High genetic diversity of hepatitis E virus in swine in São Paulo State, Brazil / Diversidade genética do vírus da hepatite E em suínos no estado de São Paulo, Brasil

A hepatite E é uma zoonose emergente que afeta diversas espécies de mamíferos, inclusive o ser humano. É ocasionada por um vírus da espécie Orthohepevirus A que possui diversos genótipos e subgenótipos. No Brasil é descrito o genótipo HEV-3, cujo principal reservatório é o porco doméstico. Testes moleculares e sorológicos demonstram o HEV-3 em diferentes estados, tanto em animais quanto em humanos. No estado de São Paulo, existem diversos estudos sobre a epidemiologia da hepatite E em humanos, mas faltam informações sobre o HEV-3 em suínos. Assim, o objetivo deste trabalho foi verificar a ocorrência de HEV por meio da técnica de RT-PCR e posterior sequenciamento em um banco de amostras de fezes de suínos colhidas entre 2008 e 2009, na região metropolitana de Campinas. Das 89 amostras analisadas, foi possível detectar o HEV-3 em sete e, pela reconstrução filogenética, foram encontrados os subgenótipos HEV-3b, HEV-3h, e HEV-3j. Uma amostra disponível no GenBank, proveniente de São Paulo, que ainda não havia sido subgenotipada, foi agrupada ao HEV-3i. Os subgenótipos HEV-3j e HEV-3i ainda não tinham sido relatados no país. O estudo demonstra uma grande diversidade genética do HEV no estado de São Paulo e reforça o caráter zoonótico da HEV-3.(AU)

Justifying CT prior to MRI in cases of suspected occult hip fracture. A proposed diagnostic protocol.

INTRODUCTION: Patients with clinical suspicion of hip fracture, but negative radiographs are suspected of having an occult hip fracture (OHF). Different diagnostic modalities are available for investigating OHF and various protocols have been suggested. MRI has the highest sensitivity and specificity, however availability is limited in many institutes. CT is readily accessible in the large majority of hospitals throughout the world but has lower sensitivity and may miss some fractures. In this article we investigate a protocol that balances these issues providing a practical and cost-effective solution. METHODS: During a four-year period between 2012 and 2016 a strict diagnostic protocol was followed at our Medical Center for patients suspected of OHF. This MRI selective protocol consisted of CT initially being performed and only when negative for fracture, followed by an MRI. Retrospective analysis of all patients who followed the protocol was performed. The patients were divided into two groups: those diagnosed by CT alone and those diagnosed by MRI after having a negative CT scan. Diagnostic performance, time to diagnosis and the cost of this protocol were evaluated. RESULTS: 103 patients were treated under the protocol. In 50 patients (49%) hip fracture was diagnosed by CT alone. In the remaining 53 patients (51%) no definitive diagnosis was reached by CT and MRI was subsequently performed. 12 of these 53 patients (23%) were diagnosed with hip fracture necessitating surgery. In the CT only group mean time from admission to diagnosis was 3 hours, in the CT + MRI group this rose to 40 hours. Cost analysis showed that this protocol was more cost-effective than performing MRI in all patients, saving an estimated 66,805 Euro during the study period. CONCLUSION: The clinical challenge of diagnosing OHF can be minimised by implementing a diagnostic protocol. The protocol should take into consideration the diagnostic sensitivity, availability and cost of different imaging modalities. An MRI selective strategy with initial CT scanning is recommended, as it reduces time to diagnosis and lowers overall costs.

The distribution of heat in bone during radiofrequency ablation of an ex vivo bovine model of osteoid osteoma.

Osteoid osteoma is treated primarily by radiofrequency (RF) ablation. However, there is little information about the distribution of heat in bone during the procedure and its safety. We constructed a model of osteoid osteoma to assess the distribution of heat in bone and to define the margins of safety for ablation. Cavities were drilled in cadaver bovine bones and filled with a liver homogenate to simulate the tumour matrix. Temperature-sensing probes were placed in the bone in a radial fashion away from the cavities. RF ablation was performed 107 times in tumours < 10 mm in diameter (72 of which were in cortical bone, 35 in cancellous bone), and 41 times in cortical bone with models > 10 mm in diameter. Significantly higher temperatures were found in cancellous bone than in cortical bone (p < 0.05). For lesions up to 10 mm in diameter, in both bone types, the temperature varied directly with the size of the tumour (p < 0.05), and inversely with the distance from it. Tumours of > 10 mm in diameter showed a trend similar to those of smaller lesions. No temperature rise was seen beyond 12 mm from the edge of a cortical tumour of any size. Formulae were developed to predict the expected temperature in the bone during ablation.

Benefits of a simple glycaemic protocol in an orthopaedic surgery ward: a randomized prospective study.

BACKGROUND: Hyperglycaemia and diabetes mellitus are common in patients hospitalized in the orthopaedic surgery ward. However, glycaemic control obtained during hospitalization is often suboptimal. No method for achieving adequate glycaemic control in this population has been validated in an in-hospital setting. INTERVENTION: An intervention including an intensive subcutaneous insulin protocol in the orthopaedic department. METHODS: All diabetic patients admitted to the Department of Orthopaedic Surgery were prospectively randomized during a 6-month period. One group (n = 30) received standard care with sliding scale insulin and the other group (n = 35) received the intervention protocol. During the intervention period, the staff was briefed on the importance of glucose monitoring and control. An intensive multiple-injection protocol consisting of four daily regular/neutral protamine hagedorn (NPH) insulin injections was initiated in diabetic patients. The programme was followed up by a consulting diabetologist. RESULTS: Mean blood glucose levels throughout the hospitalization were 161.48 ± 3.8 mg/dL in the intervention group versus 175.29 ± 2.3 mg/dL in the control group (p < 0.0005). Hospitalization was shorter by 2 days in the intervention group (p < 0.05). The number of severe hyperglycaemic events (blood glucose level above 400 mg%) was significantly lower (p < 0.05) in the intervention group. There was no significant difference in the number of hypoglycaemic events. CONCLUSIONS: The suggested four-step intervention regimen improved glycaemic control of hospitalized patients in the orthopaedic department and simplified the 'in-house' treatment of the diabetic patient. Hospital stays were reduced on average by two days (p < 0.05).

Retrospective--systematic study and quantitative analysis of cellular proliferation and apoptosis in normal, hyperplastic and neoplastic perianal glands in dogs.

Neoplasms in the perianal region are frequently diagnosed in dogs. The aetiology is unknown, and most of them are benign. In this study, 240 neoplasms of the perianal glands of dogs were retrieved from the Department of Pathology archives of the Faculty of Veterinary Medicine and Zootechny of University of São Paulo (FMVZ/USP), from 1984 to 2004. All 240 cases were re-examined by two pathologists. Nine cases (4%) were diagnosed as hyperplasia, 49 (20%) as group I adenoma, 81 (34%) were classified as moderately differentiated adenomas of the group II, 46 (19%) were poorly differentiated adenomas of group II, 48 (20%) were carcinoma of the group III according to the classification proposed by Berrocal, and 7 (13%) were other kind of tumours. Males over 8 years of age were predominantly affected. Cell proliferation was quantified by counting proliferating cell nuclear antigen (PCNA) positive nuclei, and apoptosis was quantified by counting fluorescent eosin-stained apoptotic corpuscles (AC) in normal tissue, hyperplasia and in different histologic types of neoplasia of these glands. A parallel pattern of increase in both parameters (cell proliferation and apoptosis) was obtained. The net growth index (NGI), represents how much a cell population is proliferating or dying and was achieved by dividing the mean PCNA count in 1000 cells by the mean AC stain count in 1000 cells. NGI was different between hyperplasia and neoplasia; group I adenomas have a much higher potential of growth, and NGI decreases from benign towards malignant lesions. These results show up the importance of studying cell proliferation and apoptosis to understand the carcinogenesis of dog perianal gland.

A prospective, randomised study comparing the percutaneous compression plate and the compression hip screw for the treatment of intertrochanteric fractures of the hip.

Limited access surgery is thought to reduce post-operative morbidity and provide faster recovery of function. The percutaneous compression plate (PCCP) is a recently introduced device for the fixation of intertrochanteric fractures with minimal exposure. It has several potential mechanical advantages over the conventional compression hip screw (CHS). Our aim in this prospective, randomised, controlled study was to compare the outcome of patients operated on using these two devices. We randomised 104 patients with intertrochanteric fractures (AO/OTA 31.A1-A2) to surgical treatment with either the PCCP or CHS and followed them for one year postoperatively. The mean operating blood loss was 161.0 ml (8 to 450) in the PCCP group and 374.0 ml (11 to 980) in the CHS group (Student's t-test, p < 0.0001). The pain score and ability to bear weight were significantly better in the PCCP group at six weeks post-operatively. Analysis of the radiographs in a proportion of the patients revealed a reduced amount of medial displacement in the PCCP group (two patients, 4%) compared with the CHS group (10 patients, 18.9%); Fisher's exact test, p < 0.02. The PCCP device was associated with reduced intra-operative blood loss, less postoperative pain and a reduced incidence of collapse of the fracture.

The use of intraoperative three-dimensional imaging (ISO-C-3D) in fixation of intraarticular fractures.

INTRODUCTION: The purpose of this study was to analyse the applicability and advantages of the intraoperative use of a mobile isocentric C-arm with three-dimensional imaging (Siremobil ISO-C-3D) in fixation of intraarticular fractures. METHODS: After the fixation was judged to be satisfactory relying on the images provided by routine fluoroscopy, intraoperative CT visualisation with ISO-C-3D was performed to evaluate the fracture reduction and implant position. Intraoperative revision was performed based on the additional information ISO-C-3D provided beyond routine fluoroscopy. ISO-C-3D was used on a series of 72 closed-intraarticular fractures in 70 patients. Fracture distribution was: calcaneus (25), tibial plateau (17), tibial plafond (12), acetabulum (11), distal radius (3), ankle Weber-C (3) and femoral head (l). The primary outcome measure was revision rate after final ISO-C-3D data acquisition and prior to wound closure. Secondary objectives were to measure the additional time required for ISO-C-3D use and to determine the rate of further re-do surgeries. RESULTS: Eight out of 72 (11%) fracture fixations were judged by the surgeon to require intraoperative revision following ISO-C-3D imaging. Prior to leaving the operating room, the surgeon was satisfied with fracture alignment in all the procedures. The mean additional operative time using ISO-C-3D was 7.5 min (8.2% of the mean total operative time). No patient required re-do surgery. CONCLUSION: : Intraoperative three-dimensional visualisation of intraarticular fractures enables the surgeon to identify inadvertent malreductions or implant malpositions which may be overlooked by routine C-arm fluoroscopy and hence may eliminate the need for re-do procedures. ISO-C-3D adds little operative time and may preclude the need for pre-operative and post-operative CT-scans in selected cases.

Osteoid osteoma: CT-guided radiofrequency ablation using a water-cooled probe.

BACKGROUND: The purpose of this study was to assess the safety and efficacy of computed tomography (CT) guided percutaneous radiofrequency (RF) ablation of osteoid osteoma by using the water-cooled probe. METHODS: During the period from July 2002 to February 2006, fifty-one patients with osteoid osteomas localized in femur (29), tibia (10), calcaneus (2), talus (2), metatarsus (2), humerus (1), sacrum (1), scapula (1), olecranon (1), patella (1) and thoracic vertebra (1) were treated with CT-guided RF ablation using the Cooltiptrade mark Tyco Healthcare probe. Mean age was 20 (range, 3.5-57 years) and male to female ratio was 36/15. Mean follow-up period was reported 2 years (range, 9-51 months). The procedures were carried out under general anesthesia and the patients were discharged from the hospital within 24 h. RESULTS: Technical failure was reported in only one procedure. Pain disappeared postoperatively in all the patients within 2-3 days and no patients needed analgesic treatment after a week. All patients were allowed fully weight bear and function without limitation after the procedure. Recurrence of the pain was observed in one patient who was treated successfully with a second ablation. Our primary and secondary clinical success rates were 98 and 100% respectively. In one case, wound infection was observed after the procedure as the only post-operative complication in our series. CONCLUSION: CT-guided percutaneous RF ablation of osteoid osteomas using the water-cooled probe is a safe, effective and minimally invasive procedure with high success rate and lack of relapses.

Delayed reaction to shrapnel retained in soft tissue.

BACKGROUND: Treatment of penetrating injuries to soft tissues does not require surgical excision of shrapnel. Metals usually remain inert and do not cause damage and are therefore left in soft tissue. OBJECTIVE: Characterization of delayed reaction to shrapnel retained for many years in soft tissue. PATIENTS: Four patients sustained penetrating injuries to the limbs with embedded shrapnel. Many years later, they experienced delayed reaction to the metals that required surgery, with very unusual findings. CONCLUSIONS: Although nonsurgical treatment of shrapnel in soft tissues is the treatment of choice in most cases, we need to be aware of the possibility of late complications requiring surgical treatment.

The floating hip injury: patterns of injury.

OBJECTIVE: To evaluate the relationship between mechanism of injury, type of femoral fracture and type of acetabular fracture in floating hip injury. DESIGN: Historical retrospective. PATIENTS: Twenty consecutive patients who sustained a floating hip injury, i.e. simultaneous ipsilateral fracture of the acetabulum and the femur. INTERVENTION: Statistical analysis of the correlation between the mechanism of injury and fracture type. RESULTS: Two main patterns of floating hip injury were observed. The first is the posterior type, which occurs due to a longitudinal force along the femur that causes first, a posterior type fracture of the acetabulum and thereafter, a midshaft femoral fracture. The second pattern is the central type, caused by a lateral blow to the greater trochanter, which then causes a central fracture-dislocation of the acetabulum and a proximal fracture of the femur. CONCLUSIONS: This observation explains the biomechanical nature of this injury and has treatment related implications.

Engineered human mesenchymal stem cells: a novel platform for skeletal cell mediated gene therapy.

BACKGROUND: Human mesenchymal stem cells (hMSCs) are pluripotent cells that can differentiate to various mesenchymal cell types. Recently, a method to isolate hMSCs from bone marrow and expand them in culture was described. Here we report on the use of hMSCs as a platform for gene therapy aimed at bone lesions. METHODS: Bone marrow derived hMSCs were expanded in culture and infected with recombinant adenoviral vector encoding the osteogenic factor, human BMP-2. The osteogenic potential of genetically engineered hMSCs was assessed in vitro and in vivo. RESULTS: Genetically engineered hMSCs displayed enhanced proliferation and osteogenic differentiation in culture. In vivo, transplanted genetically engineered hMSCs were able to engraft and form bone and cartilage in ectopic sites, and regenerate bone defects (non-union fractures) in mice radius bone. Importantly, the same results were obtained with hMSCs isolated from a patient suffering from osteoporosis. CONCLUSIONS: hMSCs represent a novel platform for skeletal gene therapy and the present results suggest that they can be genetically engineered to express desired therapeutic proteins inducing specific differentiation pathways. Moreover, hMSCs obtained from osteoporotic patients can restore their osteogenic activity following human BMP-2 gene transduction, an important finding in the future planning of gene therapy treatment for osteoporosis.

Retrograde nailing of femoral fractures distal to previous osteosynthesis.

As the proportion of elderly in the population grows, the incidence of femoral fractures distal to previous proximal osteosynthesis is increasing. When the gap between two rigid load-bearing fixations consists of osteopenic bone, the risk of further fractures increases. Herein the authors describe a load-sharing device that stabilizes the fracture and eliminates the osteopenic gap, allowing early mobilization and rapid return to the preinjury level of activity.

Combined injuries to the lower limbs.

Several patterns of severe lower limb injuries are presented. They all indicate high energy trauma and affect the immediate care of the patient. The improvement of evacuation systems and resuscitating methods in intensive care create many reconstruction challenges for the orthopaedic surgeon. Awareness of the different combinations which are presented can serve as a tool that may be helpful in these demanding injuries. Guidelines for management of combined injuries are essential to improve the outcome of these life-threatening situations.

Selective delivery of doxorubicin to patients with breast carcinoma metastases by stealth liposomes.

BACKGROUND: Stealth liposomes hold promise as a mode of delivering cytotoxic agents selectively to tumors in cancer patients. The objective of this study was to determine whether stealth liposomal doxorubicin accumulates selectively in bone metastases based on clinical material obtained from two patients with breast carcinoma. METHODS: Tumor tissue was obtained from two women (ages 33 years and 41 years, respectively) with metastatic breast carcinoma who responded to treatment with stealth liposomal doxorubicin and later underwent a surgical fixation procedure to treat a pathologic fracture of the femur. Drug levels in the tumor and adjacent muscle were examined by high performance liquid chromatography analysis in both patients and by fluorescence microscopy in one of the patients. RESULTS: Bone tumor fragments obtained during surgery performed 6 days after the administration of the 12th course of stealth liposomal doxorubicin in 1 patient and 12 days after the administration of the 16th course of stealth liposomal doxorubicin in the second patient had a 10-fold greater concentration of liposomal doxorubicin than tumor free muscle. Doxorubicin fluorescence and specific nuclear staining showed good colocalization, thus confirming the presence of the liposome-delivered drug in the nuclei of tumor cells. CONCLUSIONS: Using skeletal muscle as a comparator, stealth liposomal doxorubicin accumulates selectively in metastatic breast carcinoma cells within bone.

Focal, steroid responsive myositis causing dropped head syndrome.

The dropped head syndrome, which occurs in a variety of neuromuscular disorders, is usually not due to an inflammatory process and generally either self-limited or nonresponsive to therapy. We present an 80-year-old woman who developed progressive neck weakness over a few months due to a focal and restricted inflammatory process involving the neck extensor muscles. She responded dramatically to treatment with immunosuppressive therapy.

In vitro forces in the normal and cruciate-deficient knee during simulated squatting motion.

Three orthogonal components of the tibiofemoral and patellofemoral forces were measured simultaneously for knees with intact cruciate ligaments (nine knees), following anterior cruciate ligament resection (six knees), and subsequent posterior cruciate ligament resection (six knees). The knees were loaded using an experimental protocol that modeled static double-leg squat. The mean compressive tibial force increased with flexion angle. The mean anteroposterior tibial shear force acted posteriorly on the tibia below 50 deg flexion and anteriorly above 55 deg. Mediolateral shear forces were low compared to the other force components and tended to be directed medially on both the patella and tibia. The mean value of the ratio of the resultant tibial force divided by the quadriceps force decreased with increasing flexion angle and was between 0.6 and 0.7 above 70 deg flexion. The mean value of the ratio of the resultant tibiofemoral contact force divided by the resultant patellofemoral contact force decreased with increasing flexion and was between 0.8 and 1.0 above 55 deg flexion. Cruciate ligament resection resulted in no significant changes in the patellar contact forces. Following resection of the anterior cruciate ligament, the tibial anteroposterior shear force was directed anteriorly over all flexion angles tested. Subsequent resection of the posterior cruciate ligament resulted in an approximately 10 percent increase in the quadriceps tendon and tibial compressive force.

[CT-guided excision of osteoid osteoma].

CT-guided excision of osteoid osteoma is a new surgical technique that enables accurate resection of the nidus during 1-day hospitalization. We present 5-year results in 42 patients (26 males and 16 females, mean age 18 years, range 3-46). In 40 out of 42, complaints disappeared immediately after the procedure. The recovery period was short and the return to normal activity was faster than in the open surgical approach. Complications were minimal and transient.

Fracture of the posterior body of the talus--the hidden fracture.

Fractures of the entire posterior process of the talus are rare and may be easily missed. Both the ankle joint and the subtalar joint are involved. This leads to malunion and early degenerative changes. We describe four patients with fractures of the posterior process of the talus which were initially missed. The patients were treated conservatively, and early mobilization or cast immobilization did not change the poor late results. The pitfalls in the diagnosis and imaging evaluation of such fractures are discussed.

"Krammer splint technique" for immediate measuring of intramedullary nails.

We illustrate a simple and quick method of measuring the length of the intramedullary nail. The method utilizes the intervals between the metal ranks of the Krammer splint, and it has proved especially useful in the care of multitrauma victims.