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.

Monitoring of temperature increase and tissue vaporization during laser interstitial thermotherapy of ex vivo swine liver by computed tomography.

Laser interstitial thermotherapy (LITT) is a minimally invasive technique used to thermally destroy tumour cells. Being based on hyperthermia, LITT outcome depends on the temperature distribution inside the tissue. Recently, CT scan thermometry, based on the dependence of the CT number (HU) on tissue temperature (T) has been introduced during LITT; it is an attractive approach to monitor T because it overcomes the concerns related to the invasiveness. We performed LITT on nine ex vivo swine livers at three different laser powers, (P=1.5 W, P=3 W, P=5 W) with a constant treatment time t=200 s; HU is averaged on two ellipsoidal regions of interest (ROI) of 0.2 cm2, placed at two distances from the applicator (d=3.6 mm and d=8.7 mm); a reference ROI was placed away from the applicator (d=30 mm). The aim of this study is twofold: 1) to evaluate the effect of the T increase in terms of HU variation in ex vivo swine livers undergoing LITT; and 2) to estimate the P value for tissue vaporization. To the best of our knowledge, this is the first study focused on the HU variation in swine livers undergoing LITT at different P. The reported findings could be useful to assess the effect of LITT on the liver in terms of both T changes and tissue vaporization, with the aim to obtain an effective therapy.

Influence of FBG sensors length on temperature measures in laser-irradiated pancreas: theoretical and experimental evaluation.

Temperature distribution T(x,y,z,t) in tissue undergoing Laser-induced Interstitial Thermotherapy (LITT) plays a crucial role on treatment outcome. Theoretical and experimental assessment of temperature on ex vivo laser-irradiated pancreas is presented. The aim of this work is to assess the influence of thermometers dimensions on temperature measures during LITT. T(x,y,z,t) inside tissue is monitored by optical sensors, i.e., Fiber Bragg Gratings (FBGs): three FBGs with lengths of 10 mm and nine FBGs of 1 mm, at different distances (2 mm, 5 mm and 10 mm) and different quotes (0 mm, 2 mm and 4 mm) from the laser fiber tip are used. Theoretical punctual T(x,y,z,t) is averaged out on both 10 mm and 1 mm in order to compare numerical predictions with experimental data. Results demonstrate the influence of FBG length on T(x,y,z,t) measures. This phenomenon depends on the distance between sensor and applicator: it is particularly significant close to the applicator tip (2 mm) because of the high spatial T(x,y,z,t) gradient within the tissue. Both theoretical results and experimental ones show that just at a distance of 10 mm from the tip, differences between T(x,y,z,t) provided by FBGs of 10 mm and 1 mm are negligible.

Experimental assessment of CT-based thermometry during laser ablation of porcine pancreas.

Laser interstitial thermotherapy (LITT) is employed to destroy tumors in organs, and its outcome strongly depends on the temperature distribution inside the treated tissue. The recent introduction of computed tomography (CT) scan thermometry, based on the CT number dependence of the tissue with temperature, overcomes the invasiveness of other techniques used to monitor temperature during LITT. The averaged CT number (ROI = 0.02 cm(2)) of an ex vivo swine pancreas is monitored during LITT (Nd:YAG laser power of 3 W, treatment time: 120 s) at different distances from the applicator (from 4 to 30 mm). The averaged CT number shows a clear decrease during treatment: it is highest at 4 mm from the applicator (mean variation in the whole treatment of -0.256 HU s(-1)) and negligible at 30 mm, since the highest temperature increase is present close to the applicator (i.e., 45 °C at 4 mm and 25 °C at 6 mm). To obtain the relationship between CT numbers and pancreas temperature, the reference temperature was measured by 12 fiber Bragg grating sensors. The CT number decreases as a function of temperature, showing a nonlinear trend with a mean thermal sensitivity of -0.50 HU °C(-1). Results here reported are the first assessment of pancreatic CT number dependence on temperature, at the best of our knowledge. Findings can be useful to further investigate CT scan thermometry during LITT on the pancreas.

High dynamics of rDNA cluster location in kissing bug holocentric chromosomes (Triatominae, Heteroptera).

In this paper, we determine by fluorescent in situ hybridization the variability in the chromosomal location of 45S rDNA clusters in 38 species belonging to 7 genera of the Triatominae subfamily, using a triatomine-specific 18S rDNA probe. Our results show a striking variability at the inter- and intraspecific level, never reported so far in holocentric chromosomes, revealing the extraordinary genomic dynamics that occurred during the evolution in this group of insects. Our results also demonstrate that the chromosomal position of rDNA clusters is an important marker to disclose chromosomal differentiation in species karyotypically homogenous in their chromosome number.

Theoretical assessment of principal factors influencing laser interstitial thermotherapy outcomes on pancreas.

The influence of some therapy-relevant parameters on Laser Induced Interstitial Thermotherapy (LITT) outcomes on pancreas is assessed. The aim is to execute a sensitivity analysis for an optimal treatment strategy on in vivo pancreas. A numerical model based on Bioheat Equation has been implemented to assess the influence of laser settings (power P and energy E), applicator radius (r(f)) and optical properties (effective attenuation coefficient, µ(eff)) on temperature (T) distribution. Effects on pancreas undergoing LITT have been evaluated with a twofold approach: 1) T rise and maximum T (T(max)) in tissue; 2) injured volumes (vaporized and coagulated ones). We consider parameters range in typical LITT values (P from 1.5 W to 6 W, E from 500 J to 1500 J, r(f) from 150 µm to 600 µm) and optical values reported in literature. Our analysis shows that, among others, P and µ(eff) are the principal influencing factors of thermal effects on pancreas undergoing LITT: P should be carefully chosen by operator to obtain the desired injured volumes, while the accurate measurement of tissue optical properties is crucial to carry out a safe and controlled thermal therapy on pancreas.

Inheritance of resistance to pyrethroids in Triatoma infestans, the main Chagas disease vector in South America.

An outbreak of pyrethroid resistance was recently detected in Triatoma infestans from northern Argentina. To analyze the inheritance of the resistant phenotype, we carried out experimental crosses between resistant (R) and susceptible (S) strains captured in Argentina during 2005. The R strain was collected from sprayed houses in the north of the province of Salta while the S strain was collected in the province of Chaco. Both strains were bred in the laboratory for reciprocal crosses (F1), intercrosses (F2) and backcrosses (BC). The descendents were tested by a standard insecticide resistance bioassay. Resistance ratios were 1 for S strain, 103.36 for R strain and 18.34 for F1. The regression lines of F1 generations (R×S and S×R) showed no significant differences and were closer to that of the R parents, indicating that inheritance of deltamethrin resistance in T. infestans is autosomal and incompletely dominant (D=0.20). Chi-square analysis from responses of intercross and backcross progenies rejected the hypothesis of a single gene being responsible for resistance. The minimum number of independent segregation genes was three, as calculated with Lande's method. The genetic basis here described for the resistant phenotype indicate that, under pyrethroid selective pressure, the resistant genotypes could be easily spread to susceptible insects from resistant individuals, posing a major threat to vectorial control of Chagas disease.

Increased TRPV1 gene expression in esophageal mucosa of patients with non-erosive and erosive reflux disease.

BACKGROUND: Transient receptor potential channel vanilloid subfamily member-1 (TRPV1) may play a role in esophageal perception. TRPV1 mRNA and protein expression were examined in the esophageal mucosa of non-erosive reflux disease (NERD) and erosive esophagitis (EE) patients and correlated to esophageal acid exposure. METHODS: Seventeen NERD patients, eight EE patients and 10 healthy subjects underwent endoscopy after a 3-week washout from proton pump inhibitors or H2 antagonists. Biopsies, obtained from the distal esophagus, were used for conventional histology, for Western blot analysis and/or quantitative real-time polymerase chain reaction (qPCR). Overall 13 NERD patients, four EE patients and five controls underwent ambulatory pH-testing. KEY RESULTS: TRPV1 expression was increased in all NERD and EE patients, as measured by Western blot analysis (0.65 +/- 0.07 and 0.8 +/- 0.05 VS 0.34 +/- 0.04 in controls; P < 0.01) and by qPCR (1.98 +/- 0.21 and 2.52 +/- 0.46 VS 1.00 +/- 0.06; P < 0.01). Neutrophilic infiltration, in the mucosa, was detected only in EE patients. CONCLUSIONS & INFERENCES: Non-erosive reflux disease and EE patients presented increased TRPV1 receptors mRNA and protein, although no correlation with acid exposure was demonstrated. Increased TRPV1 in the esophageal mucosa may contribute to symptoms both in NERD and EE patients and possibly account for peripheral mechanisms responsible for esophageal hypersensitivity in NERD patients.

Cytogenetics and genome evolution in the subfamily Triatominae (Hemiptera, Reduviidae).

The subfamily Triatominae (Hemiptera, Reduviidae), vectors of Chagas disease, includes over 140 species. Karyotypic information is currently available for 80 of these species. This paper summarizes the chromosomal variability of the subfamily and how it may reveal aspects of genome evolution in this group. The Triatominae present a highly conserved chromosome number. All species, except 3, present 20 autosomes. The differences in chromosome number are mainly caused by variation in the number of sex chromosomes, due to the existence of 3 sex systems in males (XY, X(1)X(2)Y and X(1)X(2)X(3)Y). However, inter- and intraspecific differences in the position, quantity and meiotic behavior of constitutive heterochromatin, in the total genome size, and in the location of ribosomal 45S rRNA clusters, have revealed considerable cytogenetic variability within the subfamily. This cytogenetic diversity offers the opportunity to perform cytotaxonomic and phylogenetic studies, as well as structural, evolutionary, and functional analyses of the genome. The imminent availability of the complete genome of Rhodnius prolixus also opens new perspectives for understanding the evolution and genome expression of triatomines. The application of fluorescence in situ hybridization for the mapping of genes and sequences, as well as comparative analyses of genome homology by comparative genomic hybridization will be useful tools for understanding the genomic changes in relation to evolutionary processes such as speciation and adaptation to different environments.

Systematics of Mepraia (Hemiptera-Reduviidae): cytogenetic and molecular variation.

The haematophagous insects of the subfamily Triatominae (Hemiptera-Reduviidae) have great epidemiological importance as vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Mepraia was originally described as a monotypic genus comprised of Mepraia spinolai, distributed along coastal areas of northern Chile (from Region I to the Metropolitan Region). Recently, some M. spinolai populations have been ranked as a new species named Mepraia gajardoi. Several populations along the distribution range of the genus were sampled, and genetic differentiation was studied based upon the analysis of three molecular markers: cytogenetics (karyotype and chromosome behaviour during meiosis using the C-banding technique), mitochondrial DNA (a cytochrome oxidase I gene fragment), and nuclear ribosomal DNA (intergenic region including the two internal transcribed spacers ITS-1 and ITS-2 and the 5.8S rRNA gene). The data here presented indicate that populations within the Mepraia genus (excluding Region II specimens) can be divided into two separate lineages. One lineage is comprised of specimens from the northernmost Region I and represents M. gajardoi. The other includes samples from the southern III, IV and the Metropolitan Regions, and represents M. spinolai. Region II individuals deserve particular attention as their relationship to the two identified lineages is not clear-cut. While they appear to belong to M. spinolai based on cytogenetics and rDNA markers, COI results indicate a closer relationship to M. gajardoi. This disagreement can be due to mitochondrial DNA introgression or the retention of ancestral polymorphisms.

Genetic variability of Triatoma rubrovaria (Reduviidae: Triatominae) from Brazil, Argentina and Uruguay as revealed by two different molecular markers.

Randomly amplified polymorphic DNA (RAPD) and nuclear ribosomal DNA sequence analyses were used to assess the genetic population structure of the South American triatomine species Triatomo rubrovario throughout its geographical distribution. To investigate the genetic variability at both intraspecific and intrapopulational levels the RAPD profiles and the nucleotide sequences of the rDNA intergenic spacers, ITS-1 and ITS-2, were analysed and compared. The phenetic analysis based on RAPD profiles show three distinct clusters diverging by similarity coefficients ranging from 0.62 to 0.96. The ITS-1 and ITS-2 sequence variability detected may be considered very high, suggesting reproductive isolation between populations. A total of seven composite haplotypes (CH) were found, among which three are specific for Brazil, other three for Uruguay, and the last one common for the three countries studied. The population studied in Argentina does not represent an independent CH. Sequence analyses proved that the five populations studied are easily differentiable and that there is heterogeneity within each one. True mutations and indels are the responsible of sequence differences between haplotypes and populations, suggesting that divergence processes may presently go on within this species. The large intraspecific variability detected may underlie the known plasticity of T. rubrovaria, making it a potential intradomiciliary invader and consequently an appropriate vector for Chagas disease transmission. Therefore, this triatomine species must be continuously monitored throughout.

Evolutionary relationships based on genetic and phenetic characters between Triatoma maculata, Triatoma pseudomaculata and morphologically related species (Reduviidae: Triatominae).

The maculata group currently comprises two species of Triatominae, Triatoma maculata and Triatoma pseudomaculata, which share morphologic and chromatic characteristics. In order to clarify the systematic status of these two vectors of Trypanosoma cruzi and to infer their evolutionary relationships, we performed an enzymatic, morphometric and cytogenetic comparison of them, also taking into account two sister species not included in the group (T. arthurneivai and T. wygodzinskyi). According to our results, T. maculata and T. pseudomaculata belong to distinct evolutionary lineages. Similarly, T. arthurneivai topotypes from Minas Gerais form an independent isolated group by morphometrics. Our results also support the specific status of the Triatoma population from São Paulo State (formerly referred to T. arthurneivai), and suggest the possibility that it is T. wygodzinskyi. Finally, we suggest that only the arboricolous T. pseudomaculata from northeast Brazil and the rupicolous sister species originated from São Paulo State should be classified together in the same group.

Chromosomal variation and genome size support existence of cryptic species of Triatoma dimidiata with different epidemiological importance as Chagas disease vectors.

The wide geographical distribution of Triatoma dimidiata, one of the three major vectors of Chagas disease, ranges from Mexico to northern Peru. Since this species occupies a great diversity of artificial and natural ecotopes, its eradication is extremely difficult. In order to assist control efforts, we used chromosome analyses and DNA amount as taxonomic markers to study genetic variability in populations of T. dimidiata from Mexico, Guatemala, El Salvador and Colombia. We differentiated three groups or cytotypes defined by characteristic chromosome C-banding patterns and genome size measured by flow cytometry. The three cytotypes are restricted to different geographic locations. Cytotype 1 occurs in Mexico (excluding Yucatán), Guatemala (excluding Petén), El Salvador and Colombia. Cytotype 2 occurs in Yucatán and cytotype 3 occurs in Petén. Cytotype 1, commonly associated with domestic and peridomestic environments but also inhabiting sylvatic ecotopes, is the most widespread and with major epidemiological significance. In contrast, the Yucatán cytotype inhabits wild ecotopes but increasingly enters houses, while the Petén cytotype appears exclusively sylvatic. We suggest that these cytotypes represent cryptic species of T. dimidiata with different epidemiological relevance as Chagas disease vectors. Poor ability to colonize human dwellings, together with their restricted geographic distribution, indicate that the Yucatán and Petén putative species probably have much less epidemiological significance than cytotype 1. Thus, the genetic markers we describe are powerful tools to differentiate cryptic species in T. dimidiata with different epidemiological significance, contributing to planning the most effective control measures.

Origin and phylogeography of the Chagas disease main vector Triatoma infestans based on nuclear rDNA sequences and genome size.

For about half of all Chagas disease cases T. infestans has been the responsible vector. Contributing to its genetic knowledge will increase our understanding of the capacity of geographic expansion and domiciliation of triatomines. Populations of all infestans subcomplex species, T. infestans, T. delpontei, T. platensis and T. melanosoma and the so-called T. infestans "dark morph", from many South American countries were studied. A total of 10 and 7 different ITS-2 and ITS-1 haplotypes, respectively, were found. The total intraspecific ITS-2 nucleotide variability detected in T. infestans is the highest hitherto known in triatomines. ITS-1 minisatellites, detected for the first time in triatomines, proved to be homologous and thus become useful markers. Calculations show that ITS-1 evolves 1.12-2.60 times faster than ITS-2. Despite all species analyzed presenting the same n=22 chromosome number, a large variation of the haploid DNA content was found, including a strikingly high DNA content difference between Andean and non-Andean specimens of T. infestans (mean reduction of 30%, with a maximum of up to 40%) and a correlation between presence/absence of minisatellites and larger/smaller genome size. Population genetics analysis of the eight composite haplotypes of T. infestans and net differences corroborate that there are clear differences between western and eastern populations (60%), and little genetic variation among populations (1.3%) and within populations (40%) within these two groups with migration rates larger than one individual per generation corresponding only to pairs of populations one from each of these groups. These values are indicative either of a large enough gene flow to prevent population differentiation by drift within each geographic area or a very recent spread, the latter hypothesis fitting available data better. Phylogenetic trees support a common ancestor for T. infestans and T. platensis, an origin of T. infestans in Bolivian highlands and two different dispersal lines, one throughout Andean regions of Bolivia and Peru and another in non-Andean lowlands of Chile, Paraguay, Argentina, Uruguay and Brazil.

Nuclear rDNA ITS-2 sequences reveal polyphyly of Panstrongylus species (Hemiptera: Reduviidae: Triatominae), vectors of Trypanosoma cruzi.

Panstrongylus species are widely distributed throughout the Americas, where they act as vectors of Trypanosoma cruzi, agent of Chagas disease. Their intraspecific relationships, taxonomic position and phylogeny in relation to other Triatomini were explored using ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS-2) sequence polymorphisms and maximum parsimony, distance and maximum likelihood analyses of 10 populations representing six species of the genus (P. megistus, P. geniculatus, P. rufotuberculatus, P. lignarius, P. herreri and P. chinai). At the subspecific level, P. megistus appeared more homogeneous than P. rufotuberculatus and P. geniculatus (both with broader distribution). Several dinucleotide microsatellites were detected in the sequences of given species. Many of these microsatellites (GC, TA, GT and AT) showed different number of repeats in different populations and thus, may be very useful for population differentiation and dynamics analyses in future studies. The sequences of P. lignarius (considered sylvatic) and P. herreri (a major disease vector in Peru) were identical, suggesting that these species should be synonymised. Intrageneric analysis showed a clear separation of P. rufotuberculatus, with closest relationships between P. geniculatus and P. chinai, and P. megistus occupying a separate branch. Genetic distances between Panstrongylus species (0.11585-0.22131) were higher than those between Panstrongylus and other Triatomini (16 species from central and North America and South America) (0.08617-0.11039). The distance between P. megistus and P. lignarius/herreri (0.22131) was the largest so far recorded in the tribe. The pronounced differences in length and nucleotide composition suggest a relatively old divergence of Panstrongylus species. P. rufotuberculatus was closer to Mesoamerican Triatoma, Meccus and Dipetalogaster species than to other Panstrongylus. All Panstrongylus clustered with the Mesoamerican clade; P. rufotuberculatus clustered with the phyllosoma complex and T. dimidiata, with D. maxima and T. barberi in a basal position. The rest of Panstrongylus appeared paraphyletically in the tree. This is evidence suggesting polyphyly within the genus Panstrongylus, whose species may be related to the ancestors giving rise to central and North American Triatomini.

Chromosome homogeneity in populations of Triatoma brasiliensis Neiva 1911 (Hemiptera - reduviidae - triatominae).

Triatoma brasiliensis is the most important vector of Chagas disease in the semiarid zone of the Northeast of Brazil. Several authors have reported the occurrence of four chromatic patterns with morphological, ecological, and genetic differences. In order to determine the existence of cytogenetic differentiation between these chromatic forms, we analyzed their karyotypes and the chromosome behavior during the male meiotic process. Triatoma brasiliensis shows distinct and specific chromosome characteristics, which differ from those observed in all other triatomine species. However, no cytogenetic differences were observed between the four chromatic forms of T. brasiliensis. The lack of chromosome differentiation among them could indicate that the populations of this species are in a process of differentiation that does not involve their chromosomal organization.

Meiosis in holocentric chromosomes: orientation and segregation of an autosome and sex chromosomes in Triatoma infestans (Heteroptera).

The meiotic behaviour of the X chromosome and one autosomal pair of the heteropteran Triatoma infestans was analysed by means of C-banding plus DAPI staining. At first metaphase, the X univalent is oriented with its long axis parallel to the equatorial plate, which suggests a holocentric interaction with the spindle fibres. After this initial orientation, kinetic activity is restricted to one of both chromatid ends. The election of the active chromatid end is random and it is independent of the end selected in the sister chromatid. At second metaphase, the X and Y chromatids associate side by side forming a pseudobivalent. After that, the kinetic activity is again restricted to either of both chromosomal ends in a random fashion. At first metaphase, the fourth autosomal bivalent shows two alternative random orientations depending on the chromosome end showing kinetic activity (DAPI positive or opposite). At second metaphase, half bivalents are oriented with their long axis parallel to the equatorial plate. Three different segregation patterns are observed. The kinetic activity can be localised: (i) in the end with the DAPI signal (46.9%), (ii) in the opposite end (44.6%) or (iii) in one DAPI-positive end in one chromatid and in the opposite end in the other one (8.5%). The existence of the last pattern indicates that the same end can show kinetic activity during both meiotic divisions. Our results provide new information on the comparative meiotic behaviour of autosomes and sex chromosomes in holocentric systems.

[Laparoscopic appendectomy. Retrospective analysis of our experience compared with the literature]. / Appendicectomia laparoscopica. Analisi retrospettiva della nostra esperienza in rapporto alla letteratura.

BACKGROUND: The benefits of laparoscopic appendectomy (LA) remain controversial. This study reports a critical examination of our experience in a peripheral hospital. METHODS: A total of 128 appendectomies have been performed since January 1996, of which 63 (49%) were laparoscopic. The patients included 52 (82.5%) females and 11 (17.5%) males with a mean age of 20.8 years (range 11-46). Emergency surgery was required in 6 cases (9.5%) and was elective in 57 (90.5%). Appendectomy was performed during another operation in 3 cases (laparoscopic cholecystectomy). RESULTS: Mortality was nil. Morbidity was 1.7% (1 case). The index of conversion was 0%. Only one major complication occurred. This took the form of perforation of an ileal loop following accidental lesion during adhesiolysis. The intraoperative diagnosis was not confirmed in 12 (19%) cases: 9 ovarian cysts, 1 terminal ilieitis and 2 cases of acute salpingitis. In 10 cases (15%) surgery was associated with adhesiolysis, and in 6 cases (9.3%) the appendix was retrocecal. Mean operating time was 42 min (range 18-105 min). The mean hospitalisation was 3.3 days. CONCLUSIONS: The authors emphasise the numerous advantages of laparoscopic techniques in their experience, including excellent cosmetic results, reduced PO pain, rapid functional recovery, lower incidence of adhesion, wound infection and laparocele, and more cost-effective when mechanical staplers are not used. Owing to the ability to explore the entire abdominal cavity, the main advantage of this technique consisted in a correct differential diagnostic balance, especially in young women of child-bearing age, between appendectomy and pathologies of the uterus and adnexa.

[Laparoscopic cholecystectomy. Our experience]. / Colecistectomia laparoscopica. Nostra esperienza.

BACKGROUND: The many advantages and extreme versatility made laparoscopic cholecystectomy (L.C.) the gold standard for symptomatic cholelithiasis. The aim of this research is a retrospective analysis of personal experience with laparoscopic cholecystectomy in a peripheral hospital compared with the literature on the subject. METHODS: Since June 1982, a total of 989 L.C. have been performed. The patients were: 691 (68.6%) women and 298 (29.4%) men with a mean age of 56 years (range 17-84). The indications were: 791 symptomatic cholelithiasis, 142 chronic cholecystitis, 45 empyema-hydrops and 11 adenomyomatosis. RESULTS: No postoperative death have been observed and the conversion rate was of 24 cases (2.3%). The main complications were 3 cases of injury of the biliary tract and 2 cases of postoperative bleeding (1 from cystic artery and 1 from the umbilical wound). Minor complications observed were 12 cases (1.2%) of infections of the umbilical wound and 3 cases of umbilical hernia (0.3%). The elevation of stasis index was observed in 3 cases which solved spontaneously. The length of stay was 2 days in 957 cases (97%), 8 days in 24 cases and 4-5 days in 8 (0.7%). CONCLUSIONS: No major trochar's lesions occurred, contrary to the percentages quoted in the literature (0.02%-0.9%). The selective use of the open technique and of the multiuse conic section trocar in the "closed" technique is suggested. The 3 cases of bile duct lesions did not occur during the training period, contrary to what quoted in the literature. In personal opinion, a careful surgical technique with a good Calot's triangle preparation, is necessary to prevent these inconveniences. Parietal complications (umbilical wound infections and laparocele) even if lesser than in the laparotomy technique, can be reduced by using the endobag and suturing the abdominal fascia of the 10 mm trocars. Moreover, the use of a systematic subhepatic drainage during the first 24 postoperative hours is suggested, since it can be useful to reveal possible bleeding.

Sylvatic population of Triatoma infestans from the Bolivian Chaco: from field collection to characterization.

A sylvatic Triatoma infestans DM (dark morph) population detected in the Bolivian Chaco was characterized and compared with various domestic ones. The degree of differentiation of DM was clearly within the T. infestans intra-specific level. Nevertheless marked chromatic and morphometric differences as well as differences in antennal pattern, chromosome banding and randomly amplified polymorphic DNA support the hypothesis of a distinct population. Continuous exchange of insects between wild and domestic habitats seems unlikely in the Chaco.