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Biomedical subjects

R L Santos

Publications and source records attributed to R L Santos.

At least 19 recordsLinked to original sources

Direct observation of a precursor in a complex system.

A precursor is a kind of instability in a nonequilibrium system observed prior to a bifurcation. It has been observed in important natural phenomena such as earthquakes and epileptic seizures. In most cases just qualitative results were reported, with the exception of some quantitative observations of noisy precursors. We report the observation of precursors excited by a deterministic perturbation. Quantitative characterization of this type of precursor is important in predicting a bifurcation in advance. This predicting ability comes from a dynamical structure present on the perturbed system.

Journal Article↗

An atypical case of Trypanosoma cruzi infection in a young English Mastiff.

A case of Trypanosoma cruzi infection in a young English Mastiff from Texas is presented. Clinical signs and laboratory findings included subcutaneous edema, lymphadenopathy, weight loss, and hypoalbuminemia. Cytology of a lymph node revealed numerous amastigotes. No trypomastigotes were observed in buffy coat preparation of peripheral blood, and on histologic evaluation, most organs contained numerous interstitial pseudocysts. Initial serology was positive for both T. cruzi and Leishmania, and immunohistochemistry supported a diagnosis of Leishmania. However, additional serology supported a T. cruzi infection, and cultivation of organisms isolated from a lymph node revealed morphology consistent with T. cruzi. In addition, PCR analysis resulted in a 504 bp fragment with 99% homology to a flagellar protein of T. cruzi. Although uncommon, autochthonous cases of both T. cruzi and Leishmania have been reported in the United States. Clinical signs observed with both diseases can show many similarities, cytology may be indistinguishable, as in this case, and serological cross-reactivity is common. This case demonstrates an unusual presentation of T. cruzi and the use of multiple testing strategies to support its diagnosis.

Animals↗

Phenotypic, functional, and quantitative characterization of canine peripheral blood monocyte-derived macrophages.

The yield as well as phenotypic and functional parameters of canine peripheral blood monocyte-derived macrophages were analyzed. The cells that remained adherent to Teflon after 10 days of culture had high phagocytic activity when inoculated with Leishmania chagasi. Flow cytometric analysis demonstrated that more than 80% of cultured cells were positive for the monocyte/macrophage marker CD14.

Animals↗

Genital lesions associated with visceral leishmaniasis and shedding of Leishmania sp. in the semen of naturally infected dogs.

Although visceral leishmaniasis is primarily transmitted by a biological invertebrate vector, transmission in the absence of the vector has been reported, including venereal transmission in humans. Considering the possibility of venereal transmission, we studied genital lesions in dogs naturally infected with visceral leishmaniasis and shedding of Leishmania sp. in the semen. Approximately 200 dogs were serologically tested for anti-Leishmania antibodies and divided into three groups: 1) serologically negative dogs (n = 20), 2) asymptomatic serologically positive dogs (n = 20), and 3) symptomatic serologically positive dogs (n = 20). Samples from both testes, all segments of both epididymes, prostate gland, glans penis, and prepuce were histologically evaluated and processed for immunodetection of Leishmania sp. Semen samples were obtained from 22 symptomatic serologically positive dogs and processed for detecting Leishmania DNA by polymerase chain reaction. A significantly higher frequency of inflammation was observed in the epididymes, glans penis, and prepuce of dogs with visceral leishmaniasis, which was associated with a high frequency of immunohistochemically positive tissues (up to 95% of tissues from symptomatic dogs were positive by immunohistochemistry). Leishmania DNA was detected in eight of 22 semen samples from symptomatic dogs. Together these findings indicate that genital lesions and shedding of Leishmania sp. (donovani complex) in the semen are associated with visceral leishmaniasis. Additional studies should address the possibility of venereal transmission of the disease in the dog.

Animals↗

Endothelial mediators of 17 beta-estradiol-induced coronary vasodilation in the isolated rat heart.

The present study was designed to determine relaxation in response to 17 beta-estradiol by isolated perfused hearts from intact normotensive male and female rats as well as the contribution of endothelium and its relaxing factors to this action. Baseline coronary perfusion pressure was determined and the vasoactive effects of 17 beta-estradiol (10 microM) were assessed by in bolus administration before and after endothelium denudation by infusion of 0.25 microM sodium deoxycholate or perfusion with 100 microM L-NAME, 2.8 microM indomethacin, 0.75 microM clotrimazole, 100 microM L-NAME plus 2.8 microM indomethacin, and 100 microM L-NAME plus 0.75 microM clotrimazole. Baseline coronary perfusion pressure differed significantly between males (84 +/- 2 mmHg, N = 61) and females (102 +/- 2 mmHg, N = 61). Bolus injection of 10 microM 17 beta-estradiol elicited a transient relaxing response in all groups, which was greater in coronary beds from females. For both sexes, the relaxing response to 17 beta-estradiol was at least in part endothelium-dependent. In the presence of the nitric oxide synthase inhibitor L-NAME, the relaxing response to 17 beta-estradiol was reduced only in females. Nevertheless, in the presence of indomethacin, a cyclooxygenase inhibitor, or clotrimazole, a cytochrome P450 inhibitor, the 17 beta-estradiol response was significantly reduced in both groups. In addition, combined treatment with L-NAME plus indomethacin or L-NAME plus clotrimazole also reduced the 17 beta-estradiol response in both groups. These results indicate the importance of prostacyclin and endothelium-derived hyperpolarizing factor in the relaxing response to 17 beta-estradiol. 17 beta-estradiol-induced relaxation may play an important role in the regulation of coronary tone and this may be one of the reasons why estrogen replacement therapy reduces the risk of coronary heart disease in postmenopausal women.

Animals↗

Pathogenesis of Salmonella-induced enteritis.

Infections with Salmonella serotypes are a major cause of food-borne diseases worldwide. Animal models other than the mouse have been employed for the study of nontyphoidal Salmonella infections because the murine model is not suitable for the study of Salmonella-induced diarrhea. The microbe has developed mechanisms to exploit the host cell machinery to its own purpose. Bacterial proteins delivered directly into the host cell cytosol cause cytoskeletal changes and interfere with host cell signaling pathways, which ultimately enhance disease manifestation. Recently, marked advances have been made in our understanding of the molecular interactions between Salmonella serotypes and their hosts. Here, we discuss the molecular basis of the pathogenesis of Salmonella-induced enteritis.

Animals↗

Morphologic and molecular characterization of Salmonella typhimurium infection in neonatal calves.

The host response to Salmonella plays a major role in the outcome of infection. The present study was undertaken to further characterize Salmonella typhimurium infection in neonatal calves at both the morphologic and the molecular level using the ligated ileal loop model. Eight 4-5-week-old male Holstein calves underwent laparotomy, and loops were prepared in the ileum. The loops were either inoculated with an S. typhimurium strain pathogenic for cattle or injected with sterile LB broth as control. Samples for histology, transmission and scanning electron microscopy, and RNA extraction were collected at various time points between 5 minutes and 12 hours postinfection. Invasion of both M cells and enterocytes began at 15 minutes postinfection. No specific cell type was the main target for invasion. Intracellular bacteria were observed in the lamina propria after 1 hour postinfection. A severe acute neutrophilic response was associated with invasion of the Peyer's patches. Upregulated expression of CXC chemokines (interleukin [IL]-8, growth-related oncogenes, [GRO] alpha and gamma, and granulocyte chemotactic protein [GCP]2) was detected by reverse transcription polymerase chain reaction beginning at 1 hour postinfection. Expression of proinflammatory (IL-1beta, IL-18, and tumor necrosis factor [TNF]alpha) and anti-inflammatory (IL-10, IL-IRa, and IL-4) cytokines was also assessed. A marked increase in expression of IL-1beta was observed, whereas the profile of expression of IL-18 and TNFalpha did not change after infection. Upregulation of IL-1Ra and IL-4 but not of IL-10 was observed. These findings indicate that infection of bovine ligated ileal loops with S. typhimurium results in an acute neutrophilic inflammatory response that is associated with the upregulation of CXC chemokines (IL-8, GROalpha and gamma, and GCP2), IL-1beta, IL-IRa, and IL-4.

Animals↗

Salmonella enterica serovar typhimurium induces cell death in bovine monocyte-derived macrophages by early sipB-dependent and delayed sipB-independent mechanisms.

It was previously demonstrated that Salmonella enterica serovar Typhimurium induces cell death with features of apoptosis in murine macrophages. Mice infected with Salmonella serovar Typhimurium develop systemic disease without diarrhea, whereas the infection in cattle and in humans is localized and characterized by diarrhea. Considering these clinical disease expression differences between mice and cattle, we investigated whether serovar Typhimurium is cytotoxic for bovine macrophages. Macrophages infected with serovar Typhimurium grown in the logarithmic phase quickly underwent cell death. Macrophages infected with stationary-phase cultures or with a mutant lacking sipB underwent no immediate cell death but did develop delayed cytotoxicity, undergoing cell death between 12 and 18 h postinfection. Both pathways were temporarily blocked by the general caspase inhibitor Z-VAD-Fmk and by the caspase 1 inhibitor Z-YVAD-Fmk. Comparisons of macrophages from cattle naturally resistant or susceptible to intracellular pathogens indicated no differences between these two genetic backgrounds in terms of susceptibility to serovar Typhimurium-induced cell death. We conclude that Salmonella serovar Typhimurium induces cell death in bovine macrophages by two distinct mechanisms, early sipB-mediated and delayed sipB-independent mechanisms.

Amino Acid Chloromethyl Ketones↗

Salmonella-induced cell death is not required for enteritis in calves.

Salmonella enterica serovar Typhimurium causes cell death in bovine monocyte-derived and murine macrophages in vitro by a sipB-dependent mechanism. During this process, SipB binds and activates caspase-1, which in turn activates the proinflammatory cytokine interleukin-1beta through cleavage. We used bovine ileal ligated loops to address the role of serovar Typhimurium-induced cell death in induction of fluid accumulation and inflammation in this diarrhea model. Twelve perinatal calves had 6- to 9-cm loops prepared in the terminal ileum. They were divided into three groups: one group received an intralumen injection of Luria-Bertani broth as a control in 12 loops. The other two groups (four calves each) were inoculated with 0.75 x 10(9) CFU of either wild-type serovar Typhimurium (strain IR715) or a sopB mutant per loop in 12 loops. Hematoxylin and eosin-stained sections were scored for inflammation, and terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end labeling (TUNEL)-positive cells were detected in situ. Fluid accumulation began at 3 h postinfection (PI). Inflammation was detected in all infected loops at 1 h PI. The area of TUNEL-labeled cells in the wild-type infected loops was significantly higher than that of the controls at 12 h PI, when a severe inflammatory response and tissue damage had already developed. The sopB mutant induced the same amount of TUNEL-positive cells as the wild type, but it was attenuated for induction of fluid secretion and inflammation. Our results indicate that serovar Typhimurium-induced cell death is not required to trigger an early inflammatory response and fluid accumulation in the ileum.

Animals↗

[Aedes albopictus in bromeliads of anthropic environment in São Paulo State, Brazil].

OBJECTIVE: Aedes albopictus populations can breed in several kinds of containers, and its presence has also been reported in Bromeliaceae. The purpose of this study was to evaluate the epidemiological importance of the Bromeliaceae plants as potential breeding sites of Aedes albopictus and to document the associated Culicidae entomofauna found in this micro-habitat. METHODS: Collections of Culicidae larvae were carried out fortnightly in aquatic content of bromeliads during 1998 and 1999. Collections took place in urban and periurban areas of Ilhabela island and periurban area of Ilha Comprida island, Brazil. RESULTS: A total of 26,647 Culicidae larvae were collected at both study sites, 14,575 in the urban area and 10,987 in the periurban area of Ilhabela and the remaining 1,085 in the periurban area of Ilha Comprida. There was no statistical difference between the amount of larvae collected in urban and periurban areas of Ilhabela. Regarding the Ae. albopictus, there was found a higher frequency and amount of larvae in the urban area of Ilhabela, followed by the periurban area in the same site, whereas in the periurban area of Ilha Comprida, its presence was considered accidental, since it was reported only in the first three months. CONCLUSIONS: The results show that bromeliads, largely used in landscaping, may contribute to the spread of Ae. albopictus. The presence of this species in domesticated and domiciled bromeliads allows us to suggest that the Culicidae larva is part of the regional fauna and facilitate the contact between humans and etiological agents of the natural ecosystem.

Aedes↗

Prognostic significance of the distribution of neck node metastasis from oral carcinoma.

BACKGROUND: Carcinoma of the oral cavity presents a high risk for neck metastasis, which reduces the probability of regional control and survival. OBJECTIVES: The main objective of this study is to analyze prognostic implications of the distribution of neck metastasis in 513 patients with squamous cell carcinoma of the oral cavity. PATIENTS AND METHODS: All patients underwent surgery from 1970-1992. Tumor stages were I, 63; II, 120; III, 173; and IV, 157. Neck dissections were performed in 448 patients (115 bilateral). RESULTS: By use of multivariate regression techniques the level of lymph node involvement was the most important prognostic factor (relative risks from 1.8 to 2.5). The following variables were also associated with prognosis: mobility of lymph nodes, sex, T stage, age, and tumor thickness. CONCLUSIONS: The level of ipsilateral lymph node involvement was the most significant prognostic factor patients with in oral cancer who underwent surgical treatment. A significant decrease in survival also was seen with regard to the involvement of multiple contralateral lymph nodes. Our results support the indication of elective neck dissections in high-risk patients because among the cases that had metastases at follow-up, 50% were not candidates for salvage treatment.

Adult↗

Analysis of the gene expression profile of Schistosoma mansoni cercariae using the expressed sequence tag approach.

ESTs constitute rapid and informative tools with which to study gene-expression profiles of the diverse stages of the schistosome life cycle. Following a comprehensive EST study of adult worms, analysis has now targeted the cercaria, the parasite larval form responsible for infection of the vertebrate host. Two Schistosoma mansoni cercarial cDNA libraries were examined and partial sequence obtained from 957 randomly selected clones. On the basis of database searches, 551 (57.6%) ESTs generated had no homologs in the public databases whilst 308 (32.2%) were putatively identified, totaling 859 informative ESTs. The remaining 98 (10.2%) were uninformative ESTs (ribosomal RNA and non-coding mitochondrial sequences). By clustering analysis we have identified 453 different genes. The most common sequences in both libraries represented Sm8 calcium binding protein (8% of ESTs), fructose-1,6-bisphosphate aldolase, glyceraldehyde-3-phosphate dehydrogenase, cytochrome oxidase subunit 1, ATP guanidine kinase and triose phosphate isomerase. One hundred and nineteen identified genes were sorted into 11 functional categories, with genes associated with energy metabolism being the most abundant (13%) and diverse. The diversity and abundance of genes associated with the transcription/translation machinery and with regulatory/signaling functions were also marked. A paramyosin transcript was identified, indicating that this gene is not exclusively expressed in adult worms and sporocysts (as had been suggested previously). The possible physiological relevance to cercariae of the presence of transcripts with homology to calcium binding proteins of the EF-hand superfamily, Gq-coupled rhodopsin photoreceptor, rod phosphodiesterase 8 subunit and peripheral-type benzodiazepine receptor is discussed.

Animals↗

Effect of growth hormone and induced IGF-I release on germ cell population and apoptosis in the bovine testis.

Bovine growth hormone has been used in dairy cattle to increase milk production,but it also increases the twin parturition rate. This effect is mediated by insulin-like growth factor-I (IGF-I), which prevents follicular atresia by hindering apoptosis of granulosa cells. The action of GH and IGF-I on testicular function remains unclear. The goal of this study, therefore, was to verify the effects of short-term administration of GH and induced IGF-I release on the number of testicular germ cells, testicular morphology, and apoptosis in the bovine testis. Twenty Zebu bulls were split into 2 groups. The bulls in Group 1 (n = 10) were treated with 2 subcutaneous injections of bovine GH (500 mg/bull) 7 d apart. Group 2 bulls (n = 10) received placebos under the same protocol. All of the bulls were slaughtered 14 d after the start of treatment. Fragments of the testis were collected, fixed in Bouin's solution, embedded in paraffin, and the sections stained with hematoxilin and eosin. The paraffin-embedded sections were also used for in situ detection of apoptotic cells. Blood samples were collected at slaughter to measure serum levels of IGF-I, FSH and LH. Neither the number of Stage I seminiferous epithelium germ cells and the morphometric parameters (tubular diameter, seminiferous epithelium height, and volumetric proportions of structural components) nor the blood levels of FSH and LH showed a significant difference between the 2 groups. However, the treated animals showed an increase in serum IGF-I (P<0.01). Apoptotic germ cells were detected in the testis of both groups, showing the same pattern and a stage-specific apoptosis pattern. Most of the labeled cells were spermatocytes. The localization of apoptotic germ cells did not differ between groups. These results suggest that short-term administration of GH does not affect bovine spermatogenesis in adult bulls.

Animals↗

Factors influencing contralateral lymph node metastasis from oral carcinoma.

BACKGROUND: An ipsilateral neck dissection is mandatory during initial treatment stages II-IV oral carcinomas. However, no consensus exists whether or not to perform an elective contralateral neck dissection. METHODS: Five hundred thirteen consecutive cases of squamous cell carcinoma (269 tongue, 135 floor of the mouth, 44 inferior gingiva, 65 retromolar trigone) were reviewed. Tumor stages were: 69 T1, 227 T2, 217 T3-T4, 263 N0, 250 N1-N3. A total of 563 neck dissections were performed in 448 patients. Univariate and multivariate analysis of risk factors were performed using logistic regression. RESULTS: Two hundred twenty-three patients (49.8%) had positive nodes in the specimen (182 ipsilateral, 36 bilateral, 5 contralateral). Contralateral neck recurrences occurred in 38 cases (33 not submitted to a contralateral neck dissection initially). Multivariate logistic regression analysis demonstrated that clinical stage (p = .0001), tumor crossing midline (p = .0011), and floor of the mouth involvement (p = .0236) were the most important predictors of contralateral metastasis. CONCLUSION: The contralateral side of the neck is a common and potentially preventable site of recurrence in tumors of the oral cavity. The multivariate model obtained discriminates patients with low and high risk (more than 20%) of contralateral metastasis. The application of this mathematical model can be useful for the indication of contralateral neck dissections, because not all tumors crossing midline are associated to a high risk (stages I and II tumors not involving the floor of the mouth) and not all tumors not crossing midline are at low risk (stages III and IV tumors involving the floor of the mouth).

Adult↗

Histomorphometry and quantification of nucleolar organizer regions in bovine thyroid containing methylthiouracil residues.

In order to study the morphology and morphometry and to characterize and quantify the nucleolar organizer regions (NORs) of bovine thyroids containing methylthiouracil (MTU) residues, five animals were orally treated with a suspension of MTU (5 g/animal/day) for 20 days (group A). This treatment protocol was interrupted 5 days before the the animals were slaughtered. Six animals receiving placebos composed group B. A third group (group C) was composed of normal thyroids obtained from a slaughterhouse. All glands were previously assessed for detection of antithyroid residues by chromatography, and only those glands from MTU-treated animals were positive. Follicles of glands from group A showed wide variation in size and shape. There was a predominance of small follicles covered by multiple layers of columnar cells, sometimes forming papillary projections into the lumen, characterizing severe interfollicular and intrafollicular adenomatosis. Many follicles had vacuolated cells with nuclei showing karyolysis or pyknosis and reduced amounts of a low-density and very excavated colloid. They also showed higher follicular epithelia and larger proportions of their structural components when compared with glands of groups B and C. In the thyroids from group A, the argyrophilic nucleolar organizer region-associated proteins (AgNORs) were greater in number, with small ones scattered all over the nucleus. Although the size of AgNORs in thyroids from groups B and C was variable, these AgNORs were fewer and larger than were those in glands from group A. In conclusion, the MTU induces proliferation and regressive changes in follicular cells, and the AgNOR technique is efficient to distinguish different degrees of thyroid hyperplasia.

Animals↗

[Marking-release-recapture methods for determining the size of the natural population of Anopheles albitarsis l.s (Diptera:Culicidae)].

OBJECTIVE: In order to evaluate the size of the Anopheles albitarsis s.l. population in the Ribeira Valley region of the south-eastern S. Paulo State, Brazil, marking-release-recapture experiments were carried out on the Experimental Farm in Pariquera-Açu county where an artificial irrigation system for the cultivation of rice has been installed. The experiments aimed at determining the vectorial capacity of this species. METHODS: During the summer (January, February and March), three marking-release-recapture experiments with field and laboratory reared populations were conducted to evaluate the population size. Simultaneous human bait collections were also carried out to estimate the human biting rate. RESULTS: The population sizes of the three experiments were estimated to be 64,560, 50,503 and 22,684 mosquitoes. The bite/man/night rate varied from 41.5 to 524,5. DISCUSSION: The results indicated high mosquito density in the summer which allow one to infer that, even though the survival rate is low, the number of remaining females is large enough to outlive the parasite's extrinsic developmental period.

Animals↗

Erythrophagocytosis in the caprine trophoblast.

The mechanism of iron transfer by extravasation of maternal blood in the maternal-fetal interface and the subsequent phagocytosis of the erythrocytes by the trophoblast cells was described in ovine, bovine and other species. This research was performed due to the absence of studies on this process in the goat. Fragments of placentomes were obtained from 9 adult goats of an unspecified breed and were separated into Groups A, B and C, based on 90, 120 and 150 d of gestation, respectively. Fragments of 3 placentomes were obtained from each animal of all groups. The first of them was removed from the base, the second from the middle part and the third from the end of the gestational uterine horn. The fragments were fixed in Bouin solution and histologically processed according to the usual paraffin inclusion techniques. The slides were stained by hematoxilin-eosin and potassium ferrocyanide and examined under light microscopy The erythrophagocytosis (EP) process of the trophoblast was identified in all the examined histological sections, throughout the gestation and for each placentome. An accumulation of spots of blood was observed only in the maternal-fetal interface located in the arcade zone of the placentome. The main cells of the trophoblast of these areas presented an intense phagocytic activity and the observation of erythrocytes with hemossiderin pigments in their citoplasm was possible. The trophoblast epithelium of these areas was thicker and with a columnar aspect than the cubic aspect observed in other areas. The trophoblast binucleate cells do not seem to participate in EP. Our findings indicate that the phagocitosis process of the maternal erythrocites by the trophoblast also occurs in caprine species, indicating that this could be a mechanism of transplacentary transfer of iron in this species.

Journal Article↗

Lower transversal laparotomy has no appreciable effect on respiratory mechanics in guinea pigs.

Respiratory system, lung, and chest wall resistances and dynamic elastances were determined in six anesthetized, paralyzed, and mechanically ventilated guinea pigs both before and after lower transversal abdominal opening performed at the level of the spina iliaca anterosuperior. Furthermore, the resistances were also split into their two components, one reflecting the Newtonian resistances and the other representing the viscoelastic/inhomogeneous pressure dissipations in the system. The method of end-inflation occlusion during constant inspiratory flow was used. Chest wall configuration was also evaluated by measurements of lateral and anteroposterior diameters and circumferences at the 4th intercostal space and xiphoid levels both at functional residual capacity and at the end of tidal inspiration before and after surgery. After abdominal incision no statistically significant changes could be detected in any of the measured variables. It may be concluded that lower transversal abdominal opening does not alter respiratory mechanics.

Abdomen↗