Experimental mitochondrial myopathy induced by chronic intoxication by Senna occidentalis seeds.

Histochemical and electron microscopic studies of biceps femoris, pectoralis major and rectus femoris of chronically treated birds with seeds of the poisonous plant Senna occidentalis (0.2% external/internal tegment), were performed. The muscles had similar features of human mitochondrial myopathy as ragged-red fibers, cytochrome-oxidase negative fibers, and weak activity of the oxidative enzymes. Fibers with lipid storage were also present. Acid phosphatase activity in rare muscle fibers was also detected, and represents probably a secondary degenerative process. By electron microscopy, enlarged mitochondria with disrupted or excessively branched cristae were seen. The present study presents a new experimental model of mitochondrial myopathy that may be useful for the best knowledge of this group of diseases and for experimental trials of drugs that could reverse the mitochondrial impairment in the mitochondrial myopathies.

Morphometric evaluation of nucleolar organizer regions in cervical intraepithelial neoplasia.

The objective of this study was to determine whether some morphometric parameters and two different methods of counting AgNOR dots were correlated with the grade of cervical intraepithelial neoplasia. Thirty uterine cervix biopsies (8 cases of cervicitis, 9 CIN I, CIN II and 6 CIN III) were studied. Two methods were used to count AgNOR dots. The first one consisted of counting the number of epithelial cells with 1, 2, 3, 4, or more dots. The second method, based on a computer analysis system, consisted of counting the total number of dots in 100 cells, without considering the number of dots per cell. Using the same computer analysis system, the following parameters were measured: area, diameter, perimeter, roundness and length of each dot. The following parameters were found to be correlated with the grade of intraepithelial neoplasia: 1) number of cells with 1 dot, which decreased with increasing grade of cervical intraepithelial neoplasia; 2) number of cells with 4 dots or more, which increased with increasing grade of cervical intraepithelial neoplasia; 3) total number of dots per 100 cells, which progressively increased with increasing grade of intraepithelial neoplasia. We conclude that counting cells with 4 or more dots is the more trustworthy parameter for distinguishing the grade of cervical intraepithelial neoplasia.

Cerebral amebiasis in the acquired immunodeficiency syndrome.

Rare cases of cerebral amebiasis have been described in AIDS patients. We report the case of a 46 year-old homosexual man with AIDS who developed an intermittent amnesia and a right palpebral ptosis. The cerebrospinal fluid contained 169 cells (75% lymphocytes). The patient died five days after hospitalization. Necropsy revealed thrombosis of small vessels of the periventricular regions as well as necrosis and hemorrhage of the periventricular tissue, cerebellum and brainstem. The inflammatory process was scarce and composed mainly of CD-68 positive macrophages. In these regions as well as in meninges there were many trophozoites of ameba of the Acanthamoeba group. Although cerebral amebiasis is rare even in AIDS, the clinician should be attentive to this diagnosis in patients with an insidious encephalitis and cerebral cognitive abnormalities, with or without focal motor signs.

Squamous intraepithelial lesions in cervical smears of human immunodeficiency virus-seropositive adolescents.

Various authors have reported a high rate of human papillomavirus (HPV) infection and HPV-related neoplasias in human immunodeficiency virus (HIV)-seropositive women. On the other hand, young women are most susceptible to cervical infection because of immaturity of the cervix, as it appears that HPV has more access to the basal cells of the differentiating epithelium. The purpose of the present work was to study cervical smears of 82 adolescent HIV-seropositive women (13-21 years of age) to search for cytological evidence of cervical intraepithelial neoplasias. Twenty-one cases showed characteristic features of HPV infection and squamous intraepithelial lesions (SIL; 25.6%). Sixteen cases aged from 17 to 21 years (mean age 19.5 years) had low-grade SIL (LSIL; 19.5%) and five cases aged from 18 to 21 years (mean age 20.2 years) had high-grade SIL (HSIL; 6.1%). There was no significant difference between the mean age of patients with LSIL and HSIL. Two cases had atypical squamous cells of undetermined significance (ASCUS). In the present work it was found that HIV-seropositive adolescents have a high risk for preneoplastic lesions of the cervix (25.6%) as well as a high incidence of more aggressive lesions (6.1% of HSIL) when compared to the general population of adolescents. As it can be assumed that, if the age of acquisition of the infection in both groups (in the general population and HIV-seropositive women) is the same, it is probable that HIV infection in adolescents not only increases the frequency of HPV infections but also facilitates the evolution to more aggressive preneoplastic lesions of the cervix due to HPV.

Progression of cervical lesions in HIV-seropositive women: a cytological study.

It has been reported that cervical intraepithelial lesions have a more aggressive course in HIV-seropositive than in HIV-seronegative women. In the present investigation, the progression of these cervical lesions was studied in a group of HIV-seropositive women. Of 1,587 patients, 200 (12.6%) had a cytological diagnosis of squamous intraepithelial lesion (SIL) or invasive carcinoma. In 409 patients, more than one cytological smear was collected in 3(1/2) years. Progression occurred in 39 cases. In 24 (61.5%), the first diagnosis was benign cellular changes (BCC) and the second was low-grade SIL (LSIL) (1-yr interval in 21 cases); in 11 (28.2%), the first was BCC, and the second, high-grade SIL (HSIL) (1-yr interval in 9 cases); in 2 (5.0%), the first diagnosis was LSIL. and the second, HSIL (1-yr interval); in 2 (5.0%), the first was HSIL, and the second, invasive carcinoma (2-yr interval). These results point to the importance of cervical cytologic surveillance in HIV-seropositive patients.

Silver staining method for nucleolar organizer regions in cervical smears.

The distribution of nucleolar organizer regions (NORs) was studied in Papanicolaou preparations of cervical smears in order to distinguish benign from preneoplastic lesions. Destained smears (six defined as normal, six as inflammatory with squamous metaplasia, six as CIN I, six as CIN II, and five as CIN III) were submitted to the Ag-NOR method after staining with Orange G and EA36. Ag-NOR count was performed in previously outlined fields on the smears. Statistically significant differences (P < .05) were found between the normal smears, inflammatory smears with squamous metaplasia, and each grade of CIN. We conclude that the Ag-NOR technique could be useful to evaluate cervical smears of doubtful interpretation, using previous demarcation of the abnormal fields/cells.

Chlamydia trachomatis in cervical uterine irradiated cancer patients.

Twenty-two women with uterine carcinoma with morphological alterations postradiation treatment suggestive of Chlamydia trachomatis infection were analyzed by direct immunofluorescence. The diagnosis was confirmed in 12 (54.4%) cases. Discussions of the presence of Chlamydia in irradiated cells were not found in the literature. This finding may signify a posttherapeutic morbidity or a preradiation infection.

Image analysis of nuclear/cytoplasmic ratio in cervical smears to discriminate three grades of cervical intraepithelial neoplasia.

OBJECTIVE: To suggest which system of cytologic classification (Bethesda System or World Health Organization [WHO]) of cervical lesions is more adequate for the Public Health Service of São Paulo State, Brazil, based on morphometric study with the Jandel Videoanalysis (JAVA) System. STUDY DESIGN: The study comprised groups of typical smears: cervical intraepithelial neoplasia (CIN 1) separated into two subgroups: cytopathic effects of human papillomavirus associated or not with dyskaryosis, CIN 2 and CIN 3. The JAVA system of image analysis was used to determine the nuclear/cytoplasmic (N/C) ratio in abnormal cells from each group. RESULTS: Significant differences were detected between the three grades of CIN and between the two subgroups of CIN 1. CONCLUSION: Although image analysis is not applicable to large-scale population screening of cervical smears, its use in the present study favored the three-grade cytologic classification (WHO). In addition, the three-grade classification offers the clinician more options for treatment. Considering the clinical-laboratory characteristics of our public health service, the three-grade classification is more adequate.

Sarcoplasmic lipase and non-specific esterase inhibition in myofibers of rats intoxicated with the organophosphate isofenphos.

The expression of sarcoplasmic esterases, lipases as well as the lipid content in the myofibers of the diaphragm of rats intoxicated with the organophosphate isofenphos was studied. Lipid accumulation was documented at light, electron microsopic and by morphometric studies. The distribution of these lipid droplets was irregular and abundant in myofibers with numerous mitochondria (predominantly oxidative fibers). Histochemical inhibition of sarcoplasmic esterases and lipases was observed in the intoxicated animals. This sarcoplasmic inhibition of esterases occurs roughly in parallel to the inhibition of plasma cholinesterase activity. The inhibition of sarcoplasmic lipases may explain, at least partially, the accumulation of lipids. This inhibition probably makes difficult the use of lipids as fuel, especially in the oxidative fibers. In contrast to the small amount of muscle necrosis, (1.30+/-0.745), metabolic muscle impairment was intense and extensive, i.e., decreased activities of esterases and lipases in the sarcoplasm, that should contribute to muscle weakness. Therefore, because segmental necrosis was most prominent in oxidative fibers (and these fibers use lipids as the principal fuel and contain the greater amount of lipases in the sarcoplasm), it is possible that inhibition of activity of lipases is responsible for the segmental necrosis. Although the exact role of these metabolic changes is not known, it is possible that they contribute not only to the induction and evolution of muscle cell necrosis but also to the muscle weakness and clinical impairment of animals and humans in the acute intoxication by these compounds.

Mitochondrial metabolism impairment in muscle fibres of rats chronically intoxicated with Senna occidentalis seeds.

The chronic administration of S. occidentalis seeds was found to induce a mitochondrial myopathy in hens. This study was undertaken to determine if the chronic treatment with S. occidentalis seeds of rats (as a mammalian model) would induce a mitochondrial myopathy similar to those described in humans and to determine if the histological changes could be correlated with the amount of ingested seeds. Twenty-one days old rats were fed S. occidentalis seeds at different diet concentrations (1, 2, 3%). Rats fed 1% S. occidentalis seeds had only a few COX-negative muscle fibers in the pectoralis major muscle. Rats fed 3% Senna occidentalis seeds had a greater number of COX-negative fibers. Rats fed 2% had an intermediate number of COX-negative fibers. Activity of SDH and NADH-tr were decreased in rats of groups 2% and 3%. Our data indicate that a progressive mitochondrial metabolism impairment can be produced in rats fed S. occidentalis seeds and that this impairment can be correlated with the amount of ingested seeds.

Salpingitis due to Entamoeba histolytica.

We describe the pathology of a unique case of Fallopian tube amebiasis, associated with hydrosalpinx, in a 21-year-old woman. She complained of lower abdominal pain, had a foul-smelling green vaginal discharge and fever during one week. There was a discrete increase in body temperature and a painful abdominal palpation at the lower right side, with signs of local peritoneal irritation. Pathological examination showed a marked dilatation of the fallopian tube and hydrosalpinx. Microscopic examination showed a poorly formed granuloma composed of large macrophages with many Entamoeba histolytica trophozoites inside the fallopian tube. Even though it is a rare disease the correct diagnosis of female genital tract amebiasis is of great importance for the indication of proper therapy.

Changes in myelinated nerve fibers and skeletal muscle of rats exposed to high doses of permethrin.

Neurological signs and segmental demyelination in a cervical nerve were observed in rats treated orally with permethrin (300 mg/kg/day) for 5 days. Inflammatory and degenerative signals were recorded in the diaphragm muscle. These effects were more intense with the trade grade than with the technical grade product. The possible influence of the percentage of cis:trans isomers on the intensity of the observed effects is discussed.

The lysosomal enzymes acid phosphatase and cathepsin D in rats intoxicated with Senna occidentalis seeds.

Chronic administration of Senna occidentalis seeds induces an experimental toxic myopathy characterized by skeletal muscle fibers atrophy, decrease in histochemical activity of cytochrome oxidase, and increase of the acid phosphatase activity in muscle fibres at the light microscopic level. The mechanisms that lead to the increase of this lysosomal enzyme activity are not known and could be related to other biochemical disturbs than the mitochondrial function impairment. The main aim of the present study is to localize the acid phosphatase activity using a cytochemical method at transmission electron microscopy level and to quantify cathepsin D in muscle of rats chronically intoxicated with Senna occidentalis seeds by immunoblotting. Acid phosphatase was observed in lysosomes and over profiles of some organelles apparently not involved by lysosomal membrane. In addition immunoblotting demonstrated a decrease in the content of the precursor and of the mature form of cathepsin D in samples of muscles and liver of intoxicated animals. We concluded that there is a selective increase in acid phosphatase activity in muscle--and maybe in other tissues--of animals intoxicated with Senna occidentalis, that can be related to the skeletal muscle atrophy and the intense decrease in weight gain of these animals. Further studies should be performed to establish the mechanisms of selectivity in increase of lysosomal enzymes in different situations and pathological states.

Study of organizer nucleolar regions by the argyrophil technique in cervical intraepithelial neoplasias.

Thirty three biopsies of the uterine cervix were studied by the AgNOR method, that identifies the nucleolar organizer regions. These comprised 9 cases of cervicitis (with or without squamous metaplasia), 9 cases of cervical intraepithelial neoplasia grade I (CIN I), 8 CIN II and 10 CIN III. A hundred cells were counted and classified according to the number of AgNOR dots. We use a more practical and fast method of AgNOR dots counting in cervical intraepithelial neoplasia, in that we exclude the two basal layers and count only cells with 4 or more dots. Statistically significant differences for AgNOR dots were found between cervicitis or CIN I and CIN II cases (p < 0.02) and between CIN II and CIN III cases (p < 0.001). No statistical difference was found between the cases of cervicitis and CIN I. It was concluded that this method of AgNOR counting can be useful in the identification and classification of individual cases intraepithelial neoplasia and their differentiation from eventual difficult cases of cervicitis.

[Polyneuropathy caused by parathion: clinical, electrophysiologic and histologic studies of a case]. / Polineuropatia por parathion: estudo clínico, eletrofisiológico e histológico de um caso.

A case of 38 year old man who worked with organochlorinated and Parathion during 5 years is reported. His follow-up was up to 2 years. The onset of the disease was characterized by cholinergic signs, headache, loss of weight, trembling, miokimias, fasciculations, ataxia, myotonic phenomena (in hands only) and motor sensitive peripheral polyneuropathy (affecting the lower limbs symmetrically). Low concentrations of blood cholinesterases confirmed the etiology. Myotonic phenomena disappeared spontaneously 6 months after the initial observation. One year later, the concentration of erythrocyte acetylcholinesterase was found to be low and plasma cholinesterase was normal, suggesting that the patient was carrier of a congenital deficiency of acetylcholinesterase. In literature relationship between myotonia and intoxication due to organophosphorus was not found. The whole clinical picture, cholinergic symptoms, transitory phenomena and spontaneous motor activity could be explained by an excess of acetylcholine. Electromyography (EMG) in the first observation showed neuromuscular transmission blocking characterized by deficiency or absence of voluntary activity, unexcitability of fibular nerves, with fibrillations and positive peaks as described previously with Mipafox (another organophosphorus agent). During 2 years of observation numerous end-plates potentials of muscular fibres persisted in the EMG. A progressive increase in voluntary activity showed by unit motor potential of almost normal amplitude and very increased duration was observed. No potentials of reinnervation were noted. The results of EMG were explained as disturbances of neuromuscular transmission associated with moderate signs of denervation. The Eaton-Lambert's test and the stimulation of a single unit motor potential confirmed disorder of neuromuscular synapses. The histochemistry of brachial biceps showed scattered atrophic and angulated type I and II fibres. Teased-fibres preparations showed nerve fibres with B, C, and G alterations as defined by Dyck et al. indicating axonal degeneration. These results were according to velocity of sensitive conduction. The conduction velocity of fibular nerves was strongly delayed during all the evolution indicating serious disorders of motor nerves myelin.(ABSTRACT TRUNCATED AT 400 WORDS)

[Muscular biopsy in myasthenia gravis: histochemical and morphometric study of 4 cases]. / Biopsia muscular em miastenia grave: estudo histoquímico e morfométrico de 4 casos.

Myasthenia gravis is a neuromuscular disease characterized by excessive fatigability of muscle function and particularly involves muscles innervated by the cranial nerves. It is believed that the defect is in the neuromuscular junction. The authors studied histochemical and morphometric findings in 4 patients with myasthenia gravis showing that the type II fibres were significantly smaller than the type I fibres.

[Myotubular myopathy: clinical, electrophysiological and histological study of a case]. / Miopatia miotubular: estudo clínico, eletrofisiológico e histológico de um caso.

A cases of myotubular myopathy in a 10 years old girl is reported. Clinically, palpebral ptosis, ocular movements limitation, facial diplegia, positivity of Gower's test, muscular hypotrophy distal, foot drop and deep absent reflexes were found. These signals were described by most of authors, besides symptoms referred, like partial urinary incontinency and frequent vomits. Reflexes H absents and teary in our patient were observed but were not described in the other cases of the literature. Routine laboratory tests were within normal limits; only aldolase was lightly elevated. In all muscles examined it was noted a spontaneous electromyographic activity with +- of 2,64 +/- 1,33 ms, 36,87 +/- 30,87 microV and 88,13 +/- 24,82 /s of frequency without characteristics of desenervation potentials of myoneural plates. The voluntary electromyographic activity was of myopathic pattern. A curve I/D made in the motor point of braquial biceps muscle was normal. The biopsy was made in the motor point of this muscle for histochemical, electron immunofluorescence and vital stain microscopy. The biopsy showed 35% of fibras with central nuclei, predominance and hypotrophy of type I fibres, in some of them there were not myofibrils in the central zone, and poor differentiation between the fibre types in oxidative enzymes reactions. The electron microscopy confirmed the histochemical studies. The direct immunofluorescence was positive in some fibres. The vital stain showed beaded subterminal motor fibres. The clinical, electromyographic and principally histological findings suggest a innervation congenital disorder of muscle fibres.

[Organophosphate-induced myotoxicity]. / Miotoxicidade por organofosforados.

Organophosphates comprise a group of chemical compounds extensively used in farming as insecticides, which cause accidental poisoning in animals and men and are also used in suicide attempts. The toxicity of these compounds is due especially to the cardiac and respiratory impairment in consequence of autonomic nervous system disorders. However, it is known that some of these products induce a myopathy in experimental animals and humans. This myopathy is characterized by muscle cell degeneration, involving above all the respiratory muscles. Based on the fact that this involvement certainly enhances the respiratory impairment, this study offers an experimental method for routine evaluation of organophosphate myotoxicity, using a minimal and sufficient battery of stains and histochemical reactions, for muscle necrosis quantification. For this purpose, albino rats (Wistar) treated with the organophosphate paraoxon, were used both with and without antidotes (atropine or pralidoxime). Muscle fiber necrosis in the diaphragm of the rats treated with paraoxon or paraoxon and atropine, that affected about 15% of the fibers in some areas, was detected. In the group treated with paraoxon and pralidoxime, a minimal necrosis was seen, revealing a protective role of this later antidote during the development of myopathy.

Muscle atrophy induced in broiler chicks by parts of Senna occidentalis seeds.

Senna occidentalis (formerly Cassia occidentalis) is a common contaminant of agricultural commodities. It is toxic to cattle and poultry, reportedly being responsible for skeletal myodegeneration in these animals. All parts of the plant present toxicity, but the seeds are the most toxic. The toxin(s) responsible for the myodegeneration have not been definitively identified, nor is it known which part of the seeds is most toxic. Intoxication by this plant leads to weight loss with considerable economic repercussions. The effects of the whole seed and of parts of S. occidentalis seeds (1% in commercial feed) were compared on the pectoralis major muscle of broiler chicks intoxicated from birth until 22 days of life. There were severe clinical signals and reduced body weight in birds that received the external tegment of the seed, whereas no adverse effects were observed in birds that received the whole seed or other parts of the seed. Histological and morphometric studies showed an intense muscle fibre atrophy (both type 1 and type 2 fibres were affected) in the group that received 1% external tegment. This study may be the first step to identifying the substance(s) involved in this pathological process.

Muscle pathology in juvenile dermatomyositis.

OBJECTIVE: To study muscle biopsies, using histochemistry, on ten children with infantile dermatomyositis. DESIGN: Series of ten patients (of whom eight patients had received treatment and two had not) were submitted to muscle biopsy in order to diagnose possible inflammatory myopathy or to detect recurrences. PLACE OF DEVELOPMENT OF THE STUDY: Public Health Service of São Paulo State. PARTICIPANTS: Children with clinical features of inflammatory myopathy. INTERVENTION: Biopsies were performed on the vastus lateralis using local anesthetic. Histochemistry was performed according to standardized methods. RESULTS: Architectural changes of the muscle fibers, necrosis of variable intensity and accentuated evidence of regeneration were observed in patients who had not received treatment (2 cases) and in one case where muscular weakness persisted in spite of corticosteroid therapy. Necrosis and regeneration were minimal or absent in cases treated for one year or more (4 cases). In 3 cases with clinical and laboratorial recurrences, muscle necrosis and architectural changes were detected. CONCLUSIONS: It was concluded that muscle biopsy could aid in diagnosing infantile dermatomyositis as well as in detecting recurrences even in cases without clinical activity of the disease.