Detection of visual-field deterioration by Glaucoma Progression Analysis and Threshold Noiseless Trend programs.

BACKGROUND: To compare the ability of Glaucoma Progression Analysis (GPA) and Threshold Noiseless Trend (TNT) programs to detect visual-field deterioration. METHODS: Patients with open-angle glaucoma followed for a minimum of 2 years and a minimum of seven reliable visual fields were included. Progression was assessed subjectively by four masked glaucoma experts, and compared with GPA and TNT results. Each case was judged to be stable, deteriorated or suspicious of deterioration RESULTS: A total of 56 eyes of 42 patients were followed with a mean of 7.8 (SD 1.0) tests over an average of 5.5 (1.04) years. Interobserver agreement to detect progression was good (mean kappa = 0.57). Progression was detected in 10-19 eyes by the experts, in six by GPA and in 24 by TNT. Using the consensus expert opinion as the gold standard (four clinicians detected progression), the GPA sensitivity and specificity were 75% and 83%, respectively, while the TNT sensitivity and specificity was 100% and 77%, respectively. CONCLUSION: TNT showed greater concordance with the experts than GPA in the detection of visual-field deterioration. GPA showed a high specificity but lower sensitivity, mainly detecting cases of high focality and pronounced mean defect slopes.

[Nomogram for ocular hypertension progression risk based on the ocular hypertension treatment study]. / Nomograma de riesgo de progreso de hipertensión ocular basado en el ocular hypertension treatment study.

INTRODUCTION: A practical nomogram has been designed in order to present the results obtained from the Ocular Hypertension Study (Gordon et al. Arch Ophthalmol 2002; 120: 714-720), where the relation between intraocular pressure (IOP) and corneal thickness becomes apparent, involving the risk of evolution from ocular hypertension into glaucoma within a 6 year period. MATERIAL AND METHODS: We used a multiple logarithmic regression for the nine parameters shown in figure 1 of the above mentioned paper. RESULTS: A correlation coefficient of 0.91 (p<0.001) permits to establish the following equation: Probability of evolution (%) = 13539.5 x (1.1385IOP) x (0.9818(CORNEAL THICKNESS)). This implies that a variation of 10 microns on corneal thickness leads to an IOP's modification of 1.5 mmHg in the same sense. From these data, we designed the nomogram included in this paper. CONCLUSIONS: IOP and pachymetry together allow an estimation of the risk of evolution from ocular hypertension into glaucoma in a graphical practical way. From this indirect estimation, the influence of corneal thickness on IOP's measure seems to be much higher than previously estimated.

Nomograma de riesgo de progreso de hipertensión ocular basado en el Ocular Hypertension Treatmetn Study / Nomogram for ocular hypertension progression risk based on the Ocular Hypertension Treatment study

Introducción: Se ha construido un nomograma práctico para representar los resultados del Ocular Hipertensión Study (Gordon et al. Arch Ophthalmol 2002;120: 714-720) que relacionan la presión intraocular y el espesor corneal con el riesgo de evolucionar de hipertensión ocular a glaucoma en un plazo de 6 años.Material y Métodos: Se ha aplicado una regresión logarítmica múltiple a los 9 datos mostrados en la figura 1 del trabajo señalado anteriormente.Resultados: Un coeficiente de correlación de 0,91 (p<0,001) permite definir la siguiente ecuación:Probabilidad de evolución (%) = 13539,5 x (1,1385 PIO) x (0,9818 espesor corneal)Esto significa que una variación de 10 micras en el espesor corneal equivale a una variación de 1,5 mmHg en el mismo sentido de la presión intraocular. Con estos datos se ha construido un nomograma que se incluye en el trabajo.Conclusiones: La PIO y la paquimetría permiten estimar el riesgo de evolución de hipertensión ocular a glaucoma de una manera gráfica práctica. Estimada de esta manera indirecta, la influencia del espesor corneal sobre la medida de la presión intraocular parece muy superior a la que se había estimado hasta el momento

Introduction: A practical nomogram has been designed in order to present the results obtained from the Ocular Hypertension Study (Gordon et al. Arch Ophthalmol 2002; 120: 714-720), where the relation between intraocular pressure (IOP) and corneal thickness becomes apparent, involving the risk of evolution from ocular hypertension into glaucoma within a 6 year period.Material and Methods: We used a multiple logarithmic regression for the nine parameters shown in figure 1 of the above mentioned paper.Results: A correlation coefficient of 0.91 (p<0.001) permits to establish the following equation:Probability of evolution (%) = 13539.5 x (1.1385 IOP) x (0.9818 CORNEAL THICKNESS)This implies that a variation of 10 microns on corneal thickness leads to an IOP’s modification of 1.5 mmHg in the same sense. From these data, we designed the nomogram included in this paper.Conclusions: IOP and pachymetry together allow an estimation of the risk of evolution from ocular hypertension into glaucoma in a graphical practical way. From this indirect estimation, the influence of corneal thickness on IOP’s measure seems to be much higher than previously estimated

Subcellular localization of Dp71 dystrophin isoforms in cultured hippocampal neurons and forebrain astrocytes.

It has been suggested that the absence or altered structure of Dp71, a C-terminal dystrophin gene encoded protein, is responsible for mental alterations observed in about 30% of Duchenne muscular dystrophy patients. Most of these patients have premature translational termination or point mutations at the C-terminal domain of this gene. In brain, Dp71 is the major protein product of the dystrophin gene. To determine the function of Dp71 isoforms in this organ, it is important to document their presence and intracellular localization in brain cells. Extracts from cultured hippocampal neurons and forebrain astrocytes and 5F3 and Dys 2 monoclonal antibodies were thus used for western blots. In these conditions, two Dp71 isoforms spliced or not at exon 78 were detected in both cells (Dp71f and Dp71d, respectively). By immunocytochemistry, we mapped Dp71f and Dp71d in the Golgi complex (GC) and in neuronal nuclei. Only Dp71d was found in cytoplasmic neurofilaments. In astrocytes, these isoforms were detected in the GC. These cell localization data suggest that these Dp71 isoforms may have different functions in the same cell or organelle, as well as in the two different cells analyzed.

Biochemical and histochemical analysis of 71 kDa dystrophin isoform (Dp71f) in rat brain.

Dp71 is a member of the dystrophin family and the most abundant dmd gene product in the brain. In the present study, we focused on a short dystrophin transcript named Dp71f, which is alternatively spliced when exon 78 is absent The topographic localization of this protein in the encephalon has not been properly described yet, nor its cellular or subcellular localization, and even less its functions. Dp71f was found to be a cytoplasmic 70 kDa protein and localized in all encephalon regions studied. Double labeling using specific markers for various cell types confirmed Dp71f distribution in the cytoplasm of all cell types studied. Labeling was more conspicuous near the nucleus and diminished towards the periphery of cells. In some cases, we observed cells that were positive for actin and Dp71f in regions corresponding to lamellipodia-like structures. Dp71f and Dp71d isoforms were differently distributed. Our study is the first specific and unambiguous description of the topography and cellular localization patterns of Dp71f in brain, suggesting that Dp71f is a ubiquitous protein.

Mitochondrial expression of a short dystrophin-like product with molecular weight of 71 kDa.

In the brain, Dp71 is the most abundant protein product of the DMD gene and by alternative splicing of exon 78 two isoforms can be expressed, Dp71d and Dp71f. To explore the subcellular distribution of these Dp71 isoforms, specific monoclonal antibodies were used. Dp71d (with exon 78) was found in microsomes, while Dp71f (without exon 78) was detected in mitochondria. To determine the alterations which the absence of dystrophin proteins induces, we compared the expression of Dp71d in microsomes and Dp71f in mitochondria from mdx and mdx(3CV) mice. Dp71d in microsomes of mdx was similar to that of wild-type mice and, as expected, in mdx(3CV) this protein was undetectable. However, in mitochondria from mdx(3CV), Dp71f was overexpressed in comparison to mitochondria from mdx mice. Because in mdx(3CV) mice all the dystrophin proteins are mutated or diminished, we concluded that the protein detected in mitochondria is not a Dp71f but a novel product named Dp71f-like protein.

Changes in rat brain muscarinic receptors after inhibitory avoidance learning.

It is widely accepted that cerebral acetylcholine is necessary for learning and memory, but little is known about the type of muscarinic receptors involved in these functions. To investigate this problem, [3H]-N-methyl-scopolamine which binds to different types of muscarinic receptors, [3H]-Pirenzepine an M1 receptor antagonist, and [3H]-Oxotremorine-M which binds mainly to M2 receptors, were used as ligands to look for possible changes in muscarinic receptor density in neostriatum (NEO), hippocampus (HIP), amygdala (AMY), and temporo-parietal neocortex (CTX), after testing for retention of inhibitory avoidance, trained with high or low footshock intensities. After low reinforcement there was an M1 postsynaptic receptor up-regulation in NEO, HIP, and CTX, and an M2 presynaptic receptor down-regulation in HIP, which suggests a concerted pre- and postsynaptic cholinergic activation in this area. An up-regulation of both M1 and M2 receptors was detected in CTX of low and high footshocked animals, which indicates the presence of a cortical postsynaptic M2 receptor.

[Liver changes in protein malnutrition. An experimental study in rats]. / Cambios hepáticos en la malnutrición proteica. Estudio experimental en ratas.

It is known that protein malnutrition conditions the development of liver steatosis, and may be accompanied by fibrosis. Ito cells intervene in the fibrogenesis, converting to transitional cells and myofibroblasts. Some trace elements, such as copper (Cu), iron (Fe) and manganese (Mn) act as co-factors, and zinc (Zn) acts as an inhibitor of a variety of enzymes involved in the collagen synthesis. This study analyzes the effects on the livers of 12 mate Wistar rats following two months administration of a hypoproteic (6%) isocaloric diet, comparing histomorphometric parameters (hepatocyte and nucleum area, total fat and fibrosis) and the liver content in Fe, Cu, Zn and Mn, with those in 12 control rats of similar age and sex. The experimental group revealed a significant reduction in hepatocyte nucleum area (p < 0.001), an increase in the ratio of hepatocyte cytoplasmatic and nucleum hepatocyte area, pronounced steatosis and slight fibrosis. No differences were found in Ito cell percentages. The experimental group showed a significant increase in liver content of Fe (p = 0.01) and a significant drop in Mn content (p < 0.01), Zn (p = 0.05) and Cu (p < 0.01). Liver iron content correlated significantly with total fat level (p = 0.03).

Relative and combined roles of ethanol and protein malnutrition on muscle zinc, potassium, copper, iron, and magnesium.

The aim of the present study is to analyse the relative and combined effects of ethanol and protein malnutrition on muscle zinc, copper, iron, potassium, and magnesium in ethanol-fed rats. The study was performed in 32 animals divided into four groups, fed with the Lieber-DeCarli control, 36% ethanol, 2% protein, and 36% ethanol 2% protein containing diets, respectively. Right gastrocnemius muscle was removed 2 months later, and was studied both chemically and histochemical-morphometrically. Both muscle zinc and potassium, but not copper nor iron nor magnesium, were significantly decreased in the protein-deprived, ethanol-fed animals, the main effect of these variations being attributable to ethanol rather than to protein deprivation. However, coexisting protein deprivation aggravated the decrease in both muscle zinc and potassium. Both muscle zinc and potassium were significantly related to serum albumin, weight loss, and type IIb fibre atrophy; and muscle zinc, in addition, to the decrease in type IIb fibre proportion. Therefore, a decrease in muscle content of both elements is related to histochemical-morphometrical changes observed in alcoholic myopathy. In addition, both ethanol and protein deficiency exerted independent, highly significant effects both on type IIb fibre atrophy and proportion.

Combined effects of ethanol and protein deficiency on hepatic iron, zinc, manganese, and copper contents.

The present study has been performed in order to establish the relative and combined roles of ethanol and malnutrition on liver Fe, Zn, Cu, and Mn alterations in alcoholic male adult Wistar rats, and also the relationships between these alterations and histomorphometrically determined hepatocyte and nuclear areas, perivenular fibrotic rim area, and total amount of fat present in the liver. Four groups of 8 animals each were fed: (1) a nutritionally adequate diet (C); (2) a 36% ethanol-containing (as percent of energy), isocaloric diet (A); (3) a 2% protein-containing, isocaloric diet (PD); and (4) a 36% ethanol, 2% protein-containing, isocaloric diet (A-PD), respectively, following the Lieber-DeCarli model. Ethanol-fed, protein-deficient animals showed the highest liver Fe, and the lowest Zn and Cu values, although differences in liver Zn, Mn, and Cu values were not significantly different between PD and A-PD groups. Statistically significant differences of these parameters were observed between the A and the A-PD groups, and between the A and PD groups, except for liver iron. Except for liver Mn, differences between C and A groups were statistically significant. These alterations correlated with liver fibrosis and steatosis, serum albumin, and weight loss, except for liver Mn, which was not correlated with fibrosis or steatosis. Thus, protein deficiency seems to enhance ethanol-induced liver Fe, Zn, and Cu alterations, whereas protein deficiency, but not ethanol, seems to play a major role on liver Mn alterations.

Effect of ethanol on hepatic iron, copper, zinc and manganese contents in the male albino mouse.

Liver Fe and Cu contents (determined by atomic absorption spectrophotometry) were found to be higher in alcoholized male albino mice than in controls. Alcoholized animals killed at the 180th day of life also showed higher liver Fe and Cu contents than the alcoholized animals sacrificed at the 85th day of life. A significant correlation was established between liver Fe content and size of the pericentral hepatocytes and their nuclei. No differences between alcoholic and controls were obtained regarding liver Zn content, whereas Mn showed a clear tendency to be higher in the alcoholics.

Effects of ethanol and propylthiouracil on hepatic iron and copper contents in the male albino mouse.

With the aim to analyze whether propylthiouracil (PTU) alters ethanol-induced changes in liver iron and copper contents, 40 male albino mice were equally divided into a control group, an ethanol-treated group, a PTU-treated group and an ethanol + PTU-treated group. Twenty animals were killed at the age of 85 days and 20 at the age of 180 days. Liver iron and copper contents showed a progressive, statistically non-significant increase both in the controls and in ethanol-treated animals. Liver iron contents was significantly higher in the 180-day-old alcoholic mice as compared with controls. Animals treated either with PTU or with PTU + ethanol showed liver iron levels in the normal range, markedly different from the ethanol-treated animals (P less than 0.005). Liver copper content of the ethanol-treated animals was higher (but not significantly) than that of the controls. Liver copper levels of the PTU + ethanol-treated animals were in the range of the ethanol-treated animals. Thus, PTU seems to revert an overload of ethanol-mediated iron of copper.

Ultrastructural alterations in caudate nucleus, cerebral cortex and hippocampus produced by morphine.

1. Chronic morphine administration to rats produced diverse ultrastructural alterations in nerve cells and neuropile of caudate nucleus, cerebral cortex and hippocampus through a short term abstinence period. 2. All areas studied showed increasing damage related to time elapsed between the last morphine injection and animal sacrifice. 3. Intracytoplasmic neuronal membranous organelles mainly suffered severe swelling, membrane disarrangement and eventually cell disruption in all areas studied. 4. Hippocampus was the most affected area throughout the study, followed by caudate nucleus and cerebral cortex, where focal damage was seen. 5. Susceptibility to morphine cytotoxic effect in the three areas studied appears to be unrelated to their opiate receptor density. 6. Mitochondrial alterations produced by morphine could be related to interference of the intracellular energy production system and consequent unrestricted cell membrane permeability to water and solutes.

Structural changes in caudate nucleus, cerebral cortex and hippocampus induced by morphine. Light microscopy study.

Intraperitoneal chronic morphine administration to rats produced structural alterations in nerve cells of caudate nucleus, cerebral cortex and hippocampus. All areas studied showed increased alterations related to time elapsed between last morphine injection and fixation of tissue samples. Lesions mainly consisted in vacuolar degeneration of nerve cells. The most extense and intense lesions were observed in hippocampus. Sensorimotor cortex and caudate nucleus showed focal damage. Neurocytotoxic effect of morphine appears to be unrelated to the opiate receptor density in brain areas, and the brain regions susceptible to that effect appears to be different from brain areas related to adaptation to morphine. Effect of morphine on integrity and function of nerve cells mitochondria, interfering with energy production could play an important role in the pathogenesis of neuronal lesions.

Insulin-independent controlled physiological morphogenesis of chick muscle from fusion-capable myoblasts.

Thigh myogenic cells from 11-12-day-old chick embryos were cultured continuously in the presence of medium containing no chick embryo extract (CEE). It is known that CEE contains a muscle-inducing protein of 35,000 daltons. In spite of the absence of embryo extract and provided that calcium, starting at a concentration as low as 3 X 10(-4) M, was present in the tested media, typically aligned myotubes with 20 or more nuclei per fiber or abnormal myosymplasts were produced at will. In the first case, the result was systematically obtained when the media were unchanged. Consequently, the cell microenvironment remained undisturbed and therefore was autoconditioned throughout the 7 days of culture. In the second case, the result depended on the feeding schedules. Conversely, no myotubes were formed in cultures in embryo extract-free medium without calcium, irrespective of the frequency of medium changes. Insulin, a serum factor believed to be involved in syncytium formation process in vitro, was present in all tested media. Undialyzed or dialyzed fetal calf serum (FCS), used for the preparation of the media, contained 11 mu units of insulin per milliliter. The insulin content in all tested media was diluted, however, to one tenth the physiological serum concentration. The hormone did not promote any kind of myoblast fusion in any experiment in which calcium was deleted as a component of the tested media, regardless of the feeding schedule followed.(ABSTRACT TRUNCATED AT 250 WORDS)

Effect of different convulsant drugs on some seizure parameters in morphine-dependent mice.

An investigation was made of the effect of morphine dependence on the characteristics of seizures induced in mice by convulsant drugs with differing mechanisms of action. Morphine dependence was induced in 90-day-old mice (weighing 29 to 32 g) by a 6-day schedule of twice daily i.p. injections of increasing doses of morphine (5, 32.5, 58, 82.5, 100, and 135 mg kg-1). Thirty minutes after the last morphine administration, convulsant drugs (4-aminopyridine 8 mg kg-1, pentylenetetrazol 50 mg kg-1, bicuculline 2.5 mg kg-1, strychnine 2.0 mg kg-1, and beta-mercaptopropionic acid 50 mg kg-1) were injected. 4-Aminopyridine (4-AP) and pentylenetetrazol (PTZ) increased both the number of animals with convulsions and death and in the case of 4-AP the period of convulsion latency was also increased. Naloxone at 1.0 mg kg-1 blocked the 4-AP effects, indicating that this action was mediated through an opioid receptor. Strychnine and beta-mercaptopropionic acid had an effect opposite 4-AP and PTZ in the number of animals with convulsions and death. On the other hand bicuculline had an effect more like 4-AP and PTZ than other inhibitory synapse-blocking drugs. We conclude that chronic morphine treatment modified the response of convulsant drugs depending on their mechanisms of action.

Immunological and ultrastructural study of the surface of isolated rat testis germinal cells.

In the present study, cross absorption tests and indirect immunofluorescence detected antigenic differences between isolated testicular cell populations from 21 and 38 day old rats, respectively. Qualitatively, spermatids were the only cell type difference between these populations. Spermatozoa from adult rats were also studied. Our findings suggest that spermatocytes, spermatids and spermatozoa shared on or more antigens; spermatids and spermatozoa seemed to share one or more antigens not observed in spermatocytes by indirect immunofluorescence. Various patterns of immunofluorescence were encountered, suggesting differences in amount and distribution of antigens among the cells studied. The supramolecular appearance of the surface of spermatocyte enriched populations and of spermatid enriched populations were compared by scanning electron microscopy (SEM) and no striking difference was observed between both cell populations.