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M Sandoval

Publications and source records attributed to M Sandoval.

At least 37 records · Page 2Linked to original sources

Genistein and gut inflammation: role of nitric oxide.

Genistein, a principal soy isoflavone, has been identified as a protein kinase inhibitor that possesses immunosuppressive and anti-inflammatory properties. The aim of the study was to determine if genistein modified chronic ileitis in guinea pigs induced by the hapten trinitrobenzene sulfonic acid (TNBS), and the activity index of cultured macrophages (RAW 264.7 cells) stimulated with lipopolysaccharide (LPS). Genistein at low doses (0.1 mg/kg, s.c.) had mild anti-inflammatory effects in TNBS ileitis. Therapeutic benefit included a reduction in nitric oxide production, granulocyte infiltration and improved mucosal architecture. Genistein, at low doses, also appeared to attenuate immunohistochemical staining for inducible nitric oxide synthase (iNOS) and nitrotyrosine. The beneficial effects of genistein were not apparent at doses above 0.1 mg/kg. We found that genistein also inhibited LPS-induced nitrite production by cultured macrophages and protected against LPS-induced necrosis despite its ability to cause apoptosis. These results indicate that genistein displayed mild anti-inflammatory properties which may, in part, involve an attenuation of nitric oxide release via inducible nitric oxide synthase, and the formation of peroxynitrite.

Animals↗

Peroxynitrite-induced apoptosis in epithelial (T84) and macrophage (RAW 264.7) cell lines: effect of legume-derived polyphenols (phytolens).

Peroxynitrite (ONOO-) has been proposed as a mediator of gut inflammation and as an inducer of cell death by apoptosis. Phytolens (PHY), a water-soluble extract of polyphenolic antioxidants from nonsoy legumes (Biotics Research Corp, patent pending), was evaluated as a cytoprotective agent in human colonic (T84) and murine macrophage (RAW 264.7) cell lines. In the antioxidant testing, PHY showed a significant free radical scavenging ability against 1,1-diphenyl-2-picrylhydrazyl radical (DPPH) and superoxide (O2.) radicals with an IC50 of 4.44 and 5.87 microg/ml against DPPH and O2., respectively. Apoptosis (DNA fragmentation) was measured by an ELISA technique. Cells were exposed to oxidative stress by treating them with peroxynitrite (100-300 microM) for 4 h in the presence and absence of PHY. Peroxynitrite elicited a dose-dependent increase in DNA fragmentation in both cell lines compared to the control group receiving decomposed ONOO-. PHY (10, 30, or 50 microg/ml) significantly attenuated the degree of apoptosis in T84 cells induced by ONOO- (P < 0.05). PHY (10-100 microg/ml) did not directly affect T84 cell viability or induce apoptosis after 4 h or overnight exposure. RAW 264.7 cells exposed to PHY alone (>30 microg/ml) for 4 h displayed decreased cell viability (P < 0.05) and increased apoptosis (P < 0.05). Phytolens may have beneficial effects on inflammation by attenuating peroxynitrite-induced apoptosis. The sparing of epithelial cells while compromising the viability of macrophages suggests that PHY may be beneficial in autoimmune disorders.

Animals↗

Peroxynitrite-induced apoptosis in T84 and RAW 264.7 cells: attenuation by L-ascorbic acid.

The free radicals nitric oxide and superoxide react to form peroxynitrite (ONOO-), a potent cytotoxic oxidant. This study was designed to evaluate whether addition of L-Ascorbic acid (AsC) into the culture medium decreases peroxynitrite-induced apoptosis in human intestinal epithelial (T84) and murine macrophage (RAW 264.7) cell lines. In Experiment 1, T84 and RAW 264.7 cells were divided in two protocols: (1) treated with 100-300 microM ONOO- and incubated for 4 h, and (2) treated with 10-100 microM ONOO- and incubated overnight (14 h). In Experiment 2, T84 and RAW 264.7 cells were treated with 300 microM ONOO- and 500 microM AsC and incubated for 4 h. In Experiment 3, T84 and RAW 264.7 cells were preincubated for 2 h with 500 microM AsC then exposed to 300 microM ONOO- for 4 h. Cell viability (necrosis) was assessed by trypan blue dye exclusion. Apoptosis was quantified with a cell death detection ELISA assay. In the 4 h protocol, ONOO- induced apoptosis in T84 and RAW 264.7 cells, at levels of 100-300 microM. Concentrations of ONOO- greater than 300 microM caused necrosis. In contrast, extension of the protocol to 14 h indicated that ONOO- induced apoptosis at lower concentrations (50;-75 microM), with concentrations > 75 microM resulting in necrosis. AsC administered to the media or with preincubation plus washout, decreased peroxynitrite-induced apoptosis in T84 and RAW 264.7 cells. These results indicate that ONOO- may contribute to the pathophysiology of gut inflammation by promoting cell death and ascorbic acid may protect against peroxynitrie-induced damage.

Animals↗

Peroxynitrite-induced apoptosis in human intestinal epithelial cells is attenuated by mesalamine.

BACKGROUND & AIMS: Peroxynitrite (PN), a potent oxidant, has been implicated in the pathogenesis of gut inflammation and epithelial cell apoptosis. The aim of this study was to investigate mesalamine, a standard therapy for inflammatory bowel disease, to see if it attenuates PN-induced cytotoxicity in human intestinal epithelial cells and if mesalamine directly interacts with PN or its precursor, nitric oxide. METHODS: T84 and HT29 cells were divided in several protocols: mesalamine was administered 2 hours before, simultaneously, or 30 minutes after PN. T84 cells, grown in filter chamber inserts, were used to determine if basolateral or apical administration of PN initiated apoptosis and epithelial barrier function. The effects of mesalamine on PN decomposition and NO half-life were determined. RESULTS: Mesalamine resulted in a dose-dependent decrease in PN-induced apoptosis, whether mesalamine was administered before (>10 micromol/L; IC50, 16 micromol/L), simultaneously (25-200 micromol/L; IC50, 24 micromol/L), or 30 minutes (200 micromol/L) after PN exposure. Mesalamine protected the epithelial barrier function of T84 cells against PN. Mesalamine rapidly degraded PN, whereas the half-life of NO was not affected. CONCLUSIONS: The beneficial effects of mesalamine in the treatment of inflammatory bowel disease may involve an attenuation of PN-induced cell injury and apoptosis through direct and indirect mechanisms without affecting NO levels.

Anti-Inflammatory Agents, Non-Steroidal↗

Relative bioavailability of supplemental inorganic zinc sources for chicks.

Three experiments were conducted to investigate the relative bioavailability of reagent-grade (RG) and feed grade (FG) Zn sources for 1-d-old broiler chicks. In Exp. 1, 13 treatments included a basal corn-soybean meal diet (63 ppm Zn) or the basal diet supplemented with 400, 800, or 1,200 ppm Zn from RG sulfate, basic carbonate, oxide, or metal and fed for 20 d. Using multiple regression slope ratios with Zn sulfate set at 100%, bioavailability estimates were 78, 77, and 46% for carbonate, oxide, and metal, respectively. In Exp. 2, chicks were allotted randomly to 16 treatments that included a basal corn-soybean meal diet (75 ppm Zn) or basal diet supplemented with 300, 600, or 900 ppm Zn as either RG sulfate, FG sulfate-A, FG sulfate-B, FG oxide-A, or FG oxide-B and fed for 21 d. Multiple linear regression slope ratios gave relative estimates of 99, 81, 78, and 54% for sulfate-A, sulfate-B, oxide-A, and oxide-B sources, respectively, with RG sulfate set at 100%. In Exp. 3, chicks were fed a basal corn-soybean meal diet (35 ppm Zn) or the basal diet supplemented with 40, 80, or 120 ppm Zn from RG Zn sulfate, FG sulfate, or FG oxide and fed for 20 d. Multiple regression slope ratios with RG sulfate set at 100% gave relative bioavailability estimates of 94 and 74% for the FG sulfate and oxide, respectively. Bioavailability estimates were similar when Zn was supplemented to diets at high or low concentrations.

Animals↗

Inducible nitric oxide synthase and the regulation of central vessel caliber in the fetal rat.

BACKGROUND: The purpose of this study was to evaluate the possibility that inducible nitric oxide synthase (iNOS) regulates the fetal circulation. METHODS AND RESULTS: Positive evidence for iNOS gene expression was noted in heart central vessels and placenta of untreated rat fetuses. Rats in the last week of pregnancy were treated for 5 days with L-NG-(1-Iminoethyl)lysine (L-NIL), a selective inhibitor of iNOS, at 1, 10, and 100 micrograms/mL in the drinking water. To raise NO levels, lipopolysaccharide (LPS) 30 micrograms/kg was given by intraperitoneal injection, and sodium nitroprusside (SNP) was placed in mini-osmotic pumps to deliver 10 micrograms/kg per minute. Control animals were undisturbed. On day 21 of gestation, dams were anesthetized and fetuses were delivered by cesarean section and rapidly frozen in isopentane chilled in liquid nitrogen. Frozen sections (10 microns) were used to reconstruct a computer-generated three-dimensional image of the great vessels and ductus arteriosus. Significant constriction of the great vessels and ductus arteriosus was observed with L-NIL, whereas both LPS and SNP dilated these vessels. The vasorelaxant effect of LPS was blocked by L-NIL. NO release from placental explants was 633 +/- 41 nmol/L under basal conditions, increasing to 4.0 +/- 0.4 mumol/L with LPS administration, although placental iNOS message and protein levels were unchanged. CONCLUSIONS: We suggest that nitric oxide, generated by iNOS, plays a significant role in control of major vessel and ductus arteriosus caliber in the rat fetus. In regard to the nitrergic regulation of the circulation, the fetus is clearly different from the adult.

Animals↗

Role of inducible nitric oxide synthase expression and peroxynitrite formation in guinea pig ileitis.

BACKGROUND & AIMS: Inflammatory bowel disease is characterized by increased synthesis of nitric oxide. The aim of this study was to determine if inducible NO synthase (iNOS) was responsible for tissue injury, potentially via peroxynitrite formation, in the guinea pig model of gut inflammation. METHODS: Inflammation was induced in guinea pig ileum by intraluminal administration of the hapten trinitrobenzene sulfonic acid in 50% ethanol. iNOS gene expression was assessed by reverse-transcriptase polymerase chain reaction and Western blotting, immunohistochemistry was determined by its localization, and activity was inhibited with the specific inhibitor aminoguanidine administered via the drinking water for 7 days. Nitration of tyrosines was assessed by immunohistochemistry. RESULTS: In control animals, iNOS gene expression was minimal to absent, whereas, in hapten, inflammation-marked iNOS gene expression was evident from day 1 to 7. Nitrotyrosine and iNOS immunohistochemistry were colocalized, and positive staining was most intense in epithelia and neurons. Inhibition of NO formation prevented nitrotyrosine formation. Aminoguanidine inhibited the inflammatory response and restored morphology. CONCLUSIONS: The colocalization of tyrosine nitration with iNOS immunoreactivity suggests that iNOS may be responsible for tissue injury and the formation of NO-dependent nitrating species, potentially peroxynitrite. Inhibition of iNOS may afford a new therapeutic approach to the treatment of inflammatory bowel disease.

Amino Acid Sequence↗

Role of metallothionein and cysteine-rich intestinal protein in the regulation of zinc absorption by diabetic rats.

Hyperzincuria and low Zn absorption in diabetic animals and humans have prompted speculation that diabetics are more susceptible to Zn deficiency. There is little information, however, describing the effects of diabetes on the biochemical mechanisms of intestinal Zn transport. We evaluated Zn absorption in streptozotocin-induced diabetic rats based on a model of Zn transport in which cysteine-rich intestinal protein serves as an intracellular carrier that is inhibited by metallothionein (MT). Apparent absorption and retention of Zn and Cu in rats fed a purified diet were measured in a balance study 15-17 d after induction of diabetes. The rate of 65Zn absorption from isolated intestinal segments, molecular distribution of 65Zn in mucosal cytosol, and tissue MT levels were measured on d 20-22. Food consumption, and thus Zn and Cu intake, by diabetic rats was twice that of controls. Although fractional absorption (percent) of Zn and Cu was lower in the diabetic rats, net absorption (micrograms/100 g body weight/d) was higher. The higher net absorption in the diabetic group was offset, however, by higher urinary excretion, so that Zn and Cu retention was similar in both groups of animals. Low fractional absorption is attributable to the down-regulation of intestinal Zn transport, as indicated by the lower rate of 65Zn absorption from isolated intestinal segments in the diabetic rats. Down-regulation of intestinal transport is in turn attributable to higher concentrations of intestinal MT, which resulted in more 65Zn in the mucosal cytosol bound to MT, an inhibitor of Zn transport, and less to cysteine-rich intestinal protein.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Prognostic value of the cellular expression of carcinoembryonic antigen (CEA) in gastric adenocarcinoma].

Fifty resected gastric adenocarcinomas were studied retrospectively to evaluate survival factors. Eleven different criteria were assessed using multivariate analysis. Only two criteria were significant: 1--cellular expression of carcinoembryonic antigen (p less than 0.0001 - CCR = 1.48) 2--intravascular or lymphatic tumor emboli (p less than 0.006 - CCR = 2.06). Thus, it is suggested that in addition to the various prognostic factors available to the surgical pathologist, the presence of cellular expression of carcinoembryonic antigen in tumor tissue will aid in predicting the prognosis of patients with gastric carcinoma.

Adenocarcinoma↗

New approaches in the diagnosis of Taenia solium cysticercosis and taeniasis.

Taenia solium cysticercosis is now recognized as a priority in Mexico and a number of other developing countries, both in public health and in economic terms. Recognition of the problem has been greatly aided in recent years by new developments in molecular diagnostics. In this paper data are presented on ELISA for the detection of anti-cysticercus antibodies and of parasite antigens in patients with neurocysticercosis and in cysticercotic pigs. Also, several biological fluids were evaluated: cerebrospinal fluid (CSF), serum and saliva, all of which have proved useful. CSF is, however, the most appropriate for detection of antibodies and antigens in patients and serum in pigs. In addition, saliva may be especially used in epidemiological surveys. The electroimmuno-transfer blot technique (EITB) for antibody detection in patients and pigs has also proved highly sensitive and due to the use of an enriched fraction of glucoproteins, EITB is also highly specific. Cloned cDNA sequences from T. solium are now being assessed as an alternative source of antigens for immunodiagnosis. Two methods for the diagnosis of the adult stage of T. solium are also undergoing standardization. These are an ELISA for the detection of parasite antigens in fecal samples and DNA hybridization techniques for the detection of eggs in stools. Both assays have promising results and should now be assessed in larger numbers of clinical and epidemiological samples.

Animals↗

Long-term correction of genetic defect of liver function in rat by transplantation of liver cells after ultraviolet irradiation.

Allograft rejection limits the survival and function of transplanted hepatocytes obtained from allogeneic donors. Presentation of donor antigens by antigen-presenting cells that express class II major histocompatibility antigens (MHC) carried in the graft has been implicated in allograft rejection. Ultraviolet-B (u.v.-B, 280 to 320 nm) irradiation and short-term culture have been shown to modify the function of antigen-presenting cells. In this study, we used u.v.-B irradiation (600 J/m2) of donor hepatocytes isolated from normal inbred Wistar RHA rats, followed by 16 to 20 hours of culture prior to transplantation into histoincompatible genetically analbuminemic rats (Nagase analbuminemic rats, NAR) by intraportal infusion of 10(7) viable hepatocytes. Isolated Wistar RHA hepatocytes without u.v.-B irradiation and/or culture, or NAR rat hepatocytes, were used as control. Survival and function of the transplanted hepatocytes were monitored by serial immunoassay of serum albumin in the recipients. Serum albumin concentration did not increase after transplantation of NAR rat hepatocytes. In NAR rats that received Wistar RHA hepatocytes, serum albumin levels increased from pre-transplantation levels of 0.025 to 0.05 mg/ml to 8 to 10 mg/ml. When freshly isolated liver cells were transplanted with or without u.v.-B-irradiation, but without culturing, serum albumin levels reached a maximum in two weeks and then progressively declined to pretransplantation levels. When the cells were not irradiated but cultured for 16 to 20 hours, elevated serum albumin levels persisted for six weeks, and then progressively declined to baseline concentrations. In contrast, when the isolated normal liver cells were u.v.-B-irradiated and cultured for 16 to 20 hours before transplantation, serum albumin levels persisted at 8 to 10 mg/ml throughout the duration of this study (32 weeks). Immunocytochemistry using anti-rat serum albumin (rabbit) immunoglobulin of liver tissue from NAR rats without transplantation or after transplantation with NAR liver cells showed immunostaining in less than one in 1000 cells. Four to 16 weeks after transplantation of u.v.-B-irradiated and cultured normal allogeneic hepatocytes, one to two per 100 hepatocytes stained positive for albumin. The results indicate that u.v.-B-irradiation followed by short-term culture of allogeneic rat hepatocytes prior to transplantation results in prolonged and perhaps permanent allograft acceptance by NAR rats.

Animals↗

[Infections by anaerobes in children: frequency of isolation and antimicrobial resistance].

We studied the frequency of isolation and antimicrobial susceptibility of strict anaerobic bacteria isolated in a paediatric hospital. A total of 1,753 samples from purulent material and hemocultures were processed. One hundred and thirty strains were isolated from 95 children: 44 from 48 cases of peritonitis (91.7%), 2 from 8 brain abscesses (25%), 23 from 124 soft tissue abscesses (18.5%), 4 from 64 empyemas or lung abscesses (6.3%) and 22 from 1,509 hemocultures (1.5%). Mixed infection (anaerobic and aerobic bacteria) was detected in 38.5% of the cases; however, in peritonitis 81.3% of the cases showed mixed infection. The more frequently isolated bacteria were: Bacteroides (44.6%), Clostridium (19.2%), Fusobacterium (7.7%) and gram positive cocci (7.7%). Propionibacterium acnes was isolated in 15 specimens; however, most of them were considered as contamination. Bacteroides was isolated more frequently from patients with peritonitis (51.6%). The susceptibility to five antibiotics was tested in 124 strains using the method of serial dilutions in agar plates. The genus Bacteroides showed a high resistance to penicillin (73.1%), moderate to clindamycin (11.5%) and low to cefoxitin, chloramphenicol and metronidazole (6.8%, 2% and 5.8%). The rest of the anaerobic strains tested (other gram negatives and grampositives) were highly sensitive to all the antimicrobials tested, except for clindamycin (26.7% and 36.9% resistance, respectively). Cefoxitin, chloramphenicol and metronidazole had the lower CMI50 and CMI90 for the genus Bacteroides; and for the rest of the bacteria, penicillin had the highest activity (CMI90 less than 0.025). Since the frequency of isolation of anaerobic bacteria in children with severe infections, presumably originated from the digestive tract, paranasal sinus and middle ear is high, anaerobic cultures must be practiced in these patients. Empiric antimicrobial treatment should also be started. This treatment should be with penicillin when the probability of isolation of Bacteroides is low (infections in the face, neck, thorax and soft tissues) and with metronidazole in intraabdominal infections. Because of the severity and probable etiology of brain abscesses, treatment with penicillin and chloramphenicol or metronidazole is recommended in these cases.

Anti-Bacterial Agents↗

Comparative analysis of human and porcine neurocysticercosis by computed tomography.

Human and porcine cases of neurocysticerosis were compared by computed tomography in regard to the morphological characteristics of the parasites and the impact of the infection on the brain. In pigs and young people many parasites were usually found, but this was seldom the case in adult humans. These parasites were small and located mainly in the parenchyma and the subarachnoid space; no calcified cysticerci were found. In contrast, in adult humans cysticerci were heterogeneous in appearance and distribution. The brain of the pigs showed no evidence of inflammatory reaction while young people had oedema, enhancement and hydrocephalus; in contrast, adults showed diverse tomographic characteristics of brain response. Thus, the disease has at least 2 different forms. It may be mild with short development and parasites of similar appearance, as occurs in young hosts. In contrast, a long lasting disease occurs in adults, with diverse tomographic, neurological and parasitological features.

Adolescent↗

Reliable serology of Taenia solium cysticercosis with antigens from cyst vesicular fluid: ELISA and hemagglutination tests.

New levels of reproducibility and sensitivity have been achieved in the detection of anticysticercus antibodies in human sera by using cysticercus vesicular fluid as the source of antigens for both ELISA and hemagglutination assays. Reproducibility both between tests on a serum and between similar sera was significantly improved over typical results using antigens from whole parasite extracts. Sera collected from uninfected individuals in endemic areas gave somewhat elevated values over those collected in nonendemic areas. This necessitated the use of a higher threshold in endemic areas to avoid false positives. With the threshold appropriate for a nonendemic area, both ELISA and hemagglutination were sensitive enough to detect infection in 95% of cases. With the threshold value for sera from an endemic area, these sensitivities were reduced to 80%-90%. A prominent 103-Kd protein of vesicular fluid, not related to antigen B, elicited the strongest antibody response in neurocystercotic patients.

Antibodies↗

Intraventricular cysticercosis.

The authors review their experience with 21 cases of intraventricular cysticercosis, which corresponded to 28% of all cases of neurocysticercosis in their department during an 18-month period. The surgical approach depends on the cyst's location; one uses the transcortical microsurgical approach to reach the lateral ventricles, the transcortical or transcallosal approach to reach the 3rd ventricle, and direct exploration to reach the 4th ventricle. Cysticercus cysts should be removed because they may produce acute or chronic hydrocephalus and, if the parasite dies within the ventricles, it will generate an inflammatory reaction with local and generalized ventriculitis, which produces irreversible neurological damage.

Adult↗

The seagull incision in posterior cervical lift: indications, limitations, and surgical technique.

A seagull incision in the posterior scalp as used by us for the past 15 years for the correction of flaccidity in the posterior and anterior lower neck regions through a posterior cervical lift has been described in detail with its advantages, indications, and limitations. In our experience, the posterior cervical lift has little or no effect in correcting flaccidity in the upper anterior neck region, and therefore, this procedure should not be routinely performed in all cases of rhytidoplasty. One has to be highly selective in performing this procedure.

Esthetics↗