Whole genome sequencing and characteristics of extended-spectrum beta-lactamase producing Escherichia coli isolated from poultry farms in Banaskantha, India.

Worldwide dissemination of extended-spectrum -lactamase (ESBL)-producing Escherichia coli constitutes an emerging global health issue, with animal food products contributing as potential reservoirs. ESBL E. coli infection is associated with the high mortality and mobility rate in developing countries due to less susceptibility to antibiotics. The present study aimed to elucidate the molecular characteristics and sequence-based analysis of ESBL E. coli in the Gujarat state of India. This study included 108 E. coli strains were isolated from different poultry farms (broiler and layer) in the Banaskantha District. PCR was employed to identify genotypic ESBL-producing antimicrobial resistance genes. Overall, a high occurrence of ESBL genes was found in poultry farms due to the high usage of antimicrobials. The PCR analysis revealed that 79.62% of isolates were detected positive with one or more ESBL genes. Among them, bla TEM (63.88%) was found to be the predominant genotype, followed by bla SHV (30.55%) and bla OXA (28.70%). In the bla CTX-M group, a higher occurrence was observed in bla CTX-M-9 (23.14%), followed by bla CTX-M-2 (24.07%) and bla CTX-M-1 (22.22%). We used the whole-genome sequencing (WGS) method to evaluate the antimicrobial resistance genes, virulence factors, single nucleotide polymorphisms (SNPs), plasmid replicons, and plasmid-mediated AMR genes of one ESBL E. coli isolated. We examined the genetic relatedness of a human pathogenic E. coli strain by comparing its sequence with the broad geographical reference E. coli sequences. Escherichia coli ST 681 was determined using multi-locus sequence typing. We compared our findings to the reference sequence of Escherichia coli str. K- 12 substr. MG1655. We found 24,937 SNPs with 21,792 in the genic region, 3,145 in the intergenic region, and six InDels across the genome. The WGS analysis revealed 46 antimicrobial resistance genes and seven plasmid-mediated AMR genes viz., tetA, qnrS1, dfrA14, sul2, aph(3")-lb, aph(6)-ld, and Aph(3')-la. The ST 681 was found to have Cib, traT, and terC virulence factors and two plasmid replicons, IncFII(pHN7A8) and IncI1-I(Alpha). This study revealed a higher occurrence of ESBL E. coli detected in poultry.

Retrospective Validation of a Computer-Assisted Quantification Model of Intracerebral Hemorrhage Volume on Accuracy, Precision, and Acquisition Time, Compared with Standard ABC/2 Manual Volume Calculation.

BACKGROUND AND PURPOSE: Intracerebral hemorrhage accounts for 6.5%-19.6% of all acute strokes. Initial intracerebral hemorrhage volume and expansion are both independent predictors of clinical outcomes and mortality. Therefore, a rapid, unbiased, and precise measurement of intracerebral hemorrhage volume is a key component of clinical management. The most commonly used method, ABC/2, results in overestimation. We developed an interactive segmentation program, SegTool, using a novel graphic processing unit, level set algorithm. Until now, the speed, bias, and precision of SegTool had not been validated. MATERIALS AND METHODS: In a single stroke academic center, 2 vascular neurologists and 2 neuroradiologists independently performed a test-retest experiment that involved repeat measurements of static, unchanging intracerebral hemorrhage volumes on CT from 76 intracerebral hemorrhage cases. Measurements were made with SegTool and ABC/2. True intracerebral hemorrhage volumes were estimated from a consensus of repeat manual tracings by 2 operators. These data allowed us to estimate measurement bias, precision, and speed. RESULTS: The measurements with SegTool were not significantly different from the true intracerebral hemorrhage volumes, while ABC/2 overestimated volume by 45%. The interrater measurement variability with SegTool was 50% less than that with ABC/2. The average measurement times for ABC/2 and SegTool were 35.7 and 44.6 seconds, respectively. CONCLUSIONS: SegTool appears to have attributes superior to ABC/2 in terms of accuracy and interrater reliability with a 9-second delay in measurement time (on average); hence, it could be useful in clinical trials and practice.

Copper deficiency myeloneuropathy resembling B12 deficiency: partial resolution of MR imaging findings with copper supplementation.

Copper deficiency has been associated with a clinical syndrome, myeloneuropathy. Radiographic changes resembling B(12) deficiency in the cervical spinal cord have been described. We present a case of copper deficiency myeloneuropathy, with cervical MR imaging findings resembling B(12) deficiency, which partially reversed following copper supplementation. This is, to our knowledge, the first described case of radiographic improvement with copper supplementation.

Complete Genome Sequence of Brucella abortus SKN 13 Isolated from Placenta of Aborted Cattle in Gujarat, India.

Brucella abortus is generally known to cause brucellosis in cattle and buffalo. Here, we report the draft genome sequence of Brucella abortus SKN 13, isolated from aborted cattle placenta in the area of Gujarat, India, providing precious resources for comparative genomic analyses of Brucella field strains.

Comparison of molecular and microscopic technique for detection of Theileria annulata from the field cases of cattle.

AIM: Tropical theileriosis is fatal hemoprotozoal disease of dairy animals caused by Theileria annulata. The aim of the present study was to detect the T. annulata and comparison of results of molecular and microscopic techniques. MATERIALS AND METHODS: A total of 52 blood samples were collected from the cattle suspected for theileriosis across the Banaskantha district. All the samples were screened for theileriosis using Giemsa's staining technique and polymerase chain reaction (PCR). RESULTS: Total of 17 (32.69%) and 24 (46.15%) samples were found positive for theileriosis by microscopic examination and PCR test, respectively. It revealed that the study area is endemic for theileriosis, and the microscopic technique has 70.83% sensitivity and 100% specificity with respect to PCR technique. CONCLUSION: It may be concluded from the present study that the PCR is comparatively sensitive technique than microscopic examination and may be recommended to use in the field for screening of theileriosis in the study area, where a high prevalence of diseases have been reported due to intensive dairy farming.

Spondyloepimetaphyseal dysplasia: clinical and radiologic investigation of a large kindred manifesting autosomal dominant inheritance, and a review of the literature.

Spondyloepimetaphyseal dysplasias (SEMDs) are a heterogeneous group of skeletal disorders that can have a genetic basis, but their classification and prognostication suffer because few families have been extensively studied. We describe a large kindred affected by a unique type of SEMD that is transmitted as an autosomal dominant trait. The propositus and his affected brother and first cousin were evaluated as inpatients. Other kindred members were screened by telephone interviews and lateral thoraco-lumbar spine radiographs, and then, in most cases, investigated by additional x-ray studies. Of the 29 living members of the kindred, 22 were studied radiologically. Among the 22 subjects investigated, 15 were affected, and the status of 1 individual with minor changes on x-ray was indeterminate. The deceased patriarch was presumed to be affected. These 16 affected subjects could usually, but not invariably, be distinguished from their unaffected sex-matched siblings by their smaller heights. Nevertheless, it was only affected children who had short stature; the heights of all affected adults were normal. Often, affected individuals had rhizomelic shortening, especially of the lower extremities, and genu varum (not always evident clinically, but present on radiographs). Occasionally, they also manifested limited extension of their upper limbs. Radiologic study showed abnormal metaphyses, epiphyses, and vertebrae in affected children, but these 3 skeletal regions became less remarkable by late childhood and most affected adults had normal epiphyses. One obligate affected man had only spinal changes. Despite their normal heights, severely affected adults who had bowing deformity of their legs developed disabling degenerative joint disease limited to the knees in the 7th decade of life--disease severe enough to require knee replacement surgery.

[125I]S(-)-zacopride labels a novel 5-hydroxytryptamine sensitive recognition site in rat duodenum and ileum.

Autoradiographic binding studies using [125I]S(-)-zacopride (0.1 nM) identified non-5-HT3 specific binding sites (defined by 5-hydroxytryptamine (5-HT), 1.0 microM) in the rat duodenum and ileum and some other peripheral tissues (adrenal gland, liver, stomach, kidney and spleen). In the rat duodenum and ileum, saturation studies with [125I]S(-)-zacopride indicated that the specific binding was saturable and of high affinity to an apparently homogenous population of binding sites (duodenum Bmax = 1.88 fmol/mg, Kd = 0.078 nM; ileum Bmax = 1.60 fmol/mg, Kd = 0.071 nM). Competition studies with slices of either duodenum or ileum indicated that the pharmacology of the [125I]S(-)-zacopride recognition site in both tissues was comparable but differed from all 5-HT receptors and uptake sites reported to date. However, the [125I]S(-)-zacopride recognition site displayed some pharmacological and regional similarity to the 5-HT1P recognition site: The sensitivity of the [125I]S(-)-zacopride binding in the duodenum and ileum to GTP indicates that the radiolabelled recognition site may represent a functional G-protein coupled receptor.

A novel radioligand [125I]BQ-3020 selective for endothelin (ETB) receptors.

A linear endothelin (ET) analog, N-acetyl-LeuMetAspLysGluAlaValTyrPheAlaHisLeu-AspIleIleTrp (BQ-3020), is highly selective for ETB receptors. BQ-3020 displaces [125I]ET-1 binding to ETB receptors (nonselective to ET isopeptides) in porcine cerebellar membranes (IC50: 0.2nM) at a concentration 4,700 times lower than that to ETA receptors (selective to ET-1) on aortic vascular smooth muscle cells (VSMC) (IC50: 940nM). BQ-3020 as well as ET-1 and ET-3 elicits vasoconstriction in the rabbit pulmonary artery. The ETA antagonist BQ-123 failed to inhibit this BQ-3020-induced vasoconstriction. Furthermore, BQ-3020 elicits endothelium-dependent vasodilation. These data indicate that BQ-3020 has ETB agonistic activity. The radioligand [125I]BQ-3020 binds to cerebellar membranes at single high affinity sites (Kd = 34.4pM), whereas it scarcely binds to VSMC. [125I]BQ-3020 binding to the cerebellum was displaced by BQ-3020, ET-1 and ET-3 in a nonselective manner (IC50: 0.07-0.17nM). However, the binding of [125I]BQ-3020 was insensitive to the ETA antagonist BQ-123 and other bioactive peptides. Both [125I]ET-1 and [125I]BQ-3020 show slow onset and offset binding kinetics to ETB receptors. These data indicate that the radioligand [125I]BQ-3020 selectively labels ETB receptors and that the slow binding kinetics of ET-1 are dependent on the peptide sequence from Leu6 to Trp21, but not on the structure formed by its two disulfide bridges.

The development and evaluation of a 125I radioimmunoassay for the measurement of LSD in body fluids.

A radioimmunoassay (RIA) has been developed for the direct detection of LSD in biological fluids. The radiotracer, (+)-2-[125I]iodo-LSD, allows the use of gamma-counting rather than the liquid scintillation counting currently used for existing 3H radioimmunoassays. The assay is specific for LSD and very closely related compounds. It is inexpensive, sensitive, simple to use and small volumes of samples (50 microliter) can be assayed directly without the need for any time-consuming extraction procedures. The cut-off levels are 1.2 ng/ml in blood and 3.0 ng/ml in urine. The results obtained using the 125I assay described in this work compare very favourably with those obtained using the 3H assay currently used by Home Office Forensic Science Laboratories. The advantages of the assay make it a most appropriate method for the routine screening of LSD in biological samples of forensic interest.

The 1985 national immunisation survey: diphtheria, tetanus, and pertussis (whooping cough)

In April 1985 a national immunisation survey was conducted in which sera were collected from almost 3000 randomly selected children throughout New Zealand. The sera collected were from about 1000 new school entrants (mean age 5 years), 1000 standard 3 students (mean age 10 years), and 1000 form 4 students (mean age 15 years). The sera were tested for diphtheria and tetanus immunity and antibody to pertussis by enzyme-linked immunosorbent assay (ELISA) tests. The percentage of those immune to diphtheria decreased with age from 73.1% in the 5 year olds to 53.7% in the 15 year olds. The percentage of those immune to tetanus decreased with age from 66.9% in the 5 year olds to 54.6% in the 10 year olds but then increased to 64.0% in the 15 year olds. While more than 71.0% of the 10 and 15 year olds had measurable antibody to pertussis, only a low 54.4% of the 5 year olds did. The proportion immune to diphtheria and tetanus in both Maoris and Europeans was approximately similar in all three age groups. More Maoris (78.9%) than Europeans (66.1%) had antibody to pertussis. When the data had been standardised for age and ethnic group the percentage with antibody to pertussis and immunity to diphtheria was highest in the southern region, while for tetanus immunity the percentage immune was highest in the northern and southern regions. We conclude that a sizeable pool of new school entrants (mean age 5 years) without antibody to pertussis is accumulating at a time when pertussis still persists.(ABSTRACT TRUNCATED AT 250 WORDS)

1988 New Zealand national immunisation survey: methodology.

In 1985 the Department of Health carried out a survey using a two stage stratified random sampling technique to select approximately 3000 children (made up of equal numbers of 5, 10 and 15 year olds). The principal aim was to provide a random sample of sera which could be used or stored for the future to evaluate the national immunisation programme and for screening of an ethical nature. The sampling frame was the 1983 list of public and private schools as provided by the Department of Education. Ninety primary schools and 50 secondary schools were identified, from which 3688 children were asked to participate. There was a 79% consent response rate. The survey largely succeeded in its objective of providing a nationally representative group of children, although the response rate was lower in 5 year olds (74%), in Pacific Islanders (67%), in children from upper socioeconomic status groups (75%), and in children who were reported not to have been immunised (56%). A similar survey is recommended every five years. In the mean time, laboratory analysis of the collected serum samples continues and results will be published separately.

Hepatitis B in New Zealand children: the 1985 national immunisation survey.

In April 1985 a national immunisation survey was conducted, in the course of which blood samples were collected from 3000 randomly selected children throughout the country. There were 1000 new school entrants (mean age 5 years), 1000 standard 3 pupils (mean age 10 years), and 1000 form 4 students (mean age 15 years). The sera were tested for hepatitis B surface antigen, antibody to hepatitis B surface antigen, and antibody to hepatitis B core antigen, by ELISA. The prevalence of infection rose with age until by 15 years of age 13.1% of the study population (8.2% of the European and 42.0% of the Maori children) were marker positive. At all ages, Maori children were five times more likely to be positive for any marker, and approximately thirteen times more likely to be positive for antigen (actively infected), than the European children. Even when the data had been standardised for age and race, children resident in the eastern North Island were still almost three times more at risk than children in the South Island. Children in the remaining areas of the North Island were at approximately equal degrees of risk, intermediate between the high and low endemic areas mentioned. We conclude that universal childhood immunisation is necessary to control horizontal transmission of heptatis B virus in New Zealand.

The 1985 national immunisation survey: hepatitis A.

In April 1985 a national immunisation survey was carried out, during which sera were collected from approximately 3000 randomly selected children throughout New Zealand. The sample comprised approximately equal numbers of new school entrants (mean age 5 years), standard 3 pupils (mean age 10 years) and form 4 students (mean age 15 years). This collection of sera was tested for antibody to hepatitis A virus, a marker of past infection with this virus, by means of a sensitive ELISA test. Prevalence of infection was found to be less than 1% in the 5 year olds, about 3% in the 10 year olds, and about 9% in the 15 year olds. Amongst the 10 and 15 year olds, but not the 5 year olds, Maori children were approximately three times more likely to have been infected than European children. Children resident in the eastern part of the North Island had a higher risk of infection than other children, even after controlling for ethnic distribution.

Radiation dose reduction in paranasal sinus CT using model-based iterative reconstruction.

BACKGROUND AND PURPOSE: CT performed with Veo model-based iterative reconstruction has shown the potential for radiation-dose reduction. This study sought to determine whether Veo could reduce noise and improve the image quality of low-dose sinus CT. MATERIALS AND METHODS: Twenty patients consented to participate and underwent low- and standard-dose sinus CT on the same day. Standard-dose CT was created with filtered back-projection (120 kV[peak], 210 mA, 0.4-second rotation, and 0.531 pitch). For low-dose CT, mA was decreased to 20 (the remaining parameters were unchanged), and images were generated with filtered back-projection and Veo. Standard- and low-dose datasets were reconstructed by using bone and soft-tissue algorithms, while the low-dose Veo reconstruction only had a standard kernel. Two blinded neuroradiologists independently evaluated the image quality of multiple osseous and soft-tissue craniofacial structures. Image noise was measured by using multiple regions of interest. RESULTS: Eight women and 12 men (mean age, 63.3 years) participated. Volume CT dose indices were 2.9 mGy (low dose) and 31.6 mGy (standard dose), and mean dose-length products were 37.4 mGy-cm (low dose) and 406.1 mGy-cm (standard dose). Of all the imaging series, low-dose Veo demonstrated the least noise (P < .001). Compared with filtered back-projection low-dose CT using soft-tissue and bone algorithms, Veo had the best soft-tissue image quality but the poorest bone image quality (P < .001). CONCLUSIONS: Veo significantly reduces noise in low-dose sinus CT. Although this reduction improves soft-tissue evaluation, thin bone becomes less distinct.

Localization of a rapid CSF leak with digital subtraction myelography.

A 53-year-old woman with superficial siderosis underwent spinal MR imaging, which demonstrated a large cervicothoracic epidural fluid collection compatible with a CSF leak. Conventional and dynamic CT myelography failed to localize the dural tear because of rapid equilibration of myelographic contrast between the thecal sac and the extradural collection. The superior temporal resolution of digital subtraction myelography precisely localized the CSF leak preoperatively and led to the successful surgical correction of the dural tear.

Coverage of an ischial pressure ulcer with an interpolated contralateral posterior thigh fasciocutaneous flap.

The ipsilateral posterior thigh flap has become one of the workhorses for the closure of ischial pressure ulcers. We treated a 40-year-old paraplegic patient with a right ischial pressure ulcer, in whom all ipsilateral flap options had been expended or were precluded by scars. A contralateral left posterior thigh fasciocutaneous flap interpolated subcutaneously across the midline provided successful coverage.

Late re-opening of macular holes after initially successful treatment with vitreous surgery.

BACKGROUND: In more than 50% of eyes with idiopathic, full-thickness macular holes, vitrectomy results in clinical closure of the hole and improvement in visual acuity. Other than cataract formation, late complications of the procedure have not been reported. METHODS: Over a 4-year period, 167 eyes underwent successful vitreous surgery for idiopathic macular holes documented by clinical closure of the hole and an improvement in visual acuity of at least two Snellen lines. The authors retrospectively studied eight of these eyes. All initially had successful surgery; however, they subsequently had spontaneous late re-opening of the macular hole with acute diminution in vision. RESULTS: Late re-opening developed in these eyes between 2 and 22 months after initial surgery. The mean time for re-opening was 12.5 months. Three of eight eyes underwent a second vitrectomy, which was successful in improving visual acuity in two eyes. Another patient manifested the growth of an epiretinal membrane that apparently sealed the re-opened macular hole and improved vision. CONCLUSION: Late re-opening can complicate initially successful macular hole surgery and may occur in at least 4.8% (8 of 167) of initially successful operations. Re-opening has been documented as late as 22 months after initial surgery. Repeat vitrectomy with gas injection may result in reclosure of the hole and improvement in vision.