Antimicrobial resistance of Campylobacter isolates from retail meat in the United States between 2002 and 2007.

The emergence of antimicrobial resistance in Campylobacter spp. has been a growing public health concern globally. The objectives of this study were to determine the prevalence, antimicrobial susceptibility, and genetic relatedness of Campylobacter spp. recovered by the National Antimicrobial Resistance Monitoring System (NARMS) retail meat program. Retail meat samples (n = 24,566) from 10 U.S. states collected between 2002 and 2007, consisting of 6,138 chicken breast, 6,109 ground turkey, 6,171 ground beef, and 6,148 pork chop samples, were analyzed. A total of 2,258 Campylobacter jejuni, 925 Campylobacter coli, and 7 Campylobacter lari isolates were identified. Chicken breast samples showed the highest contamination rate (49.9%), followed by ground turkey (1.6%), whereas both pork chops and ground beef had <0.5% contamination. The most common resistance was to doxycycline/tetracycline (46.6%), followed by nalidixic acid (18.5%), ciprofloxacin (17.4%), azithromycin and erythromycin (2.8%), telithromycin (2.4%), clindamycin (2.2%), and gentamicin (<0.1%). In a subset of isolates tested, no resistance to meropenem and florfenicol was seen. C. coli isolates showed higher resistance rates to antimicrobials, with the exception of doxycycline/tetracycline, than those seen for C. jejuni. Pulsed-field gel electrophoresis (PFGE) fingerprinting resulted in 1,226 PFGE profiles among the 2,318 isolates, with many clones being widely dispersed throughout the 6-year sampling period.

The prevalence of BRCA1 mutations among young women with triple-negative breast cancer.

BACKGROUND: Molecular screening for BRCA1 and BRCA2 mutations is now an established component of risk evaluation and management of familial breast cancer. Features of hereditary breast cancer include an early age-of-onset and over-representation of the 'triple-negative' phenotype (negative for estrogen-receptor, progesterone-receptor and HER2). The decision to offer genetic testing to a breast cancer patient is usually based on her family history, but in the absence of a family history of cancer, some women may qualify for testing based on the age-of-onset and/or the pathologic features of the breast cancer. METHODS: We studied 54 women who were diagnosed with high-grade, triple-negative invasive breast cancer at or before age 40. These women were selected for study because they had little or no family history of breast or ovarian cancer and they did not qualify for genetic testing using conventional family history criteria. BRCA1 screening was performed using a combination of fluorescent multiplexed-PCR analysis, BRCA1 exon-13 6 kb duplication screening, the protein truncation test (PTT) and fluorescent multiplexed denaturing gradient gel electrophoresis (DGGE). All coding exons of BRCA1 were screened. The two large exons of BRCA2 were also screened using PTT. All mutations were confirmed with direct sequencing. RESULTS: Five deleterious BRCA1 mutations and one deleterious BRCA2 mutation were identified in the 54 patients with early-onset, triple-negative breast cancer (11%). CONCLUSION: Women with early-onset triple-negative breast cancer are candidates for genetic testing for BRCA1, even in the absence of a family history of breast or ovarian cancer.

Use of high-frequency ultrasound in the assessment of injectable dermal fillers.

BACKGROUND: The use of dermal fillers for enhancing lips and reducing wrinkles is currently one of the fastest growing sectors of the cosmetic surgery market. There are numerous fillers available, some are synthetic others are isolated from biological material. Once injected the fillers have a varied lifespan ranging from months to years depending upon the material, site of injection and individual response. Current assessment techniques of filler performance are mostly limited to evaluations of the skin surface topography, and not to what is happening to the filler beneath the skin surface. The aim of this work was to see if high-frequency ultrasound could be used to image and measure filler dimensions in situ. METHOD: This was a pilot study of six healthy female volunteers aged 36-53 visiting the surgical outpatients department of a hospital in Glasgow. Volunteers had been injected with filler material into their upper lip 6 months before the visit. The patients all had their upper lip scanned using high-frequency ultrasound. The subsequent images were then assessed using the scanner software to assess the dimensions of the filler. RESULTS: The filler material was clearly visible with the ultrasound and subsequently measurable in each scan. Each scan procedure was completed within a short time period meaning quantitative data could be acquired with minimum trauma to the volunteer. The scan images and data also provided valuable information for the volunteers and reinforced their perception of the fillers effect on their features. CONCLUSIONS: High-frequency ultrasound scanning provides a non-invasive, convenient and rapid technique for the assessment of filler performance. This pilot study produced three valuable pieces of information: The ultrasound can image the filler material from which quantitative measurements can be made. The technique is rapid and cost effective ...This investigation helped to reinforce the volunteer's perception of the filler effect.

Resistance to Bacillus thuringiensis toxin Cry2Ab in a strain of Helicoverpa armigera (Lepidoptera: Noctuidae) in Australia.

Transgenic cotton, Gossypium hirsutum L., expressing the crylAc and cry2Ab genes from Bacillus thuringiensis (Bt) Berliner variety kurstaki in a pyramid (Bollgard II) was widely planted for the first time in Australia during the 2004-2005 growing season. Before the first commercial Bollgard II crops, limited amounts of cotton expressing only the crylAc gene (Ingard) was grown for seven seasons. No field failures due to resistance to CrylAc toxin were observed during that period and a monitoring program indicated that the frequency of genes conferring high level resistance to the CrylAc toxin were rare in the major pest of cotton, Helicoverpa armigera (Htibner) (Lepidoptera: Noctuidae). Before the deployment of Bollgard II, an allele conferring resistance to Cry2Ab toxin was detected in field-collected H. armigera. We established a colony (designated SP15) consisting of homozygous resistant individuals and examined their characteristics through comparison with individuals from a Bt-susceptible laboratory colony (GR). Through specific crosses and bioassays, we established that the resistance present in SP15 was due to a single autosomal gene. The resistance was recessive. Homozygotes were highly resistant to Cry2Ab toxin, so much so, that we were unable to induce significant mortality at the maximum concentration of toxin available. Homozygotes also were unaffected when fed leaves of a cotton variety expressing the cry2Ab gene. Although cross-resistant to Cry2Aa toxin, SP15 was susceptible to CrylAc and to the Bt product DiPel.

The detection of Mycobacterium leprae protein and carbohydrate antigens in skin and nerve from leprosy patients with type 1 (reversal) reactions.

Type 1 (reversal) reactions are the most common immunological complications of leprosy. These episodes of delayed hypersensitivity produce severe local immunopathology and ultimately nerve damage. To date, the Mycobacterium leprae antigens associated with type 1 reactions have not been identified. Using monoclonal antibodies to defined protein and carbohydrate M. leprae epitopes (65, 35 and 28 kd and lipoarabinomannan [LAM]) in a two-step immunoperoxidase staining technique, M. leprae antigens were demonstrated in skin and nerve biopsies from patients in reversal reaction. Antigen presence and staining patterns were similar in skin and nerve lesions, implying that the pathological processes are similar in the two sites. Antigens were present both in macrophages and Schwann cells but also as a diffuse extracellular infiltrate associated with the inflammatory infiltrate. The 28-kd antigen was present most strongly and may be a potential candidate antigen for initiating type 1 reactions. LAM also stained strongly and persisted after treatment. The possible roles of LAM and 65 kd in the cellular events of type 1 reactions are discussed.

The use of high-frequency diagnostic ultrasound to investigate the effect of hormone replacement therapy on skin thickness.

BACKGROUND/AIMS: Previous investigations have suggested that hormone replacement therapy (HRT) could have a positive effect on the maintenance of skin thickness post-menopause. Previous skin measurement devices have proved variable in their accuracy and ease of use. This investigation assessed the effect of HRT on the skin in a noninvasive way, using high-frequency diagnostic ultrasound. METHOD: The study was a cross-sectional observational study, carried out at a menopause and gynaecology outpatient's clinic. A total of 84 women (comprising 34 HRT users, 25 post-menopausal controls, and 25 premenopausal controls) took part in the study. Each volunteer was scanned using diagnostic ultrasound on the arm. Skin thickness measurements were made from each scan using computerised image analysis. RESULTS: Skin thickness was shown to be greater in the HRT group than in the post-menopausal controls (P<0.01). CONCLUSIONS: High-frequency diagnostic ultrasound proved to be a useful clinical tool and showed that HRT appears to help maintain skin thickness in menopause.

High-frequency ultrasound imaging of the skin during normal and hypertensive pregnancies.

BACKGROUND/AIMS: Diagnosis of preeclampsia is currently made from blood pressure measurements taken at antenatal visits (either at the hospital or in the community). The aim of this work was to see whether the presence of underlying hypertensive diseases is accompanied by changes in the skin of pregnant women, which can be visualized using high-frequency diagnostic ultrasound. METHODS: This was a prospective study of pregnant and non-pregnant, hypertensive and non-hypertensive patients visiting the outpatient department of a central London Teaching Hospital. The study group consisted of 93 women, of which 30 were non-hypertensive in the second trimester of pregnancy, 26 were non-hypertensive in the third trimester of pregnancy, 9 were hypertensive in the second trimester of pregnancy, and 14 were hypertensive in the third trimester of pregnancy. Fourteen non-pregnant women of comparable age were recruited as controls. Changes in abdominal skin thickness and also skin structure, as analysed by fractal image analysis, was assessed in each patient. RESULTS: In a normal pregnancy, abdominal skin gets thinner as pregnancy progresses. In hypertensive patients, the skin thickness did not appear to alter. Image analysis of abdominal skin scans showed that the skin of non-hypertensive pregnant women and non-pregnant women are different. Whereas the analysis of hypertensive pregnant women and non-pregnant women showed they were the same. CONCLUSIONS: The data used to compare the groups indicates that if the abdominal skin of the patient does not get thinner as the pregnancy progresses there is an indication that the patient may be hypertensive. The fractal data comparing the groups indicates the following when comparing a patient's fractal signature with the non-pregnant control data: If the abdominal fractal for a pregnant woman is similar to the control group, there is an indication that the patient is hypertensive. It is difficult to predict hypertension in patients, and it is possible that a patient could develop severe preeclampsia between visits to the antenatal clinics. Therefore, if the high-frequency ultrasound scanner can pick up potential hypertensives early in pregnancy, these women could be identified as potentially high risk.

Gene Amplification of c-erbB-2 Detected by FISH.

In 1981, a novel transforming gene called neu, related to, but distinct from, the c-erbB protooncogene, was identified (1-3). In 1985, two groups independently isolated identical erbB-related genes from human DNA that they called HER-2 (4) and c-erbB-2 (5) located at chromosome band 17q11.2 and encoding a 185,000 Dalton tyrosine kinase (2,4). Studies conducted on human breast cancers showed 28% to have amplified HER2/neu present (7) and it was suggested that amplification or increased protein expression might confer a selective advantage (6). Over the last ten years, many studies have confirmed the value of using overexpression or amplification of the HER2/neu gene as a predictor of poor outcome in cases of breast cancer (7-9). Clinical trials are currently underway using anti-HER2/neu in an attempt to destroy malignant breast cells (10,11).

Simultaneous intracellular recording and calcium imaging in single neurons of hippocampal slices.

Fluorescent Ca2+ indicator dyes can be introduced into cells through the same microelectrode used for intracellular voltage recording. Simultaneous measurement of cell membrane potential and intracellular Ca2+ concentration can be very helpful in interpreting the mechanisms of Ca2+ increases. This chapter describes fluorescence image acquisition using a CCD camera and a computer program that also records a synchronized membrane potential trace. The same program allows for preliminary data analysis. More elaborate analyses can be accomplished with commercial programs. We also describe quantitative evaluations of sources of error in the use of the statistic deltaF/F as an indicator of Ca2+ concentration. Especially important errors to minimize are changes in background fluorescence and inappropriate autofluorescence corrections. Some improvement of fluorescence images of cells deep within slices may be accomplished by masking. One method is described for making a mask based on the raw fluorescence image. With another method, highly detailed cell morphologies may be conveyed by using masks based on neurobiotin injections and camera lucida drawings.

Long-term maintenance of mature hippocampal slices in vitro.

Cultures of primary neurons or thin brain slices are typically prepared from immature animals. We introduce a method to prepare hippocampal slice cultures from mature rats aged 20-30 days. Mature slice cultures retain hippocampal cytoarchitecture and synaptic connections up to 3 months in vitro. Spontaneous epileptiform activity is rarely observed suggesting long-term retention of normal neuronal excitability and of excitatory and inhibitory synaptic networks. Picrotoxin, a GABAergic Cl(-) channel antagonist, induced characteristic interictal-like bursts that originated in the CA3 region, but not in the CA1 region. These data suggest that mature slice cultures displayed long-term retention of GABAergic inhibitory synapses that effectively suppressed synchronized burst activity via recurrent excitatory synapses of CA3 pyramidal cells. Mature slice cultures lack the reactive synaptogenesis, spontaneous epileptiform activity, and short life span that limit the use of slice cultures isolated from immature rats. Mature slice cultures are anticipated to be a useful addition for the in vitro study of normal and pathological hippocampal function.

Fluorescence in situ hybridization analysis of HER-2/neu, c-myc, and p53 in endometrial cancer.

Our objective was to evaluate the association between HER-2/neu, c-myc, p53, and clinicopathologic variables in endometrial cancer using fluorescence in situ hybridization (FISH) cytogenetic analysis. FISH analysis for HER-2/neu, c-myc, and p53 was performed on 47 endometrial cancer specimens. Amplification of HER-2/neu was seen in 4/47 (8.5%) cases and amplification of c-myc was seen in 7 of 47 (15%) cases; neither was associated with adverse clinicopathologic variables or survival. Deletion of p53 was seen in 31/47 (66%) cases and was associated with poor histologic grade (P = 0.008). There was no impact of genetic alterations on overall survival or disease-free interval. Grade 3 tumor was associated with poor overall survival (P = 0.032). This study found that p53 deletion is a common genetic alteration in endometrial cancer and is associated with poor-grade tumors.

c-myc and chromosome 8 centromere studies of ovarian cancer by interphase FISH.

Forty tumor specimens from patients with ovarian cancer were studied for amplification of the c-myc oncogene relative to chromosome 8 centromere number using dual-color FISH. Interphase cytogenetic analysis showed amplification of the c-myc oncogene in 40% (16/40) of tumors using the standard oncogene:centromere ratio method of analysis. Eleven of these showed moderate amplification of c-myc, and 5 samples showed high amplification. Eight of the sixteen (50%) amplified tumors were polysomic centromere 8 as were 14 of the 24 (58%) non-amplified tumors. In previously reported work with these samples, the oncogene HER-2/neu, the chromosome 17 centromere, and the tumor suppressor gene p53 had been studied. When using the standard oncogene:centromere ratio criteria, 5 samples had amplification of both the c-myc and the HER-2/neu oncogenes, 5 samples had HER-2/neu amplification but not c-myc, 11 samples had c-myc amplification but not HER-2/neu, and 19 samples had neither oncogene amplified. The p53 gene was found to be deleted in 22.5% (9/40) of samples. The loss of the p53 gene did not appear to have any clinical correlation. The presence of an extra centromere 8 also did not appear to have any clinical correlation. The Kaplan-Meier survival curve for those patients who have c-myc amplification, while not statistically significant, appears to show a trend toward poorer survival. The survival curve for patients whose tumors have HER-2/neu amplification shows no clinical significance. It is of great interest, however, that the Kaplan-Meier plot of survival for patients whose tumors have amplification of both c-myc and HER-2/neu shows a significant difference (P = 0.047). The median survival times of the doubly amplified patient group and the non-doubly amplified groups were 12 and 43 months, respectively. This is the first study of the oncogene c-myc using FISH. The results suggest that the amplification of c-myc may indicate a poorer patient survival and that the amplification of both c-myc and HER-2/neu in combination may be a better prognostic indicator of poor patient survival.

Mus and Peromyscus chromosome homology established by FISH with three mouse paint probes.

Fluorescence-labeled DNA probes constructed from three whole house mouse (Mus domesticus) chromosomes were hybridized to metaphase spreads from deer mouse (Peromyscus maniculatus) to identify homologies between the species. Mus Chr 7 probe hybridized strongly to the ad-centromeric two-thirds of Peromyscus Chr 1q. Most of Mus 3 probe hybridized principally to two disjunct segments of Peromyscus Chr 3. Mus Chr 9 probe hybridized entirely to the whole Peromyscus Chr 7. Three Peromyscus linkage groups were assigned to chromosomes, based on linkage homology with Mus. The data also are useful in interpretation of chromosomal evolutionary history in myomorphic rodents.

Group I mGluR activation causes voltage-dependent and -independent Ca2+ rises in hippocampal pyramidal cells.

Application of the metabotropic glutamate receptor (mGluR) agonist (1S, 3R)-1-aminocyclopentane-1,3-dicarboxylic acid (ACPD) or the selective group I mGluR agonist (S)-3,5-dihydroxyphenylglycine (DHPG) depolarized both CA3 and CA1 pyramidal cells in guinea pig hippocampal slices. Simultaneous recordings of voltage and intracellular Ca2+ levels revealed that the depolarization was accompanied by a biphasic elevation of intracellular Ca2+ concentration ([Ca2+]i): a transient calcium rise followed by a delayed, sustained elevation. The transient [Ca2+]i rise was independent of the membrane potential and was blocked when caffeine was added to the perfusing solution. The sustained [Ca2+]i rise appeared when membrane depolarization reached threshold for voltage-gated Ca2+ influx and was suppressed by membrane hyperpolarization. The depolarization was associated with an increased input resistance and persisted when either the transient or sustained [Ca2+]i responses was blocked. mGluR-mediated voltage and [Ca2+]i responses were blocked by (+)-alpha-methyl-4-carboxyphenylglycine (MCPG) or (S)-4-carboxy-3-hydroxyphenylglycine (4C3HPG). These data suggest that in both CA3 and CA1 hippocampal cells, activation of group I mGluRs produced a biphasic accumulation of [Ca2+]i via two paths: a transient release from intracellular stores, and subsequently, by influx through voltage-gated Ca2+ channels. The concurrent mGluR-induced membrane depolarization was not caused by the [Ca2+]i rise.

A non-comparative multi-centre clinical evaluation of a new hydropolymer adhesive dressing.

This study evaluates the performance of a new hydropolymer adhesive dressing in the treatment of chronic or acute wounds, in the presence of blood and/or low levels of exudate, in 74 patients, in a five-site multi-centre non-comparative clinical trial of four weeks' duration. The primary efficacy variable was defined as the incidence of central island dressing adherence to the wound bed in the final stages of healing. The hydropolymer dressing performed extremely well with 98.4% (240 out of 244) of the dressings not adhering to the wound bed.

Basic principles of cancer genetics.

All cancer can be described as genetic, that is, due to altered DNA. Many of these mutations will be accumulated during the normal division of cells. However, some people may inherit abnormal genes, which predispose those individuals to high risk of certain malignancies. These individuals can sometimes be identified as having a family history of affected individuals, some of which might have early age of onset or multiple malignancies. Specific genes have been identified as being associated with certain of these malignancies. The hereditary cancers include (but are not limited to) ovary, breast, colon, endometrium, and to a lesser extent, prostate, skin and pancreas. Some of these cancer predisposing genes are highly penetrant with up to 80 to 90 percent of gene carriers developing the associated malignancy within a 70 year life expectancy. Molecular testing for the presence of cancer predisposing genes is available for many of the hereditary syndromes. While there currently is no way to correct a mutant gene, early detection and some techniques of chemoprevention are of clinical value. People who fear that they are at high risk only learn that they are not, can benefit from the relief of anxiety through the genetic counseling process.

The effects of ovarian hormone deficiency on wound contraction in a rat model.

OBJECTIVE: To demonstrate the effect of a deficiency of ovarian hormones on the process of wound contraction, using the oophorectomised rat model of the human menopause. DESIGN: A randomised controlled trial. POPULATION: Ninety-six adult Wistar rats were randomly allocated into either an oophorectomised group or a sham-oophorectomised control group. METHODS: Having confirmed a significant reduction in plasma oestradiol levels in the oophorectomised rats, full-thickness excised lesions were made in the flank skin of the adult rats at either two weeks or four months after oophorectomy, so that the effects of two different durations of hormone deficiency could be assessed and compared with the sham-oophorectomised controls. Following wounding, the rats were left for 3, 5, 10 or 22 days; wound contraction was assessed from photographs of the wounds taken at these intervals after injury. RESULTS: In the rats wounded four months after oophorectomy there was a slower rate of wound contraction, resulting in larger wounds at days 3, 5, 10 and 22, compared with control rats. No significant difference was observed in rats wounded two weeks after oophorectomy, indicating that the effects of ovarian hormone deficiency on this process are delayed. CONCLUSION: Due to the pivotal role of wound contraction in the process of wound healing these findings may be of clinical relevance and could have an important impact on the administration of hormone replacement therapy.

Developing genetic privacy legislation: the South Carolina experience.

The availability of presymptomatic and predisposition genetic testing has spawned the need for legislation prohibiting health insurance discrimination on the basis of genetic information. The federal effort, the Health Insurance Portability and Accountability Act (HIPAA) of 1996, falls short by protecting only those who access insurance through group plans. A committee of University of South Carolina professionals convened in 1996 to develop legislation in support of genetic privacy for the state of South Carolina. The legislation prevents health insurance companies from denying coverage or setting insurance rates on the basis of genetic information. It also protects the privacy of genetic information and prohibits performance of genetic tests without specific informed consent. In preparing the bill, genetic privacy laws from other states were reviewed, and a modified version of the Virginia law adopted. The South Carolina Committee for the Protection of Genetic Privacy version went a step further by including enforcement language and excluding Virginia's sunset clause. The definition of genetic information encompassed genetic test results, and importantly, includes family history of genetic disease. Our experience in navigating through the state legislature and working through opposition from the health insurance lobby is detailed herein.

Comparisons of intensity measures and their stability in male and female speakers.

The purpose of the present study was to provide data on the intensity characteristics of young adult speakers in terms of conversational intensity level, conversational intensity range, and available intensity range. Subjects included 20 males and 20 females, ages 20-30 years. Each subject was asked to read the Rainbow Passage at a conversational intensity level, as softly as possible without whispering, and as loudly as possible, on 2 separate days 1 week apart. The second and third sentences of the three readings on both days were analyzed for various intensity parameters. Results revealed a conversational intensity level of 70.42 dB for males and 68.15 dB for females. When male and female intensity measures were compared, few statistically significant differences were found. Further, when intensity measures for the first and second readings were compared, few significant differences were found.