A low-cost towed video camera system for underwater surveys: comparative performance with standard methodology.

Technological advances in the field of underwater video have led to an exponential increase in the use of drifting cameras (DC) and remotely operated vehicles (ROVs) to monitor the diversity, abundance, and size structure of marine life. Main advantages of DCs relative to ROVs are their lower costs and the much simpler logistics required to operate them. This study compares the performance of a new low-cost DC system equipped with a novel measuring device with that of a standard DC bearing an array of laser pointers. The new DC, which can be operated from a small boat, carries a pair of parallel steel "whiskers" that are dragged on the seabed within the field of view of the camera, providing a scale for measuring and estimating the density of benthic biota. An experiment conducted using an array of objects of known sizes laid on the bottom showed that its performance in terms of both size and density estimation was similar to that of the standard technique based on laser pointers. Measurement errors had a negligible negative bias (- 2.3%) and a standard deviation that ranged between 13 and 8% for objects from 25 to 110 mm in size. The whiskers offered a simplified method for density estimation that avoids the need to calculate the width of the field of view, thus reducing the video processing time by around 60% with respect to the standard method. Briefly, the new system offers an efficient low-cost alternative for benthic ecology studies conducted on soft or non-irregular bottoms.

Distribution of additional virulence factors related to adhesion and toxicity in Shiga toxin-producing Escherichia coli isolated from raw products in Argentina.

UNLABELLED: A total of 73 Shiga toxin-producing Escherichia coli (STEC) isolates, belonging to 25 serotypes and isolated from raw products in Argentina, were examined for the occurrence of genes responsible for bacterial adhesions to intestine, ehaA (EHEC autotransporter), lpfAO113 (long polar fimbriae), sab (STEC autotransporter [AT] contributing to biofilm formation), ecpA (E. coli common pilus), hcpA (haemorrhagic coli pilus), elfA (E. coli laminin-binding fimbriae), sfpA (sorbitol-fermenting EHEC O157 fimbriae plasmid-encoded) and of the toxigenic gene cdt-V (cytolethal distending toxin). Our study showed different adhesin profiles that are not linked to one specific serotype and that all analysed isolates possess, besides stx genes, some adherence genes. Several of the isolates contained also multiple toxin genes. The results of the present work alert the presence of genes coding for additional adhesins and cdt-V toxin in LEE-negative STEC strains that occur in foods, and this traits could increase their pathogenic potential. SIGNIFICANCE AND IMPACT OF THE STUDY: Meat products are one of the main vehicles of Shiga toxin-producing E. coli, and the presence of genes coding for additional adhesins and toxins could increase their pathogenic potential. There is a need for a more detailed characterization of the strains in regard to these extra virulence factors.

Seasonal variation of HUS occurrence and VTEC infection in children with acute diarrhoea from Argentina.

In order to study the seasonality of haemolytic uraemic syndrome (HUS) and verotoxigenic Escherichia coli (VTEC) infection in children, 437 patients under 6 years of age with acute diarrhoea were studied, 8% of whom progressed to HUS. VTEC was found in 10% of all of the stool samples analysed and seasonal occurrence of HUS (p < 0.01) was confirmed. VTEC infection was more prevalent in warm months, although the differences were not statistically significant. Moreover, a significant difference in the detection of O157:H7 serotype and in the vt profile between cold and warm months (autumn and winter; spring and summer, respectively) was established. The O157:H7 serotype was isolated more frequently during warm months. Moreover, a predominance of vt (2) was noted, which was partially replaced by the combination of vt (1) with vt (2) in the cold season. The results of this study indicate the seasonal variation of the disease and the presence of serotype O157:H7 and the vt types. They also reinforce the need to develop prevention programmes considering the seasonal pattern of the disease, which would generate an impact on public health. Control strategies of the pathogen in cattle in the most risky season of the year would also be of benefit.

Short communication: characterization of Shiga toxin-producing Escherichia coli isolated from newborn, milk-fed, and growing calves in Argentina.

Shiga toxin-producing Escherichia coli (STEC) cause foodborne pathogenic disease that is shed in the feces of cattle. The aim of this study was to evaluate how early young calves are colonized by STEC strains, potentially pathogenic for humans, and the prevalence in different calf categories. From 808 rectal swabs analyzed by PCR, 38% were stx positive. The prevalence in newborn (<24 h from birth), milk-fed (<2-mo-old), and growing calves (2-8 mo old) were 25, 43, and 58%, respectively. Forty different STEC serotypes were found among isolates from newborn, milk-fed, and growing calves that shed STEC strains potentially pathogenic for humans. The STEC strains could be acquired early from mothers, enabling the infection of other animal categories and confirming the risk to public health.

Multiplex PCR assay for the detection of five putative virulence genes encoded in verotoxigenic Escherichia coli plasmids.

The aim was to perform a pentavalent PCR assay for the detection of putative virulence genes encoded in VTEC plasmids, katP, espP, subA, stcE, and ehxA. The five-specific primer pairs used in the assay do not interfere with each other and generate amplification products of 914, 774, 556, 399, and 262 bp. It was selected at random 39 strains belonged to 20 serotypes in order to evaluate the multiplex in a wide variety of strains. The results of this study indicate that it is possible to perform simultaneous amplification and search for recognized plasmid-encoded virulence markers from different E. coli serotypes and apply this technique to the genetic characterization of E. coli strains isolated from reservoirs, foods or patients. This complementary technique is a useful tool to detect interstrain differences for epidemiological studies and to provide information that could be related to the risk of human infection.

Enteropathogenic Escherichia coli contamination at different stages of the chicken slaughtering process.

Enteropathogenic Escherichia coli is a foodborne pathogen that produces potentially fatal infant diarrhea, noticeably in developing countries. The aim of this study was to detect EPEC contamination by PCR at different stages of the chicken slaughtering process. We collected swabs from chicken cloacae and washed carcasses (external and visceral cavity) during the slaughtering process in 3 sampling occasions. Unwashed eviscerated carcasses were also sampled (at the visceral cavity) in the second and third sampling occasions. Enteropathogenic Escherichia coli was detected in 6 to 28% of cloacal samples, 39 and 56% of unwashed eviscerated carcasses, and 4 to 58% of washed carcasses. None of the samples were positive for bfpA, suggesting contamination with atypical EPEC. The detection of EPEC at different stages of the chicken slaughtering process showed that the proportion of contaminated samples remained or even increased during processing. In addition, the high proportion of contaminated carcasses during chicken processing represents a risk for the consumers and a challenge to improve procedures for those working in the sanitary control service.

Characterization of Shiga toxin-producing Escherichia coli isolated from dairy cows in Argentina.

AIMS: To feno-genotypically characterize the Shiga toxin-producing Escherichia coli (STEC) population in Argentinean dairy cows. METHODS AND RESULTS: From 540 STEC positive samples, 170 isolates were analyzed by multiplex PCR and serotyping. Of these, 11% carried stx1, 52% stx2 and 37% stx1/stx2. The ehxA, saa and eae were detected in 77%, 66% and 3%, respectively. Thirty-five per cent of strains harboured the profile stx1, stx2, saa, ehxA and 29% stx2, saa, ehxA. One hundred and fifty-six strains were associated with 29 different O serogroups, and 19 H antigens were distributed among 157 strains. STEC O113:H21, O130:H11 and O178:H19 were the most frequently found serotypes. The STEC O157:H7 were detected in low rate and corresponded to the stx2(+) , eae(+) , ehxA(+) virulence pattern. CONCLUSIONS: We detected a diversity of STEC strains in dairy cattle from Argentina, most of them carrying genes linked to human disease. SIGNIFICANCE AND IMPACT OF THE STUDY: The non-O157 STEC serotypes described in this study are associated worldwide with disease in humans and represent a risk for the public health. For this, any microbiological control in dairy farms should be targeted not only to the search of O157:H7 serotype.

Seasonal variation of Shiga toxin-encoding genes (stx) and detection of E. coli O157 in dairy cattle from Argentina.

AIMS: To study the seasonal variation of Shiga toxin-encoding genes (stx) and to investigate the presence of Shiga toxin-producing Escherichia coli (STEC) O157 in cattle belonging to five dairy farms from Argentina. METHODS AND RESULTS: Rectal swab samples were collected from 360 dairy cows in each season and 115 and 137 calves in autumn and in spring, respectively. The stx were investigated by multiplex PCR and it was used as the indicator for STEC. Samples positives for stx were tested by PCR for eae-gamma1 of E. coli O157 and then subjected to IMS (immunomagnetic separation). In positive animals significant differences in the prevalence of stx between warm and cold seasons were detected. In warm seasons, stx1 + stx2 increased and stx1 decreased, independently of the animal category. The prevalence of STEC O157 in cows and calves were 0.2% and 0.8%, respectively. CONCLUSIONS: This work provides new data about the occurrence of stx and STEC O157 in dairy herds from Argentina and suggests a relationship between the type of stx and season of year. SIGNIFICANCE AND IMPACT OF STUDY: The detection of STEC O157 and the seasonality of stx and its types provide an opportunity to improve control strategies designed to prevent contamination of food products and transmission animal-person.

Food partitioning and spatial subsidy in shelter-limited fishes inhabiting patchy reefs of Patagonia.

The diets of the most conspicuous reef-fish species from northern Patagonia, the carnivorous species Pseudopercis semifasciata, Acanthistius patachonicus, Pinguipes brasilianus and Sebastes oculatus were studied. Pinguipes brasilianus had the narrowest diet and most specialized feeding strategy, preying mostly on reef-dwelling organisms such as sea urchins, limpets, bivalves, crabs and polychaetes. The diet of A. patachonicus was characterized by the presence of reef and soft-bottom benthic organisms, mainly polychaetes, crabs and fishes. Pseudopercis semifasciata showed the broadest spectrum of prey items, preying upon reef, soft-bottom and transient organism (mainly fishes, cephalopods and crabs). All S. oculatus guts were empty, but stable-isotope analyses suggested that this species consumed small fishes and crabs. In general, P. brasilianus depended on local prey populations and ate different reef-dwelling prey than the other species. Pseudopercis semifasciata, A. patachonicus and probably S. oculatus, however, had overlapping trophic niches and consumed resources from adjacent environments. The latter probably reduces the importance of food as a limiting resource for these reef-fish populations, facilitating their coexistence in spite of their high trophic overlap.

Comment on "Tracking the global footprint of fisheries".

Kroodsma et al (Reports, 23 February 2018, p. 904) mapped the global footprint of fisheries. Their estimates of footprint and resulting contrasts between the scale of fishing and agriculture are an artifact of the spatial scale of analysis. Reanalyses of their global (all vessels) and regional (trawling) data at higher resolution reduced footprint estimates by factors of >10 and >5, respectively.

Pheohyphomycotic soft tissue disease caused by Alternaria alternata in a kidney transplant patient: a case report and literature review.

A 61-year-old Italian man, who underwent a renal transplantation 8 years ago, receiving azathioprine, prednisone, and cyclosporine for immunosuppressive therapy, presented with a large reddish indurated plaque with a central ulcer, which was slowly enlarged, on the right knee. From the diseased tissue biopsy, a dematiaceous fungus matching Alternaria alternata in all essential characters was isolated in pure culture. This is an uncommon fungal complication in a kidney transplant patient. A detailed morphological description of the isolate is provided as well as review of the literature.

In-stent neointimal proliferation correlates with the amount of residual plaque burden outside the stent: an intravascular ultrasound study.

BACKGROUND: The aim of this study was to evaluate the relationship between residual plaque burden after coronary stent implantation and the development of late in-stent neointimal proliferation. METHODS AND RESULTS: Between January 1996 and May 1997, 50 patients underwent intravascular ultrasound (IVUS) interrogation at 6+/-1.2 months after coronary stent implantation in native coronary arteries. IVUS images were acquired with a motorized pullback, and cross-sectional measurements were performed within the stents at 1-mm intervals. The following measurements were obtained: (1) lumen area (LA), (2) stent area (SA), (3) area delimited by the external elastic membrane (EEMA), (4) percent neointimal area calculated as (SA-LA/SA)x100, and (5) percent residual plaque area calculated as (EEMA-SA)/EEMAx100. Volume measurements within the stented segments were calculated by applying Simpson's rule. In the pooled data analysis of 876 cross sections, linear regression showed a significant positive correlation between percent residual plaque area and percent neointimal area (r=0.50, y= 45.03+0.29x, P<0.01). There was significant incremental increase in mean percent neointimal area for stepwise increase in percent residual plaque area. Mean percent neointimal area was 16.3+/-10.3% for lesions with a percent residual plaque area of <50% and 27.7+/-11% for lesions with a percent residual plaque area of >/=50% (P<0.001). The volumetric analysis showed that the percent residual plaque volume was significantly greater in restenotic lesions compared with nonrestenotic lesions (58.7+/-4.3% versus 51.4+/-5.7%, respectively; P<0.01). CONCLUSIONS: Late in-stent neointimal proliferation has a direct correlation with the amount of residual plaque burden after coronary stent implantation, supporting the hypothesis that plaque removal before stent implantation may reduce restenosis.

Human T-cell lymphoblastic lymphoma expressing the T-cell receptor gamma/delta established in immune-deficient (bg/nu/xid) mice.

A small subgroup of human CD3-positive T-cell lymphoblastic lymphoma (T-LL) has been recently identified to express the T-cell receptor (TCR) gamma/delta heterodimer. Moreover peculiar clinical and histologic patterns of spleen and liver involvement have been associated with the TCR gamma/delta phenotype of tumor cells. In this paper we describe a human T-LL cell line (LL-DP) established in beige-nude-xid (BNX) mice, that by immunophenotype, molecular, and karyotype analyses, maintained most of the features of the patient. After serial transplants in BNX mice, LL-DP acquired quite a stable phenotype, producing a visible tumor in about 5 weeks in all the intravenously injected animals. The minimum number of transplanted cells that produce a tumor in all mice is 1 x 10(6). BNX mice bearing LL-DP lymphoma cells presented marked abdominal distension and splenomegaly. Diffuse lymphadenopathy with large tumor deposits in various lymph nodes that produce architectural effacement with a diffuse growth pattern was documented. The bone marrow was completely replaced, and spleen, liver, and kidneys were involved. Invasion of the central nervous system was leptomeningeal and perivascular. Overall this model might be useful for understanding mechanisms supporting lymphoma growth and progression as well as for testing new therapeutic strategies.

Isolation of cDNA clones encoding proteins of complex structures: analysis of the Trypanosoma brucei cytoskeleton.

We have adapted a group of well-known procedures in order to devise a simple method that allows the isolation of specific cDNAs encoding proteins located in different regions of the Trypanosoma brucei cytoskeleton. cDNA clones were isolated by screening a lambda gt11 expression library with a polyspecific, polyclonal antiserum against a complex immunogen, in this case the complete cytoskeleton. The fusion proteins produced by the clones were then used as an affinity immunoadsorbant to select monospecific polyclonals. The monospecific antisera isolated were used as probes to identify and localize different cytoskeleton proteins by Western blotting and immunofluorescence. This method proved particularly useful for the molecular identification of minor components in a complex structure. It should prove applicable to the molecular analysis of other organelles or protein complexes.

Desmoplastic small round cell tumor of the pleura.

Three cases of desmoplastic small round cell tumor (DSRCT) with multiphenotypic differentiation, primary in the pleura, are presented. This is a previously unrecognized site for this tumor type. Two patients were male and one female aged 29, 24, and 17 years. All presented with chest pain and were found to have pleural-based tumors associated with pleural effusion. Abdominal involvement was not present in any of the cases. Histologically, the tumor showed the characteristic features of intra-abdominal DSRCT, including angulated nests of small cells embedded in a vascular fibroblastic stroma, focal rhabdoid phenotype, and areas of central necrosis. The neoplastic cells showed evidence of epithelial, mesenchymal, and neural differentiation with characteristic dot-like positivity for vimentin and desmin topographically corresponding to perinuclear aggregates of intermediate filaments identified on electron microscopy in one case. Two patients died of disease 2 years and 15 months after presentation, respectively, and one patient is alive with disease 18 months after presentation. The histogenesis of DSRCT is unknown. Most previously reported cases involved the peritoneum or tunica vaginalis, suggesting a histogenetic relationship to the mesothelium. The occurrence of these tumors in the pleura lends further support to this theory.

Early translaryngeal tracheostomy in patients with severe brain damage.

OBJECTIVES: To describe the effects of early translaryngeal tracheostomy on intracranial pressure (ICP), cerebral perfusion pressure (CPP), and jugular bulb saturation (SjO2); to identify the main mechanisms affecting ICP during tracheostomy; and to evaluate the long-term effects of tracheostomy on tracheal anatomy and function. DESIGN: Prospective, observational, clinical study. SETTING: Neurosurgical intensive care unit in a teaching hospital. PATIENTS: 20 patients admitted to the ICU because of head injury, subarachnoid hemorrhage, or brain tumor with a Glasgow Coma Scale less than 8. INTERVENTIONS: Patients underwent translaryngeal tracheostomy under strict neuromonitoring. MEASUREMENTS AND RESULTS: ICP rose significantly (p < 0.05) at the critical time of cannula placement while all other parameters remained stable. At this time five patients suffered intracranial hypertension (ICP > 20 mmHg). In one of them CPP dropped below 60 mmHg. Arterial CO2 tension (PaCO2) did not rise significantly. No other major complications were recorded during the procedures. Three months after tracheostomy normal findings were detected by tracheoscopy in all cases (11 patients could be examined). CONCLUSIONS: Translaryngeal tracheostomy, performed in selected patients when the risk of intracranial hypertension was reduced to the minimum, was well tolerated in the majority of cases and did not induce persistent intracranial disorders. However, ICP is affected by tracheostomy, and careful monitoring and patient selection is necessary. At follow-up no severe anatomical or functional damage was detected.

Losartan improves diastolic ventricular filling of hypertensive patients with diastolic dysfunction.

To evaluate the role of losartan on left ventricular (LV) function of hypertensive patients. Hypertensive patients (n = 19) underwent evaluation of systolic and diastolic LV function, using radionuclide ventriculography (RVG), before and at 3 mo into the treatment with the angiotensin II antagonist losartan. All patients underwent a baseline 12 lead ECG and an echocardiogram (ECHO), which was also repeated at 3 mo into treatment. Results are expressed as mean +/- SEM and statistics were performed using paired t-test. A p value < or = 0.05 was considered significant. Treatment with losartan for 3 mo had no effect on LV mass measured by echo (141+/-5 vs. 139+/-6 g/m2). The LV ejection fraction, measured by RVG, was unchanged by treatment when compared to the baseline study (58+/-2% vs. 57+/-2%, respectivelly, p = 0.49). Considering all patients involved in the study (n = 19), the LV "Peak Filling Rate" (PFR), a parameter of diastolic function measured by RVG, was also unchanged by treatment when compared to baseline (2.5+/-0.2 EDV/s vs. 2.5+/-0.3 EDV/s, respectively, p = 0.9). However the analysis of those patients with evidence of diastolic dysfunction (n = 12) on the baseline RVG (PFR < 2.5 EVD/s), demonstrated significant improvement of LV filling after therapy with losartan (PFR = 1.8+/-0.1 EDV/s vs. 2.3 +/-0.2 EDV/s, respectively, p = 0.05). This change was associated with improvement of symptoms. Our results demonstrated that hypertensive patients with diastolic dysfunction on radionuclide ventriculography have significant improvement of ventricular filling at 3 mo into treatment with losartan.

Intravesical electromotive administration of drugs for treatment of superficial bladder cancer: a comparative Phase II study.

OBJECTIVES: To evaluate the efficacy of electromotive administration (EMDA) of intravesical mitomycin-C (MMC) in patients with superficial bladder tumors and to evaluate the toxicity of the treatment. METHODS: Thirteen patients with multifocal Stages Ta-T1 and G1-G2 transitional cell carcinoma (TCC) of the bladder, primary or recurrent (group A), received MMC 40 mg (retained in the bladder for 2 hours) once a week for 8 weeks. Fifteen patients with the same characteristics (group B) were treated with EMDA/MMC at a current of 15 mA for 20 minutes once a week for 8 weeks. All lesions in the bladder except one (marker) were resected in each patient. RESULTS: In group A, 5 of 12 patients (41.6%) demonstrated complete macroscopic and histologic disappearance of the marker lesion (complete response [CR]). In group B, 6 of 15 patients (40%) had a similar CR. Recurrence rate in responders was 60% in group A versus 33% in group B after 7.6 and 6 months, respectively. Disease-free interval was 14.5 months in the EMDA/MMC group compared to 10.5 months in the MMC group. Side effects were few. CONCLUSIONS: In intermediate risk patients with TCC of the bladder, EMDA/MMC was not superior to MMC alone with a CR rate of 41% versus 41.6%. In responders, a lower recurrence rate and a longer disease-free interval were observed in the EMDA/MMC group.

Developmental profile of neuron-specific (NSE) and non-neuronal (NNE) enolase.

Neurons and glia of mature brain can be distinguished by their isoenzyme content of the glycolytic enzyme enolase. Neurons contain neuron-specific enolase (NSE) and glial cells have non-neuronal enolase (NNE). Measurement of each isoenzyme by specific radioimmunoassay during the course of brain development in rat shows that NSE levels are very low in embryonic brain and increase at a time coincident with the morphological and functional maturation of neurons. NNE levels are high in embryonic brain and decrease when NSE first appears, followed by a gradual increase to adult levels. NSE levels rise at a slower rate in brain areas known to develop over a more protracted period (forebrain, cerebellum) compared to areas that develop more rapidly (brain stem). The data are consistent with a hypothesized switch from NNE to NSE during neuronal development. In E60 and E100 monkey brain tissue NSE/NNE ratios are higher in regions containing older neurons. This suggests that a similar switch from NNE to NSE also occurs during neuronal development in monkey.

Neuron specific protein (NSP) in neuroblastoma cells: relation to differentiation.

The spectrum of enolase enzyme forms has been examined in several lines of neuroblastoma cells and compared to those present in whole brain. The neuron specific enolase (NSP) is greatly decreased in the cultured cells as judged by activity profiles and radioimmunoassay. The synthesis of neuronal enolase appears to be extensively depressed in these cells while the total enolase activity is not affected. The non-neuron form of enolase (NNE) apparently compensates for the lack of the neuronal forms in the cultured cells. The preponderance of NNE in cultured neurons suggests that this enzyme is present in immature neurons, and that neuroblastoma cells are not fully differentiated with respect to the enolase function. Dibutyryl cyclic adenosine monophosphate treatment does increase NSP levels in mouse neuroblastoma cells, but not to the levels expected for fully differentiated neurons. The results indicate that NSP is a molecular correlate of fully differentiated neurons.