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The objective of the present investigation was to study the protection afforded by a single administration of pralidoxime against the muscle necrosis induced by the organophosphate compound metamidophos at different times after intoxication. The fiber necrosis of the diaphragm muscle was quantified by a morphometric technique, comparing the area fraction occupied by necrotic muscle fibers in animals that received pralidoxime at different times after intoxication, i.e., 0, 1, 3, 6, and 12 h. Pralidoxime administration protected metamidophos-induced muscle necrosis in all groups studied except for the 12-h group. The earlier the administration of pralidoxime the greater the protection against muscle necrosis. This protection was not accompanied by complete reactivation of plasma cholinesterase activity. Results support the current opinion that pralidoxime should be administered as soon as possible after organophosphate intoxication, because in addition to reversing the muscarinic effects, early administration of pralidoxime also prevents muscle necrosis--which could impair muscular function and respiratory condition. The time difference between recovery of plasma cholinesterase activity and muscle necrosis protection indicates that this method is not completely trustworthy for patient follow-up, since some improvement may occur in spite of the low plasma cholinesterase activity.

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.

Purpose: The incidence of anogenital squamous-cell carcinoma was observed to have increased since the beginning of the human immunodeficiency virus infection epidemic among male homosexuals, both with acquired immunodeficiency syndrome and without acquired immunodeficiency syndrome. It seems that immunosuppression is the most important risk factor for the progression of anogenital lesions, recurrences of anal condyloma, and development of anal carcinoma, in particular in acquired immunodeficiency syndrome. High-grade anal intraepithelial neoplasia was predominantly observed in the human immunodeficiency virus-positive men. We have also observed a high rate of recurrences of anal lesions in cases of high-grade anal intraepithelial neoplasia. However, there are many cases of recurrences of low-grade anal intraepithelial neoplasia that cannot be predicted by routine histologic studies. By using immunohistochemical methods, we studied the expression of Ki-67 in epithelial cells of low-grade anal intraepithelial neoplasia of patients with acquired immunodeficiency syndrome to try to predict recurrence of these lesions., Methods: Anal biopsies of 38 patients were studied retrospectively. Of these patients, 14 had no recurrences (Group 1), and 24 patients had recurrence of the anal lesions before one year of follow-up (Group 2)., Results: The median percentage of Ki-67-positive cells in Group 1 was 6.3 +/- 7.03 and in Group 2 was 24.1 +/- 16.72. The difference between Groups 1 and 2 was statistically significant (P < 0.001)., Conclusions: Our results showed a high correlation between the percentage of Ki-67-positive cells and recurrences. We concluded that Ki-67 counting in low-grade anal intraepithelial neoplasia can aid in predicting recurrences and therefore aid in the follow-up of these patients.

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., (Copyright 2001 Wiley-Liss, Inc.)

The objective of the present investigation was to study the expression of the proliferation marker Ki-67 in cervical intraepithelial lesions in women with AIDS. We studied 18 low-grade cervical intraepithelial lesions (Lo-CIN) and 8 high-grade cervical intraepithelial lesions (Hi-CIN ) in AIDS patients and 18 Lo-CIN and 14 Hi-CIN in patients from the general population. Positive Ki-67 nuclei were counted. A significantly higher number of Ki-67-positive cells (p < 0.001) was found in Lo-CIN of AIDS patients (mean 29.18 +/- 10.44) as compared with Lo-CIN of general-population women (mean 17.08 +/- 7.40), whereas no significant difference in positive Ki-67 nuclei was observed in the Hi-CIN of AIDS patients and of patients from the general population (p > 0.05). We conclude that the proliferative potential of cervical cells in Lo-CIN - as studied by the expression of the proliferation marker Ki-67 - is higher in AIDS patients than in the general population., (Copyright 2001 S. Karger AG, Basel.)

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.

Synthetic pyrethroid insecticides have been used in the last two decades largely because of their high activity as an insecticide and low mammalian toxicity. Some studies have demonstrated that these products, especially compounds with an alpha-cyano group, are toxic to the mammalian central nervous system (CNS) in acute intoxications. However, morphological studies are scarce. In the present work the histopathologic changes of the sciatic and tibial nerves of rats submitted to acute intoxication with the cyanopyrethroid deltamethrin were studied. For 3 consecutive days male Wistar rats received by oral gavage deltamethrin at a dose of 45 mg/kg body wt. On the 4th day fragments of sciatic and tibial nerves were studied by transmission electron microscopy (TEM) and teasing of individual nerve fibers. In addition, another group of rats were allowed to recover until the 10th day. Teasing of nerves of animals sacrificed on the 4th day revealed myelin ovoids, which are indicative of axonal damage. TEM demonstrated rare degenerated axons completely filled with organelles, in particular mitochondria, and with electron-dense lamellar bodies that resemble myelin figures. In addition, great cytoplasmic vacuolization caused by proliferation and dilation of the rough and smooth endoplasmic reticulum and Golgi apparatus was observed in some Schwann cells. No lesion was found 7 days after discontinuation of the treatment (group2). Since these histologic changes are transitory and scarce, the question arises: Are they related to the changes in NA(+), K(+)-ATPase activity or Na(+) channels caused by pyrethroid compounds?, (Copyright 2000 Academic Press.)

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.

Skeletal muscle can be the site of inflammatory diseases that lead to muscle weakness, pain, and increased myogenic serum enzymes. Most of these inflammatory myopathies are idiopathic. In some cases inflammatory myopathies are due to infectious agents. We describe the pathological aspects of muscle biopsies of 2 Brazilian siblings who acquired toxoplasmosis at the same time and in similar conditions. One developed a tetraplegia that was confirmed to be due to inflammatory myositis due to toxoplasma. The other developed myocarditis, with heart failure, without skeletal muscle weakness. In both cases many toxoplasma organisms were observed in the muscle biopsies, but in case 1 only was there an inflammatory myopathy with myofiber necrosis; the inflammatory cells were predominantly macrophages with some CD4+ cells and rare CD20+ cells. In case 1, expression of CD54 was observed in many inflammatory cells as well in endothelial cells, but only in endothelial cells in case 2. After treatment with clindamycin and corticosteroids both cases had only partial improvement, case 1 with a residual muscle weakness and case 2 with residual cardiac insufficiency (requiring digoxin). These cases show that the presence of the parasite in myofibers is not enough to induce an inflammatory myositis with muscle cell necrosis. This suggests that immunological disturbances may contribute to the development of inflammatory myositis due to toxoplasma.

The degenerative process of the myofibers of the diaphragm of rats intoxicated with the organophosphate isofenphos, a compound that inhibits esterases, was studied at different intervals of intoxication. Early disorganization of the intermyofibrillar network and of the myofilaments, as well as dilatation of organelles, were observed by use of transmission electron microscopy. These changes precede macrophage invasion of the muscle fibers. Early expression of ubiquitin was observed in segments of muscle fibers by immunohistochemistry. Bands of polyubiquitin complexes in muscle homogenates were observed by immunoblotting. These bands disappeared in later stages of intoxication. A 42.5-kDa band corresponds to actin, as observed by immunoblotting using antisarcometric actin. This indicates relatively large amounts of polyubiquitin complex associated with sarcomeric actin in muscle fibers in early stages of intoxication. Based on these results it seems that actin is an important target in organophosphate-induced myofiber degradation and that the degradation of this protein-by the polyubiquitin pathway-may play an important role in the early disorganization of the sarcomere, as observed by electron microscopy. A possible role of the ubiquitin proteolytic pathway is that of trying to eliminate proteins modified in the early phases of muscle fiber degeneration, which is a necessary step for regeneration of the posterior segmental muscle., (Copyright 1999 Academic Press.)

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.

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.

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.

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.

Patients with AIDS can present clinical involvement of the peripheral nervous system due to different causes. In the present work, it was studied the histopathological changes in sural nerve biopsy of fifteen patients with AIDS with this clinical involvement. It was observed the presence of a polyarteritis nodosa-like vasculitis of small arteries with fibrinoid necrosis in the sural nerve of 3 patients, one of them associated to polyradiculitis due to cytomegalovirus infection (CMV). Six patients presented mild axonal loss by light microscopy. Three other patients had a more important axonal neuropathy with myelin ovoids by teasing. By the electron microscopy in these patients were observed some fibers with axonal damage. The other three patients had normal sural nerves. We concluded that sural nerve biopsy may be important in peripheral neuropathies or myelo-radiculo-polyneuropathies in AIDS especially to search for nerve vasculitis, because it can change the therapeutic approach.

Human papilloma viruses (HPVs), particularly types 16 and 18 have a key role in the development of preneoplastic and neoplastic lesions of the uterine cervix. We studied, by non isotopic in situ hybridization using probes to HPV 6, 11, 16 and 18, cervical biopsies from AIDS patients with condilomata or cervical intraepithelial neoplasia. There were 32 biopsies which showed low-grade cervical intraepithelial neoplasia (Lo-CIN); 5 biopsies showed high-grade cervical intraepithelial neoplasia (Hi-CIN). Of 32 Lo-CIN biopsies, 18 (56.3%) were positive for HPV; 7 for HPV 6 and/or 11 (21.9%), 11 for HPV 16 and/or 18 (34.4%) and one for HPV 6 and 18. Of 5 Hi-CIN biopsies 3 were positive for HPV: one for HPV 6 and 2 for HPV 16 or 18. The total positivity was 56.8% (21/37). This result was similar to those obtained by various other authors studying the general population.

The purpose of the present study was to ascertain the frequency of Pap smears abnormalities (SIL/carcinoma) in two populations in the State of S. Paulo (Brazil) who were screened for cervical cancer during the last nine years: adolescents (up to the age of 21) and adults (over 21). Statistical analysis showed a tendency to linear increase in frequency of abnormal Pap smears in both groups (adolescent: slope = 0.118 +/- 0.56; adult: slope = 0.079 +/- 0.021), being greater in the adolescent group (slope 0.039 +/- 0.038) than in the adult group. Between 1987 and 1995 the prevalence of abnormal smears tripled in the adult group (from 0.37 to 1.18) and almost quadruplicated in the adolescent group (from 0.64 to 2.10). Low grade squamous intraepithelial lesions (SIL) were the most frequent findings in cervical cytology in both groups: 92.3% in the adolescent group and 69.8% in the adult group. Few cases of high grade squamous intrapeithelial lesions (HSIL) (7.7%) and no cases of carcinoma were found in adolescent population. We suggested that sexually active adolescents should be included in cervical screening programs.

Objective: To determine the protective effect of pralidoxime on muscle fiber necrosis induced by organophosphate acute intoxication in rats., Design: Adult male Wistar rats were given oral organophosphate compounds dissolved in glycerol formal: dichlorvos, isofenphos, metamidophos, and diazinon. Half of the animals also received pralidoxime mesylate (20 mg/kg, intraperitoneal). Control animals received only the solvent. Twenty-four hours after treatment, the diaphragm muscle was collected for histological counts of necrotic muscle fibers in transverse sections., Results: Metamidophos- and isofenphos-treated animals showed the highest percentage of necrotic muscle fibers: 1.66 +/- 1.112 and 1.34 +/- 0.320, respectively. Diazinon-treated animals had a lower percentage of necrotic fibers: 0.40 +/- 0.032 (p < 0.05) compared to the first 2 products, and dichlorvos-treated animals showed the smallest: 0.05 +/- 0.021 (p < 0.05) when compared to the other 3 products. Pralidoxime reduced necrotic fibers about 20 times in metamidophos-treated animals, 10 times in isofenphos-treated animals and 6 times in diazinon-treated animals. Pralidoxime administration did not increase plasma cholinesterase activity in any group, although symptoms were reduced., Conclusions: Oxime reduced diaphragmatic muscle necrosis in experimental organophosphate intoxication, despite little effect on plasma cholinesterase. Since respiratory insufficiency is an important cause of mortality and morbidity in organophosphate intoxications, early oxime administration may be particularly beneficial.

Acute intoxication with Senna occidentalis seeds was studied in chicks. Seven-day-old chicks were fed ground dried seeds of this plant mixed with regular chicken ration at a concentration of 4% by weight for 15 days. Feed intake and body weight were markedly affected and a high level of lethality was observed. Necropsy examination of chicks from the experimental group revealed paleness and atrophy of thoracic muscles. Degenerative and necrotic fibres were observed in skeletal muscle by histological examination. Muscle histochemistry showed accumulation of lipids and numerous acid phosphatase-positive muscle fibres. Electron microscopy revealed atrophic muscle fibres, lipid storage, dilatation of the sarcoplasmic reticulum and abnormal mitochondria.

Plants of the genus Senna (formerly Cassia) are poisonous to livestock and other laboratory animals, leading to a syndrome of a widespread muscle degeneration, incoordination, recumbence, and death. The main histologic lesion is necrosis of skeletal muscle fibers. Recently, a mitochondrial myopathy with ragged-red and cytochrome oxidase (COX)-negative muscle fibers was recognized in hens chronically intoxicated with parts of seeds of S. occidentalis. The purpose of the present work was to investigate if there was peripheral nerve involvement in the acute intoxication of chicks with S. occidentalis seeds. Teasing of individual fibers revealed signs of extensive axonal damage with myelin ovoids. Ultrathin sections confirmed the axonal damage. Axons were filled with membranes, some residual disorganized filaments, and enlarged mitochondria. In some instances the axon disappeared and there was secondary degeneration of the myelin sheath. The present work is the first description of the neurotoxic effect of S. occidentalis intoxication. Future work should attempt to determine the mechanisms involved in this neuropathy.

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.

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.

Plants of the genus Senna (formerly Cassia) have been recognized as the cause of a natural and experimental syndrome of muscle degeneration frequently leading to death in animals. Histologically, it demonstrated skeletal and cardiac muscle necrosis, with floccular degeneration and proliferation of sarcolemmal nuclei. Recently, it was described as an experimental model of mitochondrial myopathy in hens chronically treated with Senna occidentalis. Currently, skeletal muscles of chicks intoxicated with seeds of the poisonous plant S. occidentalis were studied by histochemistry and electron microscopy. Since birth, the birds were fed ground dried seeds of this plant with a regular chicken ration at a dose of 4% for 11 days. Microscopic examination revealed, besides muscle-fiber atrophy, lipid storage in most fibers and a moderate amount of cytochrome oxidase-negative fibers. By electron microscopy, enlarged mitochondria with disrupted or excessively branched cristae were seen. This picture was characteristic of mitochondrial myopathy. These findings have hitherto remained unnoticed in skeletal muscle of young birds treated with S. occidentalis.

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.

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.

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.

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.

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.

Objective: Systemic sclerosis (SSc) is characterized by abnormal deposition of collagen in the skin and by visceral involvement. Muscle weakness is a relatively frequent complication of SSc, although severity varies. We studied muscle pathology in patients with SSc with progressive muscle involvement., Methods: We performed histochemical and immunohistochemical investigations to detect neural cell adhesion molecule (NCAM)., Results: Five of the 6 cases of SSc expressed NCAM in atrophic angulated fibers (some fibers stained heavily with oxidative enzymes)., Conclusion: Neurogenic involvement in SSc is more frequent than reported.

Rhodococcus equi (Corynebacterium equi) is an aerobic actinomycetes, well described as a cause of pulmonary infection in different animals as horses, pigs and cows. This pathogen has a coccobacillar aspect and a variable acid-fast stain in tissues. Rare cases of human infection by Rhodococcus species were described, the majority by Rhodococcus equi, especially in patients with immunodeficiency syndrome (AIDS) in advanced stages of the disease. Usually the diagnosis of infections by Rhodococcus species is performed by positive blood or bronchoalveolar lavage cultures. Here we described a case of a pleuro-pulmonary infection by Rhodococcus equi, with malakoplakic-like lesions, that was the first manifestation of AIDS, whose diagnosis was performed by pleural biopsy (acid-fast bacteria with a variable coccobacillar aspect inside macrophages) and pleural fluid culture.

Limb-girdle muscular weakness and wasting could be caused by different diseases (inflammatory and hereditary myopathies, muscular dystrophies and neurogenic atrophies). Among these, Limb-Girdle Muscular Dystrophy (LGMD) is an heterogeneous group of pathologies that have progressive proximal limbs and girdle weakness, with some dystrophic features by the muscle biopsy. We studied a case of LGMD in an adult man with a typical histological and histochemical profile, associated to a mitochondrial deficit characterized by presence of ragged-red fibers, a histochemical Cytochrome Oxidase deficiency and abnormal mitochondria by ultrastructure.

A 21-year-old female patient with psychotic symptoms developed hyperthermia, muscular rigidity and hypertension after administration of haloperidol. A muscle biopsy showed some atrophic and necrotic fibers, and a great number of fibers with central cores in the oxidative enzyme preparations. A related syndrome, Malignant Hyperthermia (MH), is sometimes associated with central core disease. The present case shows an association of a hyperthermic syndrome related to haloperidol with central core disease.

Sputum samples from AIDS patients with and without pulmonary tuberculosis were analyzed morphologically and immunocytochemically to determine the frequency of occurrence of Candida sp. Mycobacterial infection was detected by bacterioscopy and/or culture and cytological evaluation was performed using Papanicolaou and Toluidine Blue staining. Immunoreaction for Candida sp was performed using polyclonal antibody in selected cases with fungal structures in smears stained by the Papanicolaou or Toluidine Blue method. An increased frequency of Candida sp (2.5 times) was observed in the tuberculous group compared to the group of AIDS patients without tuberculosis. The Toluidine Blue stain showed good results for the detection of Candida sp in sputum. Due to the increased risk of this opportunistic infection among more severely immunocompromised patients. Toluidine Blue staining of sputum samples submitted to analysis seems to be a reliable screening method.

Three members of a family with a hereditary neuropathy were studied. Light, electron microscopy and teasing of isolated fibres were performed. The findings confirmed the clinical and electrophysiological hypothesis of hypertrophic form of Charcot-Marie-Tooth disease. Hypertrophy of Schwann cells with the formation of onion bulb figures as the most evident ultrastructural feature, besides demyelination, remyelination and mild axonal degeneration. Recent data about the genetic transmission and pathogenesis of the hereditary motor and sensory neuropathies (HMSN) are discussed.

We studied the incidence of cervical Chlamydia trachomatis (CT) in 65 adolescents aged between 14 and 19 years and 65 adults aged 20 years or more. All subject were more than three months advanced in a normal pregnancy. Two samples for cytology were obtained from each patient, one ecto- and endocervical, with Ayre's spatula and cytobrush, and one only endocervical with cytobrush. The first one was examined after Papanicolaou staining and the second one was examined with the easily performed immunofluorescence reaction to CT (IF)--Microtrak, SYVA. Twenty seven adolescent patients (41.5%) and fourteen adults (21.5%) had a positive IF test for CT; the rate in adolescents was significantly higher (P < 0.01) than in adults. The Papanicolaou (Pap) stained slides, examined blind for evidence of CT infection showed a sensitivity of 70.7%, a specificity of 95.5% and positive and negative predictive values of 87.8% and 87.6% respectively. Seven adolescents showed cytological signs of Papillomavirus (HPV) infection, and six of them were also CT positive; five had signs of HPV infection and 4 of them were CT positive.

A comparison between two sampling methods of cervicovaginal material--cytobrush and Ayre's spatula--was performed to verify the endocervical cell concentration in a group of 163 asymptomatic women. Age ranged from 18 to 57 years, with an average of 30. Smears of both methods showed satisfactory cellularity for study. Cytobrush showed endocervical cells (columnar and/or metaplastic) in 100 cases (61.3%) and Ayre's spatula in 39 (20.2%). Cytobrush showed more efficient results in terms of endocervical cell concentration; according to literature, this can be an important factor in the sensitivity of the cytologic method to detect intraepithelial neoplasia.

Two methods of staining (Papanicolaou versus direct immunofluorescence) and two methods of collection of the samples (Ayre's wooden spatula versus cervical Cytobrush) were compared in order to verify the efficiency in detecting Chlamydia trachomatis (CT) infections in the female genital tract. Out of 166 asymptomatic patients, 59 were positive for CT by means of direct immunofluorescence: 36 were detected in Cytobrush samples, 16 in Ayre's spatula samples, and 7 in the samples collected by both methods. Papanicolaou smears showed "moth-eaten" features suggestive of CT infection in a great number of metaplastic cells present in 35 cases: 24 collected by Cytobrush, 4 by Ayre's spatula and 7 by both methods. Our data show that Cytobrush is more efficient that Ayre's spatula in concentrating cellular material. It is thus possible to detect CT infection with more accuracy by means of direct immunofluorescence, and to suspect CT infection in smears collected by means of Cytobrush and stained by Papanicolaou's method.

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.

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.

One hundred and sixty-one cervicovaginal smears of women seen at the Instituto Adolfo Lutz, Laboratory I, in Taubaté (SP) were examined cytologically by the Papanicolaou method and by direct immunofluorescence (IF) for Chlamydia trachomatis (Ct). Thirty-three cases (20.5%) had positive results for Ct in the IF. With the Papanicolaou method, those cases presenting fine vacuolation in endocervical metaplastic cells according to recent criteria were suggestive of Ct infection. Compared to IF, the Papanicolaou method showed high sensitivity (91.6%) in the group of smears containing endocervical cells. The IF is a high sensitivity and specificity method, but its cost cannot be afforded by a great portion of our population. In view of the high incidence of Ct infection in the female genital tract, the authors concluded that the Papanicolaou method in smears containing endocervical cells may be considered the first option for the detection of this agent.

Some authors recently suggested the role of Chlamydia trachomatis as a potentiating agent of female genital tract neoplasias. Cervicovaginal smears of women with uterine carcinoma treated with radiotherapy are analyzed in the present work. A series of such smears showed morphological changes suggestive of Chlamydia infection with the Papanicolaou stain. Confirmation of the presence of such agent was done using an avidin-biotin-peroxidase technique. A higher prevalence of Chlamydia in the group of patients with carcinoma of the uterus treated with irradiation than in the population without ionizing treatment was verified. The presence of cells with radiotherapic changes infected by Chlamydia is a new finding in cytology.

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)

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.

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.