High chromosomal mobility of rDNA clusters in holocentric chromosomes of Triatominae, vectors of Chagas disease (Hemiptera-Reduviidae).

The subfamily Triatominae (Hemiptera-Reduviidae) includes more than 150 blood-sucking species, potential vectors of the protozoan Trypanosoma cruzi, causative agent of Chagas disease. A distinctive cytogenetic characteristic of this group is the presence of extremely stable chromosome numbers. Unexpectedly, the analyses of the chromosomal location of ribosomal gene clusters and other repetitive sequences place Triatominae as a significantly diverse hemipteran subfamily. Here, we advance the understanding of Triatominae chromosomal evolution through the analysis of the 45S rDNA cluster chromosomal location in 92 Triatominae species. We found the 45S rDNA clusters in one to four loci per haploid genome with different chromosomal patterns: On one or two autosomes, on one, two or three sex chromosomes, on the X chromosome plus one to three autosomes. The movement of 45S rDNA clusters is discussed in an evolutionary context. Our results illustrate that rDNA mobility has been relatively common in the past and in recent evolutionary history of the group. The high frequency of rDNA patterns involving autosomes and sex chromosomes among closely related species could affect genetic recombination and the viability of hybrid populations, which suggests that the mobility of rDNA clusters could be a driver of species diversification.

Genetics and evolution of triatomines: from phylogeny to vector control.

Triatomines are hemipteran bugs acting as vectors of the protozoan parasite Trypanosoma cruzi. This parasite causes Chagas disease, one of the major parasitic diseases in the Americas. Studies of triatomine genetics and evolution have been particularly useful in the design of rational vector control strategies, and are reviewed here. The phylogeography of several triatomine species is now slowly emerging, and the struggle to reconcile the phenotypic, phylogenetic, ecological and epidemiological species concepts makes for a very dynamic field. Population genetic studies using different markers indicate a wide range of population structures, depending on the triatomine species, ranging from highly fragmented to mobile, interbreeding populations. Triatomines transmit T. cruzi in the context of complex interactions between the insect vectors, their bacterial symbionts and the parasites; however, an integrated view of the significance of these interactions in triatomine biology, evolution and in disease transmission is still lacking. The development of novel genetic markers, together with the ongoing sequencing of the Rhodnius prolixus genome and more integrative studies, will provide key tools to expanding our understanding of these important insect vectors and allow the design of improved vector control strategies.

High Trypanosoma cruzi (Kinetoplastida: Trypanosomatidae) prevalence in Triatoma sanguisuga (Hemiptera: Redviidae) in southeastern Louisiana.

From May through November 2007, intensive weekly surveys at the site of a previously reported autochthonous human case of Chagas parasite infection resulted in the collection of 298 Triatoma sanguisuga (Leconte) specimens, of which 60.4% (180) were polymerase chain reaction positive for Trypanosoma cruzi Chagas. All were adults, in a ratio of approximately 1:1 female to male, indicating that the domicile was not colonized, but was a destination for these host-seeking adults. We report on seasonal activity pattern, T. cruzi prevalence in T. sanguisuga, and attempts at insect exclusion and control at the case residence.

Video thoracoscopic surgery used to manage tuberculosis in thoracic surgery.

BACKGROUND: The aim of this study was to evaluate the indications and results of video-assisted thoracic surgery (VATS) for the management of tuberculosis in 10 patients with unusual clinical and radiologic presentation for the disease. METHODS: From March 2000 to March 2002, 96 diagnostic VATS operations for unclear thoracic lesions were performed at the authors' institution. Their final diagnosis for 10 (10.4%) of these patients was tuberculosis. The suspected preoperative diagnoses were pancoast tumour (n = 1), pericardial effusion (n = 1), pleural mesothelioma (n = 1), pleural empyema (n = 2), mediastinal lymphoma (n = l), and lung cancer (n = 4). RESULTS: For all the patients, the diagnosis of tuberculosis was achieved by VATS. The duration of drainage was 2.5 days. There have been neither morbidity nor mortality since surgery. The hospital stay was 3 to 5 days. CONCLUSION: Thoracoscopy is a safe and effective procedure for the management of tuberculosis. Tuberculosis should be kept in mind during the differential diagnosis of unknown thoracic lesions, and also for patients who live in economically well developed countries and are not immune compromised.

Blocking the epithelial-to-mesenchymal transition pathway abrogates resistance to anti-folate chemotherapy in lung cancer.

Anticancer therapies currently used in the clinic often can neither eradicate the tumor nor prevent disease recurrence due to tumor resistance. In this study, we showed that chemoresistance to pemetrexed, a multi-target anti-folate (MTA) chemotherapeutic agent for non-small cell lung cancer (NSCLC), is associated with a stem cell-like phenotype characterized by an enriched stem cell gene signature, augmented aldehyde dehydrogenase activity and greater clonogenic potential. Mechanistically, chemoresistance to MTA requires activation of epithelial-to-mesenchymal transition (EMT) pathway in that an experimentally induced EMT per se promotes chemoresistance in NSCLC and inhibition of EMT signaling by kaempferol renders the otherwise chemoresistant cancer cells susceptible to MTA. Relevant to the clinical setting, human primary NSCLC cells with an elevated EMT signaling feature a significantly enhanced potential to resist MTA, whereas concomitant administration of kaempferol abrogates MTA chemoresistance, regardless of whether it is due to an intrinsic or induced activation of the EMT pathway. Collectively, our findings reveal that a bona fide activation of EMT pathway is required and sufficient for chemoresistance to MTA and that kaempferol potently regresses this chemotherapy refractory phenotype, highlighting the potential of EMT pathway inhibition to enhance chemotherapeutic response of lung cancer.

Inhibition of protein synthesis results in super-induction of procyclin (PARP) RNA levels.

Procyclin is an abundant surface antigen found exclusively on the procyclic forms of African trypanosomes. We are interested in the induction of procyclin gene expression during differentiation from bloodstream forms. We find that increased levels of procyclin RNA are evident as early as 15 min after triggering differentiation. The increase in procyclin RNA levels requires the temperature shift from 37 degrees C to 27 degrees C and is aided by addition of the tricarboxylic acid cycle intermediate cis-aconitate. Maximal induction is observed with a combination of three triggers of differentiation: citrate, cis-aconitate and the temperature shift. Protein synthesis does not appear to be required for induction of procyclin RNA during differentiation. In fact, addition of protein synthesis inhibitors results in super-induction of procyclin RNA levels, even under conditions where no induction is normally observed (i.e., at 37 degrees C in the absence of citrate and cis-aconitate). This super-induction was observed with four different protein synthesis inhibitors that affect different stages of translation. Thus, the accumulation of procyclin transcripts may be under the control of a negative regulator whose effective levels are reduced during differentiation from bloodstream to procyclic forms.

Adaptation of distortion product otoacoustic emission in humans.

Previous studies of animals observed a phenomenon of adaptation of distortion product otoacoustic emission (DPOAE) and found that the phenomenon was mediated to a large extent by the medial olivocochlear (MOC) reflex. The present study investigated DPOAE adaptation in humans. The following stimuli were used: f2/f1 = 1.2; f2 = 2, 4, or 5.65 kHz; L2 = 50-65 dB SPL re 20 microPa rms, L1 - L2 = 0-15 dB, where L1 and L2 represent levels of the f1 and f2 tones, respectively; duration of two-tone burst = 5.5 s; interburst gap = 20 or 30 s; number of repetitions = 40 or 64. We analyzed the 2f1 - f2 DPOAE as a function of time using a method of heterodyne envelope detection. The subjects were 20 humans aged from 15 to 54 years (median = 21 years) with normal hearing. We observed that (1) humans exhibited DPOAE adaptation phenomenon; (2) the time course of DPOAE level was characterized by a 2-exponential function; (3) distributions of the fast and slow time constants were well separated with their median values being 69 ms and 1.51 s, respectively; (4) distributions of the magnitudes of the fast and slow adaptation components were largely overlapped with their median values being 0.65 and 0.40 dB, respectively; and (5) the combined magnitude of the adaptation ranged from 0.4 to 3.0 dB with a median of 1.10 dB. To our knowledge, the present study is the first published article to describe adaptation of DPOAE in humans. These results should help advance the basic knowledge of human cochlear mechanics operating under the control of the MOC feedback system and contribute to the development of practical applications such as identifying people at high risk of acoustical injury and a clinical test of the functional status of the MOC system.

Prognosis of asymptomatic ulcerating carotid lesions.

To determine the proper approach to asymptomatic carotid bifurcation ulcerated plaque (UP), 79 patients with 91 asymptomatic UPs were identified angiographically, and a 96% follow-up was obtained with a mean duration of three years. The cumulative stroke rate by life-table analysis was 1% at seven years. Sixty-three UPs in 55 patients were classified as small, and of these patients, transient ischemic attacks (TIAs) that were appropriate to the lesion developed in three and stroke in one (7% cumulative symptom rate). Twenty-four UPs in 21 patients were classified as large, and a TIA developed in one patient (9%), but no strokes were observed in this group. The cumulative mortality was 17% at three years and 52% at seven years. Life-table curves of several subgroups were compared and showed no significant differences in either stroke rate or mortality between any of these groups. On the basis of these data, and particularly the seven-year stroke rate of 1%, prophylactic carotid endarterectomy is not justified for asymptomatic carotid bifurcation ulcerations.

Utility of the polymerase chain reaction in detection of Trypanosoma cruzi in Guatemalan Chagas' disease vectors.

For effective control programs, accurate assessment of Trypanosoma cruzi infection in vectors is essential and has traditionally been performed by microscopic examination. For particular vectors and not others, polymerase chain reaction (PCR) analysis of fecal samples recently has been shown to be an effective means of detection. The sensitivities of the PCR and microscopy for detection of T. cruzi in different anatomic sites were compared in the two major vectors of Guatemala, Triatoma dimidiata and Rhodnius prolixus. Preliminary studies established that T. cruzi can be detected by the PCR in the presence of 90% T. rangeli. One hundred thirty-five vectors were collected, and samples were obtained from the rectum, intestines, and stomach and analyzed by microscopy and the PCR. For Triatoma dimidiata rectal samples, the PCR sensitivity (39.1% T. cruzi positive) and the microscopic sensitivity (24.6% positive) was not significantly different. However, in R. prolixus, the PCR proved significantly more sensitive than microscopy: 57.6% positive by PCR compared with 22.7% by microscopy. Rectal samples showed the highest rates of infection followed by intestine and stomach samples. However, 10.5% of the Rhodnius infections would have been missed if only the rectal sample had been analyzed. Thus, the PCR is significantly more sensitive than microscopy for detection of T. cruzi in R. prolixus. Analysis of anatomic sites in addition to the rectal sample may be necessary for accurate assessment of infection in particular vectors.

Histological development of lesions in the bursa of Fabricius of chickens with inclusion body hepatitis.

Twenty chickens, 4 weeks old, from a flock with spontaneous inclusion body hepatitis were observed for five weeks following the outbreak of disease. Biopsies were made of the bursae of Fabricius on days 1, 3 or 7, and 14 after housing, and the chickens were killed on day 35. Histologically, severe lesions in bursal follicles developed within a few days. At the end of the observation period the bursal structures had completely recovered.

Lesions in chickens with spontaneous or experimental infectious hepato-myelopoietic disease (inclusion body hepatitis) in Germany.

A group of 83 two-to-eighteen-week-old chickens with acute infectious hepato-myelopoietic disease (a German form of inclusion-body-hepatitis) were observed to have the following histologic lesions: panmyelophthisis, small foci of liver necrosis, often with intranuclear inclusion bodies in hepatocytes (15 to 20% of chickens), involution-like atrophy of the bursa of Fabricius and thymus, loss of lymphatic tissue in spleen and cecal tonsils, and nonpurulent myocarditis. In 18 survivors 6 to 8 weeks after clinical signs of disease, nonpurulent myocarditis but normal lymphatic organs and bone marrow were present. A group of 75 chickens were infected after hatching with the field isolant "1942." Between the 3rd and 9th weeks postinoculation the same histologic changes-though mostly milder-were demonstrated. This syndrome differs somewhat from the syndrome described as inclusion body hepatitis in America and Europe.

Temporal ecological assessment of oil contaminated soils before and after bioremediation.

Ecotoxicity methods were used to assess different soil and oil combinations before, during and after laboratory bioremediation with associated hydrocarbon analysis. Heavy, medium and light crude oil (API gravity 14, 30, and 55) was spiked (ca. 5% w/w) into two sandy soils in the laboratory having organic carbon concentrations of 0.3 (Norwood) and 4.7% (Norwood/Baccto). The earthworm (Eisenia fetida) 14-d lethality assay, the modified Microbics Microtox Solid-Phase assay, and the 14-d plant seed germination and growth assays using corn, wheat and oats, were spiked and tested during a 360-d laboratory remediation. Eisenia was the most sensitive of the three methods utilized with survival increasing throughout bioremediation with fastest toxicity reduction in the high carbon Norwood/Baccto soils where LC50's were 100% or greater at the end of 90-d whereas, > 150-d were required to achieve a similar result in the low carbon soil. Analysis of the undiluted treatments with oily soil alone showed that earthworm survival was high after 90-d in all high organic carbon soils, and after eight months in the low carbon soils, except for the Norwood soil-light oil treatment, which required 360-d to achieve 100% survival. The Microtox assay was less sensitive with EC50's 100% or greater observed after 90-d in high carbon soils and after 240-d for all low carbon soils. After bioremediation, no effects on seed germination were observed, although some plant growth inhibition effects remained. There was no direct correlation between total petroleum hydrocarbon concentrations and toxicity.

A review of the environmental fate, effects, and exposures of bisphenol A.

Bisphenol A (CAS 85-05-7) may be released into the environment through its use and handling, and permitted discharges. BPA is moderately soluble (120 to 300 mg/L at pH 7), may adsorb to sediment (Koc 314 to 1524), has low volatility, and is not persistent based on its rapid biodegradation in acclimated wastewater treatment plants and receiving waters (half-lives 2.5 to 4 days). BPA is "slightly to moderately" toxic (algal EC50 of 1000 micrograms/L) and has low potential for bioaccumulation in aquatic organisms (BCFs 5 to 68). The chronic NOEC for Daphnia magna is > 3146 micrograms/L. Surface water concentrations are at least one to several orders of magnitude lower than chronic effects, with most levels nondetected.

Bisphenol A concentrations in receiving waters near US manufacturing and processing facilities.

Bisphenol A (BPA) (CAS 80-05-7) was analyzed in receiving waters upstream and downstream of US manufacturers (1996 and 1997) and processors (1997) during seasonal low flow periods. BPA was not detected (< 1 microgram/l) in any surface water sample in 1996 or at six of seven sites in 1997. Concentrations near the seventh site ranged from 2 to 8 micrograms/l; however, its receiving stream had no measurable flow and concentrations represent undiluted effluent. All surface water concentrations from this and other studies were less than the freshwater predicted no effect concentration (PNEC) of 64 micrograms/l, suggesting that BPA discharges from manufacturing and processing facilities to surface water do not pose an environmental concern.

Effects of time and an educational presentation on student attitudes toward wife-beating.

To determine if a public outreach educational presentation on domestic violence would produce change in attitudes toward wife-beating and if the attitude change would persist over time, 36 female and male university students were quasi-randomly assigned to groups. Half of the subjects received a 20-minute presentation on domestic violence (Group P), and half did not receive a presentation (Group NP). Groups were then tested on their responses on the Inventory of Beliefs about Wife Beating (IBWB). Three weeks later, groups were retested on their responses to the scale. MANOVA revealed that attitudes of Group P were significantly more negative toward wife-beating than attitudes of Group NP on three of the five IBWB subscales and on three of the 31 IBWB items subsumed under the subscales. Data supported the prediction that public outreach efforts would produce more negative attitudes toward wife-beating and that attitude change would not deteriorate over time.

[Vaccination against infectious anemia of poultry (CAA)--results of field studies]. / Impfungen gegen die Infektiöse Anämie des Geflügels (CAA)--Ergebnisse von Feldversuchen.

Infectious anemia of poultry is a disease of high economical significance. Connatal infection of chicks with the chicken anemia agent (CAA) via the embryonated egg causes anemia along with severe immunosuppression, thus rendering the chicken susceptible for secondary infections. In order to prevent infection of young chicks, it is necessary to induce immunity against CAA in parent flocks, with the aim to prevent connatal spread of the infection and provide maternal protection for baby chicks. In this publication, the efficacy and use of a live CAA vaccine is reported. From autumn 1986 until summer 1990, 3 experimental vaccine charges were applied in 85 broiler parent flocks with totally 3.1 million chickens. In addition, totally 293,000 broiler breeder and 171,000 layer breeder chicken were vaccinated in 1989/90. The vaccine was administered between the 13th and 19th week of life by drinking water without adverse effect to the birds. Chicken anemia symptoms were observed only at the begin of laying period in two parent flocks. These flocks had been vaccinated in the 17th and 19th week, respectively. The offsprings of all other vaccinated parent flocks remained free of chicken anemia. Day-old chicks derived from vaccinated parent flocks were protected against CAA challenge infection. It is emphasized, that vaccination should be performed within the 13th to 15th week of life, because according to our observations, this will lead to an immediate seroconversion.

[Use of toltrazuril in pullet breeding flocks raised on floors with anticoccidial-free feed]. / Einsatz von Toltrazuril bei Junghennenaufzuchten in Bodenhaltung mit antikokzidiafreier Fütterung.

In 28 replacement pullet flocks with flock size of 4,000 to 25,000 birds Toltrazuril (experimental preparation, 2.5% solution, approx. 7 mg/kg body weight) was administered for a period of two days in the drinking water for metaphylactic purpose and therapeutic use, respectively. All flocks were reared without anticoccidial feed additives. Exposition to Eimeria spp. was recorded by examination of faecal samples and intestinal scrapings. Clinical flock control was achieved by vets at eight to ten day intervals. Toltrazuril was characterized by short application period, rapid mode of action, significant reduction of oocyst shedding, good compatibility and acceptance thus proving superior to conventional chemotherapy of coccidiosis. The results suggest that, even without administration of anticoccidial feed additives. Toltrazuril is an effective agent for therapeutic use in acute coccidiosis. Under insufficient management and rearing conditions repeated metaphylactic application has to be considered.

High genetic diversity in a single population of Triatoma sanguisuga (LeConte, 1855) inferred from two mitochondrial markers: Cytochrome b and 16S ribosomal DNA.

Chagas disease remains endemic across much of Latin America, but is largely enzootic--restricted to wild mammals and triatomine vectors in the United States. Within the United States, there are ten recognized species of triatomines and 18 mammals reported naturally infected with Trypanosoma cruzi, the causative agent of Chagas disease. However, only six cases of autochthonous vector-borne transmission of T. cruzi to humans have been reported in the United States. As a follow-up to the sixth reported case, triatomine insects were collected from the index case site, in a rural area of New Orleans, LA, USA. During the summer months of 2006 and 2007, 344 Triatoma sanguisuga were collected and showed a T. cruzi infection prevalence of 56%. A subset of these insects was analyzed to infer intraspecific genetic variation from a 606 bp fragment of cytochrome b (n=54) and a 340 bp fragment of 16S ribosomal DNA (n=17). From the 54 samples, 37 cytb haplotypes (H(d)=0.978) were observed and 14.7% of positions were polymorphic. Phylogenetic analysis divides the population into two distinct groups with an average pairwise genetic distance of ~5%. The 16S rDNA sequences revealed 6 haplotypes among 17 specimens (H(d)=0.713) with 1.2% of the positions exhibiting polymorphisms. 16S polymorphism data support the concept of two groups within this single population. The genetic distance of Group 1 from Group 2 suggests that Group 1 could represent a sub-species as this level of divergence is similar to that observed among other triatomine subspecies.