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The advent of PCR has improved the identification of viruses in patients with community-acquired pneumonia (CAP). Several studies have used PCR to establish the importance of viruses in the aetiology of CAP.We performed a systematic review and meta-analysis of the studies that reported the proportion of viral infection detected via PCR in patients with CAP. We excluded studies with paediatric populations. The primary outcome was the proportion of patients with viral infection. The secondary outcome was short-term mortality.Our review included 31 studies. Most obtained PCR via nasopharyngeal or oropharyngeal swab. The pooled proportion of patients with viral infection was 24.5% (95% CI 21.5-27.5%). In studies that obtained lower respiratory samples in >50% of patients, the proportion was 44.2% (95% CI 35.1-53.3%). The odds of death were higher in patients with dual bacterial and viral infection (OR 2.1, 95% CI 1.32-3.31).Viral infection is present in a high proportion of patients with CAP. The true proportion of viral infection is probably underestimated because of negative test results from nasopharyngeal or oropharyngeal swab PCR. There is increased mortality in patients with dual bacterial and viral infection., (Copyright ©ERS 2016.)

A synthesis of 1- O -acetyl-3- O -(4-methoxybenzyl)-4- N -(9-fluorenylmethoxycarbonyl)-4- N -methyl-L-pyrrolosamine ( 7 ), which constitutes a protected form of the N , N -dimethyl-L-pyrrolosamine residues found within the antiproliferative bacterial metabolites (-)-lomaiviticins A and B ( 1 and 2 , respectively), is reported. The synthetic route to 7 proceeds in eight steps and 13% overall yield from ( E )-crotyl alcohol. The protected carbohydrate 7 is envisioned to be a useful derivative for syntheses of 1 and 2 .

Purpose: A Web-based application for coordinating medication-use evaluation (MUE) initiatives within the Veterans Affairs (VA) health care system is described., Summary: The MUE Tracker (MUET) software program was created to improve VA's ability to conduct national medication-related interventions throughout its network of 147 medical centers. MUET initiatives are centrally coordinated by the VA Center for Medication Safety (VAMedSAFE), which monitors the agency's integrated databases for indications of suboptimal prescribing or drug therapy monitoring and adverse treatment outcomes. When a pharmacovigilance signal is detected, VAMedSAFE identifies "trigger groups" of at-risk veterans and uploads patient lists to the secure MUET application, where locally designated personnel (typically pharmacists) can access and use the data to target risk-reduction efforts. Local data on patient-specific interventions are stored in a centralized database and regularly updated to enable tracking and reporting for surveillance and quality-improvement purposes; aggregated data can be further analyzed for provider education and benchmarking. In a three-year pilot project, the MUET program was found effective in promoting improved prescribing of erythropoiesis-stimulating agents (ESAs) and enhanced laboratory monitoring of ESA-treated patients in all specified trigger groups. The MUET initiative has since been expanded to target other high-risk drugs, and efforts are underway to refine the tool for broader utility., Conclusion: The MUET application has enabled the increased standardization of medication safety initiatives across the VA system and may serve as a useful model for the development of pharmacovigilance tools by other large integrated health care systems.

Purpose: The results of a survey assessing Medicare Part D enrollment, the use of pharmacotherapies for chronic diseases, and other medication-use issues in a population of elderly military veterans are presented., Methods: Medicare-eligible (i.e., ≥65 years of age) patients with documented recent service use at a single Veterans Affairs (VA) medical center were targeted for a mail survey. Women were oversampled (20%) to ensure an adequate sample size; the sample was weighted to adjust for this oversampling. Usable survey data were received from 458 survey respondents., Results: Nearly all respondents (93.2%) reported having one or more chronic conditions; of those, 93.3% reported regular use of multiple drug therapies, and 30.1% reported using medications prescribed by both VA and non-VA providers for the same chronic condition. About half of the survey respondents reported at least one office visit with a non-VA physician during the previous year, and 55.8% reported obtaining medications from non-VA pharmacies. More than half (54.1%) of the respondents reported non-VA medication coverage, with 21.2% indicating they were enrolled in Medicare Part D. Among the respondents who reported obtaining medications from non-VA pharmacies, substantial proportions reported discussing those medications with VA physicians never (38.4%) or infrequently (15.7%)., Conclusion: Although large proportions of Medicare-eligible veterans take multiple medications and use non-VA health care services and pharmacies, many do not discuss medications obtained outside the VA system with VA physicians, suggesting that increased efforts to enhance provider-patient communication and medication reconciliation across VA and non-VA systems of care may be warranted.

Objectives: To examine the change in use of high-risk medications for the elderly (HRME), as defined by the National Committee on Quality Assurance's Healthcare Effectiveness Data and Information Set (HEDIS) quality measure (HEDIS HRME), by older outpatient veterans over a 3-year period and to identify risk factors for HEDIS HRME exposure overall and for the most commonly used drug classes., Design: Longitudinal retrospective database analysis., Setting: Outpatient clinics within the Department of Veterans Affairs (VA)., Participants: Veterans aged 65 by October 1, 2003, and who received VA care at least once each year until September 30, 2006., Measurements: Rates of use of HEDIS HRME overall and according to specific drug classes each year from fiscal year 2004 (FY04) to FY06., Results: In a cohort of 1,567,467, high-risk medication exposure fell from 13.1% to 12.3% between FY04 and FY06 (P<.001). High-risk antihistamines (e.g., diphenhydramine), opioid analgesics (e.g., propoxyphene), skeletal muscle relaxants (e.g., cyclobenzaprine), psychotropics (e.g., long half-life benzodiazepines), endocrine (e.g., estrogen), and cardiac medications (e.g., short-acting nifedipine) had modest but statistically significant (P<.001) reductions (range -3.8% to -16.0%); nitrofurantoin demonstrated a statistically significant increase (+36.5%; P<.001). Overall HEDIS HRME exposure was more likely for men, Hispanics, those receiving more medications, those with psychiatric comorbidity, and those without prior geriatric care. Exposure was lower for individuals exempt from copayment. Similar associations were seen between ethnicity, polypharmacy, psychiatric comorbidity, access-to-care factors, and use of individual HEDIS HRME classes., Conclusion: HEDIS HRME drug exposure decreased slightly in an integrated healthcare system. Risk factors for exposure were not consistent across drug groups. Future studies should examine whether interventions to further reduce HEDIS HRME use improve health outcomes., (© 2011, Copyright the Authors. Journal compilation © 2011, The American Geriatrics Society.)

Background: Celiac disease (CD) is one of the most common lifelong disorders in western countries. However, most cases remain currently undiagnosed in North America, mostly due to poor awareness of CD by primary care physicians., Objectives: The aims of this study were (a) to determine whether an active case-finding strategy in primary care could increase the frequency of CD diagnosis and (b) to determine the most common clinical presentations of the condition., Methods: This was a multicenter, prospective study involving adult subjects during the years 2002-2004, attending one of the participating practices. All individuals with symptoms or conditions known to be associated with CD were tested for immunoglobulin A anti-transglutaminase (tTG) antibodies, and those with elevated anti-tTG were subsequently tested for IgA antiendomysial antibodies (EMA). All subjects who were positive for EMA were advised to undergo an intestinal biopsy and HLA typing., Results: The study group included 737 women and 239 men, with a median age of 54.3 yr. A positive anti-tTG test was found in 30 out of 976 investigated subjects (3.07%, 95% CI 1.98-4.16). CD was diagnosed in 22 patients (18 women, 4 men). The most frequent reasons for CD screening in these 22 cases were bloating (12/22), thyroid disease (11/22), irritable bowel syndrome (7/22), unexplained chronic diarrhea (6/22), chronic fatigue (5/22), and constipation (4/22). The prevalence of CD in the serologically screened sample was 2.25% (95% CI 1.32-3.18). The diagnostic rate was low at baseline (0.27 cases per thousand visits, 95% CI 0.13-0.41) and significantly increased to 11.6 per thousand visits (95% CI 6.8-16.4, P < 0.001) following active screening implementation., Conclusions: This study demonstrates that an active case-finding strategy in the primary care setting is an effective means to improve the diagnostic rate of CD in North America.

Objective: The U.S. Department of Veterans Affairs (VA) Pharmacy Benefits Management Strategic Healthcare Group (PBM-SHG) and its Medical Advisory Panel developed national criteria (guidelines) for the appropriate use of tamsulosin in the VA. Drug use evaluation (DUE) was performed to (a) determine the prescribing patterns (indications and patient follow-up monitoring as measured by a clinician.s note regarding evaluation of therapeutic response or report of adverse drug event) of tamsulosin, (b) assess the impact of the availability (not active dissemination) of national criteria for nonformulary use of tamsulosin, and (c) project the cost avoidance if generic terazosin was substituted for tamsulosin in those patients who were prescribed tamsulosin outside of appropriate use criteria., Methods: Geographically dispersed VA medical centers were identified for which tamsulosin utilization was significantly above and below the national average (4.8% of all prescriptions for alpha [alpha]-blockers) in January 2001. A data collection form for medical record abstraction was designed to capture the patient.s diagnosis, reported indication for tamsulosin, history of previous alpha-blocker use, tamsulosin follow-up evaluation, and the individual facility.s method of implementation of criteria for nonformulary use. Patients receiving a prescription for tamsulosin during a 3-month period preceding the posting of national criteria, and patients with a first-time prescription for tamsulosin during a 3-month period after the national criteria were posted were randomly selected by the PBM and assigned for chart review at each site., Results: Data for 332 patients were collected from 6 different sites over a 6-month period for each pregroup and postgroup beginning August 2001 and January 2002, respectively. Tamsulosin was prescribed for appropriate indications in 66% of patients, potentially appropriate indications in 4% of patients, and inappropriate indications in 30% of patients. Of the 206 patients (62%) who were prescribed tamsulosin as a result of a reported adverse event with a previous alpha-blocker, only 29% of the cases (N = 59) showed evidence that an attempt was made to reduce the dose of the first alpha-blocker to abate the side effects. Followup monitoring was conducted in 78% of tamsulosin patients, of whom 55% reported effectiveness of tamsulosin, and 6% reported side effects attributable to tamsulosin. No meaningful differences in prescribing patterns were found between the pregroups and postgroups relative to the posting of the criteria for nonformulary use. Two sites had some form of the criteria made available to physicians, while 4 sites had not implemented the national criteria. Extrapolation of the results to the VA system-wide yielded a conservative estimate of 480,993 dollars in potential cost avoidance for 1 quarter (July to September 2002) when corrected for patients prescribed tamsulosin outside of the criteria for nonformulary use., Conclusions: Despite the availability of national criteria for nonformulary use of tamsulosin, the results reveal that these criteria were not followed by prescribers. The DUE reinforced the need for more effective implementation and dissemination of criteria for appropriate use of tamsulosin. A formal education process is necessary to encourage appropriate use of formulary alpha-blockers and to attenuate the increased cost associated with the inappropriate prescribing of the nonformulary drug.

Lymnaea stagnalis were exposed to hypoxic and chemical challenges while ventilation, heart rate and metabolism were monitored. Hypoxia increased ventilatory behavior, but this response was eliminated by immersion in 0.75 mM nitric oxide synthase (NOS) inhibitor, 7-nitroindazole (7 NI). 7 NI also suppressed ventilatory behavior under normoxia. 10.0 mM L-arginine (ARG, the NOS substrate) increased ventilatory behavior under normoxia, but dampened the hypoxic response. The heart-rate response to NOS inhibition exhibited dose-dependent contradictory characteristics. Under both normoxia and hypoxia 0.25 mM 7 NI increased heart rate, while 0.75 mM 7 NI suppressed it. The effect of 0.50 mM 7 NI depended on whether normoxia or hypoxia was coincident; under normoxia 0.50 mM 7 NI increased heart rate, while under hypoxia this concentration suppressed heart rate. Exposure to ARG did not elicit dose-dependent contradictory responses. Heart rate increased when treated with 10.0 mM ARG under normoxia and hypoxia, while 1.0 mM ARG increased heart rate only under hypoxia. Metabolic responses to NOS inhibition also exhibited dose-dependent contradictory changes. V.O2 decreased over 60% in response to 0.75 mM 7 NI, and baseline V.O2 was restored when exposure ceased. In contrast, 0.25 mM 7 NI increased V.O2 10%, and the increase continued after exposure ceased. 0.50 mM 7 NI decreased V.O2 40%, but V.O2 increased when exposure ceased. ARG had only the effect of increasing V.O2, and only at 10.0 mM concentration. Based on these results and on NO's known role as a neuromodulator, we conclude that the cardio-respiratory responses to hypoxia are, in part, mediated by NO., (Copyright 2003, Wiley-Liss, Inc.)

Our laboratory is a leading pioneer in the enrichment of fetal cells from maternal blood with the aim of developing a non-invasive risk free form of prenatal diagnosis. By using the then novel Magnetic Activated Cell Sorting (MACS) we were among the first to detect fetal aneuploidies. The efficacy of this methodology is currently being explored in the large scale so-called, NIFTY study conducted under the auspices of the NIH, in which our group is participating. We have extended the scope of these investigations by analysing single micromanipulated erythroblasts by single cell PCR. These studies have shown that fetal genetic loci such as sex and rhesus D status can be identified with great reliability non-invasively. This analysis also irrevocably demonstrated that a large proportion of the erythroblasts in maternal circulation are of fetal origin. This aspect has now been incorporated into the next phase of the NIH study. Recent research from our laboratory has indicated that the traffic of fetal cells into the maternal periphery is significantly enhanced in preeclamptic pregnancies. We have now investigated whether this perturbation takes place early in pregnancy prior to the onset of disease symptoms, by performing a prospective study in which close to 100 pregnant women were recruited at around 20 weeks of gestation. By correlating the number of enriched erythroblasts with subsequent pregnancy outcome, we were able to show that the traffic of fetal cells was indeed significantly elevated in those pregnancies which developed preeclampsia, but not those which were affected by fetal growth retardation. We have also explored the new finding of free fetal DNA in maternal plasma. By the use of sensitive quantitative PCR we were recently able to show that the levels of this circulatory fetal DNA are elevated in pregnancies with certain aneuploidies, thereby opening the prospect of a new additional screening tool.

Lateralized human cortical activity for shifting visuospatial attention and initiating saccades. J. Neurophysiol. 80: 2900-2910, 1998. The relation between shifts of visual attention and saccade preparation was investigated by studying their electrophysiological correlates in human scalp-recorded electroencephalogram (EEG). Participants had to make saccades either to a saliently colored or to a gray circle, simultaneously presented in opposite visual hemifields, under different task instructions. EEG was measured within the short interval between stimulus onset and saccade, focusing on lateralized activity, contralateral either to the side of the relevant stimulus or to the direction of the saccade. Three components of lateralization were found: 1) activity contralateral to the relevant stimulus irrespective of saccade direction, peaking 250 ms after stimulus onset, largest above lateral parietal sites, 2) activity contralateral to the relevant stimulus if the stimulus was also the target of the saccade, largest 330-480 ms after stimulus onset, widespread over the scalp but with a focus again above lateral parietal sites, and 3) activity contralateral to saccade direction, beginning about 100 ms before the saccade, largest above mesial parietal sites, with some task-dependent fronto-central contribution. Because of their sensitivity to task variables, component 1 is interpreted as the shifting of attention to the relevant stimulus, component 2 is interpreted as reflecting the enhancement of the attentional shift if the relevant stimulus is also the saccade target, and component 3 is interpreted as the triggering signal for saccade execution. Thus human neurophysiological data provided evidence both for independent and interdependent processes of saccade preparation and shifts of visual attention.

Objective: To review the recently published medical literature for the practical and efficient use of muromonab-CD3 (OKT-3) and antithymocyte globulin (ATG) in renal transplantation., Data Sources: MEDLINE and EMBASE were searched (1985-February 1996). Key words used were antithymocyte globulin (ATG, Atgam), muromonab-CD3 (OKT-3, Orthoclone), and kidney transplantation. Thereafter, the search was restricted to English-language articles, clinical trials, and human studies., Study Selection and Data Extraction: The search was reviewed for articles of interest, and pertinent references from these articles were further reviewed to supplement the initial search. The review focused on antibody therapy as induction and/or rejection therapy in renal transplantation., Data Synthesis: Although ATG and OKT-3 are effective in delaying and reducing the occurrence of acute rejection, their impact on long-term graft survival has not been established. Improved graft survival has, however, been demonstrated in patients at high risk for rejection. These risks are described in the review. As first-line or steroid-resistant rejection therapy, ATG and OKT-3 have proven efficacious. Some studies have shown improved graft survival with OKT-3. Although serious infections may occur, OKT-3 has been shown to be effective in reversing rejections resistant to both steroids and ATG. Therefore, reserving OKT-3 for steroid- or ATG-resistant rejections may be preferred over the first-line use of OKT-3, which is limited by the development of antimurine antibodies with subsequent uses. However, the benefits of first-line antibody therapy may outweigh the risks of developing these antibodies in patients for whom high-dose steroids may not be the most appropriate treatment. Other factors that need to be considered are adverse effects, which appear to be lower with ATG, cost, and total hospital charges. The accuracy of treatment outcomes analysis among these studies is limited by variations in the immunosuppressive regimens of the study centers, doses of concomitant therapies, use of prophylactic antibiotics, and time to follow-up., Conclusions: While important benefits are realized from using antibody therapies in renal transplantation, their use is often associated with excess immunosuppression and increased treatment costs. Despite encouraging results from published trials, questions regarding the extent of their prophylactic use and impact on long-term outcomes need to be answered. The current literature contains no prospective, controlled, randomized comparisons of OKT-3 and ATG with standardized regimens of conventional immunosuppressive agents and antirejection protocols. The majority of studies use OKT-3 as part of the treatment protocol. Well-designed studies using ATG are lacking. Further research is needed to refine treatment protocols for ATG and OKT-3 to determine the optimal timing and dosing for these agents.

Cytarabine is intracellularly activated and correlations have been established between the pharmacokinetic behaviour of active metabolites and their antileukemic effect. Recently, a good response to high-dose treatment of leukemias has additionally been attributed to a so-called low deamination phenotype of cytarabine inactivation. Consequently, these findings would support plasma level monitoring of cytarabine and its metabolite uracil arabinoside in high-dose cytarabine regimens. This pharmacokinetic study presents data attempting to reevaluate these observations. Thirty-seven patients were treated by 3-h high-dose cytarabine infusions (9 patients 1000 mg/m2, 28 patients 3000 mg/m2) as part of their treatment for acute leukemia. Serial blood samples during and post infusion were analysed for cytarabine (araC) and its deamination product uracil arabinoside (araU) using HPLC with UV-detection. Considerable interindividual variation was observed in end-infusion plasma concentrations of araC (1000 mg/m2: 2.1-fold, 3000 mg/m2: 5.5-fold) and araU (1000 mg/m2: 2.7-fold, 3000 mg/m2: 2.9-fold). The median ratio of end infusion concentrations araU/araC (on a molar basis) was 5.6 (S.D. 3.0), extreme ratio values were 2 and 14. No differences of the araU/araC ratio were found between the two dosages used. Minimum plasma araC concentrations at the end of infusion were 10.5 micromol/l and 22.0 micromol/l at a dose of 1000 and 3000 mg/m2, respectively. In our European study population a "fast" deamination phenotype of cytarabine (araU/araC ratio > 14) was not be observed.

Algorithms for the differential diagnosis of anemias are commonly based on suspected incidences and simple laboratory parameters. Especially microcytic anemias are diagnosed using algorithms created in Mediterranean or Northern American regions. In a West German region we observe relatively high diagnostic uncertainty regarding common forms of anemias. As a hypothesis, this may be a result of inadequate algorithms not designed for regions with high incidences of anemias of chronic disease and low incidences of thalassemias. To further elucidate diagnostic problems we here report the frequencies of anemias in university hospital outpatients. Based on these data, the diagnostic values of different erythrocyte indices and of red cell distribution width in the differential diagnosis of anemias were calculated. 4525 patient files were reviewed retrospectively. 872 patients presented with anemia, 107 of which were hereditary forms and 765 of other forms. In hereditary anemias the frequency of thalassaemias (50 patients) and corpuscular hemolytic anemias (49 patients) was the same. Nearly half of the other anemias were covered by anemias of chronic disease and true iron deficiency anemias. Several indices intended to separate thalassemias from other microcytic anemias were tested for relevance. Sensitivity was between 0.75 and 0.85. Specificity was between 0.78 and 0.95. Red cell distribution width was not significantly different between thalassemias and iron deficiency. Only a red cell distribution width above 17.0 resulted in a specificity for iron deficiency of 0.91. Red cell distribution width is considered to be an unreliable screening test in a population with a low incidence of thalassemias. The high incidence of anemias of chronic disease in the region investigated should lead to more complex diagnostic procedures than offered by blood count values alone.

Background: Bacillary angiomatosis is a recently described vascular disorder that is associated with infection by Bartonella henselae (formerly known as Rochalimaea henselae) and Bartonella quintana (formerly known as Rochalimaea quintana); this disorder usually occurs in patients with human immunodeficiency virus infection. We report a case of cutaneous bacillary angiomatosis that occurred in a patient with chronic lymphocytic leukemia., Observations: A 55-year-old man with chronic lymphocytic B-cell leukemia, Rai stage IV, presented with multiple angiomatous papules that clinically resembled pyogenic granulomas. Histopathologic examination revealed circumscribed lobules of small vessels with plump endothelial cells, numerous neutrophils, and abundant nuclear dust; these features were diagnostic for bacillary angiomatosis. The diagnosis was confirmed by the Grocott-Gomori methenamine-silver nitrate stain that revealed argyrophilic bacteria and by ultrastructural demonstration of bacillary structures with trilaminar walls. Treatment with clarithromycin led to complete resolution of the lesions within 4 weeks., Conclusions: This case emphasizes that (1) bacillary angiomatosis must be considered in the differential diagnosis of vascular lesions in immunocompromised patients without human immunodeficiency virus infection, (2) Grocott-Gomori methenamine-silver nitrate stain is a simple and satisfactory alternative to the Warthin-Starry stain for the demonstration of bacilli in this condition, and (3) clarithromycin is an effective oral antibiotic for the treatment of this disease.

A significant aspect of holistic primary care nursing is that it seeks to assess and understand clients' health beliefs and behaviors in the context of culture. This article examines implications for primary care nursing practice based on the cultural beliefs and health behaviors of pregnant Mexican-American women. Giger and Davidhizar's transcultural assessment model is used to evaluate six cultural phenomena present in all cultural groups. Culturally sensitive primary care by certified nurse-midwives in a community health birth center setting is described.

Calcium phosphate (CaP) has been detected in the majority of urinary stones containing predominantly calcium oxalate (CaOx). Therefore, crystal phases of CaP might play an important role with respect to the formation of urinary calcium stones in general. Very often, CaP found in stones or tissue of human kidney occurs in the shape of small spherulites. In this paper, we report on a new flow model of crystallization (FMCG), which has been used to generate spherulites of CaP in a gel matrix of 1% agar-agar at 37 degrees C from a supersaturated, metastable solution continuously flowing over the gel surface. Scanning electron microscopy (SEM), X-ray diffraction and microscopic Fourier transformed infrared spectroscopy (FTIR) revealed that the particles formed (diameter: up to 200 microns) consisted of a poorly crystalline core of carbonatoapatite which was partly surrounded by a well-crystallized shell of octacalcium phosphate (OCP) showing radially oriented sheet-like structures. Subsequently, CaOx was grown on these spherulites from a flow of a correspondingly supersaturated solution conducted over the gel matrix. It could be shown by SEM that growth of calcium oxalate monohydrate (COM) was characteristically induced by the OCP shell. Radial sheet-like forms of OCP were directly continued by COM showing a certain radial orientation. The model of crystallization in gel matrices applied here should be well-suited to simulate the process of urinary stone formation under in vitro conditions.

An ion-pair high pressure liquid chromatographic method is described for the determination of cytarabine (CAS 147-94-4, araC) in human plasma. Complete separation is achieved within 10 min using a reversed stationary phase and an isocratic eluent containing 0.4 mmol/l heptane sulfonic acid as modifier. Detection by UV-absorption occurs at 270 nm. Quantification of cytarabine and of its main plasma metabolite uracil arabinoside (araU) is achieved by means of internal standardisation using adenine arabinoside (araA). Retention times of araU, araC, and araA are 3.9, 5.9 and 9.4 min, respectively. Detection limits of araC and araU are 10 and 15 ng/ml, resp. During a pharmacokinetic study of high-dose cytarabine treatment no interferences could be observed in plasma samples.

This study determined the in situ detection rate of polymerase chain reaction (PCR)-amplified human immunodeficiency virus type 1 (HIV-1) DNA and RNA in lymph nodes and peripheral blood CD4+ cells in six patients with asymptomatic HIV-1 infection and from six people who died of advanced AIDS. The lymph nodes of patients with asymptomatic infection showed expanded germinal centers where, on average, 20% of the CD21+ dendritic cells contained HIV-1 DNA. From 5 to 80% of the CD4+ cells in these lymph nodes contained HIV-1 DNA, as compared with 1-11% of the CD4+ peripheral blood mononuclear cells. The infection in most cells was latent in the asymptomatic group. In contrast, the lymph nodes of patients with advanced AIDS showed marked depletion of both dendritic and CD4+ cells. The majority of the remaining CD4+ cells in the lymph nodes and blood showed PCR-amplified viral DNA and cDNA sequences suggesting the presence of genomic and multiple spliced transcripts. It is concluded that asymptomatic HIV-1 infection is associated with a wide range of latent to active viral-positive CD4+ lymphocytes and dendritic cells in the lymph nodes. Progression to AIDS is characterized by active viral replication in many of the remaining CD4+ cells in the lymph nodes and blood.

Enantioselective catalysts produce organic compounds in enantiomerically enriched form. They are highly efficient tools for the synthesis of biologically active materials, such as pharmaceuticals and crop-protection chemicals, in which enantiomeric purity can be critical. The design of chiral ligands is the key to developing new enantioselective catalysts. Three unusual families of ligands have been used to develop practical technology for enantioselective hydrocyanation of olefins, ring-opening of epoxides, and hydrogenation of various compounds.

Potentiation of narcotic analgesics with phenothiazines have been used for preoperative and postoperative analgesia for a number of years by the podiatric community. The agents most frequently cited in studies are the combination of meperidine with either the phenothiazines chlorpromazine, trimeprazine, or promethazine. Due to the fact that the double-blind clinical studies of pain are mainly subjective, and difficult to quantify, the usefulness of phenothiazines for the sole purpose of potentiation of analgesia is questionable, especially in view of the many side effects of this group of drugs.

Stingrays, found predominantly in warm coastal waters, are responsible for at least 1,500 human injuries in this country each year. These occur most often in the lower extremity. The authors discuss the symptoms and treatment of stingray injuries and present two case reports.

In vitro lymphocyte stimulation by mitomycin-blocked tumor cells can be used to measure tumor-specific immune responses. In order to determine the responding cell type(s) in this reaction, lymph node and spleen cell populations were specifically depleted of thymus- or bone marrow-derived cells by the use of the appropriate antisera and complement or by immunoadsorption of the Fc receptor-bearing cells to antibody-coated sheep red blood cell monolayers. The compositions of both the original and the modified lymphocyte populations were determined by (a) viability counting following treatment with antisera and complement, (b) direct and indirect immunofluorescence, (c) antibody-coated erythrocyte rosette formation, and (d) response to thymus- and bone marrow-derived cell mitogens. In the lymph node cell populations, only the thymus-derived cells were stimulated by the tumor cells. However, both bone marrow- and thymus-derived cells from tumor-immune spleens underwent stimulation when exposed to tumor cells in culture.

Many human melanoma cell lines express HLA-D antigens in addition to HLA-A, -B, and -C specificities. Hybridoma antibodies produced after sensitization with melanoma cells are directed not only against tumor-associated antigens but against HLA and Ia-like specificities. Paired human melanoma cell lines and autologous lymphoblast lines were used to identify those monoclonal hybridoma antibodies that define human melanoma-associated antigens. Autologous lymphoblast lines were used to exclude hybridoma antibodies directed against HLA and Ia-like specificities. Using this method, we have identified several cell surface antigens that are expressed on the majority of human melanoma cell lines tested.

Forty-four evaluable patients with metastatic malignant melanoma confined to the skin, subcutaneous tissues, lymph nodes, and/or lung were randomly assigned to receive either 1 X 10(6), 3 X 10(6), or 9 X 10(6) units of partially purified human leukocyte (alpha) interferon by daily im injection for 42 days. One patient achieved a partial response, two had minor responses, and three others had mixed responses. The only partial response was observed at the lowest dose of interferon. Toxicity increased in frequency and intensity with increasing interferon dose. This preparation of interferon at the doses, route, and schedule used appears to have little efficacy in metastatic malignant melanoma.

The synthesis and structure of the primary translation product of apo AII in a human liver poly(A+) mRNA primed cell-free system and its cotranslational modification was studied parallel to studies in vivo with Hep G2 cells, a human hepatoma cell line. The primary translation product is a preproprotein containing 100 amino acid residues, which is cleaved by the signal peptidase of endoplasmic reticulum to pro-apo AII with the loss of the N-terminal pre-sequence consisting of 18 amino acid residues. Hep G2 cells contain about equal amounts of the proform of apolipoprotein AII and of mature apo AII. Approximately in the same ratio pro- and mature apo AII are secreted into the medium. Determination of the partial amino-acid sequence by automated Edman degradation of the labelled prepro- and proforms of apo AII led to the segmentation of the N-terminus of the primary translation product, consisting of 23 amino acid residues, into the pre-sequence (18 residues) and the pro-sequence (5 residues) with terminal Arg-Arg-residues at the cleavage site to apo AII. We must therefore correct our previously postulated 17 and 6 residues containing segmentation. So far no information has been obtained in which compartment and at what stage of posttranslational events the dimerization occurs by formation of the single disulfide bond at position Cys6 in the mature apo AII structure, leading to the symmetrical molecule.

With the advent of hybridoma technology, monoclonal antibodies (MAbs) can be produced against specific human tumor cell-surface antigens. 44X14, an MAb produced in our laboratory by the immunization of BALB/c mice with colon carcinoma cells, exhibits high affinity for breast, lung, and colon carcinomas. However, this MAb shows little evidence of in vitro cytotoxicity. To enhance the potency of this MAb, it was coupled to the A chain (RTA) of the castor-bean protein ricin. Ricin is composed of two subunits--RTA, which binds to ribosomes and inhibits protein synthesis, and the B chain (RTB), which binds to galactose residues on all human cells and facilitates entry of RTA into the cell. By chemically separating RTA from RTB, RTA can then be coupled to MAb 44X14 so as to redefine its specific toxicity. This immunotoxin 44X14-RTA was assayed for protein-synthesis inhibition in HT-29 colon carcinoma and HT-1080 sarcoma cells by [3H]leucine uptake. Intact ricin (RTA + RTB) inhibited protein synthesis by 50% at concentrations of 0.76 and 4.8 ng/ml in HT-29 and HT-1080 cells, respectively. MAb 44X14 showed the same level of inhibition on HT-29 cells at 6.4 micrograms/ml, whereas the immunotoxin MAb 44X14-RTA showed 50% inhibition at 0.15 micrograms/ml. No effect of either MAb 44X14 or MAb 44X14-RTA at concentrations up to 200 micrograms/ml was seen on HT-1080 cells. Thus, the coupling of RTA to carcinoma-specific MAb 44X14 increased its potency 50-fold without increasing its nonspecific binding to cells that do not contain the appropriate antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

The advent of hybridoma monoclonal antibody technology has opened a new frontier in the biomedical sciences. Human tumor biology and immunology are fields that should particularly benefit from its application. Many advances already have been made in the definition of human melanoma-associated antigens. Most work has been done using mouse monoclonal antibodies, but it now appears that the production of human monoclonal antibodies against human tumor antigens is an achievable goal. Mouse and human monoclonal antibodies should provide us with the reagents to catalogue and characterize both biochemically and functionally the entire range of molecules associated with human melanoma cells. These same reagents will be essential to the development of immunodiagnostic and immunoprognostic assays, and also may be used in the future as highly specific probes for the delivery of radioisotopes, chemotherapeutic agents, and toxins to metastatic tumor deposits.

In vitro lymphocyte stimulation by mitomycin-C-blocked tumor cells has been used to demonstrate tumor-specific antigens in methylcholanthrene-induced sarcomas and in syngeneic murine systems, and to follow the evolution of tumor immunity in the tumor-bearing state. Lymphocytes from normal mice, from mice bearing small methylcholanthrene-induced tumors (less than 1 cm in diameter) and from tumor-immunized mice (which had been sensitized by tumor cell inoculation and subsequent tumor (removal) were stimulated by mitomycin-C-blocked tumor cells to undergo increased DNA synthesis in culture. However, lymph node cells from mice with large methylcholanthrene-induced tumors (greater than 1 cm in diameter) had quite different characteristics. The background DNA-synthetic activity of lymph node cells from tumor-bearing mice was much higher than that of lymph node cells from normal mice or from preimmunized, non-tumor-bearing mice, and progressively increased with the duration of tumor-bearing and the size of the tumor. Lymph node cells from mice bearing large tumours appeared to be maximally stimulated by the tumor in vivo and were incapable of further stimulation by the same tumor cells in vitro, but exhibited normal responses to other antigens and nonspecific mitogens. Lymph node cells from mice bearing large tumors depleted of the adherent cells had lower background levels of DNA synthetic activity per 10(6) cells and underwent stimulation similarly to lymphoid cells from preimmunized, non-tumor-bearing mice. Spleen cells from mice bearing large tumors also had increased background levels of DNA synthetic activity, but contrary to the observations made with the lymph node cells, they were able to undergo further stimulation when cultured with the same tumor cells. Studies on the background incorporation of tritiated thymidine by lymph node cells from tumor-bearing mice revealed that the spontaneous uptake of reconstituted populations was nearly that of the original population and more than twice that of the arithmetical sum of the nonadherent and the adherent subpopulations. This was not found with normal lymph node cells or with spleen cells from normal or tumor-bearing mice. The differential effects observed with lymph node cells compared to spleen cells from tumor-bearing mice are emphasized and possible explanations for these findings are discussed.

With the use of data from the Spring 1977 sample of the USDA Nationwide Food Consumption Survey 1977-78, 1- and 3-day nutrient intakes for 8,779 individuals in 22 sex-age groups in the 48 conterminous states were compared. For most nutrients studied, 3-day mean intakes and day 1 mean intakes showed little difference. Mean intakes for day 1 and for days 2 and 3 combined were not significantly different for most sex-age groups for most nutrients. The few statistically significant differences observed for energy, fat, and carbohydrate could have been the result of chance and therefore have little practical significance. Regression analysis indicated that day 1 intake was strongest as a predictor of intake on subsequent days for energy, carbohydrate, calcium, magnesium, and phosphorus; less strong for protein, fat, iron, thiamin, riboflavin, niacin, vitamin B-6, and vitamin C; and poorest for vitamin A.

Two monoclonal antibodies produced by hybridomas were identified by an indirect 125I-protein A binding assay that define cell surface antigens expressed on cultured human melanoma cells but not on autologous lymphoblastoid cells. The first antibody, 705F6 (an IgG2b immunoglobulin), bound to 14/14 melanoma lines, 6/9 carcinomas and sarcomas, 7/7 gliomas and neuroblastomas, 2/2 fetal cell lines, 0/8 lymphoblastoid cell lines, and weakly to 2/4 leukemia lines. The second monoclonal antibody, 436G10 (IgG1), reacted with 10/14 melanomas 5/13 carcinomas and sarcomas, 2/7 gliomas and neuroblastomas, and weakly with the fetal cells, but not with the leukemic or lymphoblastoid cell lines. Comparison of 705F6 and 436G10 with 28 other monoclonal antibodies from different laboratories identified several with similar binding patterns to a panel of tumor and nontumor cell lines. Crossblocking of 125I-labeled 436G10 was not observed by R23, I12 or L10 antibodies. However, 705F6 was completely blocked by monoclonal 376.96S, showing that these two antibodies bind to the same antigenic determinant. The 705F6 antibody immunoprecipitated a 95 kd (kilodalton) membrane protein and the 436G10 antibody bound a 125 kd protein from 125I-labeled melanoma cells. The broad distribution of these two proteins on melanomas and other solid tumors suggests that they define common oncodevelopmental antigens expressed on proliferating cells.

Of 1,014 human solid tumors of various histologic types, 690 (68%) showed evidence of colony formation within 2 to 4 weeks. Tumors which grew particularly well were colon carcinoma (104/175), melanoma (134/155), lung carcinoma (62/85), breast cancer (100/140), ovarian carcinoma (50/67), and sarcoma (72/122). Histologic examination indicated that the colony-forming cells retained functional and morphologic features similar to those of the original tumor. Plating efficiencies varied between 0.01% and 0.2%, and the numbers of colonies observed formed a direct linear correlation with the number of cells plated. Recovery of viable tumor cells was increased when enzymatic tumor dissociation was used rather than a mechanical method. A simplified, supplemented medium resulted in improved cloning efficiencies when compared to previously reported methods (Hamburger and Salmon, 1977 b).