Genetic diversity of bovine viral diarrhea viruses from the Galicia region of Spain.

This study examined the frequency and diversity of bovine viral diarrhoea viruses (BVDVs) infecting cattle in Galicia (northwestern Spain). A total of 86 BVDV strains were typed in samples of serum from 79 persistently infected animals and 3 viraemic animals and of abomasal fluid from 4 fetuses. Samples came from 73 farms participating in a voluntary BVDV control programme. Typing was based on a 288-bp sequence from the 5' untranslated region amplified using primers 324 and 326. Of the 86 strains, 85 (98.8 per cent) belonged to species BVDV-1 and 1 (1.2 per cent) belonged to BVDV-2; 73 strains (84.9 per cent) were typed as BVDV-1b, 2 as BVDV-1e and 6 as BVDV-1d. One strain each was typed as belonging to 1a, 1h, 1k and 1l. The sole BVDV-2 strain was classified as 2a. These results identify BVDV-1b as the predominant species, and they indicate the presence of viral types not previously described anywhere in Spain. This is also the first report of BVDV-2 in Galicia and only the second report of BVDV-2 in Spain.

Specific inhibition of protein kinase Cbeta expression by antisense RNA affects the activation of Jurkat T lymphoma cells.

Antisense RNA technology was employed to specifically inhibit the expression of the protein kinase Cbeta (PKCbeta) isoform in Jurkat cells, to explore its influence on the expression of surface antigens (CD69) and the cytokines interleukin-8 (IL-8), tumour necrosis factor (TNF)-alpha and beta, and to characterise its controversial involvement in the expression of IL-2 and its receptor (IL-2R). Transfection of cells with an antisense PKCbeta construct (as-PKCbeta-pREP3) significantly increased IL-2R/CD25 expression in phorbol 12-myristate 13-acetate (PMA)-stimulated as-PKCbeta-pREP3 transfectants, in contrast to Jurkat cells transfected with a control as-PKCalpha-pREP3 plasmid. IL-2 production, in contrast, was strongly inhibited in both transfectant populations stimulated by PMA plus the calcium ionophore ionomycin. Three clones (asb1/asb2/asb3), selected from as-PKCbeta-pREP3 transfectants, showed decreased PKCbeta protein levels (40 percent, 50 percent and 60 percent, respectively, as determined by western blotting) and mRNA levels. The specific inhibition was confirmed in immunoblots for other PKC (alpha, delta, epsilon, gamma, theta, and lambda lambda/tau) isoforms and in immunoprecipitates from representative (c2/asb2) clones. Stimulation of PKCbeta-depleted clones significantly increased CD25 expression but decreased IL-2 production (similarly to as-PKCbeta-pREP3 transfectants) and IL-2 message levels. CD69 expression and IL-8 secretion were significantly decreased, but TNFbeta message levels were highly increased in asb2/asb3 clones, without affecting TNFalpha secretion. Analysis of the mitogen-activated protein kinase (MAP Kinase) signalling pathway showed unaltered extracellular signal regulated kinase 1/2 (ERK1/2) and p38 phosphorylation but increased activation of c-Jun N-terminal kinase (JNK1) and its substrate, the transcription factor ATF-2 (activated transcription factor-2), which are involved in IL-2 gene expression. Our results revealed new PKCbeta functions, affecting CD69 expression and IL-8 production, and support the requirement for PKCbeta in IL-2 secretion/transcription and IL-2R regulation.

[Analysis of the aetiologies of headaches in a paediatric emergency service]. / Análisis etiológico de las cefaleas desde un servicio de emergencia pediátrica.

INTRODUCTION: Headaches are a frequent cause of children's visits to different health care services but the epidemiological profile varies widely according to whether the sample is taken from among visits to hospital clinics, neuropaediatric units or emergency services. AIMS: The aim of this study was to analyse the aetiologies of the cases of headache attended in a paediatric emergency service. PATIENTS AND METHODS: We conducted a retrospective analysis of patient records with headache as the main reason for visiting over a six month period at the Paediatric Emergency Department at the Centro Hospitalario Pereira Rossell. Aetiologies were classified according to International Headache Society (IHS) criteria and the information was completed by telephone if the medical records contained insufficient data. RESULTS: The records of 185 children were studied, which represented 0.58% of the total number of visits. The most common aetiologies were non-cranial infections (43.1%), migraines (14.6%), tension-type headaches (9.2%), traumatic injury (8.1%) and sinusitis (5.4%). Serious causes constituted 4.9% of the total, with a predominance of tumours and infections of the central nervous system. The low percentage of viral meningitis in our series (1%) leads us to suspect an underdiagnosis of this disorder, given the scant number of cases in which the cerebrospinal fluid was examined. CONCLUSIONS: Our study confirmed the existence of a wide and varied range of aetiologies causing headaches, with a clear predominance of extracranial disorders and a low percentage of serious intracranial causes. Non-classifiable headaches made up 10.8% of the sample. Brain imaging studies were performed in 9.7% of cases, which is a considerably lower percentage than that of other series and demonstrates a rational use of this resource.

[The usefulness of Ki67 expression in the biopsy specimens, to predict the biochemical progresion of the prostate cancer after radical prostatectomy]. / Utilidad de la expresión de Ki67 en las biopsias preoperatorias para predecir la recidiva bioquímica del cáncer de próstata después de la prostatectomía radical.

OBJECTIVE: To evaluate the usefulness of Ki67 expression in the biopsy specimens, to predict the biochemical progression of the prostate cancer after radical prostatectomy. MATERIAL AND METHODS: We analyse the Ki67 expression in the biopsy specimens of 103 patients treated with radical prostatectomy. The mean follow up is 3.4 years (1.3-8.8 years). We correlate the biochemical progression with traditional prognostic factors as the PSA (> 10/< or = 10), Gleason (> or = 7/< 7), pT classification (pT3/pTO-2) and immunohistochemical factor Ki67 (> 3%/< or = 3%). RESULTS: Of all 103 patients, in 71 (69%) biochemical progression was not detected and in 32 (31%) biochemical progression was detected. The mean of preoperative PSA is 10.07 ng/ml in the patients without progression and 20.90 ng/ml in the patients with biochemical progression (p=0.0001). The mean of Gleason score in 6.03 in the patients without progression and 6.75 in the patients with biochemical progression (p=0.0001). The percentage of Ki67 expression is 3.95% in the patients without progression and 5.05% in the patients with biochemical progression (p=0.030). The tumors pT0-2 progressed 12/67 (17.9%) and the tumors pT3 progressed 20/36 (55.6%) (p=0.0001). Multivariant regression analysis indicate that it does not exist a statistically significant relation between Ki67 (> 3%/< or = 3%) expression in the biopsy specimens and the biochemical progression of the prostate cancer after radical prostatectomy (p=0.204). CONCLUSIONS: The immunohistochemical factor Ki67 (> 3%/< or = 3%) in the biopsy specimens, is less effective than the classic factors, PSA (> 10/< or = 10), Gleason (> or = 7/< 7) and pT classification (pT3/pT0-2), to predict the biochemical progression of the prostate cancer after radical prostatectomy.

[Orthognathic surgery. Study of nerve injuries]. / Chirurgia ortognatica. Studio sulle lesioni nervose.

The correction of oral and maxillo-facial mal-formations causes lesions to sensitive nerves due to the invasive and traumatic surgery techniques required. Generally neuro-sensory and cutaneous-mucosal defects are reversible, with full recovery within 6-12 months. In a follow-up study (2 and 8 years) we found altered tactile and pain sensitivity in 43% of patients at subjective evaluation, with objective alteration in 17% of the group. Dental sensitivity evaluated by a pulp test showed an increase in for all teeth tested; values were higher for superior than for inferior teeth. Despite the increased pulp sensitivity threshold, due to the surgical damage, no signs of pulp disease were detected. Quality of life was not affected even by such a high subjective change in sensitivity. We therefore believe that patients undergoing orthognathic surgery must be fully informed; they must be made to understand the risks involved, and that damage may occur to sensitive nerves following surgery. All surgeons, including those with experience in the field, should continually seek to improve their skill and learn new techniques, in order to reduce complications to a minimum.

Differential expression of thymosins beta(4) and beta(10) during rat cerebellum postnatal development.

The beta-thymosins are a family of actin monomer-sequestering proteins widely distributed among vertebrate classes. The most abundant beta-thymosins in mammalian species are thymosin beta(4) (Tbeta(4)) and thymosin beta(10) (Tbeta(10)), two small peptides (43 amino acids) sharing a high degree of sequence homology. In the present work, we have analyzed the distribution of Tbeta(4) and Tbeta(10) in the developing and adult rat cerebellum using in situ hybridization and immunohistochemistry techniques. Our results show that the temporal and cellular patterns of expression of both beta-thymosins are different. In the young (7 and 18 postnatal days) and adult (1 and 4 months old) rat cerebellum, Tbeta(4) was mainly expressed in the glia (microglia, Golgi epithelial cells and oligodendrocytes), neurons (granule cells and Purkinje cells), and in the capillaries. In 14-month-old rats, the Tbeta(4) immunoreactivity was only detected in some microglia cells. In young and adult animals, most of the Tbeta(10) immunoreactivity was localized in several types of neuronal cells including granule cells, Golgi neurons and Purkinje cells. In old animals, a faint Tbeta(10) signal could be detected in a few Purkinje cells. Our results suggest that each beta-thymosin could play a different function in the control of actin dynamics.

[Psychiatric approach to the clinical aspects of drug dependence: psychiatric comorbidity in drug addicts]. / L'approccio psichiatrico alla clinica delle tossicodipendenze: la comorbidità psichiatrica nel paziente tossicodipendente.

The authors analyze the most important theoretical models in the field of drug addiction: the moral, the medical, the psychodynamic and the psychiatric model, with particular reference to the concepts of comorbidity and dual diagnosis. It is pointed out as these approaches invariably lead to a dichotomy between the "defect/process" model and the "conflict/development" model. The proposed hypothesis is an attempt to reconcile the opposite points of view in an unitary vision: on the basis of the fundamental psychopathological elements (drive dyscontrol and dependence), the addicts are so distributed into three areas, different for nosography and treatment.

Role of persistent amenorrhea in bone mineral metabolism of young hemodialyzed women.

BACKGROUND: Chronic renal failure in women is frequently associated with endocrine disturbances leading to menstrual disorders. However, most studies on renal osteodystrophy have not taken into account the possible role of these hormonal disturbances on the pathogenesis of bone alterations seen in these patients. In the present study, we evaluated bone mineral metabolism in a group of young hemodialyzed women with persistent amenorrhea and compared them with similar women with regular menstruation. METHODS: We studied 74 women who were further subdivided into 43 women with regular menstrual periods and 31 women with persistent amenorrhea, defined as the absence of menstrual bleeding for more than six months. In all patients, we performed a bone mineral density (BMD) analysis and simultaneously evaluated different biochemical parameters, intact parathyroid hormone (iPTH), sexual hormone determinations that included total estradiol, follicle-stimulating (FSH), and luteinizing hormone and markers of bone resorption such as the procollagen type 1 cross-linked carboxy-terminal telopeptide (ICTP). RESULTS: Serum calcium, phosphorus, and iPTH were similar in both groups. Serum alkaline phosphatase was higher in amenorrheic women. Although the total serum estradiol concentration was normal in the amenorrheic women when compared with nonuremic women, the values were significantly lower than those in regularly menstruating women. Serum FSH and ICTP values were significantly higher in the amenorrheic women. Trabecular BMD in the lumbar spine was also significantly lower in the amenorrheic women compared with regularly menstruating dialysis patients. Lumbar spine BMD and total estradiol levels correlated significantly in the amenorrheic group. CONCLUSIONS: These studies show that persistent amenorrheic young women on dialysis have lower trabecular BMD and evidence of increased bone resorption when compared with normal menstruating women on dialysis. The possible impact of these results in the natural history of the uremic osteodystrophy remains to be determined.

Distribution of choline acetyltransferase immunoreactivity in the brain of an elasmobranch, the lesser spotted dogfish (Scyliorhinus canicula).

Although the distribution of cholinergic cells is remarkably similar across the vertebrate species, no data are available on more primitive species, such as cartilaginous fishes. To extend the evolutionary analysis of the cholinergic systems, we studied the distribution of cholinergic neurons in the brain and rostral spinal cord of Scyliorhinus canicula by immunocytochemistry using an antibody against the enzyme choline acetyltransferase (ChAT). Western blot analysis of brain extracts of dogfish, sturgeon, trout, and rat showed that this antibody recognized similar bands in the four species. Putative cholinergic neurons were observed in most brain regions, including the telencephalon, diencephalon, cerebellum, and brainstem. In the retrobulbar region and superficial dorsal pallium of the telencephalon, numerous small pallial cells were ChAT-like immunoreactive. In addition, tufted cells of the olfactory bulb and some cells in the lateral pallium showed faint immunoreactivity. In the preoptic-hypothalamic region, ChAT-immunoreactive (ChAT-ir) cells were found in the preoptic nucleus, the vascular organ of the terminal lamina, and a small population in the caudal tuber. In the epithalamus, the pineal photoreceptors were intensely positive. Many cells of the habenula were faintly ChAT-ir, but the neuropil of the interpeduncular nucleus showed intense ChAT immunoreactivity. In the pretectal region, ChAT-ir cells were observed only in the superficial pretectal nucleus. In the brainstem, the somatomotor and branchiomotor nuclei, the octavolateral efferent nucleus, and a cell group just rostral to the Edinger-Westphal (EW) nucleus contained ChAT-ir neurons. In addition, the trigeminal mesencephalic nucleus, the nucleus G of the isthmus, some locus coeruleus cells, and some cell populations of the vestibular nuclei and of the electroreceptive nucleus of the octavolateral region exhibited ChAT immunoreactivity. In the reticular areas of the brainstem, the nucleus of the medial longitudinal fascicle, many reticular neurons of the rhombencephalon, and cells of the nucleus of the lateral funiculus were immunoreactive to this antibody. In the cerebellum, Golgi cells of the granule cell layer and some cells of the cerebellar nucleus were also ChAT-ir. In the rostral spinal cord, ChAT immunoreactivity was observed in cells of the motor column, the dorsal horn, the marginal nucleus (a putative stretch-receptor organ), and in interstitial cells of the ventral funiculus. These results demonstrate for the first time that cholinergic neurons are distributed widely in the central nervous system of elasmobranchs and that their cholinergic systems have evolved several characteristics that are unique to this group.

Atmospheric aerosol optical properties: a database of radiative characteristics for different components and classes.

A database management system has been realized that, by taking physical and chemical properties (the complex refractive index and the size distribution) of basic components as its starting point, allows the user to obtain optical properties of default as well as user-defined aerosol classes. Default classes are defined in accordance with the most widely known and used aerosol models. We obtain user-defined classes by varying the mixing ratio of components, creating new mixtures of default components, or by defining user components, thereby supplying the size distribution and the refractive index. The effect of relative humidity (RH) on the refractive index and the size distribution is properly accounted for up to RH = 99%. The two known mechanisms of obtaining classes from components are allowed (internal or external mixing).