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BACKGROUND: Proficiency in laparoscopy is gradually achieved. After initial simulation, it is safe to move to real patients. Simulation improves the basic attributes of laparoscopy, and its non-availability hampers training. Virtual reality and commercial simulators are exorbitantly expensive. Cheaper non-commercial latest, mobile phone-based simulators appear ergonomically unsuitable. A need for a no-cost, home-based laparoscopic endotrainer was felt by authors. METHODS: The authors proposed the concept of smart TV and smart phone-based laparoscopy trainer (STELA), an almost zero cost, lightweight indigenous, cable-less box-type endotrainer, with a smart phone housed on the model, projecting to smart TV via Wi-fi direct. The simulation timings on STELA were compared with Universal Beetel endotrainer by a group of surgeons and residents using identical tasks like object transfer (OT) and knot making (KM). RESULTS: Data were analysed using SPSS, version 23.There was no significant difference in the mean timings of the residents (p > 0.05) on two endotrainers, for both tasks, and of surgeons for OT. Surgeons took significantly longer time (p < 0.05) in KM on STELA. Highest correlation (r = +.848) (<.05) was seen for KM on both devices by residents. CONCLUSION: STELA is a viable, technologically advanced, no cost alternative to the non-commercial cumbersome simulators especially for beginners.
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BACKGROUND: Ureteroscopic management of ureteral calculi using topical anaesthesia has been described. Most studies topically anaesthetized the urethra or urinary bladder by instilling 2% of plain lignocaine. In addition to the success rate, the pain perception in these studies was reported subjectively using non-standard criteria. Topical anaesthesia of the ipsilateral ureter and the pelvicalyceal system (PCS), in addition to urethra and urinary bladder, and use of alkalinised lignocaine, for enhanced effect, has not been reported for ureteroscopy. Using these conceptual alterations, we tested the safety and efficacy of performing ureteroscopy and laser lithotripsy under our technique of total intraluminal topical anaesthesia (TILTA). METHODS: One hundred sixty-eight patients underwent ureteroscopic laser lithotripsy by topical instillation of alkalinised lignocaine into the urinary bladder and ipsilateral ureter and PCS. Self-assessed pain perception and changes in vital parameters were objectively scored at various times. The success rate, reasons for failure, maximum pain scores, complications, and willingness to undergo repeat procedure were recorded. RESULTS: The success rate was 91.3% with a mean duration of 14.7 minutes Double J Stent (DJS) was placed in 38.1%. 61.3% patients did not need any postoperative injectable analgesic, with 11.3% requiring more than a single dose. The intraoperative changes in vitals initially and at the height of pain were not statistically significant (p>0.05). Self-assessed median pain scores between intravenous access placement and ureteroscopy were significantly different (p<0.05). CONCLUSIONS: Ureteroscopy, effectively performed under TILTA, is safe with a high success rate. The acceptable pain tolerance and the shortened convalescence in addition makes the procedure's success worth the pain.
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AIMS: The frequency of the use of intratympanic steroids (ITS) as a treatment for idiopathic sudden sensorineural hearing loss (ISSNHL) in Europe is still unknown and remains a contentious issue amongst otolaryngologists. We undertook a survey of otolaryngologists in Germany and Austria to establish if there is any professional consensus with which to form a protocol for its use. METHODS: A survey of 21 questions was distributed electronically to otolaryngologists in Germany and Austria and data on demographics, indications for intratympanic treatment, procedure, follow-up, and outcomes were analysed. RESULTS: We received 908 responses. 49.1% of otolaryngologists used ITS for ISSNHL. Of those otolaryngologists who use ITS, 73.7% do not use it as primary treatment. 20.6% use ITS in conjunction with oral steroids for primary treatment and only 5.8% use ITS as monotherapy for primary treatment. 90.5% use ITS as salvage therapy. 81.1% do not consider the use of ITS after 2 weeks from the onset of symptoms. 8.3% used a tympanostomy tube and while the most commonly used steroid was dexamethasone at a concentration of 4 mg/ml (61%), a wide variety or other steroids and concentrations were used. CONCLUSIONS: This survey illustrates wide variation of current practice of intratympanic corticosteroid injection for ISSHL in Germany and Austria. In the absence of high-level evidence, knowing what current practice is allows clinicians to assess what they do against what their colleagues are doing, and if they do something very different, make them question their practice. Moreover, the obtained data will help to direct future clinical trials with the aim to compare the outcomes of more commonly used protocols.
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Anti-Inflamatórios/uso terapêutico , Dexametasona/uso terapêutico , Perda Auditiva Neurossensorial/tratamento farmacológico , Perda Auditiva Súbita/tratamento farmacológico , Padrões de Prática Médica/estatística & dados numéricos , Áustria , Esquema de Medicação , Alemanha , Humanos , Injeção Intratimpânica , Otolaringologia , Inquéritos e Questionários , Resultado do TratamentoRESUMO
BACKGROUND: The prevalence of stress urinary incontinence (SUI) in the middle-aged Indian women is around 16%. The use of transvaginal tapes (TVTs) has revolutionised the surgical management of SUI. Patients who undergo placement of the tape via the transobturator route often complain of persistent thigh pain at the site of trocar insertion. The use of minimally invasive tapes with a single suburethral incision reduces surgical trauma by eliminating thigh incisions, while maintaining the cure achieved by conventional TVTs. The study was conducted to test the efficacy and safety of minimally invasive TVT-Secur tape placement for treatment of SUI in women. METHODS: 20 women with stress incontinence were implanted with TVT-Secur tapes and followed up for a year. RESULTS: The objective cure rate of SUI was 85% at the end of a year. The improvement in the patient satisfaction and Incontinence-specific QOL scores, of both Urogenital Distress Inventory (UDI-6) and Incontinence Impact Questionnaire-7 (IIQ-7), was statistically significant at 95% and 99% confidence levels. There were no complaints of thigh pain; however, there were intraoperative complications in the form of bladder perforation in 5% (n = 1), urethral injury in 5% (n = 1) and urethral tape exposure in 10% (n = 2), at 3 months requiring tape sectioning. CONCLUSIONS: These cure rates and complications are comparable to the standard TVT implantations at the end of a year, without thigh pain; however, a greater number of patients and a longer follow-up is required to see whether the long-term cure is maintained or not, before recommending the same as a standard of treatment.
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OBJECTIVE: To describe the surgical technique, complications, and outcomes after anterior pelvic exenteration with total vaginectomy (AETV) for recurrent or persistent genitourinary malignancies. METHODS: We reviewed the medical records of all patients who underwent AETV between 12/2002 and 07/2011. Relevant demographic, clinical, and pathological information was collected. Postoperative complications and rates of readmission and reoperation (up to 180 days after surgery) were examined, and preliminary survival data were obtained. RESULTS: We identified 11 patients who underwent AETV. The median age at the time of the surgery was 55 years (range, 36-71). The median tumor size was 0.9 cm (range, microscopic - 4). Primary tumor sites included: cervix, 6; uterus, 3; vagina, 1; and urethra, 1. Complete surgical resection with negative pathologic margins was achieved in all 11 patients. Major postoperative complications occurred in 4 patients (36%). Six patients (55%) required readmission to the hospital. No operative mortalities were observed, and none of the patients required a re-operation. With a median follow-up after the procedure of 25 months (range, 6-95), none of the patients developed a pelvic recurrence. Ten patients (91%) were alive without evidence of disease and one patient (9%) developed a pancreatic recurrence. CONCLUSION: AETV sparing the rectosigmoid and anus is feasible in highly selected patients with central pelvic recurrences. Compared to previously reported studies on total pelvic exenteration, data from this case series suggest that AETV may be associated with a lower rate of complications without compromising the oncologic outcome, while also preserving rectal function.
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Adenocarcinoma/cirurgia , Carcinoma de Células Escamosas/cirurgia , Recidiva Local de Neoplasia/cirurgia , Exenteração Pélvica/métodos , Neoplasias Urogenitais/cirurgia , Vagina/cirurgia , Adenocarcinoma/patologia , Adulto , Idoso , Perda Sanguínea Cirúrgica , Transfusão de Sangue , Carcinoma de Células Escamosas/patologia , Quimioterapia Adjuvante , Feminino , Humanos , Pessoa de Meia-Idade , Readmissão do Paciente , Exenteração Pélvica/efeitos adversos , Radioterapia Adjuvante , Estudos Retrospectivos , Fatores de Tempo , Neoplasias Urogenitais/patologiaRESUMO
OBJECTIVE: To update our report on the outcome of patients who underwent extended pelvic resection (EPR) for recurrent or persistent uterine and cervical malignancies. METHODS: We reviewed the records of all patients who underwent EPR between 6/2000 and 07/2011. EPR was defined as an en-bloc resection of a pelvic tumor with sidewall muscle, bone, major nerve, and/or major vascular structure. Complications up to 180 days post surgery were analyzed. Survivals were estimated using the Kaplan-Meier method. RESULTS: We identified 22 patients. Median age at the time of EPR was 58 years (range, 36-74). Median tumor diameter was 5.4 cm (range, 1.5-11.2). Primary tumor sites included: uterus, 13; cervix, 7; synchronous uterus/cervix, 1; and synchronous uterus/ovary, 1. The EPR structures were: muscle, 13; nerve, 10; bone, 8; vessel, 5. Complete gross resection with microscopically negative margins (R0 resection) was achieved in 17 patients (77%). There were no perioperative mortalities. Major postoperative complications occurred in 14 patients (64%). The two most common morbidities were pelvic abscesses and peripheral neuropathies. Median follow-up time was 28 months (range, 6-99). The 5-year overall survival (OS) for the entire cohort was 34% (95% CI, 13-57). For the 17 patients who had an R0 resection, the 5-year OS was 48% (95% CI, 19-73). In patients with positive pathologic margins (n=5), the 5-year OS was 0%. CONCLUSION: EPR was associated with prolonged survival when an R0 resection was achieved. The high rate of postoperative complications remains a hallmark of these procedures and properly selected patients should be extensively counseled preoperatively.
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Exenteração Pélvica/métodos , Neoplasias do Colo do Útero/cirurgia , Neoplasias Uterinas/cirurgia , Adulto , Idoso , Feminino , Humanos , Pessoa de Meia-Idade , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/cirurgia , Estadiamento de Neoplasias , Exenteração Pélvica/efeitos adversos , Taxa de Sobrevida , Resultado do Tratamento , Neoplasias do Colo do Útero/patologia , Neoplasias Uterinas/patologiaRESUMO
BACKGROUND & OBJECTIVES: The present study was carried out on stored rice variety PAU 201 in Punjab that was not permitted for milling and public distribution due to the presence of damaged grains at levels exceeding the regulatory limits of 4.75 per cent. The aim of the study was to determine fungal and aflatoxin contamination in the rice samples to assess hazard from the presence of damaged grains. Presence of iron in discoloured rice grains was also assessed. METHODS: Stored samples of paddy of PAU 201 rice variety were collected from six districts of Punjab, milled and analysed for presence of fungal and aflatoxin contamination. Scanning electron microscopy (SEM), energy dispersive X-ray (EDX) analysis and Prussian blue staining was used to determine fungal spores and presence of iron, respectively. RESULTS: Aflatoxin analysis of rice samples indicated that none exceeded the Food Safety and Standards (Contaminants, Toxins and Residues) Regulations, 2011 tolerance limit of 30 µg/kg and majority of the samples had levels <15 µg/kg. The proportion of damaged grains exceeding the limit of 5 per cent was observed in 85.7 per cent of the samples. SEM and Prussian blue staining and EDX analysis of black tipped and pin point damaged rice grains did not show presence of fungal structures and presence of iron. INTERPRETATION & CONCLUSIONS: The results of the study indicated that the stored rice samples did not pose any health concern with respect to aflatoxin contamination as per the criteria laid down by the Food Safety and Standards Authority of India.
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Aflatoxinas/análise , Contaminação de Alimentos/análise , Microbiologia de Alimentos/estatística & dados numéricos , Oryza/química , Oryza/microbiologia , Esporos Fúngicos/isolamento & purificação , Ferrocianetos , Microbiologia de Alimentos/normas , Índia , Microscopia Eletrônica de Varredura , Espectrometria por Raios XAssuntos
Doenças do Labirinto/diagnóstico por imagem , Doenças do Labirinto/patologia , Canais Semicirculares/diagnóstico por imagem , Canais Semicirculares/patologia , Osso Temporal/diagnóstico por imagem , Osso Temporal/patologia , Cadáver , Humanos , Reprodutibilidade dos Testes , Síndrome , Tomografia Computadorizada por Raios XRESUMO
In this Letter the authors introduce a wide-field transmission ultrasound approach to breast imaging based on the use of a large area acousto-optic (AO) sensor. Accompanied by a suitable acoustic source, such a detector could be mounted on a traditional mammography system and provide a mammographylike ultrasound projection image of the compressed breast in registration with the x-ray mammogram. The authors call the approach acoustography. The hope is that this additional information could improve the sensitivity and specificity of screening mammography. The AO sensor converts ultrasound directly into a visual image by virtue of the acousto-optic effect of the liquid crystal layer contained in the AO sensor. The image is captured with a digital video camera for processing, analysis, and storage. In this Letter, the authors perform a geometrical resolution analysis and also present images of a multimodality breast phantom imaged with both mammography and acoustography to demonstrate the feasibility of the approach. The geometric resolution analysis suggests that the technique could readily detect tumors of diameter of 3 mm using 8.5 MHz ultrasound, with smaller tumors detectable with higher frequency ultrasound, though depth penetration might then become a limiting factor. The preliminary phantom images show high contrast and compare favorably to digital mammograms of the same phantom. The authors have introduced and established, through phantom imaging, the feasibility of a full-field transmission ultrasound detector for breast imaging based on the use of a large area AO sensor. Of course variations in attenuation of connective, glandular, and fatty tissues will lead to images with more cluttered anatomical background than those of the phantom imaged here. Acoustic coupling to the mammographically compressed breast, particularly at the margins, will also have to be addressed.
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Acústica/instrumentação , Técnicas de Imagem por Elasticidade/instrumentação , Dispositivos Ópticos , Transdutores , Ultrassonografia Mamária/instrumentação , Desenho Assistido por Computador , Desenho de Equipamento , Análise de Falha de Equipamento , Imagens de Fantasmas , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
BACKGROUND & OBJECTIVE: The number of diabetes in India is increasing at an alarming rate. The effects of physical activity in the form of resistance training or aerobic exercises on type 2 diabetes have not been studied in Indian population. The objective of this study was to analyse the effects of eight weeks of progressive resistance training (PRT) compared with aerobic exercise (AE) on glycaemic control, metabolic profile, cardiovascular fitness parameters and general well being in adults with type 2 diabetes. METHODS: Thirty adults (14 females and 16 males mean; age 53.8 +/- 8.8 yr) with type 2 diabetes were randomly assigned to 8 wk supervised PRT (n=10) or AE (n=10) or control group (n=10). Glycosylated haemoglobin (HbA1c), lipid profile, blood pressure, heart rate, body mass index (BMI) and general well being were measured before training (i.e. 0 wk) and after 8 wk of training period. RESULTS: Plasma glycosylated haemoglobin levels decreased significantly (P<0.05) both in the PRT group (7.57 +/- 2.4% to 6.23 +/- 0.8%) and in AE group (8.11+/-0.9% to 6.66 +/- 0.9%).Total cholesterol levels decreased significantly (P<0.05) by 13.3 per cent in PRT group and by 6.1 per cent in AE group. Both exercise groups showed significantly reduction in systolic blood pressure (P<0.05). General well being improvement was much more in PRT (8.6%) as compared to AE group (2.7%). INTERPRETATION & CONCLUSION: Our findings showed that both PRT and AE were effective in improving metabolic profile of adults with type 2 diabetes but the percentage improvement in triglycerides, total cholesterol levels and general well being with PRT was more compared to AE. Further studies on a larger sample need to be done to confirm these findings.
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Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/metabolismo , Exercício Físico , Metaboloma/fisiologia , Treinamento Resistido , Adulto , Antropometria , Metabolismo Basal , Pressão Sanguínea , Diabetes Mellitus Tipo 2/etnologia , Feminino , Hemoglobinas Glicadas/análise , Frequência Cardíaca , Humanos , Índia/epidemiologia , Masculino , Pessoa de Meia-IdadeRESUMO
The present study deals with the analysis of body composition components of 86 randomly selected patients with confirmed pulmonary tuberculosis aged 16- 70 years collected from the District Tuberculosis Centre and Hospital, Amritsar. An adequate number of controls were also taken for comparison. The findings of the present study showed highly significant differences (p < or = 0.001) in all the twelve kinanthropometric variables except height. When the data was further analyzed between sputum negative and sputum positive patients with pulmonary tuberculosis, highly significant differences (p < or = 0.001) were found only in total body fat and statistically significant differences (p < or = 0.05) were observed in weight, chest circumference in normal position, abdominal circumference, right calf and buttock circumferences. It may be concluded that TB patients were more malnourished than normal people. Thus, special nutritional care should be taken to the TB patients to give them a better quality of life.
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Composição Corporal , Tuberculose Pulmonar , Adolescente , Adulto , Idoso , Feminino , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Hemodialysis patients are at higher risk of cardiovascular disease due to traditional and dialysis-related risk factors. Our aim was to study the effects of hemodialysis on the corrected QT interval (QTc) and QTc dispersion in chronic kidney disease (CKD) without clinically manifest heart disease. Two hundred cases of CKD on chronic intermittent hemodialysis of >3 months' duration were included in the study. Twelve-lead electrocardiography and samples for serum creatinine, potassium, calcium, and magnesium were taken before and after dialysis. The mean age of patients was 52.4 ± 17 years with male-to-female ratio of 3:1. QTc interval and QTc dispersion were prolonged in 47% and 59% before and 50% and 89% of patients after hemodialysis, respectively. The mean values of QTc were 433.4 ± 36.9 ms before and 451.4 ± 39.6 ms after hemodialysis (p = 0.001) and the mean values of QTc dispersion were 60.5 ± 19.3 ms before and 81.5 ± 24.4 ms after hemodialysis (p = 0.001). Similar pattern was observed in all etiological groups of CKD, except for QTc dispersion in malignancy-related CKD (p = 0.216). After hemodialysis, there was a significant fall in the mean values of serum potassium (p = 0.001), rise in serum calcium (p = 0.001), and no change in magnesium (p = 0.424). Patients with post hemodialysis QTc dispersion >74 ms had significantly low mean values of serum potassium and calcium as compared to <74 ms group. Large numbers of hemodialysis patients have a prolonged QTc interval and QTc dispersion with a significant increase in the mean values after hemodialysis. There is a significant fall in serum potassium and rise in serum calcium after dialysis.
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BACKGROUND AND PURPOSE: Five to 60% of coiled brain aneurysms recanalize, generally because of coil compaction. In vitro exclusive use of complex-shaped coils allows better packing of the aneurysmal sac and the neck as compared with helical coils. We report a single-center, prospective study using complex coils. Safety, packing density, and recanalization rate were evaluated. MATERIALS AND METHODS: Seventy-seven aneurysms were embolized using complex coils alone. Aneurysms had a volume of 265 mm3 (diameter: 7.1+/-3.3 mm) and a neck size of 4.1+/-1.8 mm (range: 1.5-12 mm). Average follow-up available in 31 patients was 10.5+/-7.6 months (range: 3-36 months). Primary angiographic endpoints included aneurysmal recanalization and (re)rupture. Primary adverse events included stroke or death. RESULTS: Complete or near-complete occlusion was achieved in all of the aneurysms but required balloon assistance in 24.6%. The packing density was computed as 37%+/-13%. No rerupture was observed during the follow-up interval. Recanalization was seen in 4 (12.9%) of 31. Two basilar tip aneurysms underwent a safe and complete recoiling. Periprocedural nondevice-related neurologic deficits were seen in 2 (2.9%) of 69 patients. CONCLUSIONS: The use of complex-shaped coils allows higher packing density, which may improve the recanalization rate. Basilar tip aneurysms remain a challenge.
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Embolização Terapêutica/instrumentação , Aneurisma Intracraniano/diagnóstico por imagem , Aneurisma Intracraniano/terapia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Angiografia Cerebral , Criança , Embolização Terapêutica/métodos , Desenho de Equipamento , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Platina , Resultado do TratamentoRESUMO
The structures of the major merozoite surface antigen of Plasmodium falciparum and the gene encoding it were indistinguishable for the Wellcome strain and the Thai clone T9/94 but different for clones T9/96, T9/98, and T9/101. The central portion of the gene is subject to the greatest variation in structure. The protein from all five lines was found to be posttranslationally modified by covalent addition of both carbohydrate and fatty acid.
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Antígenos de Protozoários/genética , Antígenos de Superfície/genética , Genes , Variação Genética , Plasmodium falciparum/genética , Animais , Quimotripsina , Hibridização de Ácido Nucleico , Mapeamento de Peptídeos , Plasmodium falciparum/imunologia , Plasmodium falciparum/isolamento & purificaçãoRESUMO
Hearing loss on pure-tone and brainstem-evoked response audiometry is well documented in chronic kidney disease patients, but there are only scant data in renal allograft recipients. The aim of this study was to evaluate cochlear function on pure-tone and brainstem-evoked response audiometry among patients of chronic kidney disease to document changes after renal transplantation. The study included 20 healthy controls and 40 patients: 10 each at stage 3 and 4 and 20 with stage 5 chronic kidney disease. Patients with known causes for hearing loss were not included. Patients with stage 5 chronic kidney disease were reevaluated 1 year after renal transplantation. Compared with healthy controls, chronic kidney disease patients showed a highly significant bilateral sensorineural hearing loss at all frequencies of 0.25 to 8.0 kHz, which was more marked in higher frequencies. A highly significant delay in the latencies of waves I, III, and V and interpeak latencies of I to III and I to V was also noted on brainstem-evoked response audiometry. Compared with their pretransplant values, there was a significant improvement in the delay in absolute latencies of I, III, and V among renal allograft recipients. There was no correlation of audiometry findings with gender, degree of renal failure, and serum sodium. Hearing loss was seen in 70% of stage 3 and 4 chronic kidney disease and 60% in stage 5. The intensity of hearing loss was variable. In conclusion, chronic kidney disease patients shows definitive audiologic dysfunction with some improvement in hearing and wave latencies after successful renal transplantation.
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Cóclea/fisiologia , Doenças Cocleares/etiologia , Perda Auditiva Central/etiologia , Perda Auditiva Neurossensorial/etiologia , Nefropatias/fisiopatologia , Transplante de Rim/fisiologia , Doença Crônica , Cóclea/fisiopatologia , Audição , Perda Auditiva Central/fisiopatologia , Perda Auditiva Neurossensorial/fisiopatologia , Humanos , Nefropatias/complicações , Nefropatias/cirurgia , Estudos LongitudinaisRESUMO
Retinal evaluation was performed in 20 consecutive type 2 diabetics before and after renal transplantation. This included 19 men and 1 woman of mean age 52 years (range 30 to 60 years). Before renal transplantation, 95% of these patients showed diabetic retinopathy (50% nonproliferative and 45% proliferative diabetic retinopathy). There was no change in retinopathy at 3 months after renal transplantation. At 1 year, two patients (10%) showed deterioration in their diabetic retinopathy, while the remaining 90% did not show any change. Also 25% of renal allograft recipients developed posterior capsular cataracts.
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Nefropatias Diabéticas/cirurgia , Retinopatia Diabética/fisiopatologia , Transplante de Rim/fisiologia , Adulto , Catarata/epidemiologia , Diabetes Mellitus Tipo 2/fisiopatologia , Progressão da Doença , Feminino , Seguimentos , Humanos , Estudos Longitudinais , Masculino , Pessoa de Meia-Idade , Fatores de TempoRESUMO
An efficient and rapid procedure for the humanization of murine monoclonal antibodies (MumAb) is described. It consists of site-directed mutagenesis (SDM) to transfer the murine complementarity-determining regions (MuCDR) onto human framework regions (HuFR), followed by polymerase chain reaction (PCR) of the SDM product. Using SDM/PCR, rapid and correct humanization of MumAb heavy chains is clearly demonstrated. Compared to current protocols this method considerably reduces the time and labour required to generate humanized mAb.
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Anticorpos Monoclonais/biossíntese , Animais , Anticorpos Monoclonais/genética , DNA/biossíntese , Humanos , Cadeias Pesadas de Imunoglobulinas/biossíntese , Camundongos , Mutagênese Sítio-Dirigida , Ácidos Nucleicos Heteroduplexes , Oligodesoxirribonucleotídeos , Reação em Cadeia da PolimeraseRESUMO
The effect of elevated human IL-6 (hIL-6) production by human bone marrow (Hu-BM) stromal cells on osteoclasts in human bone was examined. Human bone was implanted into nonobese diabetic mice with severe combined immunodeficiency (Hu-Bone-NOD/SCID mice). Immunohistochemistry of bone implants and mouse spleens (at 20 weeks), showed human CD45+ cells, B cells, and macrophages in both tissues. Thus, Hu-BM cells survive human bone transplantation and infiltrate mouse tissue. Bone implants had 75 +/- 12% (mean +/- SD) human CD45+ cells, and 9 +/- 4% mouse hematopoietic cells. A retrovirus vector containing the human IL-6 gene was used to transduce Hu-BM stromal cells (IL-6/stromal) and the PA317 cell line (IL-6/PA317). IL-6/ stromal cells (secreting, on average, 17 microg of hIL-6/10(6) cells per 24 h) were injected directly into human bone implants in Hu-Bone-NOD/SCID mice. IL-6/PA317 cells (secreting 16 microg/mL of hIL-6/10(6) cells per 24 h) were injected intraperitoneally into Hu-Bone-NOD/SCID mice. Analyses of sera from both groups of animals showed elevated levels of IL-6. However, only bone implants engrafted with IL-6/stromal cells had a statistically significant increase in osteoclast-lined mineralized trabecular bone surface (BS). Thus, a high concentration of serum hIL-6 in Hu-Bone-NOD/SCID mice alone does not increase osteoclast-lined BS in bone implants. Most importantly, it is the type of human BM cell that secretes the high levels of hIL-6 that is most critical.
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Células da Medula Óssea/citologia , Células da Medula Óssea/metabolismo , Fêmur/citologia , Interleucina-6/metabolismo , Osteoclastos/fisiologia , Animais , Células da Medula Óssea/química , Transplante Ósseo/imunologia , Fêmur/fisiologia , Fêmur/transplante , Citometria de Fluxo , Hematopoese/imunologia , Humanos , Hospedeiro Imunocomprometido , Hibridização in Situ Fluorescente , Interleucina-6/análise , Interleucina-6/sangue , Antígenos Comuns de Leucócito/análise , Camundongos , Camundongos Endogâmicos NOD , Camundongos SCID , Imunodeficiência Combinada Severa , Células Estromais/metabolismo , Transdução GenéticaRESUMO
Despite previous attempts there is currently no suitable animal model available for xenogeneic graft versus host disease (XGVHD) mediated via human immunocompetent cells. Recently, we have developed an efficient protocol to engraft SCID mice with human peripheral blood lymphocytes (Hu-PBLs). The engraftment efficiency is extremely high, such that 100% of Hu-PBL-SCID mice die of XGVHD within 4 weeks after engraftment with Hu-PBLs (3-5 x 10(7) cells). A series of experiments was performed to investigate the mechanisms involved in the severe XGVHD. The results suggest that XGVHD was induced by human CD4+ T cells, antixenogeneic (antimouse) antibodies, and lymphokines. The SCID mouse model will be extremely valuable for the evaluation and development of immunosuppressive agents and transplantation protocols for human XGVHD.