Creatinina plasmática medida por método enzimático y estimulación del filtrado glomerular con la fórmula Full-Age-Spectrum, en una población pediátrica ambulatoria de la Argentina / Plasma creatinine measured by enzymatic method and estimation of glomerular filtration rate using the Full-Age-Spectrum equation in a pediatric outpatient population in Argentina

Introducción: el método recomendado para la medición de creatinina plasmática (Cr) es el enzimático, que permite obtener la tasa de filtrado glomerular estimado (TFGe) con la fórmula Full-Age-Spectrum (FAS) para todas las edades, al normalizar la TFGe con valores poblacionales de Cr. Objetivos: obtener valores poblacionales de Cr medida con un método enzimático y evaluar la fórmula FAS, en una población pediátrica ambulatoria de la Argentina, puesto que no existen publicaciones al respecto en nuestro país. Material y métodos: estudio descriptivo, retrospectivo, transversal, por muestreo consecutivo. Se consideró la población pediátrica ambulatoria de 2 a 17 años que concurrió una sola vez, entre 07/2018 y 11/2021 al laboratorio del Hospital Municipal (Bahía Blanca, Argentina) con petición médica de Cr. Se evaluó la distribución poblacional de Cr. Se comparó FAS original (FAS-Belga) con FAS normalizada con valores locales de Cr (FAS-Local). Resultados: se estudiaron 2793 individuos. Los varones tuvieron un valor de Cr superior al de las mujeres a los 16 y 17 años. La TFGe fue menor con FAS-Local que con FAS-Belga [mediana (RI) mL/min/1,73 m2 : 107,3 (22,9) vs. 117,0 (26,5); p=0,0001; rbis=0,87 (tamaño del efecto grande)]. Del análisis del gráfico de Bland-Altman y el índice de concordancia Kappa se obtuvo que FAS-Local no fue comparable con FAS-Belga. Conclusiones: los valores poblacionales de Cr, medida con un método enzimático, son los primeros en obtenerse en una población pediátrica ambulatoria argentina. Dichos valores son necesarios para aplicar FAS en la Argentina (AU)

Introduction: the recommended test for the measurement of plasma creatinine (Cr) is the enzymatic method, which allows calculating the estimated glomerular filtration rate (eGFR) with the Full-Age-Spectrum (FAS) equation for all ages, by normalizing the eGFR with population Cr values. Objectives: to obtain population Cr values measured with an enzymatic method and to evaluate the FAS equation in an pediatric outpatient population in Argentina, since there are no reports on this subject in our country. Material and methods: A descriptive, retrospective, cross-sectional, consecutive sampling study. The pediatric outpatient population aged 2 to 17 years who attended only once to the laboratory of the Municipal Hospital (Bahía Blanca, Argentina) between 07/2018 and 11/2021 with medical request for Cr measurement. The population distribution of Cr was evaluated. The original FAS (FAS-Belgian) was compared to FAS normalized with local Cr values (FAS-Local). Results: 2793 individuals were studied. Males had a higher Cr value than females at 16 and 17 years of age. The eGFR was lower with FAS-Local than with FAS-Belgian [median (IQR) mL/min/1.73 m2: 107.3 (22.9) vs. 117.0 (26.5); p=0.0001; rbis=0.87 (large effect size)]. Analysis of the Bland-Altman plot and the Kappa concordance index showed that FAS-Local was not comparable to FAS-Belgian. Conclusions: population Cr values, measured with an enzymatic method, are the first to be obtained in an Argentine pediatric outpatient population. These values are necessary to apply the FAS in Argentina (AU)

Preanalytical phase: Effects of water ingestion during fasting on routine hematological parameters in a small cohort of young women.

BACKGROUND: We analyze the effects of water ingestion before blood extraction on routine hematological parameters. METHODS: Twenty female volunteers -mean 24 y- were included. Blood was collected after a 12 h fast period (T0) and 1 h after the ingestion of 300 ml water (T1). These parameters were analyzed: white blood cell (WBC) count; WBC differential count including lymphocytes (LYM), monocytes (MONO), neutrophils, eosinophils (EOS), and basophils; red blood cell (RBC) count; hematocrit (HCT); hemoglobin (HGB); mean cell volume; mean cell hemoglobin; RBC distribution width; and platelet count (PLT). Statistical significance: P < 0.05. Mean difference % (MD%) was calculated for each parameter and was compared with reference change value (RCV). A change was considered clinically significant when MD% exceeded the RCV. RESULTS: Significant differences were observed in (medians, T0 vs T1, P): WBC ×109/l (6.51 vs 6.12, 0.002); LYM ×109/l (2.90 vs 2.19, 0.000); MONO ×109/l (0.50 vs 0.48, 0.031); EOS ×109/l (0.17 vs 0.16, 0.003); RBC ×1012/l (4.46 vs 4.40, 0.024); HCT l/l (0.38 vs 0.37, 0.036); HGB g/l (129 vs 129, 0.009). All MDs% were lower than their respective RCV. CONCLUSION: Ingestion of 300 ml water 1 h before blood extraction does not alter the hematological parameters studied.

Estimating exotic gene flow into native pine stands: zygotic vs. gametic components.

Monitoring contemporary gene flow from widespread exotic plantations is becoming an important problem in forest conservation genetics. In plants, where both seed and pollen disperse, three components of exotic gene flow with potentially unequal consequences should be, but have not been, explicitly distinguished: zygotic, male gametic and female gametic. Building on a previous model for estimating contemporary rates of zygotic and male gametic gene flow among plant populations, we present here an approach that additionally estimates the third (female gametic) gene flow component, based on a combination of uni- and biparentally inherited markers. Using this method and a combined set of chloroplast and nuclear microsatellites, we estimate gene flow rates from exotic plantations into two Iberian relict stands of maritime pine (Pinus pinaster) and Scots pine (Pinus sylvestris). Results show neither zygotic nor female gametic gene flow but moderate (6-8%) male gametic introgression for both species, implying significant dispersal of pollen, but not of seeds, from exotic plantations into native stands shortly after introduced trees reached reproductive maturity. Numerical simulation results suggest that the model yields reasonably accurate estimates for our empirical data sets, especially for larger samples. We discuss conservation management implications of observed levels of exposure to nonlocal genes and identify research needs to determine potentially associated hazards. Our approach should be useful for plant ecologists and ecosystem managers interested in the vectors of contemporary genetic connectivity among discrete plant populations.

Riesgo cardiovascular en pacientes con sobrepeso u obesidad y leve disminución del fltrado glomerular / Cardiovascular Risk in Patients with Overweight or Obesity and Mild Decrease in Glomerular Filtration Rate

El mundo occidental ha sufrido un aumento preocupante en la prevalencia de la obesidad, la cual se asocia al desarrollo de enfermedad cardiovascular (ECV) y enfermedad renal crónica (ERC). Se ha demostrado que la obesidad induce disturbios fisiopatológicos que favorecen la injuria renal y a su vez, el deterioro de la función renal se ha asociado con la aparición de eventos cardiovasculares y mortalidad. El objetivo de este trabajo fue evaluar si un leve deterioro de la función renal se asocia a un riesgo cardiovascular (RCV) aumentado en individuos con sobrepeso u obesidad. De un total de 454 individuos que concurrieron al Servicio de Medicina Preventiva del Hospital Municipal de Bahía Blanca, se incluyeron 205 (45 %) sujetos adultos, de ambos sexos, con sobrepeso u obesidad y sin otros factores de riesgo cardiovascular, de quienes se obtuvieron datos clínicos y bioquímicos. Se estimó la Velocidad de Filtración Glomerular (VFG) con la ecuación abreviada Modification of Diet in Renal Disease y el RCV con el score Reynolds. El 19 % de la población estudiada presentó leve deterioro del Filtrado Glomerular (FG), VFG entre 60 y 89 mL/min/1,73 m², con valores normales de creatinina, observándose en este grupo una mayor proporción de pacientes con RCV aumentado (23 %) comparado con el grupo sin deterioro del FG (14 %), p = 0.153. Se observó asociación significativa entre leve deterioro del FG y edad mayor a 50 años, (p = 0,000). Se concluyó en este estudio, que en estos individuos con exceso de peso es necesario optimizar el screening de ERC, incorporando rutinariamente la estimación de la VFG y completando la evaluación de la función renal con la medición de Albúmina Urinaria y otros estudios por imágenes, para clasificarlos o no en el estadio 2 de ERC, con el fin de prevenir la ERC y la ECV. Los autores declaran no poseer conflictos de interés.(AU)

The occidental world has suffered an alarming increase in the prevalence of obesity, which is associated with the development of cardiovascular disease (CVD) and chronic kidney disease (CKD). It has been demonstrated that obesity induces pathophysiologic disturbances that promote renal injury, and at the same time, renal dysfunction has been associated with the onset of cardiovascular events and mortality. The aim of this study was to evaluate if a mild impairment in renal function is associated with an increased cardiovascular risk (CVR) in subjects with overweight or obesity. From a total of 454 individuals who presented at the Preventive Medicine Department of Hospital Municipal de Bahía Blanca, 205 (45 %) adults of both genders with over-weight or obesity and with no other cardiovascular risk factor were enrolled. Clinical and biochemical data were obtained from these individuals. Glomerular Filtration Rate (GFR) was estimated with the abbreviated equation Modification of Diet in Renal Dis-ease, CVR was determined with Reynolds score. Nineteen percent of the studied population had a mild impairment in glomerular filtration (GF), GFR between 60 and 89 mL/min/1.73m², with normal creatinine values, with a higher proportion of patients with increased CVR (23 %) being found in this group as compared to the group with no GF impairment (14 %), p=0.153. A significant association was found between mild impairment of GF and age older than 50 years (p = 0.000). In this study, we concluded that it is necessary to optimize CKD screening in overweight or obese individuals by incorporating the estimation of GFR in routine practice and performing a complete renal function evaluation including urinary albumin measurement and other imaging studies, to determine whether they should be classified or not into stage 2 of CKD with the aim of preventing CKD and CVD. No financial conflicts of interest exist.(AU)

Riesgo cardiovascular en pacientes con sobrepeso u obesidad y leve disminución del fltrado glomerular / Cardiovascular Risk in Patients with Overweight or Obesity and Mild Decrease in Glomerular Filtration Rate

El mundo occidental ha sufrido un aumento preocupante en la prevalencia de la obesidad, la cual se asocia al desarrollo de enfermedad cardiovascular (ECV) y enfermedad renal crónica (ERC). Se ha demostrado que la obesidad induce disturbios fisiopatológicos que favorecen la injuria renal y a su vez, el deterioro de la función renal se ha asociado con la aparición de eventos cardiovasculares y mortalidad. El objetivo de este trabajo fue evaluar si un leve deterioro de la función renal se asocia a un riesgo cardiovascular (RCV) aumentado en individuos con sobrepeso u obesidad. De un total de 454 individuos que concurrieron al Servicio de Medicina Preventiva del Hospital Municipal de Bahía Blanca, se incluyeron 205 (45 %) sujetos adultos, de ambos sexos, con sobrepeso u obesidad y sin otros factores de riesgo cardiovascular, de quienes se obtuvieron datos clínicos y bioquímicos. Se estimó la Velocidad de Filtración Glomerular (VFG) con la ecuación abreviada Modification of Diet in Renal Disease y el RCV con el score Reynolds. El 19 % de la población estudiada presentó leve deterioro del Filtrado Glomerular (FG), VFG entre 60 y 89 mL/min/1,73 m², con valores normales de creatinina, observándose en este grupo una mayor proporción de pacientes con RCV aumentado (23 %) comparado con el grupo sin deterioro del FG (14 %), p = 0.153. Se observó asociación significativa entre leve deterioro del FG y edad mayor a 50 años, (p = 0,000). Se concluyó en este estudio, que en estos individuos con exceso de peso es necesario optimizar el screening de ERC, incorporando rutinariamente la estimación de la VFG y completando la evaluación de la función renal con la medición de Albúmina Urinaria y otros estudios por imágenes, para clasificarlos o no en el estadio 2 de ERC, con el fin de prevenir la ERC y la ECV. Los autores declaran no poseer conflictos de interés.

The occidental world has suffered an alarming increase in the prevalence of obesity, which is associated with the development of cardiovascular disease (CVD) and chronic kidney disease (CKD). It has been demonstrated that obesity induces pathophysiologic disturbances that promote renal injury, and at the same time, renal dysfunction has been associated with the onset of cardiovascular events and mortality. The aim of this study was to evaluate if a mild impairment in renal function is associated with an increased cardiovascular risk (CVR) in subjects with overweight or obesity. From a total of 454 individuals who presented at the Preventive Medicine Department of Hospital Municipal de Bahía Blanca, 205 (45 %) adults of both genders with over-weight or obesity and with no other cardiovascular risk factor were enrolled. Clinical and biochemical data were obtained from these individuals. Glomerular Filtration Rate (GFR) was estimated with the abbreviated equation Modification of Diet in Renal Dis-ease, CVR was determined with Reynolds score. Nineteen percent of the studied population had a mild impairment in glomerular filtration (GF), GFR between 60 and 89 mL/min/1.73m², with normal creatinine values, with a higher proportion of patients with increased CVR (23 %) being found in this group as compared to the group with no GF impairment (14 %), p=0.153. A significant association was found between mild impairment of GF and age older than 50 years (p = 0.000). In this study, we concluded that it is necessary to optimize CKD screening in overweight or obese individuals by incorporating the estimation of GFR in routine practice and performing a complete renal function evaluation including urinary albumin measurement and other imaging studies, to determine whether they should be classified or not into stage 2 of CKD with the aim of preventing CKD and CVD. No financial conflicts of interest exist.

Influence of updated guidelines on short- and long-term mortality in patients with non-ST-segment elevation acute coronary syndrome (NSTE-ACS).

AIM: In 2002 the ACC/AHA guidelines for the management of patients with unstable angina (UA) and non-ST-segment elevation myocardial infarction (NSTEMI) were updated. We aimed to answer whether the implementation of updated guidelines was capable of influencing short- and long-term mortality in these patients. METHODS: We analyzed data on 812 consecutive patients who were admitted with either UA or NSTEMI between 2001 and 2004. Patients admitted in the two years before the implementation of updated guidelines (UA(01/02) group and NSTEMI(01/02) group) were compared to patients admitted in the two years thereafter (UA(03/04) group and NSTEMI(03/04) group). Yearly follow-up concerning all-cause mortality was obtained up to four years. RESULTS: The rate of revascularizations, the percentage of procedures performed within 48 h of admission, and the administration of clopidogrel increased significantly. However, still many - especially high-risk - patients did not receive revascularization. Patients of both UA groups had an identical in-hospital mortality rate. Differences in mortality between groups gained statistical significance over time (four-year mortality; 15.1% for the UA(03/04) group vs. 26.5% for the UA(01/02) group, p=0.014; HR 0.49 95% CI 0.28-0.87). In patients with NSTEMI in-hospital mortality decreased from 18.4% in the NSTEMI(01/02) group to 9.6% in the NSTEMI(03/04) group (p=0.011; HR 0.47 95% CI 0.26-0.84), and 1-year mortality from 34.7% to 25.1% (p=0.038; HR 0.63 95% CI 0.41-0.98), respectively. Mortality rates beyond one year were still lower in the NSTEMI(03/04) group as compared to the NSTEMI(01/02) group but it did not reach statistical significance. Multivariate Cox-regression analysis revealed furthermore that also patients with higher age and/or renal dysfunction benefit from an early invasive strategy. CONCLUSION: The implementation of updated guidelines for NSTE-ACS had significant impact on short- and long-term mortality. However, an early invasive strategy is still withheld to a significant number of high-risk patients, who would benefit from an invasive treatment.

Tacrolimus-eluting carbon-coated stents versus sirolimus-eluting stents for prevention of symptom-driven clinical end points.

BACKGROUND: Coating of stents has been shown to minimize the interactions between platelets, stent surface and vascular response following stent implantation. The aim of our study was to compare the tacrolimus-eluting carbon-coated JANUS(®) stent with sirolimus-eluting CYPHER(®) stent for the prevention of symptom-driven clinical end points in a real world clinical setting. METHODS: This prospective registry with a follow-up period of 24 months was conducted in 90 consecutive patients undergoing coronary artery stenting receiving CYPHER(®) (n = 48) or JANUS(®) (n = 42) stents. The primary end point was a composite of death from cardiovascular causes, nonfatal myocardial infarction and target vessel revascularisation, and the secondary end point was clinically driven in-stent restenosis. RESULTS: The primary combined endpoint occurred in 38% of patients (n = 16) in the JANUS(®) group compared to 10% (n = 5) in the CYPHER(®) group. The relative risk increase of the composite end point was therefore 63% higher in patients receiving JANUS(®) stents compared to the CYPHER(®) stents (crude HR = 1.63, 95% CI = 1.17-2.28, p = 0.004; adjusted HR = 1.79, CI = 1.26-2.55, p = 0.001). Interestingly, 75% of events in the JANUS(®) group occurred during the first 6 months after stent implantation. Similarly, the rate of clinically driven in-stent restenosis was higher in patients receiving JANUS(®) stent (n = 10, 2%) compared to the CYPHER(®) stent (n = 2, 4%). Concordantly, the relative risk for clinically driven in-stent restenosis was 81% higher in the JANUS(®) group compared to the CYPHER(®) group (crude HR = 1.81, 95% CI = 1.08-3.02, p = 0.02; adjusted HR = 2.24, CI = 1.26-3.96, p = 0.006). CONCLUSION: The use of tacrolimus-eluting carbon coated JANUS(®) stent was associated with worse clinical outcome compared to the sirolimus-eluting CYPHER(®) stent in clinical routine use.

Protein kinase CK2 in health and disease: CK2: a key player in cancer biology.

Elevated levels of protein kinase CK2 (formerly casein kinase 2 or II) have long been associated with increased cell growth and proliferation both in normal and cancer cells. The ability of CK2 to also act as a potent suppressor of apoptosis offers an important link to its involvement in cancer since deregulation of both cell proliferation and apoptosis are among the key features of cancer cell biology. Dysregulated CK2 may impact both of these processes in cancer cells. All cancers that have been examined show increased CK2 expression, which may also relate to prognosis. The extensive involvement of CK2 in cancer derives from its impact on diverse molecular pathways controlling cell proliferation and cell death. Downregulation of CK2 by various approaches results in induction of apoptosis in cultured cell and xenograft cancer models suggesting its potential as a therapeutic target.

The harmonization of the regulation of blood products: a European perspective.

The development of blood products as medicines initially took place on the national level in various countries, which resulted in considerable diversity of mechanisms and stringency of regulatory oversight. The scenario changed dramatically with the catastrophic experience that severe virus infections had been transmitted by blood products world-wide. Blood products, which had been regulated differently in the member states, became subject to the European pharmaceutical legislation in 1989. A specialized directive regulating the blood transfusion sector and the collection of plasma for fractionation was enacted in 2002. The European Community, particularly the Commission and the European Medicines Agency, is continuously refining the requirements, providing detailed technical and scientific guidance. In addition, institutions of the Council of Europe play an important role in the transfusion sector, the elaboration of the European Pharmacopoeia prescriptions, and the co-ordination of Official Medicines Control Laboratory or Laboratories batch release. However, further and sustained efforts towards international harmonization are needed. There are already important mechanisms in place, such as the International Conference on Harmonization initiative, which is producing internationally recognized guidelines on central issues. Another important achievement is the common technical document format, which enables the use of uniform applications for marketing authorization. However, there is still room for progress, for example, questions regarding regulatory requirements for licensing of in vitro diagnostic devices, or mutual recognition of inspections. The World Health Organization continues to play an important role in harmonization, both substantially by the production of high-level guidance documents or the establishment of physical international standard preparations, and in a more general sense by providing a platform for international collaboration. A very important aspect is the transparency of the creation and refinement of regulatory requirements. It is currently the rule that draft legal texts, monographs and guidelines are published for a consultation period before adoption. Effort and attention are required to keep track of the developments. However, in the era of modern electronic communication tools, the necessary information can be found on websites and comments can easily be submitted. Networking and exchange of information will continue to be crucial for development and maintenance of sound and balanced regulatory requirements.

Participation in screening colonoscopy in first-degree relatives from patients with colorectal cancer.

BACKGROUND: Current guidelines recommend screening colonoscopy in first-degree relatives of patients with colon cancer. The aim of this state-wide study was to investigate the compliance for colonoscopic in first-degree relatives, who were younger than 60 years of age. METHODS: A total of 602 patients were identified from the tumor registry of the public health insurance of Lower Saxony. A questionnaire was sent to these patients, which included a number of different questions regarding their knowledge about the risk of colon cancer for their family members, as well as their participation in screening colonoscopy. RESULTS: Data from 442 patients and their first-degree relatives (1005 siblings and 354 parents) were available; 178 parents had undergone screening colonoscopy and 344 siblings. Interestingly, the percentage of siblings who underwent screening colonoscopy was significantly higher (27%) among those siblings where the index patients were aware of the increased risk for the first-degree relatives, in contrast to the siblings of the index patients who were not aware of this risk (20%). CONCLUSION: This study demonstrates that only a minority of first-degree relatives undergo screening colonoscopy and that informing patients about the potential risk for their relatives will increase participation in screening colonoscopy in first-degree relatives of the patients.

Protein kinase CK2 as regulator of cell survival: implications for cancer therapy.

Recent studies have generated sufficient information to warrant a consideration of protein kinase CK2 as a potential target for cancer therapy. CK2 is a ubiquitous and highly conserved protein serine/threonine kinase that has long been considered to play a role in cell growth and proliferation. It is essential for cell survival, and considerable evidence suggests that it can also exert potent suppression of apoptosis in cells. This is important since the cancer phenotype is characterized by deregulation of not only proliferation but also of apoptosis. In normal cells, the level of CK2 appears to be tightly regulated, and cells resist a change in their intrinsic level of CK2. However, in all the cancers that have been examined an elevation of CK2 has been observed. Further, it appears that modest deregulation in the CK2 expression imparts a potent oncogenic potential to the cells. Disruption of CK2 by treatment of cells with antisense CK2 results in induction of apoptosis in a time and dose-dependent manner. Thus, we propose that down-regulation of CK2 by employing specific strategies to deliver antisense CK2 in vivo could have a potential role in cancer therapy.

Hepatitis C virus transmission through quarantine fresh-frozen plasma.

In 1994, quarantine fresh-frozen plasma (Q-FFP) was introduced in Germany in order to reduce the risk of HIV and HCV transmission. In 1998, an acute HCV infection of a patient was reported to us. The look-back revealed that this patient had received two Q-FFP from a donor who had seroconverted for HCV in the meantime. Recipients of further plasma donations from this donor were identified. Back-up specimens of these donations were investigated in several laboratories. A total of 25 additional HCV-PCR positive plasma units had been transfused to 12 further patients. HCV infections were diagnosed in seven of these recipients, three patients had already been deceased. One of the remaining two recipients was already HCV positive prior to transfusion, in the other patient, no HCV infection was detectable. This patient had received three units of an "early" plasma donation , which was tested negative by PCR in one laboratory, but positive in the other. The subsequent, clinically infectious donation had the same discrepant PCR results. Thus, eight cases of HCV transmission were revealed and classified as "certain" with regard to causality, also due to an identical HCV genotype, i.e. 3e. Some of these infections would have been prevented by application of a different anti-HCV assay. The assay used in the respective plasmapheresis station was in-sensitive in this individual case for more than 400 days after the first PCR positive donation. This caused the release of the above mentioned infectious units. Upon re-testing the backups, three of four other anti-HCV assays revealed a positive result already 104 days after the first PCR-positive donation. The donor had increased ALAT levels (> 23 IU/L) at nine of 28 donations, two of these were higher than 2.5 times the upper normal limit, and two were higher than 68 IU/L, which is the cut-off value for male blood donors in Germany. The results of these (look-back) studies arouse several queries, i.e. differences in the diagnostic sensitivity between current anti-HCV and PCR tests, the accuracy of risk-estimates (especially when based on hemovigilance studies for Q-FFP), the value of ALAT testing, and currently practised release algorithms for Q-FFP.

A fully-automated environmental chamber for examination of long-term effects of intermittent hypoxia on medium-size animals.

We describe the design and construction of a fully-automated environmental chamber for the simultaneous exposure of up to four medium-size laboratory animals to long-term intermittent hypoxia. The air-sealed automated environmental chamber consists of a box equipped with a ventilation fan and three electrically-activated solenoid valves. Our system was used to expose four rabbits to 12 h of repetitive episodes of hypoxia (environmental O2 concentration 12-13%) lasting 45 min followed by breathing room air for 15 min. During environmental hypoxia, the mean arterial PaO2 and PaCO2 were 41 +/- 3.0 and 24 +/- 0.7 mmHg (mean +/- SEM), respectively. In this system, opening and closing of the solenoid valves is fully computerized to allow different settings of the duration and severity of hypoxia. The chamber is safe and fully automated and cost-effective for studying the effects of long-term intermittent hypoxemia in medium-size animals.

Localization of pestiviral envelope proteins E(rns) and E2 at the cell surface and on isolated particles.

The glycoproteins E(rns) of classical swine fever virus (CSFV) and E(rns) and E2 of bovine viral diarrhoea virus (BVDV) are shown to be located at the surface of infected cells by the use of indirect immunofluorescence and by cytofluorometric analysis. The positive immunostaining of the cell surface was further analysed by immunogold electron microscopy and it could be shown that only extracellular virions were labelled. Gold granules were not seen at the cellular plasma membrane. In contrast to BVDV E2, the CSFV E2 of virions sticking to the plasma membrane was not accessible to the respective monoclonal antibodies. However, CSFV particles isolated from culture supernatant were able to bind both monoclonal anti-E(rns) and anti-E2 antibodies. For CSFV and BVDV, binding of anti-E(rns) antibodies to the virions was more pronounced than that of anti-E2. This finding was unexpected since E2 is considered to be the immunodominant glycoprotein.

A wound-induced [Ca2+]i increase and its transcriptional activation of immediate early genes is important in the regulation of motility.

Upon mechanical wounding of a confluent quiescent monolayer, cells move into the denuded zone. However, it is not well known what signaling cascade connects release from contact inhibition to cell movement at the wound edge. Mechanical wounding induced an increase in the concentration of intracellular free Ca2+ ([Ca2+]i) in endothelial cells at the wound edge. The [Ca2+]i signal was required for the transcriptional activation of two immediate early genes (IEGs), c-fos and c-jun, since blocking Ca2+ influx with Gd3+ or EGTA reduced IEG transcription, while augmenting Ca2+ influx increased IEG transcription. The transcriptional activation of the IEGs depended on protein kinase C and calmodulin-dependent protein kinase since treatment with the inhibitors Calphostin C and KN-62 significantly reduced IEG expression. Briefly blocking Ca2+ influx also produced a long-term reduction of cell motility, while augmenting Ca2+ influx increased cell motility. To evaluate whether expression of IEGs might control cell movement, we microinjected sense or antisense cDNA to c-fos into cells after wounding. Antisense c-fos cDNA inhibited motility, while sense cDNA increased motility rates. These results suggested that the [Ca2+]i rise, induced by wounding, regulated the initiation of subsequent motility through the transcriptional activation of IEGs during wounding.

Quantitative assessment of leading edge adhesion: reattachment kinetics modulated by injury-derived intracellular calcium predict wound closure rates in endothelial monolayers.

Migrating cells continually develop new substrate attachments at the leading edge (LE) in order to maintain traction for movement. This study evaluates the relationship between LE adhesion and wound closure by modulating injury-derived intracellular free Ca2+ ([Ca2+]i) signaling in endothelial cell (EC) monolayers following scrape-wounding. These data show that brief treatment with increased extracellular Ca2+ ([Ca2+]e) during wounding accelerated wound area closure rates by 50-65%, while brief treatments with calcium influx inhibitors reduced rates by 30-50%. Fura-2 studies in wounded monolayers indicated supranormal [Ca2+]e during wounding increased (by 52%), while influx-inhibitors decreased (by 36%) the percentage of cells exhibiting elevated plateau [Ca2+]i levels. Quantitative time-lapse interference reflection microscopy (IRM) together with indirect alphavbeta3 integrin immunofluorescence was used to measure the effects of 100 microM Gd3+ and 5 mM [Ca2+]e treatment on fractional LE adhesion after wounding. Influx inhibition blocked development of increased injury-derived LE adhesion. Measurements indicated a linear relationship (r2 = 0.99, 0.98) between LE adhesion, development rates (quantified as an association rate constant) and steady state wound closure rates. Changes in filopodial activity, as indicated by phase contrast microscopy, did not correlate with changes in wound closure rates, but an association existed between the percentile peak [Ca2+]i response and the initiation of filopodial activity, suggesting a role for filopodia in mediating Ca2+-sensitive acceleration. Taken together, our data suggest that injury-derived [Ca2+]i signaling may regulate wound closure rates by an adhesion-mediated mechanism.

Detection of Pseudomonas aeruginosa in bronchial and tracheal aspirates by PCR by amplification of the exotoxin A gene. / Nachweis von Pseudomonas aeruginosa aus Bronchial- und Trachealsekreten mittels PCR durch Amplifikation des Exotoxin A-Genes.

For the first time, a PCR test based on the amplification of the Exotoxin A was evaluated for its ability to rapidly detect Pseudomonas aeruginosa in tracheal and bronchial aspirates from mechanically ventilated patients. The reaction is based on the amplification of a 396 bp region within the Exotoxin A gene. The results show that this PCR-method is suitable for the detection of Pseudomonas aeruginosa even in clinical samples such as aspirates. Among the 380 clinical samples tested in this way, 57 were found to be positive while only 36 were positive using routine culture. In conclusion, these results suggest that the PCR-method mentioned above can be used to provide a specific, rapid, simple, and highly sensitive detection of Pseudomonas aeruginosa in clinical samples.

Influence of maxillary constriction on nasal resistance and sleep apnea severity in patients with Marfan's syndrome.

BACKGROUND: Marfan's syndrome is associated with a high prevalence of obstructive sleep apnea (OSA). As this syndrome is associated with a characteristic constricted maxilla and high-arched palate, we reasoned that nasal airway constriction and resultant high nasal airway resistance (NAR) may contribute to the development of OSA. Therefore, the aim of this study was to measure NAR in patients with Marfan's syndrome. In addition, we aimed to examine the influence of maxillary morphology on both NAR and the severity of OSA. METHOD: We measured NAR in 13 consecutive patients with Marfan's syndrome and 13 control subjects. NAR was measured by posterior rhinomanometry, and expressed as the inspiratory resistance at a flow of 0.5 L/s. Dental impressions were taken to evaluate maxillary arch morphology, allowing measurement of the following distances: intercuspid (ICD), interpremolar (IPD), intermolar (IMD), and maximum hard palate height (MPH). Ten of the patients and four of the control subjects had previously undergone nocturnal polysomnography. RESULTS: Mean NAR for the Marfan group was more than twice that in the control group (7.7 +/- 1.2 vs 2.9 +/- 0.4 cm H2O/L/s; p < 0.005). The patients also had marked constriction of the maxillary arch compared with control subjects. Two of the lateral maxillary measurements were significantly inversely correlated with NAR. There were significant correlations between various maxillary arch measurements (MPH/ICD, MPH/IPD, MPH/IMD) and the apnea/hypopnea index. CONCLUSION: These data suggest that high NAR is a common feature of Marfan's syndrome. Maxillary constriction with a relatively high hard palate appears to be a major reason for the high NAR. The significant correlations between indexes of maxillary constriction and sleep apnea severity suggest that maxillary morphology may play an important role in the pathophysiology of OSA in Marfan's syndrome.

[A herpesvirus-caused enzootic--Pacheco's parrot disease--in a psittacine collection]. / Eine Herpesvirus-bedingte Enzootie--Pachecosche Papageienkrankheit--in einem Psittazidenbestand.

After the official confirmation of psittacosis in a collection of psittacine birds a total of 40 of them died during treatment with tetracycline. 36 of them underwent post mortem examination. From 33 birds the causative herpesvirus of Pacheco's parrot disease (PPD) was isolated and/or a non-purulent hepatitis diagnosed, the latter a characteristic for PPD. The cause of the outbreak was assumed to be a latent herpesvirus infection of individual birds which was activated by various stress factors during the psittacosis treatment. The macroscopic and histologic lesions, the results of virological investigations and the in vitro effect of acyclovir on the multiplication of the isolated herpes virus are described.