Fn14 receptor promotes invasive potential and metastatic capacity of non-small lung adenocarcinoma cells through the up-regulation of integrin α6.

UNLABELLED: Lung cancer is one of the leading cause of cancer-related death around the world with the majority of diagnoses being non-small cell lung cancer (NSCLC). Given the poor survival rate and efficacy of current therapy for NSCLC, there is a need to identify and develop new therapeutic targets for treatment. We have observed significantly up-regulated levels of Fn14 in clinical samples of lung cancer relative to normal adjacent tissue. However, the functional role of Fn14 in these tumors is not understood yet. We used RT-PCR to establish the Fn14 expression profile in various NSCLC cell lines. Using isogenic variants of H460 NSCLC cell line with low, intermediate and high Fn14 expression as a cellular model, we determined that increased levels of integrin α6 in cells over-expressing Fn14 is suggestive of an important role of α6ß1-fn14 interactions in motility of lung carcinoma and formation of metastases. Enhanced levels of Fn14 correlated with higher tumor cell migration and invasion in an MMP-1 dependent manner. Cells over-expressing Fn14 showed increased in vivo tumor formation with metastatic capacity to lymph nodes, lungs and liver. Thus, this research may be a step toward developing improved treatment strategies for NSCLC by improved detection and inhibition of metastases. KEYWORDS: Fn14, TNFRSF12A, non-small cell lung cancer, H460 cells, motility, tumor formation and metastasis, integrin α6.

Regression analysis for comparing protein samples with 16O/18O stable-isotope labeled mass spectrometry.

MOTIVATION: Using stable isotopes in global proteome scans, labeled molecules from one sample are pooled with unlabeled molecules from another sample and subsequently subjected to mass-spectral analysis. Stable-isotope methodologies make use of the fact that identical molecules of different stable-isotope compositions are differentiated in a mass spectrometer and are represented in a mass spectrum as distinct isotopic clusters with a known mass shift. We describe two multivariable linear regression models for (16)O/(18)O stable-isotope labeled data that jointly model pairs of resolved isotopic clusters from the same peptide and quantify the abundance present in each of the two biological samples while concurrently accounting for peptide-specific incorporation rates of the heavy isotope. The abundance measure for each peptide from the two biological samples is then used in down-stream statistical analyses, e.g. differential expression analysis. Because the multivariable regression models are able to correct for the abundance of the labeled peptide that appear as an unlabeled peptide due to the inability to exchange the natural C-terminal oxygen for the heavy isotope, they are particularly advantageous for a two-step digestion/labeling procedure. We discuss how estimates from the regression model are used to quantify the variability of the estimated abundance measures for the paired samples. Although discussed in the context of (16)O/(18)O stable-isotope labeled data, the multivariable regression models are generalizable to other stable-isotope labeled technologies.

A possible mechanism of macrolide resistance among multiple resistant Campylobacter jejuni and Campylobacter coli isolated from Thai children during 1991-2000.

A total of 495 Campylobacterjejuni and 122 C. coli isolated from Thai children were screened for macrolide (erythromycin and azithromycin) resistance by disk diffusion assay. Minimum inhibitory concentrations for erythromycin, azithromycin, nalidixic acid, ciprofloxacin, tetracycline, streptomycin, gentamicin and chloramphenicol were further determined for these macrolide-resistant Campylobacter isolates. Presence of known point mutations resulting in reduced susceptibility to macrolides was investigated by PCR and DNA sequencing. Seventeen percent (23/122) of C. coli and 2.4% (12/495) of C. jejuni isolates were resistant to macrolides. By sequencing domain V of the 23S ribosomal DNA from all 35 macrolide-resistant isolates, a known point mutation of 23S rRNA associated with reduced susceptibility to macrolides was detected in all isolates except one. Among the macrolide-resistant isolates, all were multiply resistant to nalidixic acid and ciprofloxacin, of which the latter is the preferred antimicrobial used for diarrheal treatment in Thailand. Furthermore, most macrolide-resistant isolates were also resistant to tetracycline and streptomycin. The spread of macrolide and quinolone resistant Campylobacter should be monitored closely in Thailand and elsewhere as these antimicrobials are preferred drugs for treatment of diarrhea.

Re-evaluation of commercially available enzyme-linked immunosorbent assay for the detection of Giardia lamblia and Cryptosporidium spp from stool specimens.

This study aimed to detect Giardia lamblia and Cryptosporidium spp infection from stool specimens. A total of 345 stool specimens were examined by microscopy (both direct smear and formalin concentration) and EIA techniques (ProSpecT Microplate Assay) for G. lamblia and Cryptosporidium spp. Of 73 tests positive for G. lamblia, 41(56.2%) were positive by microscopy, and 71(97.3%) were positive by EIA. Of 16 tests positive for Cryptosporidium spp, 5 (31.3%) were positive by microscopy, and 16(100%) were positive by EIA technique. The results demonstrate that this EIA method is quick, simple, and more sensitive than the microscopy method and should be used for the detection of G. lamblia and Cryptosporidium spp where the prevalence of these protozoan parasites is a public health problem.

Significantly increased recovery of intestinal parasites on routine stool specimen evaluation.

Three hundred thirty-six stool samples from October 2001 through October 2002 were analyzed for the presence of intestinal parasites. Fifty-six of these (16.7%) were positive for a total of 66 parasites; 65/66 (98.5%) were detected by iodine and dimethyl sulfoxide-modified acid-fast (DMSO-mAFB) stained smears of fresh and formalin-ethylacetate sedimentation concentrated samples. Saline, iodine, and DMSO-mAFB stained smears of fresh stool samples alone detected significantly fewer parasites, finding only 50/66 (75.8%) (p < 0.05). Stool samples analyzed by trichrome stained specimens preserved in Zinc sulfate polyvinyl alcohol (Zinc PVA) detected only 41/ 66 (62.2%) of the parasites. In our study population, it was necessary to perform the National Committee for Clinical Laboratory Standard (NCCLS) recommended to accurately detect intestinal parasites. The concentration technique is simple and significantly increased the detection of intestinal parasites.

Declining prevalence of HIV-1 infection in young Thai men.

OBJECTIVE: To evaluate trends in HIV-1 seroprevalence in Thailand. DESIGN: HIV-1 serosurvey of successive cohorts of young Thai men entering service with the Royal Thai Army (RTA) between November 1989 and November 1994. METHODS: In November 1989, the RTA Medical Department began routine HIV-1-antibody screening of men who were selected by lottery for conscription. Between November 1989 and November 1994, 311,108 young men were screened at induction. Demographic data were collected between November 1991 and May 1993 and again in November 1994. RESULTS: The seroprevalence of HIV-1 among conscripts nationwide increased rapidly from 0.5% in 1989 to 3.5% in 1992 and reached 3.7% in 1993. In 1994, the overall prevalence decreased to 3.0%. The decrease was greatest in the upper North (from 12.4% in 1992 to 7.9% in 1994), where the prevalence has been the highest. However, decreases were observed in men from all regions of residence in the country, from both rural and urban areas, and at all educational levels. CONCLUSIONS: The decline in prevalence suggests declining incidence and that HIV control programs in Thailand are having an impact on the HIV epidemic.

PIP: Since 1989, the Royal Thai Army has screened new conscripts for human immunodeficiency virus (HIV)-1 infection. An analysis of HIV test results for the 311,108 young men entering the Army between November 1989 and November 1994 indicates that the Thailand Government's acquired immunodeficiency syndrome (AIDS) prevention program has had a positive impact on high-risk behaviors. The HIV seroprevalence increased from 0.5% in 1989 to 3.5% in 1992 and 3.7% in 1993, but then fell to 3.0% in 1994. The prevalence in the North--the region with the highest rate--peaked at 7.5% in 1992, then declined to 5.1% in 1994. This pattern of decline was observed in all regions of residence and at all educational levels. The only exception was a slight increase (from 4.1% in 1992 to 5.0% in 1994) in seroprevalence among the 4.2% of conscripts aged 22-29 years. Condom use, less frequent visits to commercial sex workers, and decreasing rates of sexually transmitted diseases are assumed to be the factors contributing to the decline in HIV infection.

2-Acetylpyridine thiosemicarbazones. 1. A new class of potential antimalarial agents.

Based on the antimalarial properties observed for 2-acetylpyridine 4-phenyl-3-thiosemicarbazone (1), an extensive series of related thiosemicarbazones was prepared and tested against Plasmodium berghei in mice. Screening results indicated that the presence of the 2-pyridylethylidene group was critical and that certain phenyl, benzyl, phenethyl, or cycloalkyl groups at N4 of the thiosemicarbazone moiety also contribute to antimalarial activity.

2-Acetylpyridine thiosemicarbazones. 2. N4,N4-Disubstituted derivatives as potential antimalarial agents.

The most effective antimalarial agents among the N4-monosubstituted 2-acetylpyridine thiosemicarbazones recently described by us have a cyclohexyl or a phenyl substituent and produce cures in Plasmodium berghei infected mice at a dose of 160 and 320 mg/kg, respectively. We report here on a related series of N4,N4-disubstituted 2-acetylpyridine thiosemicarbazones. Several members of this group bearing alkyl or cycloalkyl substituents at N4 show activity superior to the most active monosubstituted 2-acetylpyridine thiosemicarbazones. However, the greatest improvement in potency was seen when the N4-nitrogen atom was incorporated into a six- or seven-membered ring, such as the piperidine, piperazine, or azabicyclo[3.2.2]nonane systems, to give compounds with curative properties at a dose level as low as 20 mg/kg.

Emerging fluoroquinolone and macrolide resistance of Campylobacter jejuni and Campylobacter coli isolates and their serotypes in Thai children from 1991 to 2000.

This study investigated fluoroquinolone, macrolide resistances and serotype distributions among Campylobacter jejuni and Campylobacter coli isolated from children in Bangkok and rural settings during 1991-2000. Phenotypic identification, serotyping, and susceptibility testing were performed by standard microbiological procedures. The predominant serotypes of C. jejuni were Lior 36, 2 and 4 and of C. coli were Lior 8, 29 and 55. Resistance to nalidixic acid increased significantly during 1991-2000 and the frequency of isolates resistant to both nalidixic acid and ciprofloxacin in Bangkok was significantly greater than in rural settings. In 1996-2000, a significant trend was observed in C. jejuni isolates resistant to ciprofloxacin from Bangkok but not for macrolide resistance from both settings. In summary, fluoroquinolone resistance among C. jejuni and C. coli isolates became widespread in both Bangkok and rural settings in Thailand in the 1990s while widespread resistance to macrolides was undetected.

Single breath end-tidal PCO2 measurement during high frequency jet ventilation in critical care patients.

The relationship between arterial and end-tidal carbon dioxide tensions following a single large breath was investigated in seven critically ill patients receiving high frequency jet ventilation. There was a close correlation (r = 0.989) between arterial and end-tidal carbon dioxide tensions over a wide range (3.29-8.95 kPa). Measurement of the end-tidal carbon dioxide tension following a single large breath may be useful in monitoring the efficiency of high frequency jet ventilation in the elimination of carbon dioxide.

Development of a wide bore flow through microwave digestion device for the determination of trace metals in soil.

A flow through microwave digestion device has been developed for the determination of Cd, Cr, Mn, Ni and Pb in soil by aqua regia extraction. This device differs from existing commercially available devices as it uses a double pumping action to replace the back pressure regulator traditionally used to achieve internal pressurisation. An acid front has also been included to overcome problems associated with the dilution of acid in samples due to dispersion. Recoveries between 95 and 105% of certified values were achieved with standard deviations of less than 4% for certified reference soil (BCR 143R). A sample throughput of 6 samples per hour was achieved in the optimised system. The performance of the device was tested by digesting real soil samples ground through a 250 microns sieve and slurried without the use of surfactants. A comparison of analytical performance for analysing real samples was made between the microwave flow method and a thermal method.

2-Acetylpyridine thiosemicarbazones. 10. 2-Propionyl-, 2-butyryl-, and 2-(2-methylpropionyl)pyridine thiosemicarbazones as potential antimalarial agents.

In view of the antimalarial properties observed for many 2-acetylpyridine thiosemicarbazones, a series of N4,N4-disubstituted thiosemicarbazones derived from 2-propionyl-, 2-butyryl-, and 2-(2-methylpropionyl)pyridine was prepared for evaluation against the malaria parasite, Plasmodium berghei, in the mouse. The thiosemicarbazones were made by the reaction of methyl hydrazinecarbodithioate with a 2-acylpyridine to give the intermediate methyl 3-[1-2-pyridinyl)alkylidene]hydrazinecarbodithioates. Reaction of the latter with 3-azabicyclo[3.2.2]nonane, 1-(2-pyridinyl)piperazine, or 1H-hexahydroazepine gave the requisite thiosemicarbazones. The three thiosemicarbazones derived from 3-azabicyclo[3.2.2]nonane were most effective, the greatest potency being exhibited by 3-azabicyclo-[3.2.2]nonane-3-thiocarboxylic acid 2-[1-(2-pyridinyl)butylidene]hydrazide (4) which cured 4/5 mice at a dose of 160 mg/kg. In contrast to the related thiosemicarbazones derived from 2-acetylpyridine, virtually no toxic effects were observed in the series described here.

Evaluation of time-average dispersion models for estimating pheromone concentration in a deciduous forest.

The Sutton and more recent Gaussian plume models of atmospheric dispersion were used to estimate downwind concentrations of pheromone in a deciduous forest. Wind measurements from two bivane anemometers were recorded every 12 sec and the pheromone was emitted from a point source 1.6 m above ground level at known rates. The wingfanning response of individually caged male gypsy moths (Lymantria dispar) at 15 sites situated 20 to 80 m downwind was used to monitor when pheromone levels were above threshold over a 15-min interval. Predicted concentrations from these Gaussian-type models at locations where wing fanning occurred were often several orders of magnitude below the known behavioral thresholds determined from wind tunnel tests. Probit analyses of dose-response relationships with these models showed no relationship between predicted dose and actual response. The disparity between the predictions of concentration from these models and the actual response patterns of the male gypsy moth in the field was not unexpected. These time-average models predict concentrations for a fixed position over 3-min or longer intervals, based upon the dispersion coefficients. Thus the models estimate pheromone concentrations for time intervals appreciably longer than required for behavioral response.

Evidence for a recent decrease in measles susceptibility among young American adults.

As part of an ongoing evaluation of US Army immunization policies, two serologic surveys were conducted at an army basic training center. Antibody status to measles, mumps, and rubella were determined by an ELISA in 969 soldiers in 1989 and 992 soldiers in 1990. Measles seronegativity rates, directly adjusted to the 15- to 24-year-old US population in 1980, decreased from 21.3% in 1989 to 12.5% in 1990 (P < .001). Decreases also were observed in mumps (16.2% to 14.7%) and rubella seronegativity rates (18.4% to 14.3%) from 1989 to 1990 but were not statistically significant. In subgroup analyses, significant decreases in measles seronegativity were observed in younger, white, male, and female cohorts. There is evidence that susceptibility among young adults had declined, although it remains unclear if this is due to increased immunization or exposure to natural disease.

Oral nalbuphine for the treatment of pain after dental extractions.

A randomized, double-blind comparison of nalbuphine 30 mg or 60 mg by mouth and dihydrocodeine 30 mg by mouth was conducted in 75 patients with moderate to severe pain after surgery for dental extractions under general anaesthesia. A significant reduction in pain intensity followed each treatment and persisted throughout the 4-h observation period after nalbuphine, but only for 3 h after dihydrocodeine was given. Reduction in pain intensity was significantly greater 2, 3 and 4 h after the use of nalbuphine 60 mg than following dihydrocodeine 30 mg, and the mean total pain intensity difference was greater following nalbuphine 60 mg than following dihydrocodeine. Nalbuphine 60 mg effectively provided complete or good pain relief in more than 50% of the patients and only three patients in this group required additional analgesia during the period of observation, compared with nine patients in each of the other groups. However, the patients who received nalbuphine 30 mg had a significantly higher mean pain intensity before treatment than those in the other groups. The side-effects encountered were those typical of opioid medication; there were no statistically significant differences between the groups.

Willingness to participate in HIV-1 vaccine trials among young Thai men.

OBJECTIVES: Willingness to participate in HIV-1 vaccine trials and associated factors were investigated in a sample of 2670 Royal Thai Army conscripted recruits. METHODS: Self administered questionnaires were used. Data were collected during the final visit of a longitudinal cohort study of HIV-1 epidemiology. Cross sectional analysis of data from this visit was performed. RESULTS: 32% of the respondents reported they would "definitely" join an HIV-1 vaccine trial. Greater willingness was associated with perceived risk of HIV-1 infection and a desire to help Thai society, although tangible incentives and intentions to reduce condom use in a vaccine trial also were associated with increased willingness. Concerns about physical harm and anticipated social pressure from family not to join were the most substantial impediments to willingness. Concerns about "social harm" (for example, participation would give appearance of having AIDS virus, a partner might refuse sex) also appeared to inhibit interest in joining trials and approached significance. CONCLUSIONS: Willingness to participate was somewhat greater than in other investigations of non-injection drug user (IDU) cohorts in Thailand, with fewer concerns expressed about physical harm. Motivations appear to involve tradeoffs among perceived risk, anticipated social pressure, altruism, and tangible rewards. The absence of significant problems associated with vaccine trials to date, along with the presence of educational interventions in the study may help explain the lower level of concerns here relative to other Thai studies.

The susceptibility of young adult Americans to vaccine-preventable infections. A national serosurvey of US Army recruits.

OBJECTIVE: Due to recent resurgences of measles, mumps, and rubella among young US adults, we sought to generate antibody prevalence data for national and military immunization policy evaluations. DESIGN: We used a questionnaire and serological survey of Army recruits to assess antibody status to measles, mumps, rubella, and varicella by enzyme-linked immunosorbent assay and to poliovirus types 1, 2, and 3 by microneutralization assay. SETTING: Basic training reception centers at Fort Benning, Ga., and Fort Jackson, SC. PATIENTS: The study included 1547 US Army recruits who were inducted during September and October 1989. OUTCOME MEASURES: Seronegativity by various demographic factors. RESULTS: Seronegativity rates, directly adjusted to the 15- to 24-year-old US population in 1980, were 20.7% for measles, 15.6% for mumps, 17.5% for rubella, and 6.9% for varicella. For measles, mumps, and rubella, susceptibility was less in females, blacks, and college-educated recruits, and varicella susceptibility was greater in females and blacks. Recruitment who were born after 1969 lacked measles, mumps, and rubella antibodies more often than older recruits. The adjusted seronegativity rates for poliovirus types 1, 2, and 3 were 2.3%, 0.6%, and 14.6%, respectively; trends by age, sex, and race-ethnicity were generally unremarkable. CONCLUSIONS: Among young adult Americans, susceptibility to measles, mumps, and rubella is unevenly distributed and may be substantial. Our findings support national objectives to further improve immunization coverage in school-age and adult populations and provide further impetus for legislation requiring college entrants to present evidence of having received at least two doses of measles vaccine, with one on or after entry into elementary school.

Population dynamics in echinococcosis and cysticercosis: comparison of the response of Echinococcus granulosus, Taenia hydatigena and T. ovis to control.

A comparative study has been made of the progress in the control of ovine echinococcosis caused by Echinococcus granulosus and the ovine cysticercoses caused by Taenia hydatigena and T. ovis in New Zealand. The methods of control included an educational and a dog-dosing programme. In the Styx field trial, the Otago/Southland Surveillance Programme and the national control programme, E. granulosus declined towards extinction. In contrast, this same control effort transformed the cysticercoses from hyper-endemic to endemic status with focal epidemics. In this unstable state, the density-dependent constraint was negligible and superinfection occurred.