[Validity and usefulness of the RAE-CMBD studying patients hospitalised with influenza]. / Validez y utilidad del RAE-CMBD en el estudio de los pacientes hospitalizados con gripe.

OBJECTIVE: Understanding the hospital impact of influenza requires enriching epidemiological surveillance registries with other sources of information. The aim of this study was to determine the validity of the Hospital Care Activity Record - Minimum Basic Data Set (RAE-CMBD) in the analysis of the outcomes of patients hospitalised with this infection. METHODS: Observational and retrospective study of adults admitted with influenza in a tertiary hospital during the 2017/2018 and 2018/2019 seasons. We calculated the concordance of the RAE-CMBD with the influenza epidemiological surveillance registry (gold standard), as well as the main parameters of internal and external validity. Logistic regression models were used for risk adjustment of in-hospital mortality and length of stay. RESULTS: A total of 907 (97.74%) unique matches were achieved, with high inter-observer agreement (ƙ=0.828). The RAE-CMBD showed a 79.87% sensitivity, 99.72% specificity, 86.71% positive predictive value and 99.54% negative predictive value. The risk-adjusted mortality ratio of patients with influenza was lower than that of patients without influenza: 0.667 (0.53-0.82) vs. 1.008 (0.98-1.04) and the risk-adjusted length of stay ratio was higher: 1.15 (1.12-1.18) vs. 1.00 (0.996-1.001). CONCLUSIONS: The RAE-CMBD is a valid source of information for the study of the impact of influenza on hospital care. The lower risk-adjusted mortality of patients admitted with influenza compared to other inpatients seems to point to the effectiveness of the main clinical and organisational measures adopted.

[What about the weekend effect? Impact of the day of admission on in-hospital mortality, length of stay and cost of hospitalization]. / ¿Qué hay del efecto fin de semana? Impacto del día de ingreso sobre la mortalidad intrahospitalaria, la duración de la estancia y el coste de la hospitalización.

INTRODUCTION: There is no agreement on the existence of the weekend effect in healthcare or, if it exists, on its possible causes. The objective of the study was to evaluate the differences in healthcare outcomes between patients admitted on weekdays or weekends in a high-complexity hospital. METHODS: Observational and retrospective study of patients admitted between 2016 and 2019 in a public hospital with more than 1300 beds. Hospitalization episodes were classified according to whether admission took place between Friday at 3:00 p.m. and the following Monday at 8:00 a.m. (weekend admission) or not (admission on weekdays). Mortality, length of stay and associated costs were compared, applying their respective risk-adjustment models. RESULTS: Of the total 169,495 hospitalization episodes analyzed, 48,201 (28.44%) corresponded to the weekend, presenting an older age (54.9 years vs. 53.9; P<.001), a higher crude mortality rate (5.22% vs. 4.59%; P<0.001), and a longer average length of stay (7.42 days vs. 6.74; P<.001), than those admitted on weekdays. The median crude cost of stay was lower (€731.25 vs. €850.88; P<0.001). No significant differences were found when applying the adjustment models, with a risk-adjusted mortality ratio of 1.03 (0.99-1.08) vs. 0.98 (0.95-1.01), risk-adjusted length of stay of 1.002 (0.98-1.005) vs. 0.999 (0.997-1.002) and risk-adjusted cost of stay of 0.928 (0.865-0.994) vs. 0.901 (0.843-0.962). CONCLUSION: The results of the study reveal that the assistance provided during the weekends does not imply worse health outcomes or increased costs. Comparing the impact between hospitals will require a future homogenization of temporal criteria and risk adjustment models.

Characteristics of the major structural proteins of African swine fever virus: Role as antigens in the induction of neutralizing antibodies. A review.

African swine fever virus (ASFV) has been traditionally a major animal health problem worldwide, with important economic impact for the pig industry. Recently the disease has re-emerged in large areas of the world, including China and Eastern Europe. Therefore, it seems timely to summarize our current understanding of ASFV proteins and their antigenic properties, in connection with potential vaccine formulations. Here we review the main characteristics of the major structural proteins p150, p72 and p17 of ASFV and their antigenic properties. In particular, we emphasize that p17 was detected as a specific antigen in the immunoreaction of pig sera with neutralizing antibodies. In addition, specific immunoreactions against IP97, IP27, IP25.5, IP23 and IP13 viral infection proteins were also detected in these sera. The viral structural proteins have been studied with intracellular and extracellular viruses and, therefore the differences between both classes of viruses were also reviewed.

Prevalencia del virus de la rinotraqueitis infecciosa bovina en el departamento del Caquetá, Amazonía Colombiana / Prevalence of bovine infectious rhinotracheitis virus in Caquetá department, Colombian Amazon

RESUMEN La rinotraqueitis infecciosa bovina (IBR) es una enfermedad ampliamente difundida en el mundo con gran repercusión socio-económica en las ganaderías bovinas. El objetivo del presente estudio fue determinar la prevalencia de IBR en 100 hatos del departamento del Caquetá, para lo cual, se muestrearon 960 bovinos mayores de 36 meses entre enero y marzo de 2016 en predios seleccionados a partir de los criterios: a) tamaño (50-180 hectáreas), b) con más de 10 vacas en ordeño, c) disponibilidad de los productores para cooperar y d) accesibilidad de las vías. Las muestras de suero sanguíneo se remitieron al Laboratorio de Diagnóstico Clínico Veterinario del ICA y se analizaron a través la prueba Elisa de bloqueo (BHV-1) gB. Los sueros con porcentaje de bloqueo superior al 55% se consideraron positivos a anticuerpos para IBR. Se encontró alta seroprevalencia (73,13%), mayor además en machos que en hembras (p < 0,05). A nivel municipal la seroprevalencia fue mayor en los municipios de El Doncello, Puerto Rico y San Vicente del Caguán y se encontró diferencia significativa (p < 0,05) entre los nueve municipios analizados. A nivel de hatos, la prevalencia fue del 99%. En conclusión, la prevalencia del virus de la rinotraqueitis infecciosa bovina (IBR) en bovinos de doble propósito del departamento del Caquetá fue muy alta.

ABSTRACT Infectious bovine rhinotracheitis (IBR) is a disease widely spread throughout the world with great socio-economic impact on bovine livestock. The objective of this study was to determine the prevalence of IBR in 100 herds in Caqueta department, for which 960 cattle older than 36 months between January and March 2016 were sampled in farms selected from the criteria: a) size (50-180 hectares), b) with more than 10 cows in milking, c) availability of producers to cooperate and d) accessibility of roads. Blood serum samples were sent to the ICA veterinary clinical diagnostic laboratory and analyzed using the Elisa blocking test (BHV-1) gB, where sera with blocking percentage greater than 55% were considered as positive for IBR antibodies. High seroprevalence was found (73.13%), being higher in males than in females (p < 0.05). At municipal level, seroprevalence was greater in El Doncello, Puerto Rico and San Vicente del Caguán municipalities, significant difference (p<0.05) was found between the nine municipalities analyzed. At herd level prevalence was of 99%. In conclusion the prevalence of Infectious Bovine Rhinotracheitis (IBR) virus in dual-purpose cattle of Caquetá department was very high.

Partial complementation between the immediate early proteins ICP4 of herpes simplex virus type 1 and IE180 of pseudorabies virus.

We previously described that the immediate early (IE) IE180 protein of PRV can down-regulate the transactivation of the ICP4 promoter of HSV-1, and that the d120 virus (an ICP4-deficient HSV-1 strain) can partially replicate its viral DNA in the presence of the IE180 protein. Herein, we demonstrate that this partial complementation of d120 by IE180 is sufficient for transcription of ß, γ1 and γ2 products such as DNA pol, VP16 and gC, respectively. However, expression levels are low for VP16 and even lower for the gC, such that IE180 is unable to fully substitute for ICP4 functionally. Viral progeny was not detected in PK15 cells expressing PRV IE180.

Presence of a sexually active goat buck enhances ovulation occurrence in seasonally anestrous does after ovulation and luteolysis induction in hormonally-treated goats in seasonal anestrus.

In seasonally anestrous goat does, ovulations can be induced by combining a treatment regimen including progestagen, eCG and prostaglandins. Nonetheless, ovulations occur only once and then does return to a seasonally anestrous state. This study was performed to determine whether the presence of a sexually active buck can stimulate a second ovulation after induced luteolysis using prostaglandins following the first ovulation. Three groups of seasonally anestrous does were treated to induce ovulations using an intra-vaginally inserted sponge containing a progestin combined with eCG and prostaglandin administrations. Goats that had ovulations were treated with a prostaglandin 11 days after progestin sponge removal. After the prostaglandin injection, does continued to be isolated from bucks (n = 8), were penned with a control buck (n = 9), or were penned with a sexually active buck (n = 10). The proportion of goats having ovulations after imposing the ovulation-induction protocol was greater than 80% and did not differ among treatment groups (P >  0.05). The proportion of does having ovulations after injecting prostaglandins was greater when does were penned with a sexually active buck (8/10) than does penned with a control buck (0/9) or that were isolated from bucks (0/8; P < 0.05). It is concluded that in seasonally anestrous goat does induced to have ovulations using a hormonal treatment regimen, the presence of a sexually active buck can induce a second ovulation when there is an induced luteolysis.

Sexually active bucks counterbalance the seasonal negative feedback of estradiol on LH in ovariectomized goats.

We showed previously that the permanent presence of bucks rendered sexually active by photoperiodic treatments, thereafter called photostimulated bucks, prevents the occurrence of seasonal anovulation; also, the introduction of these sexually active bucks induces ovulations during seasonal anestrus. Here, we studied the response of ovariectomized goats bearing 12-mm subcutaneous implants filled or not with estradiol to sexually active males to determine (1) whether the permanent presence of such bucks prevents the decrease of LH despite the presence of a negative feedback by estradiol mimicking that of seasonal anestrus (experiment 1) and (2) whether the introduction of photostimulated bucks increases the plasma LH concentrations in spite of this negative feedback (experiment 2). In experiment 1, one group of goats remained in contact with sexually active bucks, whereas the other group remained in contact with control bucks. Plasma LH concentrations were high and did not differ with time or between groups of females from November to February (P > 0.05), when both types of bucks were sexually active. Afterward, in goats in contact with control and sexually inactive bucks, LH concentrations decreased from March (P ≤ 0.01) and remained low until May, whereas LH levels remained high from March to May in goats in contact with the photostimulated bucks (P > 0.05). In experiment 2, 2 groups of females bearing empty subcutaneous implants, and 2 groups of goats bearing subcutaneous implants filled with estradiol, were exposed to control or photostimulated bucks. Plasma LH concentrations did not increase in goats bearing empty implants, when exposed to control or photostimulated bucks (from 2.01 ± 0.26 to 1.98 ± 0.31 ng/mL, and from 2.45 ± 0.29 to 2.42 ± 0.21 ng/mL respectively; P > 0.05). In contrast, plasma LH concentrations increased from 0.97 ± 0.41 to 2.80 ± 0.62 ng/mL in goats exposed to the photostimulated bucks and bearing estradiol implants (P < 0.05). Thus, the permanent presence of sexually active bucks prevented the decrease of plasma LH concentration in OVX + E2 goats during the seasonal anestrus, and the introduction of the photostimulated bucks increased the plasma LH concentrations in OVX + E2 goats during the seasonal anestrus. Therefore, we conclude that in both cases, the photostimulated bucks are able to reduce or counterbalance the seasonal negative feedback of estradiol on LH secretion.

Efficiency of the male effect with photostimulated bucks does not depend on their familiarity with goats.

In ewes, the ovulatory response of females exposed to familiar rams is lower than the response of those exposed to novel ones. In goats, males rendered sexually active by exposure to long days are more efficient to induce ovulation in seasonal anestrous females than untreated males. Two experiments were conducted to determine 1) whether male goats remain familiar to females after 45days of separation; and 2) whether photostimulated males are able to stimulate the sexual activity of females, independently of their familiarity with them. In Experiment 1, three groups of goats (n=10 goats per group) were put in contact with males (n=2 per group) during 10days in November (familiarization period). These males were called familiar males. After 15, 30 and 45days of separation from the males, females of each group were exposed to familiar or novel males during 10min. In each test, goats in contact with novel males displayed more distress bleats, escapes, head butts, and sniffing than those in contact with familiar males (P<0.05). In Experiment 2, we used sexually inactive (n=4 control males), and sexually active males (n=4 photostimulated males). In February, two groups of goats (n=50 each) were put in contact with control or photostimulated males (n=2 each) during 10days ("familiar" control or photostimulated male, respectively). After 45days of separation from the males, both groups of females were further divided into two groups (n=25 goats per group). In April, two groups were re-exposed to "familiar" control or "familiar" photostimulated males (n=2 per group), whereas the other two groups were exposed to "novel" control or "novel" photostimulated males (n=2 per group). The photostimulated males displayed a higher level of sexual behavior than the controls. The proportion of goats that ovulated and displayed estrus was higher when exposed to the photostimulated males than when exposed to control ones (≥80% vs. 0%; P<0.05). These proportions did not differ between groups exposed to familiar or novel photostimulated males (P>0.05). We concluded that after 45days of separation, males are still familiar to females. The photostimulated males are able to induce the sexual activity of seasonally anestrous goats independently of their familiarity with them.

Construction of recombinant pseudorabies viruses by using PRV BACs deficient in IE180 or pac sequences: Application of vBAC90D recombinant virus to production of PRV amplicons.

We describe a simple and efficient method to obtain recombinant pseudorabies virus (PRV) in mammalian cells by using the PRV BACs, PBAC80 deficient in pac sequences and PBAC90 deficient in the IE180 gene. These essential viral sequences were used as targets to obtain viable recombinant viruses. PBAC80 was constructed, confirmed to encode a copy of the IE180 gene regulated by the inducible Ptet promoter, and used to obtain recombinant attenuated PRV viruses that express the EGFP protein (PRV-BT80GF virus). PBAC90 was used to obtain the vBAC90D virus, deficient in IE180 and free of replication-competent revertants, and which can be used as a helper in the production of PRV amplicons.

HbA(1c) in adults without known diabetes from southern Europe. Impact of the new diagnostic criteria in clinical practice.

AIMS: To analyse the differences in the prevalence of diabetes and dysglycaemia using fasting plasma glucose and HbA(1c) criteria. METHODS: Analytical cross-sectional study undertaken in a random sample of 2144 individuals (age 18-80 years) without known diabetes from the primary care setting in Malaga (Spain). Dysglycaemia was defined as fasting plasma glucose 5.6-6.9 mmol/l or HbA(1c) 39-46 mmol/mol (5.7-6.4%) and diabetes as fasting plasma glucose ≥ 7.0 mmol/l or HbA(1c)≥ 48 mmol/mol (≥ 6.5%). RESULTS: The proportion of subjects who were normoglycaemic was significantly higher using fasting plasma glucose than HbA(1c) (83.5 vs. 65%) (P < 0.0001). Compared with fasting plasma glucose, HbA(1c) detects more cases of dysglycaemia (32 vs. 14.8%) (P < 0.0001) and diabetes (3 vs. 1.7%) (P < 0.0001). CONCLUSIONS: In our environment, using HbA(1c) for the diagnosis of pre-diabetes and diabetes could increase the target population for preventive and therapeutic measures. Further cost-effectiveness studies are needed before the widespread diagnostic use of HbA(1c) can be recommended.

Construction and properties of a recombinant pseudorabies virus with tetracycline-regulated control of immediate-early gene expression.

A study was carried out to determine whether altering the control of expression of the IE180 gene of pseudorabies virus (PRV), by replacing the IE180 promoter with the tetracycline-responsive promoter (Ptet), affects virus replication and virulence. This PRV-BT90 mutant virus was constructed by complementation and recombination in Hela Tet-Off cells. The virus yield produced by infection of Hela Tet-Off cells with PRV-BT90 was similar to that of the parental virus vBecker2. Viral replication of PRV-BT90 was reduced in Vero cells as reflected by a reduction of virus yield and plating efficiency compared to vBecker2. PRV-BT90 plaque formation in Hela Tet-Off cells was inhibited in the presence of doxycycline, whereas vBecker2 plaque formation was not affected. Subcutaneous infection of mice with the two viruses revealed a LD(50) higher than 10(6) TCID(50) for the PRV-BT90 mutant virus while the LD(50) was 178 TCID(50) for the vBecker2 parental virus.

Regulation of pseudorabies virus gG glycoprotein gene promoter independently of pseudorabies immediate early IE180 protein.

The pseudorabies virus (PRV) glycoprotein known as gG is generally regarded as an early protein, and the immediate early IE180 protein regulates its expression during infection. This study, however, provides evidence that although induction by IE180 is observed, the expression of a marker protein (EGFP), or gG itself, under the control of the gG promoter, can also occur independently of the expression of IE180. This result was demonstrated both with transient transfection assays using plasmids and with viral infections. In transient transfections, the expression under control of the gG promoter depends on the cell type and surprisingly, can be 1.3-fold higher than the expression under the control of the IE180 promoter in Hela Tet-Off cells. Recombinant PRV S3 was constructed by replacing gE in the PRV genome with a chimeric transgene, expressing EGFP under the control of the gG promoter. In PK15 cells infected with NIA-3 wild-type virus or with S3 recombinant virus, expression of gG PRV mRNA (or EGFP mRNA) under the control of the gG promoter in the presence of cycloheximide was detected by RT-PCR. This again indicates that some basal expression was produced in infected cells independently of IE180. This expression was augmented by IE180 protein in both plasmid transfections and viral infections.

Diagnostic difficulties in glucokinase hyperinsulinism.

Glucokinase hyperinsulinism is a rare variant of congenital hyperinsulinism caused by activating mutations in the glucokinase gene and has been reported so far to be a result of overactivity of glucokinase within the pancreatic beta-cell. Here we report on a new patient with difficulties to diagnose persistent hyperinsulinism and discuss diagnostic procedures of this as well as the other reported individuals. After neonatal hypoglycemia, the patient was reevaluated at the age of 3 years for developmental delay. Morning glucose after overnight fast was 2.5-3.6 mmol/l. Fasting tests revealed supressed insulin secretion at the end of fasting (1.4-14.5 pmol/l). In addition, diagnostic data of the patients reported so far were reviewed. A novel heterozygous missense mutation in exon 10 c.1354G>C (p.Val452Leu) was found and functional studies confirmed the activating mutation. There was no single consistent diagnostic criterion found for our patient and glucokinase hyperinsulinism individuals in general. Often at the time of hypoglycemia low insulin levels were found. Therefore insulin concentrations at hypoglycemia, or during fasting test as well as reactive hypoglycemia after an oral glucose tolerance test were not conclusive for all patients. A glucose lowering effect in extra-pancreatic tissues independent from hyperinsulinism that results in diagnostic difficulties may contribute to underestimation of glucokinase hyperinsulinism. Mutational analysis of the GCK-gene should be performed in all individuals with unclear episodes of hypoglycemia even without documented hyperinsulinism during hypoglycemia. Delay of diagnosis might result in mental handicap of the affected individuals.

Presence of functional cannabinoid receptors in human endocrine pancreas.

AIMS/HYPOTHESIS: We examined the presence of functional cannabinoid receptors 1 and 2 (CB1, CB2) in isolated human islets, phenotyped the cells producing cannabinoid receptors and analysed the actions of selective cannabinoid receptor agonists on insulin, glucagon and somatostatin secretion in vitro. We also described the localisation on islet cells of: (1) the endocannabinoid-producing enzymes N-acyl-phosphatidyl ethanolamine-hydrolysing phospholipase D and diacylglycerol lipase; and (2) the endocannabinoid-degrading enzymes fatty acid amidohydrolase and monoacyl glycerol lipase. METHODS: Real-time PCR, western blotting and immunocytochemistry were used to analyse the presence of endocannabinoid-related proteins and genes. Static secretion experiments were used to examine the effects of activating CB1 or CB2 on insulin, glucagon and somatostatin secretion and to measure changes in 2-arachidonoylglycerol (2-AG) levels within islets. Analyses were performed in isolated human islets and in paraffin-embedded sections of human pancreas. RESULTS: Human islets of Langerhans expressed CB1 and CB2 (also known as CNR1 and CNR2) mRNA and CB1 and CB2 proteins, and also the machinery involved in synthesis and degradation of 2-AG (the most abundant endocannabinoid, levels of which were modulated by glucose). Immunofluorescence revealed that CB1 was densely located in glucagon-secreting alpha cells and less so in insulin-secreting beta cells. CB2 was densely present in somatostatin-secreting delta cells, but absent in alpha and beta cells. In vitro experiments revealed that CB1 stimulation enhanced insulin and glucagon secretion, while CB2 agonism lowered glucose-dependent insulin secretion, showing these cannabinoid receptors to be functional. CONCLUSIONS/INTERPRETATION: Together, these results suggest a role for endogenous endocannabinoid signalling in regulation of endocrine secretion in the human pancreas.

Hiperinsulinismo monogénico / Monogenic hyperinsulinism

El término hiperinsulinismo monogénico se refiere a casos de hiperinsulinemia causados por mutaciones en un solo gen. Los pacientes presentan hipoglucemias de ayuno recurrentes, niveles inadecuados de insulina e incremento de la glucemia tras la administración de glucagón endovenoso. Además, no existe cetonemia, cetonuria ni acidosis. La principal causa de este cuadro clínico son las canelopatías, en las que el hiperinsulinismo está producido por alteraciones estructurales de los canales de potasio dependientes del ATP como consecuencia de mutaciones en el receptor de la sulfonilurea 1 (SUR1) o en el rectifi cador interno de los canales de potasio (Kir6.2). La segunda causa más común es el síndrome de hiperinsulinismo-hiperamonemia, originado por mutaciones activadoras de la enzima glutamato deshidrogenasa (GDH). Este síndrome se caracteriza por cuadros de hipoglucemia hiperinsulinémica con niveles elevados de amonio, que pueden ser provocados por la ingestión de una comida rica en proteínas. Otra causa de hiperinsulinismo monogénico es el hiperinsulinismo inducido por mutaciones activadoras en el gen de la glucocinasa (GGK). Finalmente, debe incluirse también la mutación en la enzima mitocondrial 3-hidroxiacil-CoA deshidrogenasa de cadena corta (SCHAD), que cataliza el tercero de los cuatro pasos de la oxidación de los ácidos grasos en la mitocondria

The term monogenic hyperinsulinism refers to cases of hyperinsulinism caused by mutations in a single gene. The affected patients show recurrent fasting hypoglycemia, inadequate serum insulin levels, and an increase in plasma glucose levels following the administration of intravenous glucagon. In addition, there is an absence of ketonemia, ketonuria and acidosis. The main causes of these syndromes are channelopathies, in which hyperinsulinism is caused by structural changes in the ATP-sensitive potassium channels due to mutations in sulfonylurea receptor 1 (SUR1) or in Kir6.2, the pre-forming subunit of this channel. The second most frequent cause is the hyperinsulinism/hyperammonemia syndrome, caused by activating mutations of the glutamate dehydrogenase (GDH) enzyme. This syndrome is characterized by episodes of hypoglycemia with hyperinsulinism and elevated levels of ammonium, which can be triggered by the ingestion of a protein- rich meal. Monogenic hyperinsulinism can also be induced by activating mutations of the glucokinase gene. Finally, mutations of mitochondrial short-chain 3-hydroxyacyl-coenzyme A dehydrogenase (SCHAD), which catalyses the third of the four steps in mitochondrial fatty acid oxidation, should also be included

Glutaminase activity is confined to the mantle of the islets of Langerhans.

Glutamatergic signalling plays an important role in the coordination of hormone secretion from the endocrine pancreas. Thus, glutamate production must be a process exquisitely regulated to ensure a proper transmitter function. Recently we have reported that the endocrine pancreas co-expresses two isoforms of the protein glutaminase (GA), denoted as kidney-type (KGA) and liver-type (LGA). However, how GA activity, and therefore glutamate production, is regulated in the islets represents a critical issue that remains unresolved. Since the purification of these enzymes from rat islets is a daunting task, in order to characterize each isoform we have taken advantage of the spatial segregation of these isoenzymes in pancreas. To assist us with this goal, we have developed a new procedure that enables us to assay GA activity in situ. The assay is highly specific for GA as indicated by its dependence on glutamine and orthophosphate. Surprisingly, LGA, which is abundantly expressed by beta-cells, did not show detectable activity under the assay conditions. All the GA activity detected in pancreatic islets was attributed to KGA and was confined to the mantle of the islets. Double labelling analyses strongly suggested that alpha-cells should be regarded as the site of glutamate production in the endocrine pancreas.

Differential analysis of donor characteristics for pancreas and islet transplantation.

Suitable selection of donors is key to the success of human islet isolation and transplantation. Although several important donor-related factors have been identified previously, they needed to be confirmed in our setting. The aims of this study were: (1) to compare the characteristics of islet donors with those of pancreas donors (national transplant registry). (2) to compare the characteristics of islet donors resulting in a successful isolation in our facility with the characteristics of pancreas donors, and (3) to compare the characteristics of islet donors at this facility, whether or not isolation was successful, with donors elsewhere whose islets were transplanted and included in the Collaborative Islet Transplant Registry. The 35 islet isolations completed at our facility were analyzed for various characteristics. Significant differences were seen in donor age body mass index (BMI), and body weight between our islet donors and our pancreas donors (P < .001). These differences were maintained in the subgroup analysis corresponding to donors of successful isolations compared to pancreas donors (P < .01). Most successful isolations in our islet isolation facility were associated with donors of BMI >25. The percentage of successful isolation (>300,000 IEq) was higher among donors with a body weight >90 kg. We concluded that there was little overlap between the donor profiles for pancreas transplantation and for islet transplantation. More specific selection criteria relative to both BMI and body weight for islet donors may result in greater success of pancreas islet isolation and transplantation.

A comparison of enhanced green fluorescent protein expression induced by immediate-early cytomegalovirus (IE-CMV) and gG pseudorabies virus (gG-PRV) promoters, using pseudorabies virus amplicons as vectors.

This study compares the expression efficiencies of the IE-CMV and gG-PRV promoters following their transfection into cultured human and monkey cells, using pseudorabies virus amplicons as vectors and enhanced green fluorescence protein (EGFP) as an expression marker. EGFP expression was similarly strong with both promoters. Pseudorabies virus amplicons appear to be useful vectors in gene expression studies due to their replication in the presence of helpers and their wide range of cellular hosts.

Internal assessment of a novel islet isolation facility in Spain.

UNLABELLED: Islet transplantation is a promising therapy in the treatment of diabetes mellitus. Herein we present the result from the first series of islet isolations carried out in our new islet isolation facility. The aims of study were to analyze the influence of various donor characteristics on the success of islet isolation and compare these outcomes with other European and American groups. Data from 22 completed islet isolation were used to compare donor and isolation variables among successful (>300,000 IEQs) versus unsuccessful isolations. The successful isolation rate from our laboratory was 31.8%. We did not see any significant differences between successful and unsuccessful groups according to donor characteristics, although age was close to significance (38.57 +/- 10.29 versus 48.33 +/- 12.39; P = .08). Donor age (1.12 [1.23; 0.99]) and body mass index (0.065 [1.32; 3.08]) were associated with isolation success in a logistic regression model. We did not find differences among intraprocedure variables with the exception of IEQ prepurification (409,073 +/- 115,041 versus 263,776 +/- 128,988; P < .05). IEQpre and IEQpost were positively correlated (P < .05). In comparison with other groups, we observed differences in some cases related to islet yield prepurification (P < .05) but not postpurification. Purity from our islet preparations was the highest from all considered groups (P < .05). Recovery was similar in all groups. CONCLUSIONS: In our experience, donor characteristics have no influence on the success rate. The digestion step is a critical factor for success. Our results with respect to IE yield were close to that of experienced groups.

Monounsaturated n-9 fatty acids and adipocyte lipolysis in rats.

To investigate the role of the monounsaturated n-9 fatty acids (MUFA) in the lipolytic activity of adipocytes, a study was carried out in which an increase in MUFA was produced in the tissues by two different methods; by the dietary enrichment of oleic acid or by producing an essential fatty acid deficiency syndrome. For this, forty-five male Sprague-Dawley rats were fed with a normal-energy diet and were subdivided into three groups. The diets varied in the type of dietary fat; palmitic acid, olive oil, or soyabean oil+palmitic acid. At the end of the study measurements were taken of weight, plasma leptin, tissue concentration of fatty acids, fat-cell size in the epididymal and the omental adipose tissues, adipocyte lipolytic activity of both tissues after stimulation with adrenaline, and the capacity of insulin to inhibit lipolysis. The baseline and adrenaline-stimulated lipolytic activity were greater and the anti-lipolytic capacity of insulin lower in the animals undergoing an increase in MUFA in the tissues (palmitic-acid and olive-oil diets). The area under the curve of glycerol, used as an indicator of lipolytic activity, was positively correlated with the concentration of MUFA and negatively with polyunsaturated fatty acids in the adipose tissues. It is concluded that an increase in tissue MUFA, however obtained, induces an increase in lipolytic activity.