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Biomedical subjects

L H Carvalho

Publications and source records attributed to L H Carvalho.

12 recordsLinked to original sources

Scaling and root planing, systemic metronidazole and professional plaque removal in the treatment of chronic periodontitis in a Brazilian population II--microbiological results.

OBJECTIVE: The current investigation evaluated changes in levels and proportions of 39 bacterial species in subgingival plaque samples after scaling and root planing (SRP) alone or in combination with systemic metronidazole and/or professional cleaning in subjects with chronic periodontitis. METHODS: Forty-four adult subjects (mean age 45+/-6 years) with periodontitis were randomly assigned in four treatment groups, a control (C, n=10) that received SRP and placebo and three test groups treated as follows: T1 (n=12): SRP and metronidazole (M, 400 mg tid) for 10 days; T2 (n=12): SRP, weekly professional supragingival plaque removal for 3 months (PC) and placebo; and T3 (n=10): SRP, M and PC. Subgingival plaque samples were taken from seven sites per subject at baseline and 90 days post-therapy. Counts of 39 subgingival species were determined using checkerboard DNA-DNA hybridization. Significance of differences over time was determined using the Wilcoxon signed ranks test and among groups using ancova. RESULTS: The mean counts of the majority of the species were reduced post-therapy in the 4 treatment groups. Counts (x 10(5)+/-SEM) of Porphyromonas gingivalis, Tannerella forsythensis and Treponema denticola were significantly reduced in groups T2 and T3. Levels of beneficial species, such as some Actinomyces species, Veillonella parvula, Streptococcus sanguis, Streptococcus oralis and Streptococcus gordonii were minimally affected in levels when the combined therapy was applied (T3). Mean proportions of red complex species decreased from 18.4% at baseline to 3% at 90 days post-therapy in group T3 (p<0.01), from 25.8% to 2.3% in group T2 (p<0.01), from 17.7% to 5.6% in group T1 (p<0.05) and from 19.4% to 8.8% in group C (NS). Proportions of the suspected periodontal pathogens from the orange complex were also markedly reduced in groups T2 and T3. CONCLUSIONS: All treatments reduced counts and proportions of red complex species. Adjunctive therapy appeared to have a greater effect and also affected members of the orange complex.

Administration, Oral↗

Scaling and root planing, systemic metronidazole and professional plaque removal in the treatment of chronic periodontitis in a Brazilian population. I. clinical results.

OBJECTIVE: The current investigation evaluated the clinical effects of scaling and root planing (SRP) alone or in combination with systemic metronidazole and/or repeated professional removal of supragingival plaque in subjects with chronic periodontitis. METHODS: Fourty-four adult subjects (mean age: 45+/-6 years) with periodontitis were randomly assigned to four treatment groups; a control (C, n=10) that received SRP and placebo and three test groups treated as follows: Test 1 (T1) (n=12) received SRP and metronidazole (400 mg t.i.d., M) for 10 days; Test 2 (T2) (n=12) received SRP, weekly professional supragingival plaque removal for three months (professional cleaning (PC)) and placebo; and Test 3 (T3) (n=10) received SRP, M and PC. Pocket depth (PD), attachment level (AL), bleeding on probing (BOP) and presence of visible plaque and suppuration were measured at six sites per tooth at baseline and at 90 days post-therapy. Significance of differences over time was determined using the Wilcoxon test, and among groups using ancova. RESULTS: A reduction in full-mouth mean clinical parameters was observed at 90 days after all therapies. Sites with baseline PD<4 mm showed an increase in mean PD in the control group and in mean AL in all treatment groups. Sites with baseline PD of 4-6 mm in subjects who received PC as part of therapy (T2, T3) showed a marked reduction in PD, AL and in the % of sites with BOP. Subjects who received metronidazole (T1 and T3) showed the best clinical response at sites with an initial PD of >6 mm. The major clinical benefit occurred when the combination of SRP, M and PC was used. Group T3 showed the least attachment loss in initially shallow pockets. This group also exhibited the greatest reduction in the % of sites with BOP and suppuration as well as in mean PD and AL at sites with baseline PD>4 mm. CONCLUSION: The data suggest a significant clinical benefit in combining SRP, systemic metronidazole and weekly professional supragingival plaque removal for the treatment of chronic periodontitis.

Adult↗

ELISPOT assay to measure antigen-specific murine CD8(+) T cell responses.

The enzyme-linked immunospot technique (ELISPOT) relies on the visualization of cytokine secretion by individual T cells following in vitro stimulation with antigen. This assay has been developed and standardized for the quantitative detection of antigen-specific CD8(+) T cells in mice subjected to different immunization protocols [J. Immunol. Methods 181 (1995) 45]. We have identified important variables that affect the efficacy of the ELISPOT assay and in this protocol we describe this methodology in detail. As a model, we used the production of interferon-gamma by CD8(+) T cells from peripheral blood, spleen and liver of mice immunized with malaria sporozoites expressing the H-2K(d)-restricted SYVPSAEQI. This protocol has also been used successfully to detect Th1 and Th2 epitope specific CD4(+) T cells.

Animals↗

Identification of human chromosome 22 transcribed sequences with ORF expressed sequence tags.

Transcribed sequences in the human genome can be identified with confidence only by alignment with sequences derived from cDNAs synthesized from naturally occurring mRNAs. We constructed a set of 250,000 cDNAs that represent partial expressed gene sequences and that are biased toward the central coding regions of the resulting transcripts. They are termed ORF expressed sequence tags (ORESTES). The 250,000 ORESTES were assembled into 81,429 contigs. Of these, 1, 181 (1.45%) were found to match sequences in chromosome 22 with at least one ORESTES contig for 162 (65.6%) of the 247 known genes, for 67 (44.6%) of the 150 related genes, and for 45 of the 148 (30.4%) EST-predicted genes on this chromosome. Using a set of stringent criteria to validate our sequences, we identified a further 219 previously unannotated transcribed sequences on chromosome 22. Of these, 171 were in fact also defined by EST or full length cDNA sequences available in GenBank but not utilized in the initial annotation of the first human chromosome sequence. Thus despite representing less than 15% of all expressed human sequences in the public databases at the time of the present analysis, ORESTES sequences defined 48 transcribed sequences on chromosome 22 not defined by other sequences. All of the transcribed sequences defined by ORESTES coincided with DNA regions predicted as encoding exons by genscan. (http://genes.mit.edu/GENSCAN.html).

Chromosomes, Human, Pair 22↗

Cellular responses to Plasmodium falciparum major surface antigens and their relationship to human activities associated with malaria transmission.

In Brazil, two types of activities have led to the worsening of malarial transmission in the Amazon region: prospecting/mining and agricultural settlements. In the present study, we analyze the cellular response of 52 of these individuals (14 gold-miners and 38 farmers) living within the same endemic area. Two Plasmodium falciparum major surface antigens (recombinant proteins) were used for cellular proliferative assays: circumsporozoite protein and merozoite surface protein-1. The frequency of these cellular responses were significantly higher among the miners (57-64%) than the farmers (10-20%) when either recombinant protein was used. Our data suggest that a higher exposure to malaria of the gold-miners contributed to their higher in vitro cellular response compared with the farmers. These findings point the way to further studies evaluating the influence of risk factors associated with the life styles of different social groups and the immune responses to these antigens.

Adolescent↗

[Infectious mononucleosis]

OBJECTIVE: To present updated aspects of the Infectious Mononucleosis caused by the Epstein-Barr Virus. MATERIALS AND METHODS: Research of bibliographic references through the Medline and direct research of selected papers. RESULTS: A concise approach to some aspects related to the epidemiology of the virus, namely the two types that are presently known,EBVtypeAandEBVtype B, and some differences that they present. It was possible to establish a relationship between what one sees in the clinical picture and the immunological changes that occur at the same time. The author also describes the physiopathology, clinical features, complications and other syndromes associated to the EBV. As far as the laboratory workup is concerned, it is important to have a complete blood count as the first step, followed by the quantitative exam of the heterophile antibodies and antibodies antiEBV analysis. CONCLUSION: The pathologies related to the EBV are important, and certainly fascinating from the immunological point of view. Among these, the Infectious Mononucleosis caused by the EBV has shown some interesting clinical and laboratorial aspects.

Journal Article↗

Cross-reactive cellular immune response to circumsporozoite proteins of Plasmodium vivax and P. falciparum in malaria-exposed individuals.

Acquired immunity against the recombinant circumsporozoite protein of P. falciparum (rPfCS) or P. vivax (rPvCS) was studied in two malarious areas of the Brazilian Amazon. Cellular responsiveness, evaluated by proliferative assays, was detected in about 45% of individuals who had recovered from recent acute malaria infections. Peripheral blood mononuclear cells of individuals whose last malaria infection was by P. vivax responded more to the rCS proteins than those who had P. falciparum. Since in P. vivax infections hypnozoites in the liver retain CS antigen, this stage may have contributed to the increased cellular response. The unexpected result was that in primoinfections by P. falciparum or P. vivax the proliferative response did not correspond to the rPfCS and rPvCS, respectively. Furthermore, among the malaria-exposed individuals, there was a positive correlation between the intensity of the responses to the two rCS proteins. Our results suggest that cross-reactive epitopes exist in the CS protein of P. falciparum and P. vivax. In the areas studied, the frequency of antibodies against rPvCS and/or rPfCS ranged from 43% to 11%. Species-specific antibodies against the CS protein were detected in the primoinfected individuals. Some individuals living in the endemic area but with no clinical history of malaria were positive by serology (8%) or by in vitro proliferation (21%). However, antibodies and cellular responses against rCS were detected only in malaria-exposed individuals, since those living outside the endemic area were all negative.

Adult↗

Molecular genetic approach to cell proliferation control and neoplasia.

A number of gene products involved in the control of cell proliferation fall into one of two classes: oncogenes and tumor suppressor genes. The same gene products have also been associated with malignant growth (tumors) caused by radiation, chemicals and tumor viruses. Here we describe our attempts to elucidate the molecular mechanisms underlying polyomavirus-induced cell transformation and the anti-tumor activity of glucocorticoid hormones. Wild type and mutant polyomavirus middle T (MT) overexpressing cell lines, generated with retroviral vector constructs, were used to investigate the role played by peptide growth factor primary response genes (fos, jun, myc, JE, KC) in viral transformation and to map the transduction pathway of the mitogenic signal of MT. Overexpression of MT leads to increased AP-1 (Fos/Jun) transcriptional complex activity. Transformation defective mutant analysis allowed the identification of sites in the MT molecule that are crucial for this activity. Two different approaches were used to investigate the molecular basis for glucocorticoids anti-tumor activity, namely: blind cloning of cDNAs and analysis of growth control genes in C6 glioma cell variants that are either hypersensitive (C6/ST1) or unresponsive to glucocorticoids (C6/P7). Four different glucocorticoid-regulated cDNA sequences were isolated using differential hybridization. A number of differentially expressed sequences were isolated from glucocorticoid-treated C6/ST1 cells by differential display (DDRT-PCR) and are currently being characterized. Expression of known growth control genes in C6/ST1 cells allowed the identification of important candidates for glucocorticoid hormone targets.

Animals↗

A method for screening drugs against the liver stages of malaria using Plasmodium gallinaceum and Aedes mosquitos.

1. The radical cure of human malaria caused by Plasmodium vivax requires two drugs, i.e., a blood schizontocide such as chloroquine to clear the circulating parasites, and primaquine aimed at the liver stages (hyponozoites) responsible for the late relapses of this parasite. Primaquine is unique as a radical curative drug but is highly toxic. The only useful model currently available for screening drugs to replace primaquine is Plasmodium cynomolgi-induced malaria in Rhesus monkeys. Because of the limited availability and cost of these animals, the development of non-primate models for such screening would be of considerable value. 2. We used a drug-screening assay for the liver stage malaria parasite based on the ability of such drugs to stop development of gametocytes in the mosquito vector. The inhibition of the sporogonic cycle of malaria in the mosquito by primaquine (15 mg/kg) was confirmed here and used for re-evaluation of the gametocyte method. 3. We observed that the level of parasitemia in the untreated control chicken used to infect mosquitos was a crucial factor affecting the subsequent development of sporogony. Thus, parasitemia was carefully controlled in the studies involving oocyst development. Parasitemias lower than 6% at the beginning of the experiment and increasing were found to be most appropriate for the production of the infectious gametocytes during a period of 8 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Aedes↗

Antimalarial chemotherapy with natural products and chemically defined molecules.

In the present work we have described the in vivo antimalarial activity of six different plants. Two of them (Vernonia brasiliana and Eupatorium squalidum) were tested in a randomic approach among 273 crude extracts from plants; four (Acanthospermum australe, Esenbeckia febrifuga, Lisianthus speciosus and Tachia guianensis) were selected after screening 22 crude extracts from different medicinal plants used in Brazil against fever and/or malaria. We also studied chemically defined molecules and some of them showed antimalarial activity in vitro. Some aspects of recent research with natural products aiming to produce drugs are discussed.

Animals↗

Antimalarial activity of crude extracts from Brazilian plants studied in vivo in Plasmodium berghei-infected mice and in vitro against Plasmodium falciparum in culture.

1. Ninety-five crude extracts obtained with either organic solvents or water from 48 Brazilian plants or parts of plants were evaluated experimentally as blood schizontocides. Seventy-three extracts were obtained from 33 plants randomly collected using an empirical approach, and 22 from 15 "medicinal" plants. 2. The crude extracts were screened in vivo at up to 1.0 g/kg, po, for 4 days in mice infected with blood forms of Plasmodium berghei and parasitemia was determined on the fifth day. 3. Six plants, 2 randomly collected, Vernonia brasiliana and Eupatorium squalidum, and 4 "medicinal" plants, Acanthospermum australe, Esenbeckia febrifuga, Lisianthus speciosus, and Tachia guianensis, were partly active against the rodent malaria, i.e., they showed 40-50% inhibition of P. berghei multiplication. Forty-two plants whose extracts presented no antimalarial activity are reported. 4. Four extracts with antimalarial activity were also tested in vitro using P. falciparum cultures and two of them, V. brasiliana and A. australe, were active. Extracts of V. brasiliana caused about 50% inhibition of parasite multiplication at relatively low doses (40 ng/ml) as compared to chloroquine (30 ng/ml) and quinine (50 ng/ml). 5. The relatively high percentage of positive results obtained here for "medicinal" plants vs randomly chosen plants demonstrates the effectiveness of the ethnopharmacological approach to drug testing.

Animals↗

In vitro activity of natural and synthetic naphthoquinones against erythrocytic stages of Plasmodium falciparum.

In an attempt to identify new antimalarial compounds we studied the blood schizonticide effect of chemically defined natural products which were isolated from plants (Bignoniaceae) or synthesized. Different concentrations of these drugs (up to 20 microM) were incubated in vitro with blood forms of Plasmodium falciparum for 72 h. A total of 12 drugs of the naphthoquinones group were tested. Eight of them were isolated from plants (NP) and 4 synthesized (S). Three of the drugs (2 NP and one S) were very active and completely inhibited parasite growth at the higher concentrations used (20 microM); 5 drugs were partially active (3S and 2NP) and 3 (NP) were totally inactive. Lapachol was among the drugs tested. Although it has been considered a potential antimalarial agent, it exhibited very low activity (20% inhibition of schizogony). The antimalarial activity of our naphthoquinones against drug-resistant strains was superior to that of chloroquine and quinine which were used as controls. Two of the P. falciparum strains used for the tests were strongly chloroquine resistant. If these naphthoquinones prove to be active in vivo and are of low toxicity, they will be promising candidates for treatment of human malaria particularly since they are easily synthesized.

Animals↗