Osteoarticular tuberculosis in an HIV-positive patient: a case report

The authors report a case of a 38-year-old HIV-positive woman, with subcutaneous nodules on the thoracic region with 3 months of evolution. Clinical, laboratory, and epidemiological features were evaluated and associated with apparent damage to the T11-T12 vertebrae, identification by imaging tests, positivity in a polymerase chain reaction-based test, and reactivity to the Mantoux tuberculin skin test (PPD-RT 23). The patient was diagnosed with osteoarticular tuberculosis and received treatment for a year, and clinical cure was achieved.

Rheumatic fever: a multicenter study in the State of São Paulo

Rheumatic fever is still the most commonly seen rheumatic disease in Brazilian pediatric rheumatology clinics. It remains a significant health problem since subsequent cardiac sequelae represent one of the most important causes of chronic heart disease in children. We reviewed the clinical manifestations of rheumatic fever in 786 patients, followed at seven pediatric rheumatology clinics in the state of São Paulo, Brazil. All patients were diagnosed according to revised Jones' criteria. Regarding major criteria, 396 (50.4%) children exhibited carditis, 453 (57.6%) polyarthritis, 274 (34.8%) chorea, 13 (1.6%) erythema marginatum, and 12 (1.5%) subcutaneous nodules. Valvular lesions documented by echocardiography in the absence of accompanying auscultatory findings were found in 144 (18.3%) patients. Migratory polyarthritis was observed in 290 (64.0%) patients with articular involvement. Documented previous streptococcal infection assessed by serum antistreptolysin (ASO) titers occurred in 531 (67.5%) patients. Even though prophylaxis with benzathine penicillin was recommended to all patients, recurrent attacks were observed in 147 (18.7%). We emphasize the high frequency of chorea, silent carditis and recurrences in our series as well as the variable clinical presentation of arthritis in rheumatic fever. Multicenter studies should be encouraged to improve our understanding of the clinical features of rheumatic diseases in children and adolescents.

Life cycle of Trypanosoma cruzi (y strain) in mice

Since 1958, we have studied experimental Chagas' disease (CD) by subcutaneous inoculation of 1,000 blood forms of Trypanosoma cruzi (Y strain) in Balb/C. mice. Evolution of parasitemia remained constant, beginning on the 5th and 6th day of the disease, increasing progressively, achieving a maximum on about the 30th day. After another month, only a few forms were present, and they disappeared from the circulation after the third month, as determined from direct examination of slides and the use of a Neubauer Counting Chamber. These events coincided with the appearance of amastigote nests in the tissues (especially the cardiac ones), starting the first week, and following the Gauss parasitemia curve, but they were not in parallel until the chronic stage. In 1997, we began to note the following changes: Parasites appeared in the circulation during the first week and disappeared starting on the 7th day, and there was a coincident absence of the amastigote nests in the tissues. A careful study verified that young forms in the evolutionary cycle of T. cruzi (epi + amastigotes) began to appear alongside the trypomastigotes in the circulation on the 5th and 7th post-inoculation day. At the same time, rounded, oval, and spindle shapes were seen circulating through the capillaries and sinusoids of the tissues, principally of the hematopoietic organs. Stasis occurs because the diameter of the circulating parasites is greater than the vessels, and this makes them more visible. Examination of the sternal bone marrow revealed young cells with elongated forms and others truncated in the shape of a "C" occupying the internal surface of the blood cells that had empty central portions (erythrocytes?). We hypothesize that there could be a loss of virulence or mutation of the Y strain of Trypanosoma cruzi.

Pheochromocytoma treated by laparoscopic surgery

OBJECTIVE: To evaluate the results of the laparoscopic technique in the treatment of adrenal pheochromocytoma. METHOD: Ten patients, 7 men and 3 women, between 10 and 67 years of age (mean 48) with pheochromocytoma underwent transperitoneal laparoscopic adrenalectomy and were evaluated retrospectively, based on clinical, laboratory, and pathological diagnosis. In all cases there was a solid unilateral adrenal tumor, 5 on the left side and 5 on the right side, whose greater diameter varied from 7 to 80 mm (mean 32). Nine of the 10 patients were chronically hypertensive or had already had hypertensive crises. One patient was normotensive, but presented metabolic alterations suggestive of adrenergic hyperfunction. RESULTS: No deaths occurred in this series. There were two (20%) conversions to open surgery, one due to venous bleeding and one due to the difficulty of dissection behind the vena cava in a patient presenting a partially retro-caval tumor. Surgical time in the 8 non-converted cases ranged from 70 to 215 minutes (mean 136). One patient (10%) received blood transfusion, and another (10%) presented two complications - acute renal failure and a subcutaneous infection. Both had been converted to open surgery. None of the non-converted cases was transfused or presented complications. Hospital discharge occurred between the 2nd and 11th post-operative day (mean 3). The pathological exam of the surgical specimens confirmed the diagnoses of pheochromocytoma in all 10 cases, one of them associated with an aldosterone-producing cortical tumor. CONCLUSIONS: Laparoscopic adrenalectomy for selected patients presenting pheochromocytoma is feasible and provides good results.

Oral manifestations of Albright hereditary Osteodystrophy: a case report

Albright hereditary osteodystrophy is a hereditary metabolic disorder of dominant autosomal etiology that is commonly characterized by short stature, round face, small metacarpus and metatarsus, mental retardation, osteoporosis, subcutaneous calcification, variable hypocalcemia, and hyperphosphatemia. In this study, we report a clinical case of a 17-year-old woman with Albright hereditary osteodystrophy, and we discuss her clinical, radiographic, and laboratory test characteristics together with the oral manifestations, and we correlate them with the characteristics found in the literature. We also discuss the odontological management of treatment of related periodontal disease and planning for corrections of related malocclusions.

Acellular Gellan-gum based bilayered structures for the regeneration of osteochondral defects: a preclinical study

 In orthopaedics, the management and treatment of osteochondral (OC) defects remains an ongoing clinical challenge. Autologous osteochondral mosaicplasty has been used as a valid option for OC treatments although donor site morbidity remains a source of concern [1]. Engineering a whole structure capable of mimicking different tissues (cartilage and subchondral bone) in an integrated manner could be a possible approach to regenerate OC defects. In our group we have been proposing the use of bilayered structures to regenerate osteochondral defects [2,3]. The present study aims to investigate the pre-clinical performance of bilayered hydrogels and spongy-like hydrogels in in vivo  models (mice and rabbit, respectively), in both subcutaneous and orthotopic models. The bilayered structures were produced from Low Acyl Gellan Gum (LAGG) from Sigma-Aldrich, USA. Cartilage-like layers were obtained from a 2wt% LAGG solution. The bone-like layers were made of 2wt% LAGG with incorporation of hydroxyapatite at 20% and 30% (w/v). Hydrogels and spongy-like were subcutaneouly implanted in mice to evaluate the inflammatory response. Then, OC defects were induced in rabbit knee to create a critical size defect (4 mm diameter and 5 mm depth), and then hydrogels and sponges implanted. Both structures followed different processing methods. The hydrogels were injected allowing in situ  crosslinking. Unlike, the spongy-like were pre-formed by freeze-drying. The studies concerning subcutaneous implantation and critical size OC defect were performed for 2 and 4 weeks time, respectively. Cellular behavior and inflammatory responses were assessed by means of histology staining and biochemical function and matrix deposition by immunohistochemistry. Additionally, both OC structures stability and new cartilage and bone formation were evaluated by using vivo- computed tomography (Scanco 80). The results showed no acute inflammatory response for both approaches. New tissue formation and integration in the adjacent tissues were also observed, which present different characteristic behaviors when comparing hydrogels and sponges response. As future insights, a novel strategy for regeneration of OC defects can be designed encompassing both, hydrogels and spongy-like structures and cellular approaches. References: 1. Espregueira-Mendes J. et al. Osteochondral transplantation using autografts from the upper tibio-fibular joint for the treatment of knee cartilage lesions. Knee Surgery, Sports Traumatology, Arthroscopy 20,1136, 2012. 2. Oliveira JM. et al, Novel hydroxyapatite/chitosan bilayered scaffold for osteochondral tissue-engineering applications: Scaffold design and its performance when seeded with goat bone marrow stromal cells. Biomaterials 27, 6123, 2006. 3. Pereira D R. et al. Gellan Gum-Based Hydrogel Bilayered Scaffolds for Osteochondral Tissue Engineering. Key Engineering Materials 587, 255, 2013.

Spontaneous healing of Mycobacterium ulcerans lesions in the guinea pig model

Buruli Ulcer (BU) is a necrotizing skin disease caused by Mycobacterium ulcerans infection. BU is characterized by a wide range of clinical forms, including non-ulcerative cutaneous lesions that can evolve into severe ulcers if left untreated. Nevertheless, spontaneous healing has been reported to occur, although knowledge on this process is scarce both in naturally infected humans and experimental models of infection. Animal models are useful since they mimic different spectrums of human BU disease and have the potential to elucidate the pathogenic/protective pathway(s) involved in disease/healing. In this time-lapsed study, we characterized the guinea pig, an animal model of resistance to M. ulcerans, focusing on the macroscopic, microbiological and histological evolution throughout the entire experimental infectious process. Subcutaneous infection of guinea pigs with a virulent strain of M. ulcerans led to early localized swelling, which evolved into small well defined ulcers. These macroscopic observations correlated with the presence of necrosis, acute inflammatory infiltrate and an abundant bacterial load. By the end of the infectious process when ulcerative lesions healed, M. ulcerans viability decreased and the subcutaneous tissue organization returned to its normal state after a process of continuous healing characterized by tissue granulation and reepethelialization. In conclusion, we show that the experimental M. ulcerans infection of the guinea pig mimics the process of spontaneous healing described in BU patients, displaying the potential to uncover correlates of protection against BU, which can ultimately contribute to the development of new prophylactic and therapeutic strategies.

Protein-based nanodevices for therapeutic applications

Tese de Doutoramento em Biologia Molecular e Ambiental (área de especialização em Biologia Celular e Saúde).

Pneumomediastino espontâneo como apresentação de infecção por Mycoplasma pneumoniae

Introdução: O pneumomediastino espontâneo (PE) define-se pela presença de ar livre no mediastino, sem causa traumática. É raro em idade pediátrica e exige elevada suspeição diagnóstica. Caso Clínico: Adolescente de 17 anos, longilíneo, fumador ocasional, sem história traumática nem patologia pulmonar, observado na Urgência por dor retrosternal intensa acompanhada por dispneia ligeira. Referência a acessos de tosse seca violenta horas antes. O diagnóstico de PE suspeitou-se pela palpação de enfisema subcutâneo supraclavicular e confirmou-se por telerradiografia do tórax mostrando ar livre mediastínico. A investigação revelou infeção por Mycoplasma pneumoniae pelo que foi medicado com macrólido para além do repouso e analgesia propostos para o PE, evoluindo favoravelmente. Discussão/Conclusôes: O PE é um diagnóstico a não esquecer perante dor torácica aguda no adolescente. A presença de sintomas, por vezes, frustres e a evolução habitualmente benigna contribuem para um provável subdiagnóstico. O tabagismo é um fator predisponente. A associação a infeção por Mycoplasma pneumoniae só raramente foi descrita.

HOXA9 as a key regulator in glioma initiation, aggressiveness and response to therapy

Tese de Doutoramento em Ciências da Saúde

Aspectos parasitários observados no local inoculado com esporozoitos de Plasmodium Gallinaceum

The following is a summary of the studies made on the development of Plasmodium gallinaceum sporozoites inoculated into normal chicks. Initially large numbers of laboratory reared Aëdes aegypti were fed on pullets heavily infected with gametocytes. Following the infectious meal the mosquitoes were kept on a diet of sugar and water syrup until the appearance of the sporozoites in the salivary glands. Normal chicks kept in hematophagous arthropod proof cages were then inoculated either by bite of the infected mosquitoes or by subcutaneous inoculations of salivary gland suspensions. By the first method ten mosquitoes fed to engorgement on each normal chick and were then sacrificed immediately afterwards to determine the sporozoite count. By the second method five pairs of salivary glands were dissected out at room temperature, triturated in physiological saline and inoculated subcutaneously. The epidermis and dermis at the site of inoculation were excised from six hours after inoculation to forty eight hours after appearance of the parasites in the blood stream and stretched out on filter paper with the epithelial surface downward. The dermis was then curretted. Slides were made of the scrapings consisting of connective tissue and epithelial cells of the basal layers which were fixed by metyl alcohol and stained with Giemsa for examination under the oil immersion lens. Skin fragments removed from normal chicks and from regions other than the site of inoculation in the infected chicks were used as controls. In these, only the normal histological aspect was ever encountered. In the biopsy made at the earliest period following inoculation clearly defined elongated forms with eight or more chromatin granules arranged in rosary formation were found. The author believes these to be products of the sporozoite evolution. Search for transition stages between these forms and sporozoites is planned in biopsies to be taken immediately following inoculation and at given intervals up to the six hour period. 1.) 6 and 12 hour periods. The bodies referred to above found in the first period in great abundance, apparently in proportion to the large numbers of sporozoites inoculated, were perceptibly reduced in numbers in the second period. 2.) 18 hour period. Only one biopsy was examined. This presented a binuclear body shown in Fig. 1, having a more or less hyaline protoplasm staining an intense blue and a narrow vacuole delimiting the cell boundaries. The two chromatin grains were quite large presenting a clearly defined nuclear texture. 3.) 24 hour period. A similar body to that above (Fig. 2) was seen in the only preparation examined. 4.) 60 hour period. The exoerythrocytic schizonts were found more frequently from this period onward. Several such were found no longer to contain the previously described vacuoles (Fig. 3). 5.) 84 hour period. Cells bearing eight or more schizonts were frequently encountered here. That these are apparently not bodies in process of division may be seen in Fig. 4. From this time onward small violet granules similar to volutine grains appeared constantly in the schizont nucleus and protoplasm. These are definitely not hemozoin. The above observations fell within the incubation period as repeated examinations of the peripheral and visceral blood were negative. Exoery-throcytic parasites also were never encountered in the viscera at this time. Exoerythrocytic schizonts searched for at site of inoculation 1, 24 and 48 hours after the incubation period were present in large number at all three times with apparent tendency to diminish as the number within the blood stream increased. Many of them presented the violet granules mentioned above. The appearance of the chromatin and the intensity of staining of the protoplasm varied from body to body which doubtless corresponds to the evolutionary stage of each. This diversity of aspect may frequently be seen in the parasites of the same host cell (Fig. 5.). These findings lend substance to the theory that the exoerythrocytic forms are the link between the sporozoites and the pigmented parasites of the red blood corpuscles. The explanation of their continued presence in the organism after infection of the blood stream takes place and their presence in cases infected by the inoculation blood does not come within the scope of this work. Large scale observations shortly to be undertaken will be reported in more detail particularly observations on the first evolutionary phases of the sporozoite within the organism of the vertebrate host.

Implantação do mixoma infeccioso do coelho na câmara anterior do ôlho da cobaia

Mammalian cancer as well as the Rous chicken sarcoma has been successfully transplanted into the anterior chamber of the eyes of guinea pigs. It was of interest, therefore, to see if the infectious myxomatosis of rabbits, another representative of the infectious tumors, could be grown in the anterior chamber of the guinea pig eye, and, if growth occurred, to compare the tumor's behaviour with that of growths of the above mentioned etiology. Forty-three full-grown guinea pigs from mixed stocks were used throughout, and seventy-eight heterologous transplantation experiments were performed. The grafts measuring less than 2 mm. in diameter were cut from the subcutaneous tissue in skin lesions of rabbits with infectious myxomatosis recently killed. The transfer to the anterior chamber was performed after the usual technique. Some degree of partial survival was found in 23,8% of the grafts, in which typical myxoma cells could be demonstrated fifteen days after the transplantation. The transplant apparently does not increase in size, differing in that respect from that of the Rous chicken sarcoma, which increases in size by 2 or 3 diameters in 2 weeks (Shrigley, Greene & Duran-Reynals, 1945). The virus was still alive in 26% of the grafts 21 days after transplantation, and was able to induce a typical disease when injected to normal rabbits. No alteration in the properties of the virus after growth in the guinea pig was noticed, and this also is different from what happens with the Rous chicken sarcoma.

Tratamento eclético da lepra: 20 anos de experiência

The author gives the clinical records of 20 patients, 14 of which were treated during 17 months, and the rest much less. The method used for treatment was the ecclectical — preferred by the A. since many years —, in which he associates electricity with minor surgery, chaulmestrol (pure or with 0.5 p. c. iodine or 4 p. c. creosote) and other auxilliary curative agents. The chaulmoogra derivatives were used daily as nostrils tamponage (2 p.c. mentholated or thymolated ester), periodically by injections inside the enlarged lymph nodes and nerve abscesses, and twice weekly by subcutaneous infiltrations (MUIR method) 5 c.c. each, and chaulmoogra soap tablets per os. The galvanocauterisation session was once per week, on active leprotic lesions, followed by painting with 30 p. c. trichloracetic acid solution. All 20 patients were bacilliferous before treatment, and became stronger positive after some months treatment. At the end 14 negativated and 6 remained positive and sometimes bacilli being very scanty. 16 out of 20 gave interesting serological reactions, viz.: Wassermann, Stern, Kahn. Rubino, Witebsky and Gaté (Formol-gel) positive in 5; Stern, Rubino. Witebsky and Gaté positive and Wassermann and Kahn negative in 2; Stern. Kahn, Witebsky and Gaté positive and Wassermann anticomplementary in 1; Wassermann. Stern, Kahn, Rubino and Gaté negative in 1. in the beginning, and a few months later Stern. Witebsky and Gaté becoming positive; Stern. Rubino. Witebsky and Gaté positive and anticomplementary W. in 1; Stern, Witebsky and Gaté positive and Wassermann. Kahn and Rubino negative in 1; Witebsky and Gaté positive and Wassermann, Stern, Kahn and Rubino negative in 1; Witebsky 3 times anticomplemantary and strongly positive Gaté in 1; Wassermann and Gaté positive in 1; Stern and Rubino positive in 1 and Stern test negative in one. In 7 cases high Formol-gel were associated with a high sedimentation index. Many cases had very high S.I. being a false measure of the severity of the disease; others remained very high notwithstanding the great improvement of the disease. All patients with more than 12 months treatment became practically symptom free. Lepra reaction amongst them was rare and always started by embolic rash, being controlled by destruction of such skin lesions by galvanocauterisation. In a few cases the lepromata infiltrated with "Subintrol" (a 3 p. c. special chaulmoogra soap prepared by Dr. ASTROGILDO MACHADO) were completely destroyd and never relapsed. In October 1946 Dr. ERNEST MUIR saw here a few cases treated by the author's method and suggested the combination of Diasone with galvanocauterisation which is being done now with satifactory results. The second part of this paper, reporting many leprosy cases treated by the so-called ecclectical method, which are symptom free and negativated since five to ten years, will be published as soon as the sulfone-therapy be summarised in some reliable scientific report to be compared with.

Transplantação de baço de coelho com mixoma infeccioso na câmara anterior do ôlho da cobaia: sôbre a degeneração mixomatosa: inclusões intranucleares

Heterotranplantability of myxoma of rabbits was formely demonstrated when grafts from subcutaneous tissue in skin were used (MARGARINOS TÔRRES & RITA CARDOSOS, 1949). Better results are reported in this paper when grafts from the spleen of infected rabbits were employed. While grafts from normal spleen are almost completely absorbed in sixteen days, those from infected rabbits give origin to full-grown and vascularised tissue in which typical myxoma cells are predominant elements. Progressive growth of heterotransplantated myxoma cells is another similarity between infectious myxoma and malignant tumors. Formation of clear areas of circular contour (interference of a diffusible substance?) associated to myxomatous degeneration is very conspicuous. Peculiar changes of the ground substance, reticular and collagenous fibers (globular swelling, rosary and bulb formation) apparently related to myxomatous degeneration are described. An unexpected finding was the presence of typical intranuclear inclusion bodies in five among forty-eight grafts examined in the sixth day.

Inhibitory effect of cyclophosphamide on the biosynthesis of a perchloro-soluble protein fraction of ehrlich ascites carcinoma cells

The perchloro-soluble mucroptotein fraction was determined in the cells of Ehrlich ascites carcinoma on the 10th and 12th days post-inoculation of the tumor. After 3 days of a single subcutaneous dose of cyclophosphamide (200 mg/kg) the mucoprotein levels were found considerable lower. This difference was highly significant statistically.