Multidrug resistance in Salmonella isolates of swine origin: mobile genetic elements and plasmids associated with cephalosporin resistance with potential transmission to humans.

The emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context requiring continuous surveillance. Resistance to ciprofloxacin and cephalosporins is of particular concern. Since pigs are a relevant source of foodborne Salmonella for human beings, we studied transmissible AMR genes and MGE in a collection of 83 strains showing 9 different serovars and 15 patterns of multidrug resistant (MDR) previously isolated from pigs raised in the conventional breeding system of Northern Spain. All isolates were susceptible to ciprofloxacin and three isolates carried blaCMY-2 or blaCTX-M-9 genes responsible for cefotaxime resistance. Filter mating experiments showed that the two plasmids carrying blaCTX-M-9 were conjugative while that carrying blaCMY-2 was self-transmissible by transformation. Whole-genome sequencing and comparative analyses were performed on the isolates and plasmids. The IncC plasmid pSB109, carrying blaCMY-2, was similar to one found in S. Reading from cattle, indicating potential horizontal transfer between serovars and animal sources. The IncHI2 plasmids pSH102 in S. Heidelberg and pSTM45 in S. Typhimurium ST34, carrying blaCTX-M-9, shared similar backbones and two novel "complex class 1 integrons" containing different AMR and heavy metal genes. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.IMPORTANCEThe emergence of foodborne Salmonella strains carrying antimicrobial resistance (AMR) in mobile genetic elements (MGE) is a significant public health threat in a One Health context. Since pigs are a relevant source of foodborne Salmonella for humans, in this study, we investigate different aspects of AMR in a collection of 83 Salmonella showing nine different serovars and 15 patterns of multidrug resistant (MDR) isolated from pigs raised in the conventional breeding system. Our findings emphasize the importance of sequencing techniques to identify emerging AMR regions in conjugative and stable plasmids from livestock production. The presence of MGE carrying clinically relevant AMR genes raises public health concerns, requiring monitoring to mitigate the emergence of bacteria carrying AMR genes and subsequent spread through animals and food.

Vaccine properties of Brucella melitensis 16MΔwzm and reactivation of placental infection in pregnant sheep.

Brucellosis, a worldwide zoonotic disease, is endemic in many developing countries. Besides causing significant economic losses for the livestock industry, it has severe consequences for human health. In endemic regions, small ruminants infected by Brucella melitensis are the main source of human brucellosis. Rev1, the only vaccine currently recommended to control the disease in sheep and goats, has several drawbacks. Rough lipopolysaccharide (R-LPS) mutants have been tested as alternatives, but most lack efficacy. Those in the Wzm/Wzt system responsible for O-polysaccharide export to the periplasm have been proposed as promising vaccine candidates, although to date they have been scarcely investigated in the natural host. In the present work, we studied the biological properties of a 16MΔwzm in-frame deletion mutant, including its safety in pregnant mice and sheep. In mice, 16MΔwzm prevented placental and fetal infections before parturition and protected against B. melitensis and Brucella ovis infections. In sheep, 16MΔwzm was equally safe in lambs, rams, and non-pregnant ewes, inducing some transient Rose Bengal reactions (<7 weeks). The serological reactions occurred earlier and more strongly in pregnant than in non-pregnant ewes and were significantly reduced when conjunctival rather than subcutaneous vaccination was used. In ewes vaccinated at mid-pregnancy, 16MΔwzm was not shed in vaginal discharges during the pregnancy and did not induce abortions/stillbirths. However, some ewes showed a transitory reactivation of infection in placentas and/or milk at parturition, accompanied by a seroconversion in smooth LPS (S-LPS) and/or R-LPS tests. Overall, 16MΔwzm can be considered as a safe vaccine for lambs, rams, and non-pregnant ewes, but its use at mid-pregnancy should be avoided to prevent vaccine dissemination at parturition. If the efficacy results against B. melitensis and B. ovis observed in mice are confirmed by further studies in the natural host, 16MΔwzm could constitute a useful vaccine.

Assessment of pharmacogenomic SLCO1B1 assay for prediction of neuromuscular pain in type 2 diabetes mellitus and cardiovascular patients: preliminary results.

OBJECTIVE: At present, several strategies for preventing neuromuscular pain in Type 2 Diabetes Mellitus (T2DM) have been investigated. Recently, findings on genetic variants associated with adverse events to statin-based therapy have been reported. The study aimed at measuring whether Pharmacogenomics (PGx) profile can affect neuromuscular pain in patients carrying T2DM and cardiovascular diseases. An extensive panel of 5 polymorphisms on 4 candidate genes, previously validated as significant markers related to Sulphonylureas and Glitinides (SU-G) plus Simvastatin neuromuscular toxicity, is herein analyzed and discussed. PATIENTS AND METHODS: We genotyped 76 T2DM patients carrying cardiovascular dyscrasia undergone anti-diabetic and anti-cholesterolemic polypharmacy. 35 subjects out of the total received concurrent SU-G and Statin-based therapy. Candidate variants consisted of drug transporters, such as Solute Carrier Organic 1B1 (SLCO1B1) Val174Ala ATP-binding cassette subfamily B member (ABCB1), subfamily C member 8 (ABCC8), and drug biotransformers of Cytochrome P450 Family (CYP) including CYP2C9*2 CYP2C9*3 CYP2C8*3, and CYP3A4*22. Moreover, we also focused on an early outline evaluation of the genotyping costs and benefits. RESULTS: 6 out of 35 patients treated with SU-G plus statins (17.1% experienced adverse neuropathy events). Pharmacogenomics analysis showed a lack of any correlation between candidate gene polymorphisms and toxicity, except for the SLCO1B1 T521C allele; 14.3% of patients had a high risk for grade >2 neuromuscular pain (Odds Ratio [OR] 2.61.95% CI 0.90-7.61, p=0.03). CONCLUSIONS: The clinical polymorphism effectiveness outlined therein will be assured by diagnostic improvements suitable for driving treatment decisions. In light of our experimental results and literature data, the analysis of the SLCO1B1 T521C variant will allow clinicians to take advantage from a better treatment planned for their patients in order to minimize neuromuscular pain and maximize benefits.

Retinopatía diabética proliferativa ilustrada por reflectancia infrarroja / Proliferative diabetic retinopathy imaged by near-infrared reflectance

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Relationship between Salmonella infection, shedding and serology in fattening pigs in low-moderate prevalence areas.

Salmonella is a major foodborne pathogen causing important zoonosis worldwide. Pigs asymptomatically infected in mesenteric lymph nodes (MLN) can be intermittent shedders of the pathogen through faeces, being considered a major source of human infections. European baseline studies of fattening pig salmonellosis are based on Salmonella detection in MLN. This work studies the relationship between Salmonella infection in MLN and intestinal content (IC) shedding at slaughter and the relationship between the presence of the pathogen and the serologic status at farm level. Mean Salmonella prevalence in the selected pigs (vertically integrated production system of Navarra, Spain) was 7.2% in MLN, 8.4% in IC and 9.6% in serum samples. In this low-moderate prevalence context, poor concordance was found between MLN infection and shedding at slaughter and between bacteriology and serology. In fact, most of shedders were found uninfected in MLN (83%) or carrying different Salmonella strains in MLN and in IC (90%). The most prevalent Salmonellae were Typhimurium resistant to ACSSuT ± Nx or ASSuT antibiotic families, more frequently found invading the MLN (70%) than in IC (33.9%). Multivariable analysis revealed that risk factors associated with the presence of Salmonella in MLN or in IC were different, mainly related either to good hygiene practices or to water and feed control, respectively. Overall, in this prevalence context, detection of Salmonella in MLN is an unreliable predictor of faecal shedding at abattoir, indicating that subclinical infections in fattening pigs MLN could have limited relevance in the IC shedding.

Diagnostic value of arterial blood gas lactate concentration in the different forms of mesenteric ischemia.

PURPOSE: The role of serum lactate measurement in patients with intestinal ischemia still remains unclear. The aim of this study was to prospectively evaluate the diagnostic performance of arterial blood gas lactate concentrations in the patients with acute mesenteric ischemia and its different forms. METHODS: All the patients reporting abdominal pain associated with risk factors for mesenteric ischemia underwent arterial blood gas and contrast enhanced abdominal computer tomography (CT). RESULTS: At CT, 201 patients (70.7%) showed a nonischemic disease (group 1) and 83 patients (29.2%) showed findings of mesenteric ischemia. Out of these, 35 patients (42.1%) showed bowel ischemia secondary to non vascular causes (group 2) and 48 (57.8%) had a vascular intestinal ischemia (group 3). Out of these, 20 showed small bowel arterial occlusion (group 3a), 13 a small bowel nonocclusive ischemia (group 3b), 7 a venous small bowel occlusion (group 3c) and 8 showed isolated colonic ischemia (group 3d). The median lactate serum level was significantly higher in patients with vascular ischemia if compared with patients with nonischemic disease and secondary mesenteric ischemia (p < 0.0001; Kruskal-Wallis test). The areas under ROC curves for the lactate serum levels in the groups 2, 3, 3a, 3b, 3c and 3d were, respectively, 0.61, 0.85, 0.93, 0.93, 0.68 and 0.67. CONCLUSIONS: Arterial blood gas lactate levels seem to show good diagnostic accuracy in diagnosing small bowel arterial and nonocclusive ischemia and poor accuracy in diagnosing secondary mesenteric ischemia, small bowel venous ischemia and ischemic colitis.

Grosor coroideo macular después de la cirugía vitreorretiniana: efecto a largo plazo de la vitrectomía con y sin cerclaje escleral / Macular choroidal thickness after vitreoretinal surgery: Long-term effect of pars plana vitrectomy with and without encircling scleral buckling surgery

Objetivo: Evaluar el espesor coroideo (EC) macular a largo plazo tras vitrectomía pars plana (VPP) con o sin cerclaje escleral (CE) para la reparación del desprendimiento de retina regmatógeno primario, después de al menos 6 meses de la cirugía. Métodos: Estudio observacional que incluyó: 15 ojos (15 pacientes) que se sometieron al CE añadido a VPP; 15 ojos (15 pacientes) que se sometieron a VPP y los 30 ojos contralaterales (OC) normales. Se obtuvo, en cada ojo, con la tomografía de coherencia óptica enhanced depth imaging, 2 escáneres de una línea de 6 mm perpendiculares, centrados en la fóvea. Se midió el EC en varios puntos maculares: subfoveal (EC-SF) y en un radio de 1, 2 y 3 mm respecto a la fóvea. El EC de los ojos sometidos a CE + VPP se comparó con el EC de los OC respectivos y con el EC de aquellos sometidos a VPP. Resultados: El EC-SF de los ojos del grupo CE + VPP fue significativamente mayor en comparación con los OC (p = 0,001). Los EC en un radio de 1, 2 y 3mm respecto a la fóvea de los ojos operados estaban significativamente aumentados en los ojos del grupo CE+VPP (p = 0,001, p = 0,005 y p = 0,001, respectivamente). El EC-SF de los ojos del grupo VPP y el EC - SF de los OC fue similar (p = 0,691). El EC-SF de los ojos del grupo CE + VPP fue significativamente mayor que el EC-SF de los ojos del grupo VPP (p = 0,019). Conclusiones: El EC de los ojos sometidos a CE + VPP estaba aumentado después de al menos 6 meses de la cirugía, en comparación con el EC de los respectivos ojos adelfos y el EC de los ojos sometidos a VPP, lo que podría deberse a una reducción del drenaje venoso causada por el CE. El EC de los ojos sometidos a CE+VPP estaba aumentado después de al menos 6 meses de la cirugía, en comparación con el EC de los respectivos ojos adelfos y el EC de los ojos sometidos a VPP, lo que podría deberse a una reducción del drenaje venoso causada por el CE (AU)

Purpose: To evaluate the macular choroidal thickness (CT) of eyes subjected to pars plana vitrectomy (PPV) whether or not combined with encircling scleral buckling (ESB) surgery for primary rhegmatogenous retinal detachment repair at 6 months or more after surgery. Methods: This observational study included: 15 eyes (15 patients) submitted to combined ESB+PPV; 15 eyes submitted to PPV and their respective 30 normal fellow eyes (FE). Two 6mm lineal perpendicular optical coherence tomography B-scans centred on the fovea with enhanced depth imaging were performed on each eye. CT was measured at several macular locations: subfoveal (SF-CT) and at a radius of 1, 2, and 3mm from the fovea. CTs of the eyes in the CE+PPV group were compared to CT in the PPV group and the CTs of all operated eyes were compared to the CTs of their FE. Results: SF-CT of the eyes in the ESB+PPV group was significantly increased compared to their FE (P = .001). CT at a radius of 1, 2, and 3 mm from the fovea of the ESB + PPV group were significantly increased (P = .001, P = .005, and P = .001, respectively). The SF-CT of the PPV group was similar to their FE (P = .691). The SF-CT of the ESB + PPV group was significantly increased compared to SF-CT of the PPV group (P = .019). Conclusions: The CT of the eyes subjected to combined ESB and PPV was significantly increased at 6 months or more after surgery compared to the CT of their FE and to the CT of the eyes subjected to PPV alone, which could be explained by a venous engorgement caused by the ESB (AU)

Macular choroidal thickness after vitreoretinal surgery: Long-term effect of pars plana vitrectomy with and without encircling scleral buckling surgery. / Grosor coroideo macular después de la cirugía vitreorretiniana: efecto a largo plazo de la vitrectomía con y sin cerclaje escleral.

PURPOSE: To evaluate the macular choroidal thickness (CT) of eyes subjected to pars plana vitrectomy (PPV) whether or not combined with encircling scleral buckling (ESB) surgery for primary rhegmatogenous retinal detachment repair at 6 months or more after surgery. METHODS: This observational study included: 15 eyes (15 patients) submitted to combined ESB+PPV; 15 eyes submitted to PPV and their respective 30 normal fellow eyes (FE). Two 6mm lineal perpendicular optical coherence tomography B-scans centred on the fovea with enhanced depth imaging were performed on each eye. CT was measured at several macular locations: subfoveal (SF-CT) and at a radius of 1, 2, and 3mm from the fovea. CTs of the eyes in the CE+PPV group were compared to CT in the PPV group and the CTs of all operated eyes were compared to the CTs of their FE. RESULTS: SF-CT of the eyes in the ESB+PPV group was significantly increased compared to their FE (P=.001). CT at a radius of 1, 2, and 3mm from the fovea of the ESB+PPV group were significantly increased (P=.001, P=.005, and P=.001, respectively). The SF-CT of the PPV group was similar to their FE (P=.691). The SF-CT of the ESB+PPV group was significantly increased compared to SF-CT of the PPV group (P=.019). CONCLUSIONS: The CT of the eyes subjected to combined ESB and PPV was significantly increased at 6 months or more after surgery compared to the CT of their FE and to the CT of the eyes subjected to PPV alone, which could be explained by a venous engorgement caused by the ESB.

PEPFAR support of alcohol-HIV prevention activities in Namibia and Botswana: a framework for investigation, implementation and evaluation.

BACKGROUND: The association between harmful use of alcohol and HIV infection is well documented. To address this dual epidemic, the US President's Emergency Plan for AIDS Relief (PEPFAR) developed and implemented a multi-pronged approach primarily in Namibia and Botswana. We present the approach and preliminary results of the public health investigative and programmatic activities designed, initiated and supported by PEPFAR to combat the harmful use of alcohol and its association as a driver of HIV morbidity and mortality from 2008 to 2013. APPROACH: PEPFAR supported comprehensive alcohol programming using a matrix model approach that combined the socio-ecological framework and the Alcohol Misuse Prevention and Intervention Continuum. This structure enabled seven component objectives: (1) to quantify harmful use of alcohol through rapid assessments; (2) to develop and evaluate alcohol-based interventions; (3) to promote screening programs and alcohol abuse resource services; (4) to support stakeholder networks; (5) to support policy interventions and (6) structural interventions; and (7) to institutionalize universal prevention messages. DISCUSSION: Targeted PEPFAR support for alcohol activities resulted in several projects to address harmful alcohol use and HIV. Components are graphically conceptualized within the matrix model, demonstrating the intersections between primary, secondary and tertiary prevention activities and individual, interpersonal, community, and societal factors. Key initiative successes included leveraging alcohol harm prevention activities that enabled projects to be piloted in healthcare settings, schools, communities, and alcohol outlets. Primary challenges included the complexity of multi-sectorial programming, varying degrees of political will, and difficulties monitoring outcomes over the short duration of the program.

Chronic Ocular Graft vs Host Disease as a Serious Complication of Allogeneic Hematopoietic Stem Cell Transplantation: Case Report.

Chronic graft-vs-host disease (cGVHD) is a serious systemic immunological complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). Ocular GVHD (O-GVHD) is frequently associated with cGVHD. Secondary corneal epithelial changes can occur in the setting of advanced chronic O-GVHD-associated keratoconjunctivitis sicca (KCS), which generally has a stable course with conventional medical treatment. Bilateral corneal ulcers and ocular perforation, although not frequent, can occur in most extreme cases. The authors describe 2 clinical cases of ocular perforation (Clinical case 1) and bilateral simultaneous corneal ulcers (Clinical case 2) due to advanced chronic O-GVHD, which can rarely occur despite treatment. A close ophthalmological follow-up and good dialogue with the multidisciplinary transplantation team are essential after allo-HSCT.

Reduction of subclinical Salmonella infection in fattening pigs after dietary supplementation with a ß-galactomannan oligosaccharide.

AIMS: To assess the efficacy of a ß-galactomannan oligosaccharide (ß-GMOS) for the control of Salmonella infection in fattening pigs. METHODS AND RESULTS: Three different doses (0.5, 3 and 2 kg ß-GMOS per ton of feed) were used during the entire period of growing in three similar and independent field trials carried out in a small fattening unit (≈ 100 pigs). Treatment was randomly assigned to half of the pens. Individual serum samples (20-25 per group) were collected at different times during the fattening period and a similar number of faecal samples during the fattening period and at slaughter. In addition, mesenteric lymph nodes were collected at slaughter. Herdcheck(®) Swine Salmonella ELISA was used for serological analyses, the ISO 6579:2002/Amd 1 : 2007 for bacteriology and the PFGE for molecular characterization of Salmonella strains. The addition of ≥ 2 kg t(-1) of ß-GMOS to the pig diet during the entire fattening period was associated with a reduction in Salmonella prevalence, shedding and seroconversion. CONCLUSIONS: Feed supplementation with ß-GMOS may be a useful complementary tool for the control of salmonellosis in fattening pigs. SIGNIFICANCE AND IMPACT OF THE STUDY: ß-GMOS may be a complementary way of reducing Salmonella shedding and infection in fattening pigs.

HIV infection and risk factors among the armed forces personnel stationed in Kinshasa, Democratic Republic of Congo.

Despite recent declines in HIV incidence, sub-Saharan Africa remains the most heavily affected region in the global HIV/AIDS epidemic. Estimates of HIV prevalence in African military personnel are scarce and inconsistent. We conducted a serosurvey between June and September 2007 among 4043 Armed Forces personnel of the Democratic Republic of Congo (FARDC) stationed in Kinshasa, Democratic Republic of Congo (DRC) to determine the prevalence of HIV and syphilis infections and describe associated risk behaviours. Participants provided blood for HIV and syphilis testing and responded to a demographic and risk factor questionnaire. The prevalence of HIV was 3.8% and the prevalence of syphilis was 11.9%. Women were more likely than men to be HIV positive, (7.5% vs. 3.6% respectively, aOR: 1.66, 95% C.I: 1.21-2.28, p < 0.05). Factors significantly associated with HIV infection included gender and self-reported genital ulcers in the 12 months before date of enrollment. The prevalence of HIV in the military appears to be higher than the general population in DRC (3.8% vs. 1.3%, respectively), with women at increased risk of infection.

[Anatomical variants of the medial calcaneal nerve and the Baxter nerve in the tarsal tunnel]. / Variantes anatómicas del nervio calcáneo medial y nervio de Baxter en el túnel del tarso.

The tarsal tunnel is composed of the posterior border of the medial malleoulus, the posterior aspect of the talus and the medial aspect of the calcaneus. The medial calcaneal nerve emerges from the posterior aspect of the posterior tibial nerve in 75% of cases and from the lateral plantar nerve in the remaining 25%. Finally, the medial calcaneal nerve ends as a single terminal branch in 79% of cases and in numerous terminal branches in the remaining 21%. To describe the anatomical variants of the posterior tibial nerve and its terminal branches. To describe the steps for tarsal tunnel release. To describe Baxter nerve release. The anatomical variants of the posterior tibial nerve and its terminal branches within the tarsal tunnel were studied. Then the Lam technique was performed; it consists of: 1) opening of the laciniate ligament, 2) opening of the fascia over the abductor hallucis muscle, 3) exoneurolysis of the posterior tibial nerve and its terminal branches, identifying the emergence and pathway of the medial calcaneal branch, the lateral plantar nerve and its Baxter nerve branch and the medial plantar nerve. Baxter nerve was found in 100% of cases. In 100% of cases in our series the nerve going to the abductor digiti minimi muscle of the foot was found; 87.5% of cases had two terminal branches. The dissections proved that a crucial step was the release of the distal tarsal tunnel.

Epidemiology of subclinical salmonellosis in wild birds from an area of high prevalence of pig salmonellosis: phenotypic and genetic profiles of Salmonella isolates.

The epidemiology of subclinical salmonellosis in wild birds in a region of high Salmonella prevalence in pigs was studied. Three hundred and seventy-nine faecal samples from 921 birds trapped in 31 locations nearby pig premises, and 431 samples from 581 birds of 10 natural settings far from pig farms were analysed for the presence of Salmonella spp. Positive samples were serotyped and analysed for antimicrobial resistance (AR). Phage typing and pulsed-field gel electrophoresis (PFGE) on Salmonella Typhimurium isolates were also carried out. The overall proportion of Salmonella-positive samples was 1.85% (95% CI=0.93, 2.77). Salmonella isolation was positively associated with samples collected from birds in the proximity of a pig operation (OR=16.5; 95% CI=5.17, 52.65), and from non-migratory (or short-distance migration) birds (OR=7.6; 95% CI=1.20, 48.04) and negatively related to mostly granivorous birds (OR=0.4; 95% CI=0.15, 1.13). Salmonella Typhimurium was the most prevalent serotype and four different XbaI PFGE patterns were observed that matched the four phage types identified (U310, U311, DT164 and DT56). Only 20% of the strains showed multi-AR. In three farms, a high degree of homogeneity among isolates from different birds was observed. These findings suggested that pig farms may act as amplifiers of this infection among wild birds, and the degree of bird density may have much to do on this transmission. Some of the Salmonella serotypes isolated from bird faeces were of potential zoonotic transmission and associated with AR. Monitoring salmonellosis in wild bird is advised.

Sensitivity of the ISO 6579:2002/Amd 1:2007 standard method for detection of Salmonella spp. on mesenteric lymph nodes from slaughter pigs.

The ISO 6579:2002/Amd 1:2007 (ISO) standard has been the bacteriological standard method used in the European Union for the detection of Salmonella spp. in pig mesenteric lymph nodes (MLN), but there are no published estimates of the diagnostic sensitivity (Se) of the method in this matrix. Here, the Se of the ISO (Se(ISO)) was estimated on 675 samples selected from two populations with different Salmonella prevalences (14 farms with a ≥ 20% prevalence and 13 farms with a <20% prevalence) and through the use of latent-class models in concert with Bayesian inference, assuming 100% ISO specificity, and an invA-based PCR as the second diagnostic method. The Se(ISO) was estimated to be close to 87%, while the sensitivity of the PCR reached up to 83.6% and its specificity was 97.4%. Interestingly, the bacteriological reanalysis of 33 potential false-negative (PCR-positive) samples allowed isolation of 19 (57.5%) new Salmonella strains, improving the overall diagnostic accuracy of the bacteriology. Considering the usual limitations of bacteriology regarding Se, these results support the adequacy of the ISO for the detection of Salmonella spp. from MLN and also that of the PCR-based method as an alternative or complementary (screening) test for the diagnosis of pig salmonellosis, particularly considering the cost and time benefits of the molecular procedure.

Identification of early induced genes upon water deficit in potato cell cultures by cDNA-AFLP.

For plant cells in the early phases of water stress exposure, the genes induced under such conditions play a key role in detecting and responding to water deficit. In this study, potato cell suspensions were used as a simplified model system to dissect early molecular changes upon low water potential. In particular, the cDNA-amplified fragment length polymorphism approach was used to capture genes rapidly activated in potato cell cultures in response to water deficit induced by short-term exposure (up to 1 h) to polyethylene glycol. Selective amplifications with 38 primer combinations allowed the visualization of about 167 transcript-derived fragments (TDFs) differentially expressed upon exposure to low water potential. The gene expression pattern of 18 up-regulated genes was further investigated by semi-quantitative reverse transcriptase polymerase chain reaction analysis. Sequencing and similarity analysis revealed that TDFs present homologies chiefly with proteins involved in chaperone activity and protein degradation (hsps, proteinase precursor), in protein synthesis (elongation factor, ribosomal proteins) and in the ROS scavenging pathway (phenylalanine ammonia-lyase, peroxidase). Our findings might contribute to describe the potential role of genes activated in the early phases of plant response to drought.

Surgical strategy for the treatment of sporadic medullary thyroid carcinoma: our experience.

BACKGROUND: Medullary thyroid carcinoma (MTC) is a rare disease which accounts for approximately 5-9% of all thyroid cancers and originates from the calcitonin-screening parafollicular C cells. MTC can be divided into two subgroups: sporadic (75%) or inherited (25%). The majority of patients with invasive MTC have metastasis to regional lymph nodes at the time of diagnosis, as evidenced by the frequent finding of persistently elevated calcitonin levels after thyroidectomy and the high rates of recurrence in the cervical lymph nodes reported in retrospective studies. OBJECTIVES: The purpose of the study is to review our single institution's experience with MTC since 1998 and to evaluate surgical strategy, patterns of lymph node metastases and calcitonin response to compartment-oriented lymphadenectomy in patients with primary or recurrent sporadic medullary thyroid carcinoma. METHODS: A retrospective review of 26 patients treated for MTC at the "Antonio Cardarelli" Hospital referral center, in Naples, between 1998 and 2012. There were 18 female and 8 male patients, median age at presentation was 55 years, and median follow-up for survivors was 5 years. Total thyroidectomy was performed in all 26 patients; central compartment (CC) node dissection (level VI) in 12 (46%) patients; central plus lateral compartment (LC) node dissection (levels II, III, and IV) in 7 (27%) patients. 4 patients (15%) underwent reoperation for loco-regional recurrent/persistent MTC. Results. After a median post-surgical follow-up of 5 years (range 1-10 years), 63 % of patients were living disease-free, 15% were living with disease and/or persistently elevated calcitonin levels after surgery, 11% were deceased due to MTC and 11 % were lost to follow-up. CONCLUSIONS: We agree with most authors advocating for a total thyroidectomy and prophylactic central neck dissection in the setting of clinically detected MTC. Lateral neck dissection may be best reserved for patients with positive preoperative imaging. Nevertheless MTC has a high rate of lymph node metastases that are sub optimally detected preoperatively in the central compartment by neck ultrasound or intra-operatively by the surgeon, and reoperation is associated with a higher rate of surgical complications. In our limited experience, patients with thyroid confined nodular pathology, without nodal disease and unknown preoperative diagnosis of MTC, underwent only total thyroidectomy with a good prognosis.

Distribution and function of polycystin-2 in mouse retinal ganglion cells.

The polycystin family of transient receptor potential (TRP) channels form Ca(2+) regulated cation channels with distinct subcellullar localizations and functions. As part of heteromultimeric channels and multi-protein complexes, polycystins control intracellular Ca(2+) signals and more generally the translation of extracellular signals and stimuli to intracellular responses. Polycystin-2 channels have been cloned from retina, but their distribution and function in retinal ganglion cells (RGCs) have not yet been established. In the present study, we determined cellular and subcellular localization as well as functional properties of polycystin-2 channels in RGCs. Polycystin-2 expression and distribution in RGCs was assessed by immunohistochemistry on vertical cryostat section of mouse retina as well as primary cultured mouse RGCs, using fluorescence microscopy. Biophysical and pharmacological properties of polycystin-2 channels isolated from primary cultured RGCs were determined using planar lipid bilayer electrophysiology. We detected polycystin-2 immunoreactivity both in the ganglion cell layer as well as in primary cultured RGCs. Subcellular analysis revealed strong cytosolic localization pattern of polycystin-2. Polycystin-2 channel current was Ca(2+) activated, had a maximum slope conductance of 114 pS, and could be blocked in a dose-dependent manner by increasing concentrations of Mg(2+). The cytosolic localization of polycystin-2 in RGCs is in accordance with its function as intracellular Ca(2+) release channel. We conclude that polycystin-2 forms functional channels in RGCs, of which biophysical and pharmacological properties are similar to polycystin-2 channels reported for other tissues and organisms. Our data suggest a potential role for polycystin-2 in RGC Ca(2+) signaling.

Salmonellosis in finishing pigs in Spain: prevalence, antimicrobial agent susceptibilities, and risk factor analysis.

A herd-based survey of Salmonella in pigs was carried in a major pig producing region of Spain. Mesenteric lymph nodes were collected from the carcasses of 25 pigs from each of 80 herds at time of slaughter. Salmonella spp. were isolated from 31% of animals and 94% of herds. Within-herd prevalence ranged from 4 to 88%, with the prevalence in most herds being greater than 10%. A large diversity of Salmonella serotypes was found, with Typhimurium, 4,[5],12:i:-, and Rissen being the most prevalent. Two or more serotypes coexisted in 73% of the herds. Salmonella Typhimurium was present in 68% of the herds. Most (82%) of the Salmonella isolates belonged to serogroups targeted by enzyme-linked immunosorbent assay tests for pig salmonellosis. Resistance to at least one antimicrobial agent was detected in 73% of the strains, and one or more resistant strains were recovered from pigs in 93% of the herds. Antimicrobial agent resistance (AR) was more frequent among the most prevalent than it was among the rarer serotypes. Twenty-five multi-AR patterns were found. Resistance to three or more families of antimicrobial agents was found in 75% of AR strains. The finding that many of the herds yielded isolates of several multi-AR patterns indicates that Salmonella infections were acquired from multiple sources. High prevalence of Salmonella in herds was associated with lack of rodent control programs, herds from farms with only finishing pigs, herds managed by more than one full-time worker, herds for which the source of drinking water was not a city supply, and relatively long fattening times.