Efficient and cost-effective extraction of genomic DNA from formalin-fixed and paraffin-embedded tissues.

Diagnostic and investigative molecular pathology frequently has to resort to extraction of DNA from formalin-fixed and paraffin-embedded tissue samples. Although many different protocols are reported for this type of material, extraction of sufficient amounts of intact DNA is still challenging. Here, the authors report a reproducible, simple, cost-effective, and efficient protocol that yields up to 140 µg of DNA from approximately 10 to 15 mg of formalin-fixed and paraffin-embedded tissue samples and compare it to available protocols. The protocol allows stable amplification of DNA fragments up to 600 bp in length in a wide variety of tissues.

Some diminutive colorectal polyps can be removed and discarded without pathological examination.

BACKGROUND AND STUDY AIMS: Pathological examination of colorectal polyps is useful if clinical management is affected (i. e. when invasive carcinoma is detected or postpolypectomy surveillance interval is guided). Our aim was to assess whether the pathological examination of some diminutive (measuring ≤ 5 mm) polyps can be omitted. PATIENTS AND METHODS: Consecutive patients undergoing a colonoscopy at Pasteur Hospital (Colmar, France) between January and August 2008 were included in this prospective study. Six senior gastroenterologists predicted the future surveillance interval without referring to the result of pathological examination. RESULTS: In all, 350 polyps from 175 patients were removed and analyzed. The endoscopist was able to predict the correct surveillance interval without referring to the result of pathological examination in 118 patients (67.4 %; 95 % confidence interval [CI] 60.5 - 74.4). The pathological examination of 18.4 % (95 % CI 13.7 - 23.1) of diminutive polyps either associated with a cancer or a polyp measuring ≥ 10 mm or removed in very old or frail patients could be omitted without any consequence for the patient. If diminutive polyps one or two in number were discarded without pathological examination in patients with a personal history of colorectal neoplasm, three patients out of 43 would have a 5-year instead of a 3-year surveillance interval. As a whole, if 44.1 % (95 % CI 38.0 - 50.1) of diminutive polyps were discarded, the surveillance interval would remain identical in 98.3 % (95 % CI 96.4 - 100) of patients. CONCLUSIONS: The pathological examination of up to 44 % of diminutive polyps (i. e. 33 % of all polyps), can be safely omitted. The pathological examination would be required only for those with suspicious gross appearance, those three or more in number, and those isolated one or two in number that are removed from people without personal history of colorectal neoplasm.

A new subgroup of immunoglobulin heavy chain variable region genes for the assessment of clonality in feline B-cell lymphomas.

In human medicine, PCR-amplification of the complementarity determining region 3 of the immunoglobulin heavy chain genes followed by polyacrylamide gel electrophoresis (PAGE) is an accepted method to assess clonality in B-cell lymphomas and thereby facilitates the differentiation of neoplasias from benign hyperplasias or reactive infiltrates. To generate a basis for the development of a PCR-based assay for the assessment of clonality in feline B-cell lymphomas we analyzed 28 transcripts (cDNA) of the feline immunoglobulin heavy chain variable region genes (IGHV). Transcripts were generated using techniques for the amplification of unknown sequences (i.e. the SMART RACE and the CapFishing technique) as well as primers derived from sequences of the NCBI Trace archive of the cat. Analysis of this archive revealed traces similar to the human IGHV-1 and IGHV-3 subgroups of genes. By identification of the subgroup-specific leader sequence within the traces, two subgroup-specific primers for this region were designed and used to amplify the heavy chain variable region genes. Using all amplification techniques, transcripts of both subgroups were created and the subgroups were denominated according to their human counterparts as feline IGHV-1 and feline IGHV-3. By aligning previously described transcripts of the feline IGHV genes to our transcripts we were able to assign these to the IGHV-3 subgroup; therefore, this study provides the first description of the feline IGHV-1 subgroup of genes. On the basis of the IGHV-1 and IGHV-3 transcripts we developed a PCR-based assay. For each of the two subgroups we used one sense primer derived from the first and one sense primer derived from the third framework region each in combination with a mixture of three antisense primers derived from the fourth framework region. With these four sets of primers, the assay was able to detect monoclonality in 7/10 (70%) cats with histologically and immunohistochemically diagnosed B-cell lymphomas. In two of these cases, monoclonal rearrangement of the IGHV genes was only detectable with IGHV-1 subgroup-specific primers. Amplification of feline hyperplastic lymphatic tissue only gave results indicative of polyclonal populations. The use of a PCR-based assay in combination with standard techniques for the diagnosis of feline lymphoma is helpful and the characterization of the additional subgroup of feline variable regions genes puts this assay on a broader basis.

Evaluations of primary metals from NHEXAS Arizona: distributions and preliminary exposures. National Human Exposure Assessment Survey.

NHEXAS AZ is a multimedia, multipathway exposure assessment survey designed to evaluate metals and other analytes. This paper reports the analyte-specific concentration distributions in each of the media examined (air, soil, house dust, food, beverage, and water), for various methodologies used (inductively coupled plasma-atomic emission spectroscopy and hydride generation-atomic absorption spectroscopy). Results are reported for the five primary metals (Pb, As, Cd, Cr, and Ni). Ingestion was the most important pathway of exposure. Metal concentrations in air were very low (ng/m3) and found only above the 90th percentile. Metals were commonly found in house dust and soil. Exposure transfer coefficients minimize the importance of this component for those over the age of 6 years. When ranked by exposure, food, beverage, and water appeared to be the primary contributors of metal exposure in NHEXAS AZ. For instance, at the 90th percentile, Pb was undetected in air, found at 131 and 118 microg/m3 in floor dust and soil, respectively, and measured at 16 microg/kg in food, 7.1 microg/kg in beverage, and 2.0 and 1.3 microg/l in drinking and tap water, respectively. We calculated preliminary estimates of total exposure (microg/day) for each participant and examined them independently by age, gender, and ethnicity as reported by the subjects in the NHEXAS questionnaire. At the 90th percentile for Pb, total exposures were 64 microg/day across all subjects (n=176); adult men (n=55) had the greatest exposure (73 microg/day) and children (n=35) the least (37 microg/day). Hispanics (n=54) had greater exposure to Pb (68 microg/day) than non-Hispanics (n=119; 50 microg/day), whereas non-Hispanics had greater exposure for all other metals reported. These results have implications related to environmental justice. The NHEXAS project provides information to make informed decisions for protecting and promoting appropriate public health policy.

Guiding language development: how African American mothers and their infants structure play interactions.

This investigation explored how African American mothers and their infants at the single-word stage of development structured their play and communicated with one another. Six mother-child dyads of low socioeconomic status (SES) and six of middle SES were observed at play. Few group differences were found, with the majority of the differences involving language behaviors. The middle-SES dyads included language goals more often in their play. Middle-SES infants initiated play verbally more frequently and produced over twice as many vocalizations as their low-SES peers. In addition, middle-SES mothers used a wider variety of words when playing with their children than their low-SES counterparts. A range of play styles was found within both groups. These were categorized into three general play styles: mothers and children actively involved in play; mothers' involvement varied; and children actively engaged and mothers attentive.

Long-term outcome of patients with very long stents for treatment of diffuse coronary disease.

OBJECTIVES: The study sought to determine the 6-month clinical outcome of patients who underwent implantation of very long coronary stents to treat diffuse disease and/or long dissections and to compare the findings with those reported in the literature for patients who underwent implantation of multiple short coronary stents. BACKGROUND: New designs of flexible stents enable the implantation of long stents rather than multiple short, older design stents. The initial experience is very promising but the long-term outcome has not been described yet. METHODS: Fifty-seven consecutive patients in whom 67 long stents (>/=30 mm) were successfully deployed were included in this study. Six-month clinical and angiographic follow-up was prospectively collected. Patients with recurrent angina underwent coronary angiography without further testing. Patients who remained asymptomatic at the 6-month follow-up visit underwent positron emission tomographic imaging, and those with results suggestive of ischemia underwent coronary angiography. A combined study end point was defined as death, myocardial infarction, and the need for target vessel revascularization. RESULTS: Only 1 patient (2%) reached a study end point at hospital discharge. An additional 20 patients (total 21 patients [37%]) reached an end point by 6 months. The outcome was not influenced by the clinical presentation (stable or unstable angina) or by the indication for stenting (elective or emergency). Predictors for adverse outcome were multiple stents per narrowing (63% vs 29%, P <. 04), and stents smaller than 3.5 mm (49% vs 22%). Narrowing and stent length were not predictive of a study end point in narrowings that were successfully treated by a single long stent. CONCLUSIONS: Elective stenting provides an effective solution for patients with diffuse coronary disease provided that a single long stent (usually <40 mm) can cover the full length of the narrowing. The results are better when vessels larger than 3 mm are treated. Compared with multiple short stents, implantation of a single long stent is probably at least as effective, and the procedure is quicker and cheaper and thus should be the preferred approach.

A first step incorporating smoking education into a Brazilian medical school curriculum: results of a survey to assess the cigarette smoking knowledge, attitudes, behaviour, and clinical practices of medical students.

Smoking-related illnesses are the leading causes of morbidity and mortality in Brazil. Despite a smoking prevalence of approximately 40%, there is limited national effort to reduce tobacco use in Brazil by means of public education and training of health care professionals to promote smoking education. In particular, the need for information about tobacco warrants increased emphasis in undergraduate medical education. An educational program on nicotine addiction during medical school could facilitate the incorporation of smoking cessation interventions into routine medical practice. As a preliminary step toward implementing a tobacco education and intervention program, this study was designed to assess knowledge and attitudes about smoking among Brazilian medical students. Five hundred thirteen (N = 513) medical students from the Federal University of Rio Grande do Sul, the southernmost state of Brazil, completed a self-reported questionnaire during the 1995-1996 academic school year. Most students recognize the adverse health effects of smoking and the importance of their professional role in promoting smoking cessation. In contradiction, however, few medical students currently provide their patients who smoke with even minimal intervention. This discrepancy supports the idea that training in nicotine addiction and smoking cessation techniques will help medical students to develop the skills and confidence needed to successfully intervene with their current and future patients.

Creating illegal immigrants.

PIP: This paper studies the prospects of implementation of guest-worker programs by governments that are based on an intended temporary presence of foreign workers. The purpose of guest- worker programs is to alleviate sector-specified labor shortages. The intention of a temporary stay is reflected in families left behind, and in the employers that often take responsibility for their workers' housing, health care, and other services during the temporary stay. Yet, some foreign workers prefer not to return home. In this case, the government and the employer ensure that the worker returns home after his legal employment. The government's intention is revealed through a policy granting the worker a temporary admission to the country. The legal employer pays a bond to bring in the foreign worker, and should the worker switch to illegal employment, the employer loses the value of the bond. Despite the intentions of the government, the legal employer can find his position to be quite precarious. If his legal workers are still with him at the end of the legal period of employment, it is only because they have rejected an illegal offer or they have never received an offer; and, if the worker leaves the legal employer just before he is to be deported, the employer has paid twice, once through the higher second-period wage and the second time by way of the forfeited bond. The anticipations for success of the intentions of temporary guest-worker programs can only be overly optimistic if the instruments used to enforce the policy are a bond and duration of permissible stay. The accumulation of an illegal population appears to be an inevitable consequence of a guest-worker program.^ieng

A contextually relevant assessment of the impact of child maltreatment on the social competencies of low-income urban children.

OBJECTIVE: A developmental-ecological model was used to explore the psychological adjustment and peer social competencies of maltreated preschool children. METHOD: Enhanced research methods were used that included verification of maltreatment status, multiple-variable matching of comparison children, and contextually relevant assessment. One hundred eight Head Start children, of whom 54 were maltreated, participated. Assessment of social functioning included measurement of peer play interactions, global social skills, peer sociometrics, and teacher and parent ratings of behavior problems. RESULTS: Findings indicated that maltreated children were significantly less interactive in peer play and overall exhibited less self-control and interpersonal skill in social interactions than nonmaltreated children. In addition, maltreated children were more likely than their nonmaltreated peers to receive low sociometric ratings and were found by both teachers and parents to display more internalizing behavior problems than comparison children. CONCLUSIONS: These findings indicate the importance of using developmentally sensitive research methods to inform contextually relevant interventions.

One-stage coronary angiography and angioplasty.

The combination of diagnostic angiography and angioplasty as a single procedure is becoming common practice in many institutions, but the feasibility and safety of this strategy have not been reported. This report describes 2,069 patients who underwent coronary angioplasty over a 3-year period at an institution where combined angiography and angioplasty is the norm. All patients were prepared before angiography for potential immediate angioplasty. In 1,719 patients, angioplasty was performed immediately after the diagnostic angiogram, while separate procedures were performed in 350 patients. Of those 350 patients, 254 were referred for angioplasty after diagnostic angiography at other hospitals. One thousand one hundred ninety-seven patients were admitted electively for treatment of stable angina pectoris, and 872 underwent procedures during hospitalization for unstable angina or acute myocardial infarction. One thousand nine hundred seven patients (92.2%) had successful angioplasties; in 130 patients (6.3%) the lesion could not be dilated, but no complication occurred, and in 32 patients (1.5%) angioplasty ended with a major complication (0.8% death, 1.0% Q-wave myocardial infarction, 0.5% emergency coronary artery bypass surgery). There was no difference between the combined and staged groups with regard to success, major and minor complication rates or in length of hospitalization after angioplasty. We conclude that routine combined strategy for angiography and angioplasty is feasible, safe, easier for the patient, and more cost-effective than 2 separate procedures.

Role of the Essential Health Services Commission in health care reform.

The New Jersey Essential Health Services Commission was established in 1992. The author reviews the role of the New Jersey Essential Health Services Commission in health care reform, including a subsidized health benefits program and payments to hospitals for charity care.

[The state of professional psychological guidance for patients with multiple sclerosis]. / Zum Stand der professionellen psychischen Betreuung Multiple-Sklerose-Kranker.

Over a period of two years, a total of 847 patients with multiple sclerosis (283 male and 564 female aged 18 to 79 years) were interviewed relative to the psychological care provided. Two patients currently participated in regular client-centered group therapy, 11 patients had regularly attended individual or group psychotherapy in previous years, and one patient had undergone family therapy several years ago. Another 26 patients reported occasional supportive counselling by their neurologist or family doctor in the framework of outpatient care. Ten patients used relaxation techniques such as autogenic training, and 42 patients reported emotional benefit from contacts within an MS self-help group. The findings of our study show that professional psychological care of persons with MS is extremely sparse, substantiating the call for better psychological services in coping with MS.

Urinary leukocyte esterase screening test for asymptomatic chlamydial and gonococcal infections in males.

We evaluated the ability of the urinary leukocyte esterase test to predict culture-verified chlamydial and gonococcal urethritis among asymptomatic adolescent males. Nine hundred forty-eight sexually active males provided first-catch urine samples for esterase screening, and 76 (8%) tested positive (greater than or equal to 1+). Among 435 boys who agreed to undergo urethral culture, the esterase was positive in 66 (15%), Chlamydia trachomatis was isolated from 39 (9%), and Neisseria gonorrhoeae was isolated from 14 (3%). The sensitivity, specificity, and positive and negative predictive values for the esterase test were 72%, 93%, and 58% and 96%, respectively. Using the esterase test to screen asymptomatic males for urethritis, we identified 38 culture-verified infections that otherwise would have remained undetected. The urinary leukocyte esterase test is a noninvasive and cost-effective screening method to detect urethritis among asymptomatic adolescent males.

Ultrasound assessment of the bladder volume after anterior colporrhaphy.

In 40 gynecological patients 44 different determinations of the bladder volume were made using ultrasonic methods. The product of bladder depth, height, and width, as determined from transverse and sagittal scans, showed the best correlation with the bladder volume measured by urethral catheter (r = 0.981). For 73% of the measurements the error was under 20% when the true bladder volume was greater than 100 cm3. This method gives a reasonable assessment of the bladder residual volume. It is quick, safe, and repeatable and, therefore, useful in postoperative clinical practice.

The computer. An orthopedic instrument.

The computer has become an important tool in orthopedic practice. Indeed, it is now invaluable and will soon be essential. It is still possible to run an orthopedic office without a computer--just as it is still possible to perform meniscectomy by arthrotomy--but that is not what most of us would do. The computer enhances efficiency, can improve business practice, assists in marketing, and helps store and retrieve medical information. The ultimate impact is not only on "business" functions but also on the quality of care.

Left ventricular function in acute myocardial infarction: assessment by nuclear angiography.

Left ventricular function was assessed in 38 patients two to six days after acute myocardial infarction using nuclear angiocardiography and the following parameters were measured: Left ventricular end-diastolic (LVEDV) and end-systolic volumes (LVESV), ejection fraction (LVEF), indices of left ventricular filling and emptying, right ventricular ejection fraction and ejection rate. Their clinical significance was assessed by their relationship to the patients site and size of infarction, functional capacity, morbidity and mortality. The most sensitive indices of depressed left ventricular function were the EF and ESV. Thus, function was preserved in patients with a small inferior infarction (LVEF = 0.57 +/- 0.07, LVESV = 69 +/- 14 ml) and in Killip Class I (LVEF = 0.48 +/- 0.13, LVESV = 80 +/- 20 ml). Function was disturbed most in patients with extensive anterior infarction (LVEF = 0.18 +/- 0.12, LVESV = 131 +/- 46 ml), Killip Class IV (LVEF = 0.13 +/- 0.07, LVESV = 160 +/- 35 ml), cardiogenic shock (LVEF = 0.14 +/- 0.07, LVESV = 160 +/- 35 ml), pulmonary edema (LVEF = 0.11 +/- 0.06, LVESV = 166 +/- 25 ml) and pulmonary capillary wedge pressure greater than 20 mm Hg (LVEF = 0.14 +/- 0.07, LVESV = 160 +/- 33 ml). Previous infarction was associated with LV dilatation and a greater LVEDV. A lower ejection fraction signified a large infarct and poor left ventricular function. If the ejection fraction was less than 0.15, the patients were unlikely to leave the hospital alive, or if less than 0.25, they were left with poor residual ventricular function and either had significant cardiac failure or high late mortality. Nuclear angiocardiography was a simple method of predicting the clinical pattern and prognosis in each patient and emphasized the importance of limiting infarct size in acute myocardial infarction.