Effect of porcine reproductive and respiratory syndrome virus 2 on angiogenesis and cell proliferation at the maternal-fetal interface.

Angiogenesis and cell proliferation in reproductive tissues are essential events for the maintenance of pregnancy, and alterations can lead to compromised fetal development and survival. Porcine reproductive and respiratory syndrome virus 2 (PRRSV-2) induces reproductive disease with negative financial and production impact on the swine industry. PRRSV-2 infection alters placental physiology through inflammatory and apoptotic pathways, yet fetal susceptibility varies. This study aimed to evaluate angiogenesis and cell proliferation in the porcine maternal-fetal interface (MFI) and determine if these physiological processes were altered by PRRSV-2 infection. Thirty-one pregnant gilts were inoculated with PRRSV-2 at gestation day 86 ± 0.4 (mean ± SD). Seven control gilts were sham-inoculated. All gilts were euthanized at 12 days postinoculation. Angiogenesis and cell proliferation were determined through the detection of vascular endothelial growth factor (VEGF) and Ki-67, respectively, using immunofluorescence of the MFI from 4 fetal resilience groups: uninfected (UNIF), high viral load-viable (HVL-VIA), and HVL-meconium-stained (MEC) from PRRSV-infected gilts, as well from sham-inoculated (CON) gilts. VEGF immunolabeling in the uterine submucosa was significantly lower in MEC compared with UNIF and HVL-VIA groups. Significantly greater Ki67 immunolabeling was detected in the trophoblasts of CON fetuses versus all other groups, and in uterine epithelium of CON and UNIF fetuses versus HVL-VIA and MEC. These results suggest that fetal resilience may be related to greater cell proliferation in uterine epithelium, and fetal compromise with reduced uterine submucosal angiogenesis, except fetuses with intrauterine growth restriction, in which inherently lower submucosal angiogenesis may be protective against PRRSV infection.

Ultrasonographic Diagnosis of A2 or A4 Flexor Tendon Pulley Injury: A Systematic Review.

A2 or A4 annular finger pulley tears are common injuries in rock climbers. This study reviews the measurement procedures used and tendon-to-bone distance data obtained on high-resolution ultrasound images when diagnosing isolated rupture of the A2 or A4 pulleys. Out of 3447 records extracted, only 7 remained after applying the exclusion criteria. In diagnosing a complete rupture, tendon-to-bone distance used varied widely from 1.9 to 5.1 mm for A2 and from 1.8 to 3.1 mm for A4. Our findings point to a lack of consensus diagnostic criteria for pulley injuries and identify technical details needing further research.

Isolation and in vitro antimicrobial susceptibility of porcine Lawsonia intracellularis from Brazil and Thailand.

BACKGROUND: Lawsonia intracellularis is an obligate intracellular bacterium which cannot be cultured by conventional bacteriological methods. Furthermore, L. intracellularis needs enriched medium and a unique atmosphere for isolation, cultivation and propagation. Because of this,there are only a few isolates of L. intracellularis available and few studies in vitro demonstrating the susceptibility of this bacterium to antimicrobial agents. The objectives of this study were to isolate South American and Southeast Asia strains of L.intracellularis and to determine the in vitro antimicrobial activity against these isolates. Tested antimicrobials included: chlortetracycline, lincomycin, tiamulin, tylosin and valnemulin(against both Brazilian and Thailand strains) and additionally, amoxicillin, zinc-bacitracin, carbadox, enrofloxacin, gentamicin, sulfamethazine, trimethoprim, spectinomycin and a combination (1:1) of spectinomycin and lincomycin were also tested against the Thai isolates. The minimum inhibitory concentration (MIC) was determined by the antimicrobial activity that inhibited 99% of L. intracellularis growth in a cell culture as compared to the control (antimicrobial-free). RESULTS: Two strains from Brazil and three strains from Thailand were successfully isolated and established in cell culture. Each antimicrobial was evaluated for intracellular and extracellular activity. Pleuromutilin group (valnemulin and tiamulin) and carbadox were the most active against L. intracellularis strains tested. Tylosin showed intermediate activity, chlortetracycline had variable results between low and intermediate activity, as well as spectinomycin, spectinomycin and lincomycin, amoxicillin, sulfamethazine and enrofloxacin. L. intracellularis was resistant to lincomycin, gentamicin, trimethoprim, colistin and bacitracin in in vitro conditions. CONCLUSIONS: This is the first report of isolation of L. intracellularis strains from South America and Southeast Asia and characterization of the antimicrobial susceptibility patterns of these new strains.

Synergic Effect of Brachyspira hyodysenteriae and Lawsonia intracellularis Coinfection: Anatomopathological and Microbiome Evaluation.

Brachyspira hyodysenteriae and Lawsonia intracellularis coinfection has been observed in the diagnostic routine; however, no studies have evaluated their interaction. This study aimed to characterize lesions and possible synergisms in experimentally infected pigs. Four groups of piglets, coinfection (CO), B. hyodysenteriae (BRA), L. intracellularis (LAW), and negative control (NEG), were used. Clinical signals were evaluated, and fecal samples were collected for qPCR. At 21 days post infection (dpi), all animals were euthanized. Gross lesions, bacterial isolation, histopathology, immunohistochemistry, and fecal microbiome analyses were performed. Diarrhea started at 12 dpi, affecting 11/12 pigs in the CO group and 5/11 pigs in the BRA group. Histopathological lesions were significantly more severe in the CO than the other groups. B. hyodysenteriae was isolated from 11/12 pigs in CO and 5/11 BRA groups. Pigs started shedding L. intracellularis at 3 dpi, and all inoculated pigs tested positive on day 21. A total of 10/12 CO and 7/11 BRA animals tested positive for B. hyodysenteriae by qPCR. A relatively low abundance of microbiota was observed in the CO group. Clinical signs and macroscopic and microscopic lesions were significantly more severe in the CO group compared to the other groups. The presence of L. intracellularis in the CO group increased the severity of swine dysentery.

Detection of PRRSV-2 alone and co-localized with CD163 positive macrophages in porcine placental areolae.

The porcine epitheliochorial placenta creates a barrier for the transplacental transfer of some nutrients from the dam to the fetus, as well as feto-lethal viruses such as porcine reproductive and respiratory syndrome virus 2 (PRRSV-2). Areolae are specialized structures within the porcine placenta with a high absorptive and substance transport capacity that facilitate embryonic development. The overarching aim of this study was to characterize the localization of PRRSV-2 in and adjacent to areolae to provide insight into whether transplacental transmission might occur through placental areolae. Control (CON) plus three phenotypic fetal groups were selected based on levels of virus in fetal placenta, sera and thymus, to determine if fetal resilience was related to differences in PRRSV-2 localization, alone or co-localized with CD163+ macrophages. These fetal groups represented a range of susceptibility: uninfected (UNINF) being resistant, infected in placenta only (PLCO) being resilient, and high viral load viable (HVL-VIA) being most susceptible. Finally, potential factors related to PRRSV-2 localization, including the severity of inflammation in endometrium and placenta, and intrauterine growth restriction, known resilience factors, were assessed. Thirty-one pregnant gilts were inoculated with PRRSV-2 at gestation day 86 ± 0.4. Seven pregnant gilts were sham-inoculated. Gilts were euthanized at 12 days post-infection. Presence of PRRSV and CD163+ macrophages were determined using immunofluorescence in cryosections of maternal-fetal interface (MFI) with and without areolae. In the maternal, fetal and cavity of areolar region PRRSV particles were found both independently and co-localized with CD163+ macrophages. Similarly, individual, and co-localized particles were observed in the maternal and fetal sides of the MFI region of infected fetuses. Weak positive correlations were observed between the counts of CD163+ macrophages and some inflammation scores in endometrial and placental tissues, but no correlations with PRRSV-2 localization. There were no differences across the four fetal groups evaluated. These results suggest that transplacental transmission of PRRSV may occur through the areolae, either as non-cell associated or in association with infected CD163 macrophages.

Experimental Infection of Pigs with a ST 245 Brachyspira hyodysenteriae Isolated from an Asymptomatic Pig in a Herd with No History of Swine Dysentery.

Swine dysentery (SD) is characterized by a severe mucohemorrhagic colitis caused by infection with Brachyspira species. In infected herds the disease causes considerable financial loss due to mortality, slow growth rates, poor feed conversion, and costs of treatment. B. hyodysenteriae is the most common etiological agent of SD and infection is usually associated with disease. However, isolated reports have described low pathogenic strains of B. hyodysenteriae. The aim of this study was to describe an experimental infection trial using a subclinical B. hyodysenteriae isolated from an animal without clinical signs and from a disease-free herd, to evaluate the pathogenicity and clinical pathological characteristics compared to a highly clinical isolate. Forty-eight 5-week-old pigs were divided into three groups: control, clinical and the subclinical isolates. The first detection/isolation of B. hyodysenteriae in samples of the animals challenged with a known clinical B. hyodysenteriae strain (clinical group) occurred 5th day post inoculation. Considering the whole period of the study, 11/16 animals from this group were qPCR positive in fecal samples, and diarrhea was observed in 10/16 pigs. In the subclinical isolate group, one animal had diarrhea. There were SD large intestine lesions in 3 animals at necropsy and positive B. hyodysenteriae isolation in 7/15 samples of the subclinical group. In the control group, no diarrhea, gross/microscopic lesions, or qPCR positivity were observed. Clinical signs, bacterial isolation, macroscopic and histologic lesions were significantly difference among groups, demonstrating low pathogenicity of the subclinical isolate in susceptible pigs.

Samples sizes required to accurately quantify viral load and histologic lesion severity at the maternal-fetal interface of PRRSV-inoculated pregnant gilts.

Porcine reproductive and respiratory syndrome virus (PRRSV) is transmitted vertically, causing fetal death in late gestation. Spatiotemporal distribution of virus at the maternal-fetal interface (MFI) is variable, and accurate assessment of viral concentration and lesions is thus subject to sampling error. Our objectives were: 1) to assess whether viral load and lesion severity in a single sample of endometrium (END) and placenta (PLC), collected near the base of the umbilical cord (the current standard), are representative of the entire organ; and 2) to compare sampling strategies and evaluate if spatial variation in viral load can be overcome by pooling of like-tissues. Spatially distinct pieces of END and PLC of 24 fetuses from PRRSV-2-infected dams were collected. PRRSV RNA quantified by RT-qPCR was compared in 5 individual pieces per fetus and in respective pools of tissue and extracted RNA. Three distinct pieces of MFI were assessed for histologic severity. Concordance correlation and kappa inter-rater agreement were used to characterize agreement among individual samples and pools. The viral load of individual samples and pools of END had greater concordance to a referent standard than did samples of PLC. Larger pool sizes had greater concordance than smaller pool sizes. Average viral load and lesion severity did not differ by location sampled, and no technical advantages of pooling tissues versus RNA extracts were found. We conclude that multiple pieces of MFI tissues must be evaluated to accurately assess lesion severity and viral load. Three pieces per fetus provided a reasonable balance of cost and logistic feasibility.

Grip Force Measurement as a Complement to High-Resolution Ultrasound in the Diagnosis and Follow-Up of A2 and A4 Finger Pulley Injuries.

The ability of finger flexors to generate force has been studied in relation to climbing performance. However, not much attention has been paid to the decrease in finger grip force in relation to annular pulley injuries. The purpose of the present study was to determine if an injured annular pulley implies a finger flexor force decrease, as well as its relation to clinical and sonographic changes. We performed an observational study in 39 rock climbers with A2 or A4 pulley injuries to the 3rd or 4th fingers. The variables considered were pain upon palpation, ultrasound tendon-bone distance, and finger grip strength decrease. Three rock climbing grip types were considered: the one finger crimp, open crimp, and close crimp. Injured rock climbers presented a decrease in finger grip strength compared to non-injured controls when performing a one finger crimp (p < 0.001). There exists a significant correlation between a tendon-bone distance at the level of the injured pulley and a decreased finger grip strength measured by performing a one finger crimp (p = 0.006). A decrease in finger grip strength could be considered in the diagnostic and follow-up process of A2 and A4 pulley injuries to the 3rd and 4th fingers.

Oral fluid for detection of exposure to Lawsonia intracellularis in naturally infected pigs.

To demonstrate the utility of oral fluid (OF) for indirect diagnostic detection of Lawsonia intracellularis (Li), 15 pig farms were studied. Serum and fecal samples were collected from 20 animals from five different age groups on each farm. OF samples were collected from animals in two pens of the same age groups. Serum and OF samples were analyzed in an immunoperoxidase in monolayer assay (IPMA) for the detection of anti-Li immunoglobulin G (IgG) and A (IgA). Compatible results were found between PCR and IgG in OF in four of the five ages evaluated. Simultaneous detection of IgG in serum and OF was mainly observed on farms showing clinical signs suggestive of porcine proliferative enteropathy (PPE). These findings demonstrate the potential usefulness of OF in detecting anti-Li antibodies as a diagnostic tool that can be used to monitor PPE in herds with clinical signs compatible with the disease.

[Prognostic factors in renal cancer with venous thrombus survival analysis.] / Factores pronósticos en cáncer renal con trombo en vena. Análisis de supervivencia.

OBJECTIVES: To analyze surgery for renal cancer with venous thrombus at different levels, perioperative complications and prognostic factors associated to overall, cancer-specific and disease-free survival. MATERIAL AND METHODS: Retrospective analysis of 42 cases of renal cancer with venous thrombus performed between 2005 and 2015. The level reached by the thrombus was established according to the Mayo Clinic classification. Postoperative complications were staged according to Clavien-Dindo classification. RESULTS: Most frequent in males. Mean age 65.7 years. 16.6% were tumors with level II thrombus. Subcostal approach was performed in 58.9%. Extracorporeal circulation with cardiac arrest and hypothermia was established in 2 patients. Resection of metastatic disease was performed in 3 patients during radical nephrectomy. Reoperation was 2.3% while, perioperative mortality was 4.7%. 30% presented with metastases at diagnosis. Twenty patients progressed at 15.5 months (3-55). Overall survival was 60 months. The cancer-specific mortality was 75%. Disease-free survival was 30% at 55 months. CONCLUSIONS: Surgical treatment of renal cancer with venous thrombus requires a multidisciplinary management. The surgical technique varies according to the level reached by the venous thrombus. Tumor stage is the most important prognostic factor. Thrombus level influences prognosis, with longer survival for patients with tumor thrombus confined to the renal vein (pT3a) in comparison to tumors with thrombus in the atrium (pT3c).

Niveles séricos de apolipoproteínas A-I y B en niños escolares de escuelas rurales / Serum levels of apolipoprotein A-I and B in rural school children

Los biomarcadores de riesgo aterogénico están presentes desdeetapas tempranas de la vida, uno de ellos son las apolipoproteínas. Estas juegan un papel fundamental en eldesarrollo de aterosclerosis según resultados de numerosos estudios epidemiológicos. El objetivo del trabajo fue evaluar los niveles de apolipoproteínas (apo) A-I y B y el coeficiente apo B/ apo A-1 en niños dislipidémicos. Se realizó un estudioobservacional, descriptivo, de corte transversal con muestreo no probabilístico. En una población de 132 niños, de ambos sexos, entre 4 a 13 años de edad, pertenecientes a 4 escuelas rurales de diferentes localidades del Paraguay, se encontraron 38 niños hipercolesterolémicos en quienes además se evaluaron los niveles de apolipoproteínas A-I y B. Se obtuvo consentimiento informado de los padres. El colesterol total fue medido por elmétodo enzimático automatizado y las apolipoproteínas A-I y B por el método inmunoturbidimétrico. Las concentraciones medias de apo A-I y B fueron de 159,89 ± 28,63 mg/dl y 99,79 ± 20,86 mg/dl respectivamente, la concentración media de colesterol fue de 192 ± 22,31. La media del cociente apo B/apo A-I fue de 0,64. Se detectaron niveles moderados a altos de apo B, en un considerable número de niños (63,5 %), y una correlación positiva entre las apolipoproteínas B y el colesterol, un 10, 53 % de los niños presentaron riesgo aterogénico alto deacuerdo al cociente apo B/apo A-I.

Biomarkers for atherogenic risk are present early in life, one such biomarker are the apolipoproteins. Apolipoproteins play a key role in the development of atherosclerosis according to results of numerous epidemiological studies. Our objective was to assess the levels of apolipoprotein (apo) A1 and B and the apo B/A-1 ratio in dyslipidemic children. We performed an observational, descriptive, cross-sectional study with nonprobabilistic sampling. In a population of 132 children aged 4 to13 years of both sexes from 4 rural schools in different parts of Paraguay, levels of apolipoproteins A-1 and B were assessed in 38 who were found to be hypercholesterolemic. Informedconsent was obtained from parents. Total cholesterol was measured by the automated enzymatic method andapolipoproteins A1 and B by the immunoturbidimetric method. Mean concentration was 159.89 ±28.63 mg/dl for apo A-1 and 99.79 ±20.86 mg/dl for apo B, while the mean cholesterol concentration was 192 ±22.31. The mean ratio of apo B/A1 was 0.64. We detected moderate to high levels of apo B in a considerable number of children (63.5%), and a positive correlation between apolipoprotein B and cholesterol. Some10.53% of children demonstrated high atherogenic risk as shown by the apo B/A1 ratio found.

Efectos térmicos centrales y cutáneos en el miembro superior de la inducción anestésica con tiopental / Core and cutaneous thermal effects on the upper limb after induction of general anaesthesia with thiopental

Objetivos: Describir y comparar los efectos térmicos (centrales y cutáneos en el miembro superior) y fotopletismográficos inmediatos a la inducción anestésica con fentanilo, tiopental sódico y vecuronio. Pacientes y métodos: Hemos estudiado las variaciones de las temperaturas central y cutáneas (del miembro superior: dedo, antebrazo y brazo), así como las variaciones del flujo sanguíneo digital, en 20 pacientes adultos, ASA-I (12 hombres y 8 mujeres), cada 5 minutos durante los 20 minutos iniciales de la anestesia general. La temperatura se midió con termosondas desechables (esofágicas y cutáneas) conectadas a un termómetro digital y el flujo sanguíneo digital mediante fotopletismografía digital. La anestesia se realizó sin premedicación con: tiopental sódico (5 mg x kg), fentanilo 0,1 mg y vecuronio 0,1 mg x kg. El mantenimiento anestésico se realizó con N2O-O2 (75/35 por ciento) y fentanilo 0,1 mg a los 10 minutos, sin anestésicos inhalatorios. Resultados: La inducción anestésica se asoció con vasodilatación digital intensa y rápida, con un incremento marcado de la amplitud de la onda fotopletismográfica (AOFP) (AOFP = 6,9 ñ 3,9 mV/V, a los 5 min, P<0,001) y de la temperatura cutánea del dedo (Ta Dedo = 6,3 ñ 2,8ºC, a los 10 min, P<0,001). Sin embargo, los cambios de la temperatura cutánea en el brazo y antebrazo fueron muy inferiores y más lentos (menores de 1ºC en los 20 min, en ambos casos). Solamente encontramos una correlación significativa entre la AOFP y la Ta Dedo: r=0,54 (P<0,001). Conclusiones: Tras inducción anestésica con tiopental sódico, fentanilo y vecuronio sucede un calentamiento cutáneo en el miembro superior, distribuido de forma desigual. El calentamiento es marcado en los dedos mientras que en el brazo y en el antebrazo las variaciones térmicas son menores. La apertura de las fístulas arterio-venosas termorreguladoras de las manos, consecuente con la inhibición simpática de la inducción anestésica, puede tener un papel destacado en la génesis de la hipotermia de redistribución de la inducción de la anestesia general.

Sindrome de Down, experiencia de 10 años (1988-1998) / Down Sindrome, experience of 10 years (1988-1998) / Annual reports 1998

Trescientos cincuenta y nueve pacientes portadores del Síndrome de Down fueron seleccionados entre 39 individuos con la enfermedad que concurrieron al Departamento de Genética del IICS para su estudio citogenético, en un periodo de 10 años. Se describen, distribución por sexo, peso, presencia o ausencia de malformaciones menores y mayores, y los tipos de alteraciones cromosómicas. Además los datos de los progenitores (edad, estado civil, consaguinidad). El 39,8 por ciento de los pacientes eran del sexo femenino y 60,2 por ciento del sexo masculino. Los tipos de alteraciones cromosómicas fueron en el 90,3 por ciento (324/359) una trisomia 21 libre simple, 7,2 por ciento (26/359) mosaico, 1.9 por ciento (7/359) translocaciones y doble aneuplodia en el 0,6 por ciento (2/326) de los casos. El 53, 2 por ciento tenia menos de 30 dias al momento de la consulta. El 100 por ciento de los pacientes presentó tres malformaciones menores y 30 por ciento malformaciones mayores, entre las que se destacan las cardíacas en un 20 por ciento. En el 60 por ciento de los casos se observo que la edad materna era mayor a los 35 años