Evaluation of coagulation profile in dogs with ACTH-dependent hyperadrenocorticism compared to healthy dogs by rotational thromboelastometry (ROTEM).

Despite their low morbidity, thromboembolic events in hyperadrenocorticism are associated with high mortality. Identifying the main hemostatic abnormalities will improve the prophylactic approach of these canine patients. The aim of this study was to evaluate hemostatic alterations related with ACTH-dependent HAC and its association with hypercoagulable state. For this purpose, 25 dogs diagnosed with ACTH-dependent HAC were compared with 28 healthy dogs as a control group. The hemostatic variables included platelet count, antithrombin, fibrinogen, D-dimer, PT, aPTT, rotational thromboelastometry (ROTEM) and platelet aggregation. Results showed a hypercoagulable state in 32% (8/25) dogs by ROTEM, which had at least 2 of the next features: decreased coagulation time (CT) or clot formation time (CFT) on INTEM (5/25) or EXTEM (4/25); increased maximum clot firmness (MCF) on INTEM (9/25), EXTEM (6/25) and FIBTEM (9/25). These same variables had a significant difference (P≤ 0.05) compared with the control group, as well as the parameters of α-angle and CT. Median fibrinogen levels (310 vs.178mg/dL), mean platelet aggregation (11.1 vs. 7.9 Ohms), median platelet count (360 vs. 225 ×103/µL) and mean antithrombin activity (140 vs. 119%) were increased in ACTH-dependent HAC dogs compared to control group. PT (7.1 vs. 8.0seconds) and aPTT (11.6 vs. 15.2seconds) were also shortened in ACTH-dependent HAC dogs. Our findings confirm the presence of a hypercoagulable tendency in dogs with HAC. Although multifactorial, fibrinogen concentration and MCF FIBTEM showed the relevance of this protein for hypercoagulability in HAC.

Therapy of Canine Hyperlipidemia with Bezafibrate.

BACKGROUND: Bezafibrate (BZF) is effective in the treatment of hypertriglyceridemia in human patients, but there are no data on its use in dogs. OBJECTIVE: To assess the safety of BZF in hyperlipidemic dogs and its efficacy in decreasing serum triglyceride (TG) and cholesterol (CHO) concentrations. ANIMALS: Forty-six dogs, 26 females and 20 males, mean (±SD) age of 9 (±3) years, with TG ≥150 mg/dL (33 dogs also were hypercholesterolemic [>300 mg/dL]). METHODS: Prospective, uncontrolled clinical trial. Dogs were treated with bezafibrate once daily, using 200 mg tablets at a dosage of 4-10 mg/kg (depending on body weight). Serum TG and CHO concentrations and alanine aminotransferase (ALT) and creatine kinase (CK) activity before and after 30 days of treatment were compared. RESULTS: Sixteen dogs (34.8%) had primary hyperlipidemia, and 30 dogs (65.2%) had secondary hyperlipidemia (including spontaneous hyperadrenocorticism [41.3%, n = 19/46], chronic treatment with glucocorticoids [10.8%, n = 5/46], and hypothyroidism [15.2%, n = 7/46]). After 30 days, serum TG concentration normalized (<150 mg/dL) in 42 dogs (91.3%) and CHO concentration normalized (<270 mg/dL) in 22 of 33 dogs (66.7%). There was no difference in baseline TG concentration between the primary and secondary hyperlipidemia subgroups, but the decrease in TG concentration after treatment was greater in the primary hyperlipidemia subgroup. No adverse effects were observed, but ALT activity decreased significantly after 30 days of treatment. CONCLUSIONS AND CLINICAL IMPORTANCE: Over 30 days, BZF was safe and effective in treatment of primary and secondary hyperlipidemia in dogs.

A new training model for robot-assisted urethrovesical anastomosis and posterior muscle-fascial reconstruction: the Verona training technique.

A training model is usually needed to teach robotic surgical technique successfully. In this way, an ideal training model should mimic as much as possible the "in vivo" procedure and allow several consecutive surgical simulations. The goal of this study was to create a "wet lab" model suitable for RARP training programs, providing the simulation of the posterior fascial reconstruction. The second aim was to compare the original "Venezuelan" chicken model described by Sotelo to our training model. Our training model consists of performing an anastomosis, reproducing the surgical procedure in "vivo" as in RARP, between proventriculus and the proximal portion of the esophagus. A posterior fascial reconstruction simulating Rocco's stitch is performed between the tissues located under the posterior surface of the esophagus and the tissue represented by the serosa of the proventriculus. From 2014 to 2015, during 6 different full-immersion training courses, thirty-four surgeons performed the urethrovesical anastomosis using our model and the Sotelo's one. After the training period, each surgeon was asked to fill out a non-validated questionnaire to perform an evaluation of the differences between the two training models. Our model was judged the best model, in terms of similarity with urethral tissue and similarity with the anatomic unit urethra-pelvic wall. Our training model as reported by all trainees is easily reproducible and anatomically comparable with the urethrovesical anastomosis as performed during radical prostatectomy in humans. It is suitable for performing posterior fascial reconstruction reported by Rocco. In this context, our surgical training model could be routinely proposed in all robotic training courses to develop specific expertise in urethrovesical anastomosis with the reproducibility of the Rocco stitch.

Surgical treatment of Peyronie's disease with small intestinal submucosa graft patch.

The objective of the study was to report our results using a porcine small intestinal submucosa graft (Surgisis ES, Cook Medical) for tunica albuginea substitution after plaque incision. We retrospectively evaluated patients surgically treated at our institution for Peyronie's disease (PD) by means of plaque incision and porcine small intestinal submucosa grafting (Surgisis) between 2009 and 2013. At the same time a literature review was conducted, searching for similar reports and results. Forty-four patients were identified who had been diagnosed with PD between 2009 and the beginning of 2013, and had been treated with corporoplasty, plaque incision and grafting with Surgisis for a severe curvature of the penis. Curvature of the penis was dorsal in 40 patients (90%) and laterally on the right in 4 patients (10%). Mean duration of surgery was 165 min (range 90-200). Mean size of the graft was 6.5 cm(2) and the mean follow-up was 19.2 months (range 11-48). In patients with severe curvature of the penis due to PD and the need for corporoplasty with plaque incision and graft placement, Surgisis represents a good option with a low risk of complications, below the rate described with previously investigated graft tissues.

Densitometric kneecap changes after unilateral knee arthroplasty.

PURPOSE: To assess bone density of kneecaps in subjects with femoro-tibial prosthesis before and after surgery by means of DEXA examination. SUBJECTS AND METHODS: We examined 34 patients with unilateral femoro-tibial prosthesis, 20 healthy subjects of the same age and non-carriers of knee replacement and 14 healthy young adult subjects. All the data sets were analysed by two radiologists (AS and AM). The coincidence of the results between the two specialists was evaluated by means of Cohen's Kappa index and the results were considered statistically significative if p value is < of 0.05. RESULTS: The values of patellar BMD in the group of 34 patients, were: a minimum of 0.386 g/cm(2) (K = 0.879, p = 0.0012), a maximum 1.707 g/cm(2) (K = 0.886, p = 0.0016). The comparison between the left and right knee showed the following data: minimum difference 0.034 g/cm2 (K = 0.901, p = 0.0015), maximum difference of 0.622 g/cm(2) (K = 0.908, p = 0.0017), the average was found to be of 0.277 g/cm(2) (K = 0.894, p = 0.0018). But this difference tends to decrease 6 months after surgery. In the group of healthy young adults, we obtained the following values: a minimum of 0.782 g/cm(2) (K = 0.907, p = 0.0025), maximum 1.503 g/cm(2) (K = 0.932, p = 0.0012). Between both knees, the difference was minimal 0.006 g/cm(2) (K = 0.951, p = 0.0035) and maximum 0.096 g/cm(2) (K = 0.926, p = 0.0007) with an average difference of 0.058 g/cm(2) (K = 0.954, p = 0.0026). In the group of healthy subjects of the same age and non-carriers of knee replacement the values were average higher. A maximum value of 1.134 g/cm(2) (K = 0.894, p = 0.0028) and a minimum value of 0.944 g/cm(2) (K = 0.892, p = 0.0023) were found; between both knees a minimum difference of 0.010 g/cm(2) (K = 0.918, p = 0.0047) and a maximum of 0.090 g/cm(2) (K = 0.937, p = 0.0017) were found, with an average difference of 0.052 g/cm(2) (K = 0.956, p = 0.0024). CONCLUSIONS: DEXA examination of the patellar is recommended as a supplementary study to the clinical and radiological standard exams because it is able to provide additional information to determine when to intervene surgically, on the basis of patellar bone density values.

Incidental discovery of testicular microlithiasis: what is the importance of ultrasound surveillance? Two case reports.

Many studies have demonstrated an association between diffuse bilateral testicular microlithiasis (TM) and gonadal and extragonadal germ cell tumors. Nevertheless, it is still uncertain whether ultrasound surveillance is really necessary in patients with TM in the absence of other risk factors such as previous testicular cancer, a history of cryptorchidism or testicular atrophy. We report the cases of a 33- and a 39-year-old man presenting with a retroperitoneal extragonadal tumor. The first patient underwent an MRI examination in order to rule out a lumbosacral hernia: MRI images showed no slipped disks but a voluminous retroperitoneal solid mass. The histological analysis revealed an immature teratoma. The second patient came to the emergency department complaining of abdominal pain, vomiting, weight loss and mild jaundice: ultrasound examination showed a large, ill-defined heterogeneous abdominal mass, confirmed by CT and MRI examination. The histology diagnosed a yolk sac tumor. In both patients, the testicular sonography was performed to rule out a focal lesion, but it displayed bilateral TM without a focal testicular mass. Based on our direct experience, we highlight the importance of annual ultrasonographic surveillance of the testis and the retroperitoneal space in patients with occasionally detected TM.

Ureterocele ectópica em cão: relato de caso / Ectopic ureterocele in a dog: a case report

Relataram-se o quadro clínico, o diagnóstico, o tratamento e o acompanhamento de um cão com ureterocele ectópica e hidronefrose/hidroureter associados. Após as informações obtidas nos exames laboratoriais e de diagnóstico por imagem, o animal foi submetido à ureterocelectomia e à neoureterostomia. O cão apresentou evolução favorável após a conduta terapêutica. Apesar de pouco frequente, a ureterocele deve ser considerada como diagnóstico diferencial em animais jovens com histórico de incontinência urinária.

The clinical features, diagnosis, treatment and monitoring of a dog with an ectopic ureterocele and concomitant hydronephrosis/hydroureter were reported. After the information obtained in laboratory tests and imaging diagnosis, the animal was submitted to ureterocelectomy and neoureterostomy. The dog presented a favorable evolution after the therapy. Although infrequent, the ureterocele should be considered a differential diagnosis in young animals with history of urinary incontinence.

Germline mutation analysis of Tpit in Poodle dogs with ACTH-dependent hyperadrenocorticism / Análise de mutação germinativa do Tpit em cães da raça Poodle com hiperadrenocorticismo ACTH-dependente

There is a high incidence of pituitary-dependent hyperadrenocorticism (PDH) in Poodle dogs, with family members being affected by the disease, suggesting a genetic involvement. Tpit is an obligate transcription factor for the expression of pro-opiomelanocortingene and for corticotroph terminal differentiation. The aim of the present study was to screen the Tpit gene for germline mutations in Poodles with PDH. Fifty Poodle dogs (33 female, 8.71±2.8 years) with PDH and 50 healthy Poodle dogs (32 females, 9.4241 2.8 years) were studied. Genomic DNA was isolated from peripheral blood, amplified by PCR and submitted to automatic sequence. No mutation in the coding region of Tpit was found, whereas the new single nucleotide polymorphism p.S343G, in heterozygous state, was found in the same frequency in both PDH and control groups. We concluded that Tpit gain-of-function mutations are not involved in the etiology of PDH in Poodle dogs.

O hiperadrenocorticismo ACTH-dependente (HACAD) apresenta elevada incidência em cães da raça Poodle, sendo que membros da mesma família têm sido acometidos pela doença, sugerindo envolvimento genético. Tpit é um fator de transcrição obrigatório para a expressão do gene da pro-opiomelanocortina e para a diferenciação terminal dos corticotrofos. O objetivo deste trabalho foi pesquisar mutações germinativas no gene Tpit em Poodles com HACAD. Cinquenta cães da raça Poodle, 33 fêmeas, média de idade de 8,71±2,8 anos, com HACAD, e 50 cães Poodles saudáveis, 32 fêmeas, média de idade de 9,4±2,8 anos, foram estudados. Mutações na região codificadora do gene Tpit não foram identificadas. Foi observado um novo polimorfismo em heterozigose, p.S343G, com a mesma frequência no grupo de cães com HACAD e no grupo-controle. Conclui-se que a mutação ativadora no gene Tpit não está envolvida na patogênese do hiperadrenocorticismo ACTH-dependente em cães da raça Poodle.

Giant lipoma of descending colon diagnosed at CT: report of a case.

Colonic lipomas are rare benign lesions, detected accidentally. These are often asymptomatic, but large lipoma may produce symptoms as abdominal pain, nausea, weight loss, diarrhea, constipation, hemorrhage, and intussusception. Colonic lipomas are more often localized in the ascending colon: literature reports less than 20 symptomatic cases situated in the descending colon. We report the case of a young man with a colonic giant lipoma diagnosed at Computed Tomography, who presented with rectum bleeding and 5-kg weight loss. The case was interesting because of the patient's young age, the tumor's location in the left side of the colon and the giant size (5.5 cm).

Magnetic resonance cholangiography: past, present and future: a review.

INTRODUCTION: Magnetic resonance cholangio-pancreatography (MRCP) is a valuable method for the evaluation of biliary and pancreatic diseases and a valuable alternative to endoscopic retrograde cholangiopancreatography (ERCP). It is noninvasive and does not require the use of contrast material or ionizing radiation. Since its introduction in 1991, this technique has significantly improved in spatial resolution, now allowing the accurate assessment of the major bilio-pancreatic diseases. STATE OF THE ART: MRCP is commonly performed with heavily T2-weighted sequences in order to highlight static fluids, as those contained in dilated pancreatic and biliary ducts. Newest MR equipments allow to perform MRCP within 10-15 minutes, due to the availability of ultra-fast sequences. Currently, MRCP is widely performed as a primary imaging modality for the assessment of obstructive jaundice and other benign or malignant bilio-pancreatic ducts abnormalities. The primary MRCP application is the evaluation of biliary obstructions due to choledocholithiasis, iatrogenic strictures, cholangiocarcinoma or pancreatic carcinoma. Other MRCP applications include the assessment of the exocrine pancreatic function, following secretin stimulation. Whenever needed, the MRCP may be completed with a conventional contrast-enhanced magnetic resonance imaging (MRI) of the upper abdomen and functional studies as well, thus providing an all-in-one mophological and functional study of the pancreas and biliary system. More recent applications include the possibility of 3D reconstructions and the use of hepato-biliary contrast agents, that provide a higher definition of the biliary tree, both in pathologic and normal conditions. The introduction of 3Tesla magnets could provide higher anatomic detail. CONCLUSIONS: In the next years the role of MRCP will further expand, due to the availability of faster sequences, 3D imaging and functional studies.

Preoperative T and N staging of gastric cancer: magnetic resonance imaging (MRI) versus multi detector computed tomography (MDCT).

BACKGROUND AND AIM: Multi Detector Computed Tomography (MDCT) is widely used in the preoperative staging of gastric cancer. MRI has an emerging role in the evaluation of intestinal diseases, although its role in the staging of gastric cancers is still to be defined. The aim of our study was to compare the diagnostic accuracy of MDCT and Magnetic Resonance Imaging (MRI) in the diagnosis and preoperative staging of gastric cancer, in comparison with histopathology. MATERIALS AND METHODS: Twenty-five patients with an endoscopic diagnosis of gastric cancer underwent preoperative contrast-enhanced MDCT and MRI, blind to the results of endoscopy. MDCT (64 slices) was performed after oral administration of 800-1000 mL of tap water and scopolamine injection five minutes before the examination. The scan was performed in the axial plane before and after intravenous injection of iodinate contrast medium. Multiplanar reconstruction images were obtained on coronal and sagittal planes. MRI was performed with a 1.5 T Magnet, using the same patient's preparation, by acquiring T2-weighted HASTE sequences, with or without fat saturation (FS), True FISP (True fast imaging with steady state precession) and T1-weighted VIBE (Volumetric interpolated breath-hold examination) sequences, with and without FS, before and after contrast agent (gadolinium) i.v. injection. Gold standards (GS) were surgical and histopathological findings. Two groups of radiologists, blind each other, analyzed MRI images and MDCT findings, and related to GS results. RESULTS: Detection rate of gastric lesions and T staging for gastric cancer were similar for MRI and MDCT (92%); MRI imaging was superior than MDCT in staging the T parameter (60% versus 48%); the accuracy of MRI imaging and 64-MDCT did not differ significantly in the evaluation of N staging (68% versus 72%). CONCLUSIONS: Both MRI and MDCT were comparable in staging gastric cancer. MRI was more accurate in evaluation of T stage than MDCT, although both imaging modalities showed low accuracy in detection of early gastric cancer and in differentiation of T2 from T3 stage.

Quaternary structure of the human Cdt1-Geminin complex regulates DNA replication licensing.

All organisms need to ensure that no DNA segments are rereplicated in a single cell cycle. Eukaryotes achieve this through a process called origin licensing, which involves tight spatiotemporal control of the assembly of prereplicative complexes (pre-RCs) onto chromatin. Cdt1 is a key component and crucial regulator of pre-RC assembly. In higher eukaryotes, timely inhibition of Cdt1 by Geminin is essential to prevent DNA rereplication. Here, we address the mechanism of DNA licensing inhibition by Geminin, by combining X-ray crystallography, small-angle X-ray scattering, and functional studies in Xenopus and mammalian cells. Our findings show that the Cdt1:Geminin complex can exist in two distinct forms, a "permissive" heterotrimer and an "inhibitory" heterohexamer. Specific Cdt1 residues, buried in the heterohexamer, are important for licensing. We postulate that the transition between the heterotrimer and the heterohexamer represents a molecular switch between licensing-competent and licensing-defective states.

Hyperhomocysteinemia is associated with cognitive impairment in multiple sclerosis.

Hyperhomocysteinemia (HHcy) has been associated with cognitive impairment in various neurological diseases. Cognitive impairment occurs early in multiple sclerosis (MS). Conflicting data have been reported regarding plasma total homocysteine (tHcy) levels in MS patients, and the impact of HHcy on cognitive impairment in MS is not known. This study investigated whether plasma total homocysteine levels are increased in MS and if HHcy is associated with cognitive impairment in MS. We compared tHcy levels in 94 patients with MS and 53 healthy age-matched controls. We used a neuropsychological test battery that included the Raven's Coloured Progressive Matrices, the Visual Search Test, the Trail Making Test A and B, the Immediate and Delayed Recall of a Short Story, the 30 Paired Word Associates, the Rey-Osterrieth Complex Figure Test, and the Semantic and Verbal Fluency Tests. Clinical (sex, age, type of MS, relapse, disease duration, coexisting disease, smoking habit, and physical disability) and laboratory variables (HHcy, low serum levels of folate and vit.B12, MTHFR genotype) were evaluated for their ability to predict cognitive impairment. The mean tHcy was higher in patients (13.19 micromol/L, SD5.58) than in controls (9.81 micromol/L, SD2.53; p < 0.001). Univariate analysis determined the following factors to be associated with cognitive impairment: higher age at observation, chronic progressive course of disease, longer disease duration,moderate or severe physical disability, and frequency of HHcy. With multivariate regression analysis, there remained a significant association only between frequency of HHcy and cognitive impairment (beta 0.262, p = 0.01). We conclude that tHcy levels are increased in MS and that HHcy is associated with cognitive impairment in this disease.

Stiff person syndrome and motor mononeuropathy with conduction block: a singular association.

The "Stiff person syndrome"(SPS) is a rare dysimmune chronic neurological disorder, sometimes paraneoplastic, characterized by progressive stiffness, painful persistent or spasmodic muscle contractions, mostly involving spine and lower extremities. In 60 to 90 percent of cases, non-paraneoplastic forms are associated to the presence of anti-glutamic acid decarboxylase (anti-GAD) antibodies in the cerebrospinal fluid and in the serum, while anti-amphiphysin antibodies are frequently associated to paraneoplastic types. The relevant treatment consists of three basic approaches: increase in the inhibitory processes in charge of muscle activity control, re-modulation of the immune response, removal of any associated neoplasia. Indications regarding the efficacy of high-dose intravenous immunoglobulin (IVIG) also in this dysimmune pathology are on the increase. We described an unusual case of autoimmune SPS associated with an exclusively motor left peroneal nerve neuropathy, with conduction block, treated with high-dose intravenous immunoglobulin (IVIG), oral cyclosporine, sodium valproate, baclofen and diazepam.

Obstructive primary bladder neck disease: evaluation of the efficacy and safety of alpha1-blockers.

OBJECTIVE: The efficacy and safety of using alpha(1)-adrenergic blockers for treating primary bladder neck obstruction in young and middle-aged men was assessed as the first therapeutic step, before surgery, in a symptomatic non-neurogenic selected group of patients. MATERIALS AND METHODS: From January 1995 to December 2001, primary bladder neck obstruction was diagnosed in 41 men whose average age was 43 years. All of them were evaluated by a complete clinical history, American Urological Association (AUA) symptom score index, physical examination, uroflowmetry, transabdominal ultrasound prostatic volume determination, ultrasound post-void residual determination, videourodynamics including pressure-flow analysis and upper urinary tract screening with renal ultrasound or an excretory urogram. A full daily dose of alpha(1)-adrenergic blockers (alfuzosin or tamsulosin) was administered for at least 6 months. Successful treatment was defined as improved symptoms, voiding diary, maximum flow rate and pressure-flow parameters. Patients who did not gain improvement of symptoms with pharmacological treatment were offered surgery. RESULTS: Overall, pharmacological treatment was successful in 29/41 patients (70.7%) whereas bladder neck endoscopic incision was mandatory in 12/41 (29.3%). CONCLUSIONS: alpha(1)-Blockers were effective and safe for treating young and middle-aged men with symptomatic bladder neck obstruction.

Pharmacokinetic study of intravesical gemcitabine in carcinoma in situ of the bladder refractory to bacillus Calmette-Guérin therapy.

INTRODUCTION: Gemcitabine, a chemotherapeutic agent, has been shown to be active against transitional cell cancer of the bladder. The aim of the study was to determine the pharmacokinetic profile of gemcitabine, administered intravesically in patients with carcinoma in situ(CIS). MATERIAL AND METHODS: Nine patients with CIS refractory to intravesical bacillus Calmette-Guérin (BCG) therapy were enrolled. Gemcitabine was given in 50 ml 0.9% NaCl by catheterization and held in the bladder for 1 h, once weekly for 6 consecutive weeks. The pharmacokinetics for gemcitabine metabolites were performed in plasma and serum. Dose levels were: 1,000, 1,250, and 1,500 mg. Clinical evaluation was repeated 4 weeks after therapy and thereafter every 6 months. RESULTS: Grade-1 neutropenia was observed only in 1 patient. Grade-1 urinary frequency and hematuria were observed in 1 and 3 patients, respectively. No grade 2-4 toxicity or clinically relevant myelosuppression were observed. Gemcitabine was detectable in serum, but with an irrelevant pharmacological effect, in only 1 patient treated with 1,500 mg of gemcitabine. With regard to activity, after 6 instillations of this drug, 4 complete responses were observed. CONCLUSION: Intravesical gemcitabine is well tolerated and safe. No systemic absorption with a clinical or pharmacological effect was detected and only slightly irritative bladder symptoms were observed. These results warrant further investigation in phase-II trials.

Cerebellar subacute syndrome due to corticosteroid-responsive encephalopathy associated with autoimmune thyroiditis (also called "Hashimoto's encephalopathy").

The corticosteroid-responsive encephalopathy associated with autoimmune thyroiditis (the so-called "Hashimoto's Encephalopathy") is a rare disorder with multiple symptomatology, breaking out with an acute or subacute onset and having a relapsing course, not correlated to thyroid hormone levels, with autoimmune pathogenesis, and usually associated with Hashimoto's thyroiditis. In this paper, we report on a case study regarding a 46 year-old woman showing a subacute course cerebellar syndrome, associated with Hashimoto's thyroiditis, diagnosed as "Hashimoto's encephalopathy". The possible pathogenesis and the major aspects of the differential diagnostic sector are discussed with particular reference to an ataxic syndrome caused by a progressive non-familial adult onset cerebellar degeneration (PNACD), associated with the thyroid disease itself.

Correlates of Mycobacterium tuberculosis infection in a prison population.

Prisons represent a crucial setting for tuberculosis control. Currently, there is scarce information concerning Mycobacterium tuberculosis (MT) infection in European prisons, and no data are available for Italy. This study aims to describe the prevalence and correlates of MT infection in an Italian prison population. In this multicentre cross-sectional study, 1,247 inmates from nine prisons were recruited and asked to undergo questioning regarding socio-economic and demographic variables, tuberculin skin testing (TST), chest radiographs and testing for HIV, hepatitis B and hepatitis C virus infection. TST was positive in 17.9% of the 448 evaluable subjects. With multivariate logistic regression (performed among male inmates), MT infection was correlated with age (adjusted odds ratio (OR) 4.12 for inmates aged 31-40 yrs; 3.78 for those aged >40 yrs), being foreign-born (OR = 4.9), education < or =5 yrs (OR = 1.88) and length of detention (increased risk per yr: 11%). As with elsewhere in the world, the prison system in Italy features a population with an increased rate of Mycobacterium tuberculosis infection and at-risk rate for Mycobacterium tuberculosis transmission. Improved tuberculosis surveillance and control measures are deemed necessary in correctional facilities nationwide, especially for subjects with the above risk factors and those who are HIV infected, in whom the tuberculin skin testing can be misleading. The screening of entrants is particularly important to avoid undiagnosed smear-positive tuberculosis cases.

Analysis of heterodimer formation by Xklp3A/B, a newly cloned kinesin-II from Xenopus laevis.

kinesin-II motor proteins are composed of two different kinesin-like motor proteins and one cargo binding subunit. Here we report the cloning of a new member of the kinesin-II superfamily, Xklp3A from Xenopus laevis, which forms a heterodimeric complex with Xklp3B. The heterodimer formation properties between Xklp3A and B have been tested in vitro using reticulocyte lysate expression and immunoprecipitation. To this end we produced a series of Xklp3A and B constructs of varying length and tested their propensity for heterodimer formation. We could demonstrate that, in contrast to conventional kinesin, the critical domains for heterodimer formation in Xklp3A/B are located at the C-terminal end of the stalk. Neither the neck nor the highly charged stretches after the neck region, which are typical of kinesins-II, are required for heterodimer formation, nor do they prevent homodimer formation. Dimerization is controlled by a cooperative mechanism between the C-terminal coiled-coil segments. Classical trigger sites were not identified. The critical regions for dimerization exhibit a very high degree of sequence conservation among equivalent members of the kinesin-II family.

Identification of the yeast mitochondrial transporter for oxaloacetate and sulfate.

Saccharomyces cerevisiae encodes 35 members of the mitochondrial carrier family, including the OAC protein. The transport specificities of some family members are known, but most are not. The function of the OAC has been revealed by overproduction in Escherichia coli, reconstitution into liposomes, and demonstration that the proteoliposomes transport malonate, oxaloacetate, sulfate, and thiosulfate. Reconstituted OAC catalyzes both unidirectional transport and exchange of substrates. In S. cerevisiae, OAC is in inner mitochondrial membranes, and deletion of its gene greatly reduces transport of oxaloacetate sulfate, thiosulfate, and malonate. Mitochondria from wild-type cells swelled in isoosmotic solutions of ammonium salts of oxaloacetate, sulfate, thiosulfate, and malonate, indicating that these anions are cotransported with protons. Overexpression of OAC in the deletion strain increased greatly the [(35)S]sulfate/sulfate and [(35)S]sulfate/oxaloacetate exchanges in proteoliposomes reconstituted with digitonin extracts of mitochondria. The main physiological role of OAC appears to be to use the proton-motive force to take up into mitochondria oxaloacetate produced from pyruvate by cytoplasmic pyruvate carboxylase.