Differential regulation of Shigella Spa47 ATPase activity by a native C-terminal product of Spa33.

Shigella is a Gram-negative bacterial pathogen that relies on a single type three secretion system (T3SS) as its primary virulence factor. The T3SS includes a highly conserved needle-like apparatus that directly injects bacterial effector proteins into host cells, subverting host cell function, initiating infection, and circumventing resulting host immune responses. Recent findings have located the T3SS ATPase Spa47 to the base of the Shigella T3SS apparatus and have correlated its catalytic function to apparatus formation, protein effector secretion, and overall pathogen virulence. This critical correlation makes Spa47 ATPase activity regulation a likely point of native control over Shigella virulence and a high interest target for non-antibiotic- based therapeutics. Here, we provide a detailed characterization of the natural 11.6 kDa C-terminal translation product of the Shigella T3SS protein Spa33 (Spa33C), showing that it is required for proper virulence and that it pulls down with several known T3SS proteins, consistent with a structural role within the sorting platform of the T3SS apparatus. In vitro binding assays and detailed kinetic analyses suggest an additional role, however, as Spa33C differentially regulates Spa47 ATPase activity based on Spa47s oligomeric state, downregulating Spa47 monomer activity and upregulating activity of both homo-oligomeric Spa47 and the hetero-oligomeric MxiN2Spa47 complex. These findings identify Spa33C as only the second known differential T3SS ATPase regulator to date, with the Shigella protein MxiN representing the other. Describing this differential regulatory protein pair begins to close an important gap in understanding of how Shigella may modulate virulence through Spa47 activity and T3SS function.

Development of an effective and rapid qPCR for identifying human ChREBPα/ß isoforms in hepatic and adipose tissues.

Most quantitative real-time PCR (qPCR) detection methods use two types of chemistries to measure the expression levels of ChREBP isoforms, hydrolysis probes for ChREBPα and SYBR Green for ChREBPß. Hydrolysis probes are not available to determine the ChREBPß isoform. The aim of this study was to develop a qPCR assay based only on hydrolysis probes for both ChREBP isoforms. Liver and adipose tissue biopsies from patients undergoing elective cholecystectomy surgery were used to perform qPCR. To validate this assay, the results were compared with sequencing and High Resolution Melting (HRM) PCR assays. Direct sequencing was used to determine the sequence showing site where ChREBPß presents its specific splicing (1 b exon/2 exon) in order to design the primers and the probe. We developed a qPCR assay to determine the ChREBP isoforms expression based on hydrolysis probes. It assays showed good efficiency (95.50%, on average), high reproducibility, and a strong linear correlation (R2 ≥ 0.99) for tissues tested. HRM analysis confirmed the specificity of the primers and the result of this assay matched (100%) with the outcomes obtained by sequencing and qPCR. Also, we obtained the ChREBPß sequence showing exon 1b spliced to exon 2, bypassing exon 1a, and retaining the remainder of the ChREBPα exons. Based on the use of hydrolysis probes, our method can efficiently identify the expression of both ChREBP isoforms. Thus, the comparability of the qPCR results using a single chemistry (hydrolysis probes) to discriminate between both ChREBP isoforms was possible.

Subtrochanteric fractures of the femur. Treatment with locking intramedullary nailing. An experience without C-Arc. The guide-wire stopping method for verify distal locking is described.

OBJECTIVE: Evaluate the results of treatment of subtrochanteric fracture with interlocked intramedullary nail and describe a technical for accurate and secure verification of distal locking position when we do surgery without arc-C. METHODS: A case series where was reviewed the results in 49 patients with subtrochanteric fracture and treated locking intramedullary nailing. The AO and Russell-Taylor Classification were used. The statistic procedure was done with SPSS program. The traumatic hip scale of Sander et al. was used for final evaluation. Technical for doing distal locking is detailed. RESULTS: This series evaluated 49 patients: 35 male and 14 female patients. The mean age was 36 years old (range: 18-86 years). Traffic accidents and gunshot wounds were injury forms most frequent. There were not transoperative complications. The surgical time was between 90 and 120min (mean: 108min). The mean follow-up was 24 months (range: 18-36 months). According to Sanders score, it was reported 22 excellent results, 20 good results and 7 regular results. All distal locking procedures were successfully performed. CONCLUSION: Although this series consists in a few numbers of patients, we recommend the placement of interlocked intramedullary nail (Closs-MB Bioimpianti® and Orthosintese®) in subtrochanteric fractures. This device allows placement of distal locking through the insertion frame with safety and precision, even in surgery rooms without arc-C. The guide-wire stopping method allows the verification of distal locking in an objective, accurate, safe and reproducible way.

Resultado a largo plazo del tratamiento quirúrgico de las fracturas del tobillo. (5 años de seguimiento) / Long-term outcome of surgical treatment of ankle fractures. (5 years follow up)

Antecedentes: Algunos autores documentan alteración funcional con molestias asociadas en un importante número de pacientes a largo plazo luego de cirugía en fracturas de tobillo. Lesiones iniciales de cartílago, así como reducción primaria deficiente predispone a resultados desfavorables. Método: serie clínica. Evaluación funcional de fracturas de tobillo luego de cinco años de la cirugía. Se utilizó las escalas de Olerud y Molander, y Baird para evaluación. La prueba Chi2 se utilizó para inferencia estadística. Resultados: 24 pacientes, 15 hombres y 9 mujeres; La edad promedio fue 28 años (16-68 años). Hubo una asociación directamente proporcional entre dolor y limitación laboral (X2: p <0.05); igualmente el dolor estuvo relacionado de manera directa con los puntajes más bajos de las escalas (X2: p <0.05). A mayor deterioro radiológico mayor intensidad de dolor (X2: p <0.05). Hubo una asociación directa entre reducción inicial inadecuada y deterioro radiológico (X2: p <0.05). 75% resultados satisfactorios. Conclusiones: El dolor fue el síntoma capital que condicionó el resultado final. El deterioro radiológico se asoció mayormente a mala reducción inicial. Un importante número de pacientes presentan algún grado de deterioro de la función 5 años después de la cirugía, aun con buena reducción inicial(AU)

Background: Some authors document discomfort associated with functional impairment in a significant number of patients after long-term surgery on ankle fractures. Initial cartilage injuries and poor primary reduction predisposes unfavorable results. Method: clinical series. Functional assessment of ankle fractures after five years of surgery. We used scales Olerud Molander, and Baird for evaluation. The Chi2 test was used for statistical inference. Results:. 24 patients, 15 males and 9 women, mean age was 28 years (16-68 years). There was an directly proportional association between pain and labor limitation (X2: p <0.05), and also the pain was related directly with the lowest scores of the scales (X2: p <0.05). A further deterioration radiological, greater pain (X2: p <0.05). There was a direct association between inadequate initial reduction and radiological deterioration (X2: p <0.05). Conclusions. Pain was the main symptom that determined the outcome. The radiological deterioration was associated mostly with poor initial reduction. A significant number of patients have some degree of impaired function 5 years after surgery, even with good initial reduction. 75% of patients had satisfactory results(AU)

Cirugía percutánea en fascitis plantar y espolón calcáneo / Percutaneous surgery plantar fascitis and heel spur

Se realizó un estudio prospectivo tipo serie clínica donde se avaluá la evolución de 12 pacientes (24 pies) quienes presentaban talalgia crónica por fascitis plantar con presencia de espolón calcáneo y fueron tratados con fasciotomía plantar percutánea. La mayoría de los pacientes son femeninos (90 por ciento) y la edad promedio es 40,5 años. Tiempo de seguimiento 12 meses. El resultado final fue valorado por escala análoga visual (EAV) para cuantificar dolor, opinión de paciente y reintegro a actividades habituales. El procedimiento estadístico se hizo con el programa SSPS 12,0 donde se calcularon medidas de tendencias central y de dispersión. El análisis inferencial se realizó con comparación de medidas a través de prueba T student y con la prueba no paramétrica Chi cuadrado. Se reportó 90 por ciento de resultados satisfactorios. La cirugía percutánea del pie en fascitis plantar con espolón calcáneo es un método sencillo y eficaz. Mejora el tiempo quirúrgico y logra el reintegro precoz del paciente a sus actividades habituales.

We realized a trial study where was evaluated 12 pacients (24 foots), 9 women and 3 male, who presented pain heel asociated with plantar fascitis and sprus and were realizaed fasciotomy percutaneus surgery to determine the efectiveness. The results was evaluated with analog visual scale (VAS) both the pain and the daily activities. The SSPS 12,0 statist program was utlized. The inferencial study was done with T student test and Chi square. Most patients were female (90 percent), mean age 40,5 years old. The follow up was twele months. The points in VAS to pain descending to 8,5 to 1,5. Pain was significantly lower at every assessment point as compare to preoperative values (P<0.05). Was reported90 percent of satisfactory results. The fasciotomy percutaneus surgery is a safe, efective and single procedure to treatment the pain heel for plantar Fascitis and calcaneal spurs. Those patients had significantly less postoperative pain and returned to regular activities early.

[Percutaneous surgery for plantar fasciitis due to a calcaneal spur]. / Cirugía percutánea en fascitis plantar por espolón calcáneo.

OBJECTIVE: Determine the efficacy of percutaneous surgical treatment for talalgia due to a calcaneal spur. MATERIAL AND METHODS: This is an observational, descriptive, clinical series analyzing the outcomes of 10 patients with a diagnosis of talalgia due to plantar fasciitis with a calcaneal spur treated with percutaneous foot surgery. The end result was assessed with a visual analog scale (VAS) to measure pain, the patients' opinion and their return to activities of daily living. Central tendency and scatter measurements were calculated. The inferential analysis was done with the non-parametric chi square (chi2) test. RESULTS: Most patients were females (90%) and mean age was 40.5 years. Follow-up was 12 months. One patient had bleeding of the approached area. Pain was reduced from 8 to 1.5 in the VAS. Nine patients returned to their activities. Two patients had occasional mild pain upon prolonged bipedestation. Ninety percent of results were satisfactory. CONCLUSIONS: Percutaneous foot surgery in talalgias caused by plantar fasciitis due to a calcaneal spur is a simple and effective method. It reduces the operative time and allows for an early return of patients to their activities of daily living.

Técnica quirúrgica antifuga en el implante de catéter de Tenckhoff para diálisis peritoneal / Antileaking surgical technique in implantation of Tenckhoff's catheter for peritonal dialisis

Se realizó un estudio experimental, prospectivo, comparativo, de selección aleatoria y ciego en los Hospitales Nuevo Civil y México Americano de Guadalajara, de enero a diciembre de 1994, en que se analizaron dos procedimientos utilizados para disminuir hiperazoemia en pacientes con insuficiencia renal crónica o aguda, con la finalidad de establecer eficiencia obtenida mediante la aplicación de catéteres de diálisis peritoneal rectos y curvos a la cavidad peritoneal. En ambas técnicas se fijo firmemente el catéter a la aponeurosis peritoneal o posterior del músculo recto anterior del abdomen. Distribuimos 90 pacientes en dos grupos de 45, para comparar la técnica quirúrgica original de colocación de catéter de diálisis frente a una técnica que denominamos "antifuga". El objetivo principal es demostrar que utilizando la técnica antifuga y clasificando la consistencia de la aponeurosis posterior del músculo recto anterior del abdomen (APRA) y utilizando el grosor de la sutura según su firmeza, pudiera existir un menor número de fugas, infecciones del túnel subcutáneo e infección en el sitio de salida de la pared abdominal, además de disminuir otras complicaciones secundarias, como son: dehiscencia, herniación e infección de la herida quirúrgica. Con base a los resultados de este estudio experimental concluimos que existe correlación entre la fuga temprana de soluciones con la técnica original y con la edad, grado de obesidad y APRA de los pacientes. Además, que el de catéteres en espiral fallan que menor frecuencia que los rectos, por lo que recomendamos la técnica antifuga y el empleo de catéteres en espiral en el manejo de diálisis peritoneal ambulatoria para pacientes con insuficiencia renal crónica