Polymerase chain reaction detection of BK virus and monitoring of BK nephropathy in renal transplant recipients at the University Hospital La Fe.

INTRODUCTION: Reactivation of BK infection occurs in immunocompromised hosts causing tubulointerstitial nephropathy (BKVN). Approximately 5% of kidney transplant recipients (KTR) develop BKVN, special half of whom lose their grafts. However, BKVN morphologic diagnosis on a renal biopsy is complicated, because the cytopathic changes can sometimes mimic rejection. Thus, BKV DNA-polymerase chain reaction (PCR) assay on serum, urine, and renal tissue is useful for early detection and monitoring of BKV. MATERIALS AND METHODS: We performed routine monthly urine cytologies looking for decoy cells as a marker of virus replication. Then, we performed a qualitative PCR on urine and serum in all recipients (independently of positive or negative cytology). We amplified 3 BK viral genome regions, LT (early transcription region) and VP1 (late transcription region) seeking a more accurate virus detection, and the TCR (control transcription region) region to perform a polymorphism sequence analysis to identify the BK genomic variant. Finally, the BKVN diagnosis was confirmed using renal biopsy. RESULTS: At present, 132 patients have been monitored. Thirteen of 40 (33%) were PCR-urine-positive cases (5 LT+/VP1- and 8 LT+/VP1+), and 10 of 132 (7.5%) were PCR-serum-positive cases (7 LT+/VP1- and 3 LT+/VP1+). When we compared PCR-urine and cytology results, 11 of 40 (27.5%) patients showed a positive cytology, 6 of whom were PCR- urine-positive (1 LT+/VP1- and 5 LT+/VP1+); whereas, 29 patients showed a negative cytology, 7 of whom were PCR-urine-positive(3 LT+/VP1- and 4 LT+/VP1+). Thus, comparison of PCR- urine and cytology results revealed false-positive and false-negative cases. Finally, TCR sequence analysis was performed in 9 patients to identify the BK genomic variants. CONCLUSION: Testing for BKV DNA in urine and serum is a noninvasive early detection assay and monitoring tool.

Cytochemical and stereological analysis of rat cortical astrocytes during development in primary culture. Effect of prenatal exposure to ethanol.

This study has investigated the effect of prenatal alcohol exposure on the qualitative and quantitative ultrastructure of proliferating and differentiated astrocytes in primary cultures as well as on the cytochemical activity of several subcellular phosphatase markers, including acid phosphatase, uridine diphosphatase, thiamine pyrophosphatase, 5'-nucleotidase and glucose-6-phosphatase. The astrocytes were obtained from 21-day-fetuses of both control and alcohol-fed rats. Our results show that several cell components, such as mitochondria, rough endoplasmic reticulum and lysosomes, exhibit qualitative and/or quantitative ultrastructural changes during the process of astrocyte maturation. In some cases these morphological changes are accompanied by variations in the cytochemical activity of enzymes located in these and other cell components, suggesting that these enzymes, and therefore the functional state of these organelles, are modulated during astrocyte development. When prenatally exposed to ethanol, both proliferating and differentiated astrocytes showed striking ultrastructural alterations compared with controls, including an increment of lysosomes as well as a decrease in the values of stereological parameters relative to mitochondria, rough endoplasmic reticulum and Golgi apparatus. Cytochemical analysis of these cells indicates that prenatal exposure to ethanol decreased the activities of all the enzymes tested, except for acid phosphatase, which was increased in both groups of treated astrocytes. These results suggest that prenatal exposure to ethanol could affect astrocytes during development in two different but probably complementary ways: a) by causing a delay in astrocyte maturation and, b) by inducing a direct toxic effect on these cells.

Neurologic complications associated with hepatitis C virus infection.

A series of nine patients with neurologic complications of hepatitis C virus infection is reported. Seven patients presented a combination of chronic sensory polyneuropathy, multineuropathy, and encephalopathy related to cryoglobulinemia. The noncryoglobulinemic symptoms consisted of an anterior optic neuropathy and a restless legs syndrome with small-fiber neuropathy. Corticosteroids and cyclophosphamide were useful in controlling vasculitic episodes. Interferon-alpha caused remission in half of the treated patients.

Effect of cholesterol and its autooxidation derivatives on endocytosis and dipeptidyl peptidases of aortic endothelial cells.

The effects of cholesterol (CHO) and cholesterol autooxidation derivatives (CAD) on the endocytosis of cationized ferritin (CF) by endothelial cells have been investigated. The effect of both substances on the activity of lysosomal enzymes dipeptidyl peptidase I (DPP I) and dipeptidyl peptidase II (DPP II) was also studied. Treatment of rats with CAD induced striking alterations in the ultrastructure of endothelial cells and makes it impossible to analyze the effect of this toxin on endocytosis processes. In contrast, CHO-treated cells displayed a good ultrastructural preservation and showed an increased ability to endocyte ferritin, as compared with controls. Both DPP I and DPP II activities increased after 3 weeks of CAD or CHO treatment. Our results indicate that although CHO damage endothelial cells, the most important effects could be attributed to CAD which usually accompanies CHO-supplemented diets.

Changes in the adrenergic mechanisms of cerebral arteries after subarachnoid hemorrhage in goats.

We have examined the effects of experimental subarachnoid hemorrhage (SAH), induced by delivering autologous blood into the subarachnoid space, on the adrenergic mechanisms of the goat cerebrovascular bed. To achieve this, the response to noradrenaline was recorded both in vivo, by measuring cerebral blood flow in unanesthetized animals, and in vitro, by recording isometric tension in isolated cerebral arteries. In addition, we checked the function of adrenergic innervation by measuring the tritium efflux evoked by electrical stimulation in cerebral arteries preloaded with [3H]-noradrenaline, and we examined this innervation by using both fluorescent and electron transmission microscopy. All studies were performed before and 3, 7, and 14 days after SAH. Injections of noradrenaline (0.1-10 micrograms) directly into the cerebro-arterial supply produced reductions in cerebral blood flow, with no concomitant changes in mean arterial blood pressure and heart rate, which were significantly enhanced (P < 0.01) 3 and 7 days after SAH and returned to control values 14 days after hemorrhage induction. In isolated cerebral arteries, noradrenaline (10(-8)-10(-4) mol/L) produced concentration-dependent contractions, which were also significantly enhanced (P < 0.05) 3 and 7 days after SAH and returned to control values in cerebral arteries obtained 14 days after SAH. On the other hand, increases in the release of tritium induced by electrical stimulation in cerebral arteries preloaded with [3H]-noradrenaline were significantly lower (P < 0.01) after SAH. Moreover, microscopical studies showed a reduction in catecholamine fluorescence and signs of sympathetic degeneration in some perivascular axons after SAH.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholesterol and 25-hydroxycholesterol retention in specimens of liver and aorta prepared for electron microscopy. I. Standard fixation methods and metabolism of the labeled sterols.

In the present work, several preparatory procedures commonly used for electron microscopy (EM) were evaluated as to their ability to preserve cholesterol (CHO) and CHO derivatives in tissue. We also determined in several rat tissues to what extent the sterols used as tracers are metabolized. Sprague-Dawley rats were injected intraperitoneally with [1 alpha,2 alpha(n)-3H]cholesterol ([3H]CHO) and 25-hydroxy-[26,27-3H]cholesterol ([3H]25-OH-CHO). Lipids of the liver, aorta and brain were extracted one and five days after injection, and the distribution of the labeled lipids was followed by thin-layer chromatography. When labeled CHO was injected as tracer, most of the radioactivity remained associated with the CHO fraction. When 25-hydroxycholesterol (25-OH-CHO) was used, we found that it was mostly metabolized to yield more polar compounds. Our results show that the loss of CHO and CHO derivatives from tissues depends not only on the preparatory procedure used for EM, but also on the type of tissue studied.

Cholesterol and 25-hydroxycholesterol retention in specimens of liver and aorta prepared for electron microscopy. II. Effect of filipin, osmium, digitonin and saponin.

Sprague-Dawley rats were injected intraperitoneally with [1 alpha, 2 alpha(n)-3H]cholesterol or 25-hydroxy-[26,27-3H]cholesterol, and one and five days later liver and aortic tissues were fixed. The extent to which these sterols were lost from the tissues during preparation for electron microscopy (EM) was examined utilizing different fixation procedures and various protective agents. Radioactive tracers, scintillation counting and standard EM techniques were used. Although most of the procedures examined caused major lipid losses, useful fixation procedures that allow retention of cholesterol or 25-hydroxy-cholesterol in liver and aortic tissues were found and are described here.

Modified internal bleaching of severe tetracycline discoloration: a 6-year clinical evaluation.

Since 1985, 534 severely discolored teeth have been treated with intentional endodontics and internal bleaching; of these, 258 were treated with the conventional technique and 276 were treated with the modified technique. Six years of clinical and radiographic evaluation revealed that the results obtained with the first group (conventional technique) remain stable and that a very noticeable improvement has been achieved with the second group of teeth, treated with the modified technique.

Dentinogenesis imperfecta: a case report.

Dentinogenesis imperfecta is a localized form of mesodermal dysplasia of the dentin affecting both the primary and permanent dentitions. Most previous reports on dentinogenesis imperfecta describe treatment with overdentures, which have several disadvantages. The present report describes a case of dentinogenesis imperfecta in an 11-year-old girl. A combination of restorative, prosthetic, and surgical treatment was used to resolve the condition.

Peripheral nerve hyperexcitability: a clinical and immunologic study of 38 patients.

OBJECTIVE: We studied a case series of peripheral nerve hyperexcitability (PNH) aiming to describe clinical characteristics, immunologic and cancer associations, antibodies against neuronal antigens (voltage-gated potassium channel antibodies [VGKC-Abs] and other), and muscle biopsy findings. METHODS: Patients presenting with clinical and electrophysiologic signs of PNH were selected. We studied clinical and electrophysiologic features; a panel of non-neuronal organ-specific antibodies, immunofluorescence on rat nervous tissues, and radioimmunoprecipitation for VGKC-Abs; and muscle biopsies. RESULTS: Thirty-eight patients were included. After the exclusion of 6 cases with axonopathy of known origin, patients were subdivided according to the presence of electrophysiologic findings of motor axonopathy and association with cancer: axonopathic-PNH (group A: 12 patients), isolated nonparaneoplastic PNH (group B: 16 patients), and isolated paraneoplastic PNH (3 with thymoma and myasthenia gravis, 1 with thyroid carcinoma). PNH clinical features were similar in groups A and B. We found an overall high prevalence of clinical autoimmunity (33% of group A and 63% of group B) and systemic non-neuronal autoantibodies (42% of group A and 75% of group B). However, VGKC-Abs were only positive in 2 patients of group B. Ten patients underwent muscle biopsy, which showed inflammatory changes in 2 cases and nonspecific myopathic features in 8. CONCLUSIONS: PNH is a heterogeneous disorder involving the peripheral nerves in patients with a high propensity for developing autoimmunity. Associated muscle diseases are frequent in the form of myositis, myasthenia gravis, or nonspecific myopathic pathologic findings. VGKC-Abs were uncommon in this series.

MYH7 gene tail mutation causing myopathic profiles beyond Laing distal myopathy.

OBJECTIVE: To describe a wide range of clinical and pathologic myopathic profiles associated with the p.K1729del mutation in the MYH7 gene, known to cause Laing distal myopathy. METHODS: A study conducted in the Safor region (Spain), setting of a large cluster of patients. Clinical, neurophysiologic, muscle imaging, and muscle biopsy studies and MYH7 gene sequencing were investigated in 32 patients from 4 kindreds. Data from 36 deceased or nonexamined patients were collected from hospital records or relatives. RESULTS: Onset ranged from congenital to the 6th decade. All patients presented weakness of great toe/ankle dorsiflexors and many had associated neck flexor, finger extensor, and mild facial weakness. In most cases, involvement of proximal and axial muscles was observed either clinically or by muscle imaging, sometimes giving rise to scapuloperoneal and limb-girdle syndromes. Disabling myalgias, skeletal deformities, and dilated cardiomyopathy in one patient were associated features. Life expectancy was not reduced but the spectrum of disability ranged from asymptomatic to wheelchair confined. Electromyographic neurogenic features were frequently recorded. Muscle fiber type disproportion, core/minicore lesions, and mitochondrial abnormalities were the most relevant pathologic alterations. All patients carried the p.K1729del mutation in MYH7. CONCLUSIONS: The p.K1729del mutation in the MYH7 gene expresses notable clinical variability and electromyographic and pathologic features that can lead to the misdiagnosis of neurogenic atrophies, congenital myopathies, or mitochondrial myopathies. Mutations in genes encoding other sarcomeric and reticulo-sarcoplasmic proteins involved in calcium regulation share pathologic characteristics with our patients, suggesting a possible pathogenetic connection.

[Occlusal rehabilitation in cranio-mandibular dysfunction syndrome with ceramometal restorations]. / Reabilitaçao oclusal em síndroma de disfunção crâneo-mandibular com restaurações ceramo-metálicas.

We present a clinical case of a temporo-mandibular joint dysfunction pain syndrome treated with occlusal rehabilitation consisting on ceramometalic prosthodontic restorations. The result obtained after five years of evolution is excellent, due to the use of adequate technical and clinical procedures together with suitable hygienic habits and the post-treatment establishment of an occlusal splint used exclusively at night.

Prenatal exposure to ethanol alters plasma membrane glycoproteins of astrocytes during development in primary culture as revealed by concanavalin A binding and 5'-nucleotidase activity.

We have investigated the effect of prenatal exposure to ethanol on the extent of binding and surface distribution of the lectin concanavalin A (con A) on rat cortical astrocytes during the periods of proliferation and differentiation in primary culture. The enzymatic activity of the plasma membrane glycoprotein 5'-nucleotidase was also assessed. The cells were obtained from control fetuses (no exposure to ethanol) and from fetuses prenatally exposed to ethanol. The main findings were: 1) both proliferating and differentiating control astrocytes showed two distinct types of surface con A receptors that could correspond to high- and low-affinity binding sites; 2) the extent of con A binding was greater in mature than in proliferating control cells; 3) the distribution of con A on cell surface components changed with differentiation; 4) the activity of 5'-nucleotidase showed a substantial increment during the period of differentiation; and 5) prenatal exposure to ethanol clearly decreased the ability of astrocytes to bind con A, altered the surface distribution of the receptors for this lectin, and decreased the activity of 5'-nucleotidase. These effects were more marked in proliferating cells. In conclusion, it is shown that the extent of con A labeling and the activity of 5'-nucleotidase in astrocytes are dependent on the stage of cell differentiation and that prenatal exposure to ethanol alters the plasma membrane structure of these cells during development.

[Radico-diagram: a technic for study of tooth root]. / El radico-diagrama: una técnica de estudio de la raiz dentaria.

The use of a simple measuring system such as the root diagram provides us with information about the morphology and the inclination of the roots of dental pieces. This system applied on x-rays taken with accurate and adequate techniques facilitates and enables one to study extensive groups of the population with common or divergent ethnic features and their subsequent comparative analysis.

[Plastic facial prosthesis]. / Prótesis plástica facial.

Patients, who present severe facial defects, become the most disturbing challenge of prosthodontia. In this cases, with extensive loose of substance, some times other therapies, are contraindicated. In this paper, we review the procedures to make facial prostheses.

[Determination of kinesiographic parameters in phonation]. / Determinación de patrones kinesiográficos en la fonacíon.

We have registered mandibular movements produced during the reading of a text in spanish with a Sirognathograph in a random sample of 71 subjects, aged between 24 and 35, on the three planes of space. The obtained curves have allowed to measure 7 variables in order to define the characteristics of mandibular movements during speech. The results demonstrate that determinant planes for this purpose are sagittal and frontal.

[Removable partial denture. A variation on the gauge used in design]. / Prótesis parcial removible. Una variación de la galga utilizada en diseño.

In this article we describe a modification of the gauge of the surveyor in such a way that we can measure the height of the retention arms' slide area over the abutment teeth, in order to obtain the same height in the reciprocal arm slide area to avoid the jiggle movements of the abutment teeth during the removal and insertion.

[Current esthetic parameters in fixed prosthodontics: a literature review]. / Parametros actuales estéticos en prostodoncia fija: una revisión bibliografica.

We show in this study the need to realize a careful selection of the color to be based on to put a single pre-fabricate guide to be outstanding the advantage and difficulties regarding to another systems. I denote the significance from the anatomic morphology of the restoration as well as the necessity of a defects control from the vision in the clinical like ceramist.

[Effective response to risperidone treatment in manic syndrome secondary to neurocysticercosis]. / Buena respuesta a risperidona en fase maniaca secundaria a neurocisticercosis.

This study presents a clinical case about a patient suffering a manic syndrome secondary to neurocysticercosis and no responding to traditional neuroleptic treatment. We administrated the risperidone treatment, that was effective.