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The mammalian innate immune system recognizes pathogen-associated molecular patterns through pathogen recognition receptors. Nod1 has been described recently as a cytosolic receptor that detects specifically diaminopimelate-containing muropeptides from Gram-negative bacteria peptidoglycan. In the present study we investigated the potential role of Nod1 in the innate immune response against the opportunistic pathogen Pseudomonas aeruginosa. We demonstrate that Nod1 detects the P. aeruginosa peptidoglycan leading to NF-kappaB activation and that this activity is diminished in epithelial cells expressing a dominant-negative Nod1 construct or in mouse embryonic fibroblasts from Nod1 knock-out mice infected with P. aeruginosa. Finally, we demonstrate that the cytokine secretion kinetics and bacterial killing are altered in Nod1-deficient cells infected with P. aeruginosa in the early stages of infection.

Introduction: Esophagojejunostomy after total gastrectomy still remains a high risk anastomosis with a considerable morbidity and mortality. The majority of these anastomoses are performed by the intraluminal stapler technique, yet stenoses are a known late complication even after an uneventful postoperative course. In the present study, the osophagojejunostomy with the biofragmentable anastomosis ring (BAR) was examined in dogs., Methods: 28 dogs were randomized into a group of manual suture (n = 14) and a BAR-group (n = 14). After gastrectomy, the esophagojejunostomy was performed by hand-suture with polypropylene 4-0 in the manual suture group, and with the 25/1.5 mm BAR in the BAR-group. In both groups the Roux-en-Y jejunojejunostomy was performed by hand-suture. The dogs were evaluated on postoperative days 4, 7 and 14 with regard to macroscopy, bursting strength, tissue hydroxyproline and histology., Results: There was one leakage without clinical effect in the hand-sewn group on postoperative day 4; there was no leak in the BAR-group. In observing fibre-free enteral feeding, neither functional disorders nor obstruction of the BAR were observed. The general anastomosis parameters were matchable between the groups., Conclusion: The infracarinal BAR-esophagojejunostomy is comparable to the hand-sewn anastomosis in the dog-model.

Objective: To analyze the outcome of neonatal pelvic ectasia (PE) and the association between this entity and vesicoureteral reflux and/or other urinary tract abnormalities., Patients and Methods: We performed a retrospective study of 255 children (205 boys, 50 girls) with an ultrasonographic diagnosis of PE in the first month of life. The initial ultrasonographic examination was indicated by urinary tract infection in 30 neonates, abnormalities in the prenatal ultrasonographic examination in 150 and by other reasons in 75. Pelvic ectasia was classified in four stages according to anteroposterior pelvic diameter: I < 1 cm, II 1-1.5 cm, III 1.6-2 cm, and IV > 2 cm., Results: Pelvic ectasia was bilateral in 153 children (60 %) and unilateral in 102 (left side in 81.4 % and right side in 18.6 %). Stage I was found in 75.49 %, stage II in 20.34 %, stage III in 3.9 % and stage IV in 0.24 %. The mean follow-up was 32.6 +/- 25.2 months. At the end of the first year, the results of renal ultrasound were normal in 70.2 % of left-sided PE and in 55.9 % of right-sided PE, but 46 patients (18 %) showed worsening of PE between the first and second ultrasound scans. Voiding cystourethrography was performed in 79.6 % of the children and some abnormalities were found in 50 (24.6 %): urethral dilatations in two patients and vesicoureteral reflux in 48. No correlation was found between vesicoureteral reflux and the degree of ectasia (74 % had an anteroposterior diameter of < or = 1 cm). Urinary tract infection was present in 24.3 % of the children and 13 required surgery (eight pyeloplasties, four urethral reimplantations and two resections of type III urethral valves)., Conclusions: Neonatal PE was more prevalent in boys (4:1) and was more frequently located on the left side in both sexes. Associated vesicourethral reflux was found in 23.64 % with no correlation between the degree of dilation and the presence or degree of reflux. Consequently, cystourethrography should be performed in any child with pelvic ectasia, regardless of stage, side or sex.

Virtual reality (VR) has been demonstrated as an effective tool to help people overcome a variety of anxiety disorders. In this case study, the use of VR as a distractor to alleviate pain and anxiety associated with an invasive medical procedure for a pediatric cancer patient was explored. An A-B-C-A design during four consecutive medical appointments in an outpatient oncology clinic compared no distraction (A), non-VR distraction on a computer screen (B), and VR distraction with a head set (C). Behavioral observations of distress by the researcher and reports of pain and anxiety by the patient, parent, and nurse were taken before and during the procedure. The child's pulse was monitored throughout the procedure. The findings from this case study suggest benefit from using VR distraction, as indicated by lower pain and anxiety ratings, reduced pulse, and fewer observed behavioral indices of distress. The need for larger scale studies and application of VR with younger children is discussed in the context of confirming effectiveness of this technique and providing more generalizable information about efficacy.

Sulfate-reducing bacteria are considered as strict anaerobic microorganisms, in spite of the fact that some strains have been shown to tolerate the transient presence of dioxygen. This report shows that membranes from Desulfovibrio gigas grown in fumarate/sulfate contain a respiratory chain fully competent to reduce dioxygen to water. In particular, a membrane-bound terminal oxygen reductase, of the cytochrome bd family, was isolated, characterized, and shown to completely reduce oxygen to water. This oxidase has two subunits with apparent molecular masses of 40 and 29 kDa. Using NADH or succinate as electron donors, the oxygen respiratory rates of D. gigas membranes are comparable to those of aerobic organisms (3.2 and 29 nmol O(2) min(-1) mg protein(-1), respectively). This 'strict anaerobic' bacterium contains all the necessary enzymatic complexes to live aerobically, showing that the relationships between oxygen and anaerobes are much more complex than originally thought.

Complex II from the thermoacidophilic archaeon Acidianus ambivalens, an archetype of an emerging class of succinate dehydrogenases (SDH), was extracted from intact membranes and purified to homogeneity. The complex contains one molecule of covalently bound FAD and 10 Fe atoms. EPR studies showed that the complex contains the canonical centres S1 ([2Fe-2S]2+/1+) and S2 ([4Fe-4S]+2/+1) but lacks centre S3 ([3Fe-4S]+1/0); these observations agree with the fact that the iron-sulfur subunit contains an extra cysteine that may allow the binding of a new centre, most probably a tetranuclear one. Succinate-driven oxygen consumption is observed in intact membranes indicating that in vivo, complex II operates as a succinate:quinone oxidoreductase, despite missing the typical anchor domain subunits. The pure complex was found to contain bound caldariella quinone, the enzyme physiological partner. An alternative membrane anchoring for this new type of SDHs, based on the amphipathic nature of the putative helices found in SdhC, is suggested.

In aquatic systems, bottom sediments have often been considered as the main methylmercury (MeHg) production site. In tropical floodplain areas, however, floating meadows and flooded forests extend over large areas and can be important Hg methylating sites. We present here a cross-system comparison of the Hg net methylation capacity in surface sediments, flooded soils and roots of floating aquatic macrophytes, assayed by in situ incubation with 203Hg and extraction of formed Me203 Hg by acid leaching and toluene. The presence of mono-MeHg was confirmed by thin layer chromatography and other techniques. Study areas included floodplain lakes in the Amazon basin (Tapajós, Negro and Amazon rivers), the Pantanal floodplain (Paraguay river basin), freshwater coastal lagoons in Rio de Janeiro and oxbow lakes in the Mogi-Guaçú river, São Paulo state. Different Hg levels were added in assays performed in 1994-1998, but great care was taken to standardise all other test parameters, to allow data comparisons. Net MeHg production was one order of magnitude higher (mean 13.8%, range 0.28-35) in the living or decomposing roots of floating or rooted macrophyte mats (Eichhornia azurea, E. crassipes, Paspalum sp., Eleocharis sellowiana, Salvinia sp., S. rotundifolia and Scirpus cubensis) than in the surface layer of underlying lake sediments (mean 0.6%, range 0.022-2.5). Methylation in flooded soils presented a wide range and was in some cases similar to the one found in macrophyte roots but usually much lower. In a Tapajós floodplain lake, natural concentrations of MeHg in soil and sediment cores taken along a lake-forest transect agreed well with data on net methylation potentials in the same samples. E. azurea, E. crassipes and Salvinia presented the highest methylation potentials, up to 113 times higher than in sediments. Methylation in E. azurea from six lakes of the Paraguay and Cuiabá rivers, high Pantanal, was determined in the 1998 dry and wet seasons and ranged from 1.8 to 35%. Methylation was lower in washed roots than in untreated roots of E. azurea and methylation in solids isolated from the roots, was higher than in sediments but lower than in untreated roots. This indicates that the methylation in roots zones occurs mainly in the root-associated solids. Floating meadows are sites of intense production of biomass and of highly bioavailable MeHg and appear to be an essential link of the MeHg cycle in tropical aquatic systems.

We report the case of a 47-year-old woman with a large keloid scar on the sternum who was submitted to a simultaneous scar removal with bilateral breast-reduction mammaplasty. Breast reduction was performed to reduce local skin tension and to provide a skin flap for the full reconstruction of the scar-removal site. The association of these surgical procedures stands as a viable alternative for the reconstruction of the sternum region, producing less keloid scarring.

Cutaneous injuries of the inferior third of the leg and dorsum of the foot represent a great challenge for plastic surgeons. The poor vascularization and subsequent poor healing encountered in these regions demand detailed knowledge of the local anatomy to select the best surgical alternative for each patient. In patients in whom local or free muscular flaps are not suitable, the fasciocutaneous flaps seem to be a good alternative. The distal pedicled fasciocutaneous flap of the calf was used to cover skin defects of the leg and foot in 22 patients. In 4 patients the authors designed an island flap to prevent a large defect in the donor area and to decrease the time of hospitalization and recovery of the patient. In a severe defect, the flap was used as a cross-leg flap. The results were satisfactory because the flap provided stable coverage for different defects with few complications. The island flap is a good option for achieving a better aesthetic result and can be used in select patients.

Plasmacytoma is an unusual disease of plasma cells, its presentation in the urethra is rare. We report the sixth known case of primary urethral plasmacytoma, which was treated by surgical excision, without recurrence for 3 years of follow-up.

The succinate dehydrogenase complex of the thermoacidophilic archaeon Acidianus ambivalens was investigated kinetically and by EPR spectroscopy in its most intact form, i.e., membrane bound. Here it is shown that this respiratory complex has an unusual iron-sulfur cluster composition in respect to that of the canonical succinate dehydrogenases known. The spectroscopic studies show that center S3, the succinate responsive [3Fe-4S]1+/0 cluster of succinate dehydrogenases, is not present in membranes prepared from aerobically grown A. ambivalens, nor in partially purified complex fractions. On the other hand, EPR features associated to the remaining centers, clusters S1 ([2Fe-2S]1+/2+) and S2 ([4Fe-4S]2+/1+), could be observed. Similar findings were made in other archaea, namely Acidianus infernus and Sulfolobus solfataricus. Kinetic investigations showed that the A. ambivalens enzyme is reversible, capable of operating as a fumarate reductase - a required activity if this obligate autotroph performs CO2 fixation via a reductive citric acid cycle. Sequencing of the sdh operon confirmed the spectroscopic data. Center S3 ([3Fe-4S]) is indeed replaced by a second [4Fe-4S] center, by incorporation of an additional cysteine, at the cysteine cluster binding motif (CxxYxxCxxxC-->CxxCxxCxxxC). Genomic analysis shows that genes encoding for succinate dehydrogenases similar to the ones here outlined are also present in bacteria, which may indicate a novel family of succinate/fumarate oxidoreductases, spread among the Archaea and Bacteria domains.

Objective: The purpose of this study was to assess the incidence of vascular complications in pediatric renal transplant patients and to evaluate the accuracy and limitations of duplex-Doppler color ultrasound (US) in their diagnosis., Patients and Methods: The US studies of 89 transplants performed in 76 patients were reviewed. Ages of the patients ranged from 11 months to 23 years (mean 13 years). The US exams were performed every 24 hours during the first week post-transplantation, once a week during the hospital stay and whenever graft dysfunction occurred. A vascular map of the graft was performed with color Doppler and maximum systolic velocity, systolic acceleration time, resistive index and flow velocity in the renal vein were determined with pulsed Doppler. An angiography was performed in all patients with US diagnosis of vascular thrombosis, stenosis or arteriovenous fistula., Results: Seventeen grafts (15%) showed vascular complications including renal artery stenosis (8 cases), renal artery thrombosis (4 cases), arteriovenous fistula (4 cases) and one renal vein thrombosis. US studies were able to detect six renal artery stenosis (false negative results in two cases), seven renal artery thrombosis (false positive results in three grafts), all of the arteriovenous fistulas and one renal vein thrombosis., Conclusions: Doppler-duplex color US has a high sensitivity and specificity in diagnosis of vascular complications and should be the first approach for their diagnosis. The presence of false positive and false negative results makes it necessary to perform angiography when symptomatic thrombosis, arteriovenous fistula or stenosis is suspected.

Objective: The characterization of drug users, especially as regards self-medication and the determination of its prevalence in the population studied., Material and Method: About 413 people that had used drugs in the previous month were interviewed., Results: From the interviewed, 69.9% used medicines and of those 76.1% were self medicated. Headache (28.8%) was the main complaint among the self-medicated group. Acetilsalicilic acid was the most frequently used medicine (25.4%). As regards the drugs utilized, 51.2% of the users had received a recommendation from a third party and 51.7% used old prescriptions, given in previous consultations., Conclusion: Age, schooling and absence of periodic medical consultation were significant statistical factors in self-medication.

A clinical case of sheep deaths due to Phytolacca decandra occurred in Southern Brazil and was experimentally duplicated by force-feeding the plant. General neurologic signs and diarrhea with rumen and reticulum hyperemia and coagulation necrosis of the mucosa were seen.

Sarcomas constitute less than one percent of laryngeal neoplasms. Fibrosarcomas are by far the most commonly reported mesenchymal tumors of the larynx. We report two cases of fibrosarcoma of the larynx and discuss clinical aspects, diagnosis and treatment.

The present investigation studies the role of multivariate statistical methods on quantitative histopathological features of cells in uterine cervix epithelium to discriminate between normal and abnormal uterine cervix samples. 143 histological specimens were included in the study involving normal cervix, cervical intraepithelial neoplasia (CIN) lesions and cervical human papillomavirus (HPV) infection with and without CIN (condyloma-CIN and condyloma-NCIN groups, respectively). Deep, middle and superficial regions of the cervical squamous epithelium were morphometrically analyzed. Identification of normal cervix from pathological cases was highly achieved with a specificity of 100%. The application of discriminant statistical method within pathological specimens showed an acceptable percentage of cases correctly classified; thus, an efficiency of 83.0% and 74.6% was obtained in order to discriminate within CIN and condyloma-CIN grades respectively. These percentages increased when differentiation between each grade of CIN versus condyloma-CIN were considered, using only 1-3 morphometrical parameters. Our findings indicate that the combination of nuclear and cytoplasmic quantitative features, specially size parameters, permit a high correct percentage classification of cervix samples. The discrimination process was better when few diagnostic categories were included; however, 100% specificity for normal samples was always reached.

We quantitatively studied the nuclear and cytoplasmic characteristics of cells in the normal cervix, squamous intraepithelial lesions (cervical intraepithelial neoplasia [CIN]) lesions and cervical human papillomavirus (HPV) infection with and without CIN (condyloma-CIN and condyloma-NCIN groups, respectively). The morphometric features of the deep, middle and superficial layers of the cervical epithelium were calculated on 143 cervical samples. The morphometric parameters selected for the analysis were area; perimeter; maximal, minimal and equivalent circle diameters; nuclear/cytoplasmic ratio; and several shape factors. Our quantitative findings support the opinion of other authors that HPV infection is associated with shape and size modifications of the nucleus and cytoplasm in CIN. In relation to cellular size, there were no differences between a normal cell, regardless of its region, and a virus-infected cell. The nuclear differences are predominantly in the deep layer areas. The CIN samples showed changes in both cellular and nuclear form and size but lacked substantial differences in the tumor grades and areas studied. The condyloma-CIN samples revealed more notable differences in both tumor grade differentiation and different cellular layers.

One hundred forty samples of normal and pathological endometrium were examined morphometrically in order to achieve a discriminant function among normal and pathological diagnoses. Samples of proliferative and secretory normal endometrium, simple and atypical hyperplasia, and well-differentiated adenocarcinoma were included in this study. Morphometrical and stereological characteristics for both glands and lumina as well as epithelium were registered. Stepwise discriminant analysis was performed. There is an increase in size of glands from proliferative endometrium to well-differentiated adenocarcinoma. The stereological measurements parallel morphometrical measurements. The accuracy of diagnosis by morphometrical and stereological means were 91% when the normal group (proliferative and secretory phase) were included. Accuracy increases to 95% when only three pathological groups were used; only one simple hyperplasia and one well-differentiated adenocarcinoma were incorrectly diagnosed. In all functions, the volume density of epithelium was selected. The reproducibility on a new data set demonstrates an efficiency of 92, 87, and 100% for three discriminant analyses (five, four, and three diagnostic groups, respectively).

Background: The stereological estimate of mean glomerular volume is an objective and reproducible method of measuring glomerular size in terms of absolute volume. This method provides objective, unbiased, and reproducible data that may be used in the description of glomeruli., Experimental Design: We have made a morphometric analysis of glomerular size in three cortical areas (superficial, midcortical, and juxtamedullary) in normal and diabetic animals, using the point-sampled intercepts method. The kidney weight and volume as well as the cortical and glomerular volume density were also calculated. Our study included 35 animals (5 control and 30 experimental). Fifteen experimental animals were treated with 50 mg/kg streptozotocin dissolved in citrate buffer at pH 4.5. The other 15 animals were sham injected with citrate buffer. In all cases the tissues were fixed in 10% formaldehyde and embedded in paraffin. Sections were cut at 4 microns and stained with periodic acid-Schiff and a systematic sampling was performed., Results: In the control and experimental groups the juxtamedullary glomeruli was significantly larger than superficial and midcortical glomeruli. Mean values of the kidney weight and volume and the volume-weighted mean glomerular volume show a significant increase at 4, 20, and 50 days after streptozotocin-induced diabetes., Conclusions: The increase in the volume-weighted mean glomerular volume was similar in the three cortical areas, thus suggesting that the effect of diabetes on glomerular enlargement is similar in all cortical areas.

The authors dated 100 normal endometrial biopsies. Only endometrial specimens with histologically confirmed subphases and no evidence of organic disease were accepted for study. The morphometry and stereologic measures of the glands, lumina and endometrial epithelium were assessed. Quantitative assessment of the normal endometrium supported the histologic events that occur in the endometrial cycle (proliferative and secretory phases). There was a progressive increase in morphometric and stereologic values of the glands, epithelium and lumina during the endometrial cycle. Using stepwise discriminant analysis, 94% of the specimens were correctly classified into the categories of proliferative and secretory phase; two quantitative parameters were required, the epithelial volume density and longest luminal diameter. When three subphases within proliferative and secretory endometrium were considered, the overall accuracy was 90% and 78%, respectively.

The lateral calcaneal artery flap is an axial flap which comprises also the saphenous vein and the sural nerve. Its use has been reported for coverage of skin loss of the malleolus and calcaneal area. The donor area should be closed by a split thickness skin graft. In the present report, eight patients who had skin loss on the foot were submitted to reconstruction with the lateral calcaneal flap. In seven there were island flaps and in one a bridge of skin similar to that described by Grabb and Argenta. The etiology was trauma in five patients and decubitus ulcer in three. The results were good. The flap was adequate for the cover on the posterior foot area even after submitted to the strain of normal walking. The advantages were simpler technique and consistency of the anatomical landmarks.

1. Since monocyte-macrophages have been recognized as HIV targets in addition to CD4+ T-lymphocytes, we have evaluated HIV infection of purified peripheral blood mononuclear cell fractions obtained from 10 seropositive asymptomatic hemophiliacs and compared it with that of 10 asymptomatic homosexual patients. 2. HIV was isolated more frequently from the lymphocytes than the monocytes of both groups of patients. 3. HIV preferentially replicated in phytohemagglutinin-stimulated lymphocytes compared with growth factor-treated monocytes. Monocytes did not preferentially harbour HIV in either group.

Experiments were carried out on 40 adult Wistar rats to determine the effect of the application of a pneumatic tourniquet on the capillary network of the tibialis anterior muscles. The tourniquet was applied for four hours to the right hind limb above the knee. During the degeneration processes (6 to 24 hours postischemia), the capillary density and capillary-to-fiber ratio decreased significantly with respect to a control group of untreated animals. In the subsequent regenerative processes, there was a greater increase of fiber density than of capillary density, resulting in a capillary-to-fiber ratio lower than that of the control group. In the last period (30 to 90 days postischemia), both capillary density and fiber density returned to normal values.

Experience with scleral buckling for exudative and tractional retinal detachments in very young premature infants who have acute retrolental fibroplasia (RLF) is reported. In this series, surgery was successful in 24 of 32 eyes (75%) of 23 infants, (age at surgery ranged from 1.75 months to 6 months, average age of 3 months). We attribute the 15% improvement in the success rate over our 1979 series in part to closer screening with emphasis on the examination at age 8 weeks, to earlier referral for treatment, and to our observation that these fragile eyes can withstand reoperation if buckle revision is necessary to combat persistent retinal traction. We encountered no serious complications either at surgery or immediately after surgery. We stress that (1) because of the high incidence of spontaneous remission, buckling surgery should not be performed until the detachment has progressed well posterior to the equator and remission is obviously unlikely (Grade IV or V); (2) excessive cryotherapy and undue surgical trauma should be avoided; and (3) because scleral erosion is certain as the eyes grow, the encircling band should be transected six months to one year after surgery.

A case of pure squamous cell carcinoma of the stomach in a 63-year-old woman has been reported. The tumor was located in pyloric antrum and histologic examination disclosed a well differentiated squamous cell carcinoma, with many horn pearls. Histogenesis of similar cases of this unusual tumor have been briefly discussed.

We report a case of the Werdnig-Hoffmann disease in a 4-month-old male infant. The morphological study revealed perimysial fibrosis, variability in the size of muscle fibers, absence of target fibers, few central nuclei and normality in vessels, nerves and neuromuscular junctions. The morphometrical examination showed the existence of normal-sized and atrophic fibers in both fibrillar types, as well as in hypertrophic type I fibers. The percentage of fibrillar types and the data obtained from the form factor are normal. Random distribution of type I and II muscle fibers were observed.

Ten patients had their vulvar and perineal areas reconstructed using musculocutaneous flaps of the gracilis muscle. Seven patients had extensive resections done for skin carcinomas, two due to traumas and one due to a sequel of Fournier's Syndrome. Reconstruction in tumor cases were performed primarily and the other secondarily. The skin island measured 12 x 8 centimeters and the muscle was taken entirely. The results were satisfactory in all cases. The muscle survived completely, but there was partial skin island loss in 8 instances. In 3 of those cases, there was need for a revision of the flap and skin grafting. The authors concluded that there is always a risk to take the skin island together with the gracilis muscle and the use of the isolated muscle with skin grafting should be considered im most cases.

The cardiovascular effects of a patent ductus arteriosus (PDA) in a premature baboon model of hyaline membrane disease treated with conventional positive-pressure ventilation (CPPV) or high-frequency ventilation (HFV) were studied. Twenty-seven (84%) of 32 infant baboons delivered at 75% gestation had positive retrograde aortograms at one hour of life. Eleven (34%) of these infant baboons had persisting evidence for retrograde flow over the first 24 h by Doppler examination. Significantly higher fluid requirements and bicarbonate requirements were present in the animals with a persisting PDA. Mean aortic BP values were significantly lower in these same animals. No significant differences were found between groups when mean airway pressure, heart rate, urine output, or form of ventilation were compared. The only significant differences found between the conventional and high-frequency ventilated animals with PDA were higher peak airway pressures and lower PEEP levels in the conventionally ventilated infants. The premature baboons can be a useful primate model to study the PDA. There appears to be no significant difference regarding the clinical effects of HFV and CPPV on PDA.