Variantes genéticas recesivas asociadas a abortos y mortalidad de terneros en ganado lechero : estudio sobre su presencia e impacto en el desempeño reproductivo y productivo de vacas holando de Uruguay

Los experimentos I y II de la presente tesis consistieron en la optimización de las técnicas PCR-HRM/melting en tiempo real para la detección de la malformación vertebral compleja (CVM, OMIA 001340-9913) y la deficiencia de colesterol (CD, OMIA 001965-9913), así como para determinar su existencia en la muestra representativa de vacas Holando (n = 279 para CVM; n = 103 para CD) de Uruguay. Las amplificaciones de los productos PCR de 79 pb de CVM y de 170 y 146 pb de CD permitieron diferenciar claramente los dos genotipos. Se encontró que la prevalencia de CVM (6,45 %) fue alta. El inserto mutante BoERVK asociado con CD no fue detectado. El experimento III consistió en: (a) identificar toros (n = 3028) con información genética para haplotipos relacionados con problemas de fertilidad que causan abortos y mortalidad de terneros en ganado lechero (HH1, OMIA 000001-9913; HH3, OMIA 001824-9913; y HH4, OMIA 001826- 9913) mediante análisis del catálogo de padres Holando 2021; y (b) determinar la presencia de estos haplotipos mencionados, JH1 (OMIA 001697-9913) y CVM con el chip bovino GeneTitan®, estimar sus frecuencias y evaluar su impacto en la fertilidad de las vacas Holando y cruzas Holando con Jersey (n = 170) pertenecientes a un tambo experimental. De un total de 1468 toros con información genética para HH1 y de 1471 para HH3 y HH4, se encontraron 90 portadores para HH1, 60 para HH3 y 6 para HH4, respectivamente. Mediante genotipado con el chip se determinaron las prevalencias para HH1 y CVM que fueron de 0,59 y 4,3 % (call rate > 0,99), respectivamente. No se encontraron alelos mutantes para HH3, HH4 y JH1. Se evaluaron los datos reproductivos y se observó la presencia de los alelos mutantes de CVM y HH1 en vacas repetidoras, que parieron terneros muertos, con un promedio de 4 repeticiones del servicio para lograr la preñez

Building monitoring system in a large social housing intervention in Northern Italy

Within the framework of the well-known problem of the performance gap, the paper demonstrates how a building monitoring system is able to provide feedback data instrumental to address the ongoing management issues of multi-family buildings in social housing: the need to have a good understanding of what works and what does not in building operation, the need of bills controlling and allocation of individual costs between the occupants, the facility and energy management requirements including the understanding of occupant’s behavior. It adopts a case study approach, discussing the case of a large environmentally friendly social housing intervention consisting in 323 flats, in which a building monitoring system was installed

HLA Haplotype Mismatch Transplants and Posttransplant Cyclophosphamide

The use of high dose posttransplant cyclophosphamide (PT-CY) introduced by the Baltimore group approximately 10 years ago has been rapidly adopted worldwide and is becoming a standard for patients undergoing unmanipulated haploidentical (HAPLO) transplants. PT-CY has been used following nonmyeloablative as well as myeloablative conditioning regimens, for bone marrow or peripheral blood grafts, for patients with malignant and nonmalignant disorders. Retrospective comparisons of HAPLO grafts with conventional sibling and unrelated donor grafts have been published and suggest comparable outcome. The current questions to be answered include the use of PT-CY for sibling and unrelated donors transplant, possibly in the context of prospective randomized trial. HLA Haplotypes and Haploidentical Transplants The short arm of chromosome 6 is home to the human major histocompatibility complex (MHC) genes, which code for human leukocyte antigens (HLA): during meiosis, the MHC region does not undergo "crossing over" (except for rare events), and we inherit one HLA haplotype from the father and one from the mother. Biology of HAPLO Transplants The double problem of a HAPLO transplant is rejection of the graft, or host versus graft (HvG), and rejection of the host, or graft versus host disease (GvHD), and this double problem was the cause of failure of initial attempts. The first successful haplotype mismatch transplants were carried out in the early 1980s in severe combined immunodeficiency (SCID) patients, in whom the risk of rejection is minimal [2]: GvHD was prevented using T cell depletion (TCD) in the absence of any additional posttransplant GvHD prophylaxis Unmanipulated Marrow Grafts following a Nonmyeloablative Conditioning Regimen (NMA) and PT-CY The John Hopkins group hypothesized 40 years ago that high dose cyclophosphamide (CY), given 3 days after infusion of mismatched grafts, would protect rats from graft versus host disease (GvHD) Unmanipulated Marrow Grafts following a Myeloablative (MA) Conditioning Regimen A study using PT-CY and myeloablative conditioning (MAC) regimens has shown a low incidence of graft failure (<5%), a low incidence of acute grades II-IV GvHD (18%), a very low rate of severe grades III-IV acute GvHD (3%), and relapse as expected with a conventional CyA+MTX GvHD prophylaxis [10]. Unmanipulated Peripheral Blood Grafts and PT-CY Several centers have elected to use exclusively peripheral blood (PB) as a stem cell source in place of bone marrow (BM), based on donor preference, inability to secure operating room hours, and outcome considerations. These centers are unwilling to use BM and have thus adopted the Baltimore protocol using unmanipulated PB instead of BM. Results have been unexpectedly encouraging: this is clear when PB is given after the original Baltimore NMA conditioning regimen. In a recent paper, unmanipulated BM and PB have been compared in the NMA setting: incidence of acute and chronic GvHD, nonrelapse mortality, relapse, and survival were quite comparabl

IDENTIFICATION OF HOLSTEIN COWS CARRIERS OF COMPLEX VERTEBRAL MALFORMATION BY HIGH RESOLUTION MELTING CURVES (HRM)

The objective of this study was the optimization and implementation of a reliable and economical molecular screening method for the detection of the mutant allele of CVM (complex vertebral malformation, c.559G>T, SLC35A3) by HRM analysis, as well as analyzing its existence in a representative sample of Holstein cows from the Milk Genomic DNA Bank of Uruguay. The optimization of the HRM methodology in the RotorGene™ 6000 equipment (Corbett Life Science, Australia) by amplification of the 79 bp PCR products clearly differentiated the two genotypes: homozygous, wild type: GG; and heterozygous, carrier for the mutation CVM: GT (c.559G>T; SLC35A3). In the analyzed sample, the frequency of the mutant allele (T) for CVM was high (q = 0.032), with a prevalence of carrier cows of 6.45%. It is concluded that the PCR-HRM analysis is a fast, easily interpretable, low cost, and highly accurate technique for the detection of this mutation in Holstein cattle, which may be implemented in genetic selection programs

Different duplex/quadruplex junctions determine the properties of anti-thrombin aptamers with mixed folding.

Mixed duplex/quadruplex oligonucleotides have attracted great interest as therapeutic targets as well as effective biomedical aptamers. In the case of thrombin-binding aptamer (TBA), the addition of a duplex motif to the G-quadruplex module improves the aptamer resistance to biodegradation and the affinity for thrombin. In particular, the mixed oligonucleotide RE31 is significantly more effective than TBA in anticoagulation experiments and shows a slower disappearance rate in human plasma and blood. In the crystal structure of the complex with thrombin, RE31 adopts an elongated structure in which the duplex and quadruplex regions are perfectly stacked on top of each other, firmly connected by a well-structured junction. The lock-and-key shape complementarity between the TT loops of the G-quadruplex and the protein exosite I gives rise to the basic interaction that stabilizes the complex. However, our data suggest that the duplex motif may have an active role in determining the greater anti-thrombin activity in biological fluids with respect to TBA. This work gives new information on mixed oligonucleotides and highlights the importance of structural data on duplex/quadruplex junctions, which appear to be varied, unpredictable, and fundamental in determining the aptamer functional properties

Ambulatory Surgery for Perianal Disease

The gold standard in the diagnosis and treatment of proctological diseases is the exploration of the anal canal and distal rectum under anaesthesia (EUA), routinely performed as day case surgery. In selected cases it can be conducted as an outpatient exploration (OE) during a specialist surgical consultation. In the outpatient setting it is possible and safe to perform rubber band ligation, sclerotherapy and infrared coagulation for the treatment of haemorrhoidal disease, excision and incision of thrombosed external haemorrhoids, abscess drainage, setonage and fistulotomy also in case of perianal Crohn’s disease, anal warts and skin tags removal. In terms of patients’ satisfaction and success rate OE is comparable to EUA. All procedures can be performed under local anaesthesia. Pain control after the procedure is provided by oral pain killers

Procedimiento para la caracterización de una celda de punto triple de agua Isotech en el LATU

La realización en el punto triple de agua es crucial para una mejor aproximación a la Escala Internacional de Temperatura (ITS-90), ya que la definición de esta última está directamente vinculada al punto fijo citado. Es necesario conocer la influencia de la presión hidrostática en el punto triple de agua para corregir los valores de resistencia medidos y obtener una mejor aproximación al valor real de la temperatura en dicho punto. Este artículo describe las dificultades encontradas al estudiar el perfil térmico correspondiente a una celda de punto triple de agua comercial bajo condiciones normales de operación con un baño Dewar termostatizado con hielo picado. Este trabajo fue llevado a cabo en dos fases: en la primera se detectaron problemas con los perfiles de temperatura, cuyo comportamiento era lejano al ideal. Para mejorar dicho comportamiento se hicieron cambios en el equipamiento, con el fin de minimizar el tiempo de estabilización de la celda y mejorar la transferencia de calor. El aislamiento del baño fue mejorado y se colocó un dispositivo ecualizador de temperatura de cobre en el pozo de la celda. Los resultados se discuten en el presente artículo. Abstract The Realization of the Triple Point of Water (TPW) is of main importance when approximating to the International Temperature Scale of 1990 (ITS-90), because it is properly involved in its definition. Therefore it is necessary to know the Hydrostatic Pressure influence at the Triple Point of Water to correct the values of resistance measured and to obtain a better approximation of the real temperature of this point. This article shows the difficulties found when studying the thermal profile of one commercial cell of the triple point of water under normal operation conditions in a Dewar thermostatic bath with crushed ice. The work was performed in two phases. In the first phase problems were detected with the temperature profiles, whose behaviors were far away from the ideals ones. To improve this behavior changes were made in the equipment. The isolation of the bath was improved and copper bushing was set up into the well of the cell. Results are discussed in this paper

Ensayo de aptitud realizado en LATU para validación del método de calibración y uso de pipetas de vidrio y pipetas automáticas

En 2008 se realizó un ensayo de aptitud para calibración de material volumétrico, con la participación de varios analistas (en su mayoría nuevos en la tarea) de los laboratorios de ensayo del Laboratorio Tecnológico del Uruguay (LATU). Para el mismo se utilizaron dos pipetas, una de vidrio de 1 ml y una pipeta automática de volumen regulable entre 0 y 1 ml, ésta última calibrada para el valor correspondiente a 1 ml. Con el fin de evaluar la repetibilidad en la medición, cada analista repitió entre 6 y 10 veces la calibración con cada pipeta. Se evaluó la media aritmética de las mediciones de cada analista para cada pipeta y se estimó su incertidumbre expandida. Aquí se presentan los gráficos con las medias y las incertidumbres obtenidas para cada tipo de pipeta y se efectúa un estudio comparativo de los resultados. A modo de conclusión se evaluó la reproducibilidad de cada tipo de instrumento, en base a lo cual se formulan recomendaciones para su uso y calibración. Abstract In 2008 several chemical analysts from LATU’s test laboratories (most of them new to their task) took part in a Proficiency Test for the use of volumetric glassware. Two pipettes were used: a glass pipette of 1 ml, class B, and an automatic pipette of variable volume between 0 ml and 1 ml. The latter was calibrated at 1 ml. All analysts were asked to repeat each calibration between 6 and 10 times to allow an estimate of the measurement repeatibility. The average value performed by each analyst for each pipette was evaluated and its uncertainty estimated. Graphs are presented showing the analysts’ means and uncertainties for each pipette and the results are compared. As a conclusion, the reproducibility of each kind of instrument is evaluated according to which recommendations are given for the calibration and use of the two types of pipette

Preoperative Immunonutrition vs. Standard Dietary Advice in Normo-Nourished Patients Undergoing Fast-Track Laparoscopic Colorectal Surgery

Immunonutrition (IN) appears to reduce infective complications and in-hospital length of stay (LOS) after major gastrointestinal surgery, but its use in normo-nourished patients is still controversial. The primary aim of this comparative observational study was to evaluate if pre-operative IN reduces in-hospital stay in patients undergoing laparoscopic colorectal resection for cancer under an enhanced recovery after surgery (ERAS) program. The influence of IN on time to first bowel movements, time to full oral diet tolerance, number and type of complications, reasons of prolonged LOS and readmission rate was evaluated as secondary outcome. Patients undergoing ERAS laparoscopic colorectal resection between December 2016 and December 2019 were reviewed. Patients who have received preoperative IN (group A) were compared to those receiving standard dietary advice (group B). Mean in-hospital LOS was significantly shorter in patients receiving preoperative IN than standard dietary advice (4.85 ± 2.25 days vs. 6.06 ± 3.95 days; p < 0.0492). No differences in secondary outcomes were observed. Preoperative IN associated with ERAS protocol in normo-nourished patients undergoing laparoscopic colorectal cancer resection seems to reduce LOS