Most of the increased protein degradation in muscle atrophy caused by starvation and denervation is due to activation of a non-lysosomal ATP-dependent proteolytic process. To determine whether expression of the ubiquitin-proteasome-dependent pathway is activated in atrophying muscles, we measured the levels of mRNA for ubiquitin (Ub) and proteasome subunits, and Ub content. After rats had been deprived of food for 1 or 2 days, the concentration of the two polyubiquitin (polyUb) transcripts increased 2-4-fold in the pale extensor digitorum longus muscle and 1-2.5-fold in the red soleus, whereas total muscle RNA and total mRNA content fell by 50%. After denervation of the soleus, there was a progressive 2-3-fold increase in polyUb mRNA for 1-3 days, whereas total RNA content fell. On starvation or denervation, Ub concentration in the muscles also rose by 60-90%. During starvation, polyUb mRNA levels also increased in heart, but not in liver, kidney, spleen, fat, brain or testes. Although the polyUb gene is a heat-shock gene that is induced in muscles under certain stressful conditions, the muscles of starving rats or after denervation did not express other heat-shock genes. On starvation or denervation, mRNA for several proteasome subunits (C-1, C-3, C-5, C-8 and C-9) also increased 2-4-fold in the atrophying muscles. When the food-deprived animals were re-fed, levels of Ub and proteasome mRNA in their muscles returned to control values within 1 day. In contrast, no change occurred in the levels of muscle mRNAs encoding cathepsin L, cathepsin D and calpain 1 on denervation or food deprivation. Thus polyUb and proteasome mRNAs increased in atrophying muscles in co-ordination with activation of the ATP-dependent proteolytic process.

Metabolic acidosis often leads to loss of body protein due mainly to accelerated protein breakdown in muscle. To identify which proteolytic pathway is activated, we measured protein degradation in incubated epitrochlearis muscles from acidotic (NNH4CI-treated) and pair-fed rats under conditions that block different proteolytic systems. Inhibiting lysosomal and calcium-activated proteases did not reduce the acidosis-induced increase in muscle proteolysis. However, when ATP production was also blocked, proteolysis fell to the same low level in muscles of acidotic and control rats. Acidosis, therefore, stimulates selectively an ATP-dependent, nonlysosomal, proteolytic process.We also examined whether the activated pathway involves ubiquitin and proteasomes (multicatalytic proteinases). Acidosis was associated with a 2.5-to 4-fold increase in ubiquitin mRNA in muscle. There was no increase in muscle heat shock protein 70 mRNA or in kidney ubiquitin mRNA, suggesting specificity of the response. Ubiquitin mRNA in muscle returned to control levels within 24 h after cessation of acidosis. mRNA for subunits of the proteasome (C2 and C3) in muscle were also increased 4-fold and 2.5-fold, respectively, with acidosis; mRNA for cathepsin B did not change. These results are consistent with, but do not prove that acidosis stimulates muscle proteolysis by activating the ATP-ubiquitin-proteasomedependent, proteolytic pathway. (J. Clin. Invest. 1994. 93: 2127-2133

Expression of HSP-27, HSP-60 and HSP-70 was estimated in the cortex, outer medulla and inner medulla (papilla) of rats undergoing water diuresis or water restriction for two days. The mRNAs for HSP-27 and HSP-60 in renal papilla were two- to threefold greater in rats during water restriction than in those excreting a dilute urine, but levels of mRNA for HSP-70 were not reduced by water diuresis and Western analysis for HSP-70 protein showed no difference between water-loaded and water-restricted animals.

The possible protective effect of heat-shock proteins (HSPs) on ischemic injury to renal cells was assessed in two different experimental models: ischemia-reflow in intact rats and medullary hypoxic injury as seen in the isolated perfused rat kidney. Heat shock was induced by raising the core temperature of rats to 42 degrees C for 15 minutes. Following this, Northern blots showed enhanced gene expression of HSP70, HSP60 and ubiquitin at one hour and reaching a maximum by six hours after heat shock in all regions of the kidney, but most prominently in medulla and papilla. The HSP70 protein in the kidney, estimated by immunohistochemical means, was detectable 24 hours following heat shock and further increased at 48 hours following heat shock. In the first set of experiments, the animals underwent uninephrectomy followed by cross clamping of the remaining renal artery for 40 minutes prior to reflow. Serum creatinine and urea nitrogen rose to 3.15 +/- 0.98 and 126.4 +/- 62.5 mg/dl at 24 hours. No significant differences were observed at 24, 48 and 72 hours after reflow between these values in control rats and rats pretreated with heat shock 48 hours earlier. Severe morphological damage to proximal tubules of the renal cortex was observed to the same extent in both groups. In a second set of experiments, the right kidney was removed either 24 or 48 hours after heat shock and perfused in isolation for 90 minutes. Functional and morphological parameters were compared with those of isolated perfused kidneys obtained from animals that had not been subjected to heat shock.(ABSTRACT TRUNCATED AT 250 WORDS)

Assay m the presence of 10% polyethylene glycol has been systematically used with potentially regulatory liver enzymes as an Indirect way to Induce aggregation of enzymes correspondmg to that which could occur at their phywologcal concentrations Pyruvate kmase L was markedly affected by polyethylene glycol, as was muscle phosphorylase a, while pyruvate kmase M as well as glucokmase, fructose-1,6-blsphosphatase and other liver enzymes examined were not affected Enzyme concentration Polyethylene glycol Liver enzyme Pyruvate kmase Glycogen phosphorylase a

Objetivo: Determinar la prevalencia de vaginitis y vaginosis bacteriana en pacientes con flujo vaginal y su asociación con características clínicas y de laboratorio. Material y métodos: Se llevó a cabo un estudio transversal. Se estudiaron 370 pacientes que acudieron a la consulta ginecológica del Hospital Nacional Arzobispo Loayza de enero a marzo de 1998. A todas las pacientes se les tomó muestras de flujo vaginal para la medición del pH, del test de amina y la identificación microscópica de “células clave”, Trichomonas vaginalis, levaduras e hifas. Resultados: La prevalencia de infección vaginal fue de 42.2%; siendo vaginosis bacteriana la infección más frecuente (23.24%), seguido de candidiasis vaginal (16.2%) y tricomoniasis vaginal (7.8%). Vaginosis bacteriana estuvo asociada a mal olor postcoital, ausencia de signos inflamatorios en vagina, flujo vaginal blanquecino, lechoso, homogéneo y fétido. La candidiasis vaginal estuvo asociada a prurito, ardor vulvovaginal, eritema vulvar y vaginal, flujo vaginal amarillento, grumoso sin olor, test de amina negativo; así como ausencia de relaciones sexuales, ningún compañero sexual en el último año, ninguna gestación, una vida sexual menor de dos años y paridad de ninguno a un hijo. La tricomoniasis vaginal estuvo asociada a eritema vaginal, flujo vaginal amarillo verdoso, espumoso, homogéneo y fétido y test de amina positivo. Conclusión: Un diagnóstico correcto y oportuno de las infecciones vaginales no debe basarse sólo en las características clínicas sino en la confirmación con métodos sencillos de laboratorio.

No abstract

To evaluate the role of atrial natriuretic factor (ANF) in chronic heart failure (HF), the biosynthesis and storage of ANF in cardiac and noncardiac tissues and the level of plasma ANF were measured in rats exhibiting minimal [2-fold rise in left ventricular end diastolic pressure; myocardial infarct (MI) scar length, 25% left ventricle (LV)] and moderate-severe (3-fold rise in left ventricular end diastolic pressure; decreased contractility (dp/dtmax); MI scar length, 47% LV) chronic HF 30 and 60 days after coronary arterial ligation. In rats with moderate-severe HF (30 days post-MI), the cardiac ANF mRNA concentration (determined by dot blot analysis) increased in three heart chambers [LV, 6-fold; left atria (LA), 3-fold; right ventricle (RV), 2-fold], cardiac immunoreactive ANF (IRANF; determined by RIA) concentration increased on the left side (LV, 7-fold; LA, 33%), but was unchanged (RV) or reduced on the right side (right atria, 33%), and plasma IRANF increased 3-fold above sham control values. Excluding the LV (used for MI scar length), the pattern and magnitude of change in ANF mRNA concentration in moderate-severe HF at 60 days were similar to those at 30 days; the cardiac IRANF concentration at this time was the same (LA) or less than (RV, 66%) sham values, and plasma IRANF increased 6-fold above respective sham values. Generally, the changes in the concentrations of cardiac ANF message and peptide and levels of circulating ANF peptide were smaller in rats with minimal HF. The minute quantities of ANF mRNA and IRANF detected in noncardiac tissues (lung, liver, pituitary, aortic arch, brain, kidney, and salivary gland) were unaltered by HF. These findings show that chronic HF, as defined by hemodynamic and histological measurements, specifically and continuously stimulates atrial as well as ventricular ANF biosynthesis; levels of plasma and cardiac ANF are increased early in HF, but with time are subject to modulation. The cardiac ANF system is the prime locus for the effects of HF, as noncardiac ANF biosynthesis and storage are undisturbed by chronic HF.