectomy. General anesthesia was induced by propofol (1.5-2 mg/kg), fentanyl (50-100 µg) and rocuronium (0.6-1 mg/kg) and maintained with sevoflurane (MAC 0.8-1) and fentanyl (1-2 µg/kg). Intraoperative analgesia was performed with ketorolac (30 mg), paracetamol (1 g) and morphine (0.1 mg/kg). Postoperative analgesia in the control group (N.=41) included morphine (0.2-0.4 mg/kg/day), whereas the OXN (N.=42) group only received oxycodone (10 mg)/naloxone (5 mg) for the first 48 hours. As rescue analgesic, both groups received paracetamol (3 mg). Bowel Function Index (BFI) and pain numeric rating scales (NRS) were measured at day 0, 1, 2, 3, 5 and 7, whereas vital parameters, rescue medication and side effects were recorded for the first three days only. RESULTS: Bowel function indices were significantly improved in OXN-treated patients at all time points compared to morphine-treated patients. Mean static pain NRS was significantly decreased at day 2 and day 3 and dynamic pain NRS at day 3 in the OXN group. Side effects, rescue analgesic and antiemetics were more frequent in the control group. CONCLUSIONS: Improved pain control, bowel function and reduced side effects were observed with OXN compared to morphine in patients who underwent hysterectomy.

Virgin Coconut Oil Supplementation Prevents Airway Hyperreactivity of Guinea Pigs with Chronic Allergic Lung Inflammation by Antioxidant Mechanism.

Asthma is a chronic inflammatory disease of the airways characterized by immune cell infiltrates, bronchial hyperresponsiveness, and declining lung function. Thus, the possible effects of virgin coconut oil on a chronic allergic lung inflammation model were evaluated. Morphology of lung and airway tissue exhibited peribronchial inflammatory infiltrate, epithelial hyperplasia, and smooth muscle thickening in guinea pigs submitted to ovalbumin sensitization, which were prevented by virgin coconut oil supplementation. Additionally, in animals with lung inflammation, trachea contracted in response to ovalbumin administration, showed a greater contractile response to carbachol (CCh) and histamine, and these responses were prevented by the virgin coconut oil supplementation. Apocynin, a NADPH oxidase inhibitor, did not reduce the potency of CCh, whereas tempol, a superoxide dismutase mimetic, reduced potency only in nonsensitized animals. Catalase reduced the CCh potency in nonsensitized animals and animals sensitized and treated with coconut oil, indicating the participation of superoxide anion and hydrogen peroxide in the hypercontractility, which was prevented by virgin coconut oil. In the presence of L-NAME, a nitric oxide synthase (NOS) inhibitor, the CCh curve remained unchanged in nonsensitized animals but had increased efficacy and potency in sensitized animals, indicating an inhibition of endothelial NOS but ineffective in inhibiting inducible NOS. In animals sensitized and treated with coconut oil, the CCh curve was not altered, indicating a reduction in the release of NO by inducible NOS. These data were confirmed by peribronchiolar expression analysis of iNOS. The antioxidant capacity was reduced in the lungs of animals with chronic allergic lung inflammation, which was reversed by the coconut oil, and confirmed by analysis of peribronchiolar 8-iso-PGF2α content. Therefore, the virgin coconut oil supplementation reverses peribronchial inflammatory infiltrate, epithelial hyperplasia, smooth muscle thickening, and hypercontractility through oxidative stress and its interactions with the NO pathway.

Infarto agudo de miocardio en mujer joven con diagnóstico de lupus eritematoso sistémico / Acute myocardial infarction in a young woman with systemic lupus erythematosus

Resumen Se presenta un caso de infarto agudo de miocardio con elevación del segmento ST en una paciente de 22 años de edad, con LES, HTA y nefropatía a la cual se le realizó angioplastia coronaria exitosa a tercio medio de arteria descendente anterior. Evolucionó sin signos de insuficiencia cardiaca, sin embargo, debido al retraso en el diagnóstico presentó deterioro grave de la función ventricular. El infarto agudo de miocardio con elevación del ST es un evento muy poco frecuente en mujeres pre menopáusicas pero, en comparación con la población general las pacientes con lupus eritematoso sistémico presentan al menos un 50% más de riesgo de padecerlo independientemente de la edad. En esta población, las etiologías más frecuentes son la vasculitis, la aterosclerosis precoz y la trombosis secundaria a síndrome antifosfolipídico. A su vez, en contexto de lupus, se han descripto condiciones como la presencia de nefritis lúpica, que favorecen aún más a la aparición del infarto de miocardio, constituyendo subgrupos de mayor riesgo. El incremento del riesgo de IAM en los pacientes con LES debe tenerse en cuenta y hay que sospechar como diagnóstico diferencial del dolor precordial aún en mu jeres jóvenes, incluso menores de 25 años, población categorizada como de bajo riesgo cardiovascular según los scores y criterios tradicionales. Esto evitaría las demoras en el diagnóstico y tratamiento con consecuencias pronósticas adversas como la necrosis miocárdica extensa y su impacto negativo sobre la función sistólica ventricular como ocurrió en esta paciente.

Abstract We repor a case of acute ST elevation myocardial infarction in a 22-year-old patient with SLE, hypertension and nephropathy who underwent successful coronary angioplasty to a middle third of the left an terior descending artery. She evolved without signs of heart failure however, due to the delay in diagnosis, she presented severe deterioration of ventricular function. ST segment elevation myocardial infarction is a very rare event in young premenopausal women, but compared to the general population, patients with lupus have at least a 50% higher risk of suffering it regardless their age. In this population, the most frequent causes are vasculitis, early atherosclerosis and secondary thrombosis to antiphospholipid syndrome. In the context of lupus, conditions such as the presence of nephritis have been described as favoring the appearance of myocardial infarction, constituting subgroups of higher risk. The increased risk of AMI in patients with SLE must be taken into account and must be suspected as a differential diagnosis of precordial pain in young women, even those under 25 years of age, a population categorized as having low CV risk according to traditional scores. This would avoid delays in diagnosis and treatment with adverse consequences such as extensive myocardial necrosis and its impact on ventricular systolic function, as occurred in this patient.

Hypercaloric Diet Establishes Erectile Dysfunction in Rat: Mechanisms Underlying the Endothelial Damage.

Obesity is characterized by an excessive increase in body mass, leading to endothelial damage that may favor the development of erectile dysfunction (ED). ED is defined as the inability to achieve or maintain a penile erection long enough to have a sexual intercourse. In this context, different ED models were developed, however the high price of special animals or the long period to establish the disease has limited studies in this field. Therefore, this study proposed to establish and characterize a novel model of ED in rats associated to a hypercaloric diet consumption. Animals were randomly divided into control group (CG), which received a standard diet, and obese group (OG), fed with a hypercaloric diet during 8 weeks. Rat's erectile function was evaluated in vivo and in vitro. Food and caloric intake of OG were reduced compared to CG, due to an increased diet energy efficiency. However, OG presented an increased body mass, inguinal, retroperitoneal and epididymal adipose tissues, as well as body adiposity index at the end of experimental protocol. In erectile function analysis, there was a decrease in the number and the latency of penile erections in OG. Additionally, the contractile reactivity of corpus cavernosum was increased in OG, favoring penile detumescence and related to a reduced nitric oxide bioavailability and an increased in contractile prostaglandins levels as a consequence of endothelial damage. Moreover, the endothelium-relaxation reactivity of corpus cavernosum was attenuated in OG associated to the oxidative stress. Thus, it was provided a model for advances in sexual dysfunction field and drug discovery for ED treatment.

Interobserver agreement among pathologists for semiquantitative hormone receptor scoring in breast carcinoma.

The American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) guidelines recommend reporting of hormone receptor test results in a semiquantitative manner. This study used 74 resected estrogen receptor (ER)-positive invasive breast cancers to determine reproducibility of semiquantitative scoring of hormone receptors using the H-score method. Four pathologists independently scored each slide. Agreement among observers was analyzed via Fleiss κ statistics on ER and progesterone receptor (PR) categorical scores. Intraclass correlation coefficient (ICC) was used to estimate the interobserver agreement for ER and PR H-scores on a continuous scale (0-300). There was 100% agreement for categorical ER results (κ = 1) and 97% agreement (κ = 0.823, P < .001) for categorical PR results. For quantitative H-scores, ICC agreement was 0.85 (95% confidence interval [CI] = 0.79-0.90) for ER and 0.87 (95% CI = 0.82-0.92) for PR. Because the H-score provides a continuous measure of tumor hormone receptor content, we suggest universal adoption of this method.

Adaptación cultural de la versión peruana de la Muscle Appearance Satisfaction Scale (MASS) para dismorfia muscular / Cultural Adaptation of the Peruvian Version of the Muscle Appearance Satisfaction Scale (MASS) for Muscle Dysmorphia

La dismorfia muscular (DM) es una forma de trastorno dismórfico corporal considerado como un trastorno obsesivo compulsivo, caracterizado por la preocupación de tener una musculatura insuficiente, lo que lleva a realizar una actividad física intensa y prolongada y al consumo de suplementos proteicos y esteroides anabólicos. Afecta principalmente a varones jóvenes de 18 a 30 años, que optan por dietas hipo-calóricas e hiperproteicas, lo que genera efectos perjudiciales como problemas renales y hepáticos a largo plazo1. Según el Instituto Nacional de Salud del Perú, los problemas de salud mental son una de las prioridades de investigación del país, porque representan una importante carga de enfermedad para la sociedad y hay escasez de información científica local2. La DM no es la excepción y, como se la ha descrito en tiempos recientes, no se ha encontrado estudios peruanos sobre su frecuencia y los factores asociados; una de las causas es que no existe un instrumento validado para este contexto cultural.

Muscle dysmorphia (DM) is a form of body dysmorphic disorder considered as an obsessive compulsive disorder, characterized by the concern of having insufficient muscles, which leads to intense and prolonged physical activity and the consumption of protein supplements and anabolic steroids . It mainly affects young men between the ages of 18 and 30, who choose hypocaloric and hyperprotein diets, which generates harmful effects such as kidney and liver problems in the long term1. According to the National Institute of Health of Peru, mental health problems are one of the country's research priorities, because they represent a significant burden of disease for society and there is a shortage of local scientific information2. DM is no exception and, as it has been described in recent times, no Peruvian studies have been found on its frequency and associated factors; one of the causes is that there is no validated instrument for this cultural context.

Intercellular adhesion molecule-1 mediates cellular cross-talk between parenchymal and immune cells after lipopolysaccharide neutralization.

The mechanisms by which parenchymal cells interact with immune cells, particularly after removal of LPS, remain unknown. Lung explants from rats, mice deficient in the TNF gene, or human lung epithelial A549 cells were treated with LPS and washed, before naive alveolar macrophages, bone marrow monocytes, or PBMC, respectively, were added to the cultures. When the immune cells were cocultured with LPS-challenged explants or A549 cells, TNF production was greatly enhanced. This was not affected by neutralization of LPS with polymyxin B. The LPS-induced increase in the expression of ICAM-1 on A549 cells correlated with TNF production by PBMC. The cellular cross talk leading to the TNF response was blunted by an anti-ICAM-1 Ab and an ICAM-1 antisense oligonucleotide. In A549 cells, a persistent decrease in the concentration of intracellular cAMP was associated with colocalization of LPS into Toll-like receptor 4 and the Golgi apparatus, resulting in increased ICAM-1 expression. Inhibition of LPS internalization by cytochalasin D or treatment with dibutyryl cAMP attenuated ICAM-1 expression and TNF production by PBMC. In conclusion, lung epithelial cells are not bystanders, but possess memory of LPS through the expression of ICAM-1 that interacts with and activates leukocytes. This may provide an explanation for the failure of anti-LPS therapies in sepsis trials.

Hemoglobina glucosilada A1c (A1c): Variación etárea y determinantes clínicos y para clínicos en población sana / Glycated hemoglobin A1c: Clinical and laboratory variations in a selected population

Objetivos. El objetivo principal de este estudio era determinar si existía alguna tendencia o patrón de comportamiento de los niveles de A1c a diferentes edades, en personas sanas. Otros objetivos eran encontrar los determinantes clìnicos y paraclínicos mas importantes del nivel de A1c en personas sanas, modelar el comportamiento de la A1c en función de la edad, y comparar los niveles de A1c entre ambos sexos. Métodos. Se estudiaron 130 personas aparentemente sanas distribuidas en 5 grupos etáreos: 7 a 12 años (n=35), 13 a 18 años (n=34), 19 a 26 años (n=20), 27 a 45 años (n=19) y 46 a 60 años (n=22). Se practicó examen físico completo y determinación de glucemia de ayuno, perfil de lípidos, ácido úrico y cuadro hemático. Los niveles de A1c se determinaron por inmunoturbidimetría. Se realizaron comparaciones múltiples entre grupos de edad, así como regresión polinomial con los grupos de edad como variable independiente y A1c como variable dependiente. Para establecer los determinantes de la A1c se realizó regresión lineal múltiple. Conclusiones. Nuestros datos sustentan la hipótesis de que los niveles de A1c varían con la edad y el sexo en población no diabética. Hacia el futuro sería recomendable ajustar las metas de A1c en el tratamiento de los pacientes con diabetes según la edad y el sexo