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Biomedical subjects

M A Sánchez

Publications and source records attributed to M A Sánchez.

At least 19 recordsLinked to original sources

Sex steroid receptor expression in the oviduct and uterus of sheep with estrus synchronized with progestagen or prostaglandin analogues.

The objective of this study was to investigate differences in the expression of estrogen receptor-alpha (ERalpha), progesterone receptor (PR) and the proliferative indexes (Ki-67), in the uterus and oviduct of sheep with estrus synchronized either by prostaglandin analogues (Group PA, n=27) or by treatment with progestagens (Group P, n=29) on days 4 and 7 (day 0=estrus), when the embryos were collected. Immunohistochemical methods were used to quantify ERalpha, PR and Ki-67 in six superficial and deep compartments in the uterus and oviduct. The expression of ERalpha was significantly (P<0.01) lower in progestagen treated ewes than in prostaglandin analogues treated group in the luminal epithelium, superficial glands and superficial stroma in the uterus on day 4. The expression of PR was significantly lower in progesterone treated ewes than in the PA Group in the superficial gland (P<0.05) in both days studied. The lowest expression of PR was observed in the luminal caruncular epithelium and superficial glands in both treatments, obtaining the lowest levels on day 4 (P<0.05). There were significant differences between days 4 and 7 in the Ki-67 immunostaining in the luminal epithelium (P<0.01) and superficial glands (P<0.05). A higher cell proliferation was observed in the uterine epithelium (P<0.05) on day 4 in the animals treated with progestagens. Results indicate that sheep with synchronization of estrus with progestagens showed a reduction of ERalpha and PR protein expression in most of oviductal and uterine cells.

Animals↗

Cyclic changes of the ovarian surface epithelium in the rat.

The ovarian surface epithelium (OSE) plays pivotal roles during ovulation and postovulatory wound repair. In this paper we describe the proliferative activity of the OSE through the estrous cycle in adult cycling rats, by immunohistochemical detection of DNA-incorporated bromodeoxyuridine (BrdU). Immunohistochemical detection of estrogen receptor alpha (ERalpha) and progesterone receptor was also performed. The cycle of the OSE consists of a proliferative phase (that lasts for two consecutive estrous cycles) and a quiescent phase of variable duration. Cyclic changes in the OSE were related to the underlying ovarian structure. OSE areas covering growing follicles entered into the proliferative phase during the transition from proestrus to estrus, with the appearance of fast-growing class 1 follicles, destined to ovulate at the end of the current estrous cycle. A labeling index (after pulse-labeling BrdU treatment) of about 7% was maintained throughout the estrous cycle in parallel to follicle growth. Cumulative BrdU-labeling (after daily BrdU treatment) indicated that about 1/3 of the total OSE cell proliferation was related to follicle growth. Following ovulation, OSE cells covering newly-formed corpora lutea showed a labeling index of about 50% that decreased through metestrus and diestrus (about 13% and 3%, respectively), returning to basal levels by proestrus. Cumulative BrdU-labeling indicated that about 2/3 of the total proliferative activity was related to ovulation repair/luteinization. The remaining OSE covering ovarian stroma or structurally regressing corpora lutea of previous cycles showed negligible BrdU labeling. The equivalent proliferative activity found in the OSE covering newly-formed corpora lutea in indomethacin-treated rats lacking rupture of the OSE at the apex, demonstrated that ovulation-triggered proliferation was not dependent on the loss of integrity of the OSE at the ovulation site. OSE cells expressed ERalpha throughout the cycle, but no differential expression was found between proliferating and quiescent OSE areas. On the contrary, OSE cells did not express PR at any time of the cycle. These data indicate the existence of a cycle of the OSE, related to the cyclic changes in the underlying ovarian structure and strongly suggest that the proliferative activity of the OSE is regulated by local microenvironmental rather than by systemic factors.

Animals↗

Loss of lineage antigens is a common feature of apoptotic lymphocytes.

The analysis of apoptosis in cell populations involves the detection of their specific lineage antigen (LAg) expression. This experimental approach relies on their assumed constant expression, but it is unclear whether such expression is actually maintained during cell death. We examined whether the loss of LAgs is a common feature of apoptotic lymphocytes and whether some might completely lose their LAgs. The changes in the expression of CD3, CD5, CD8, CD4, CD28, CD56, and CD19 were monitored in highly purified lymphocyte populations obtained by negative selection in a fluorescence-activated cell sorter. These were cultured for 24 h with or without phytohemagglutinin or staurosporin. For each LAg-positive subset studied, apoptosis was consistently more common among cells showing partial or total loss of LAg expression compared with cells maintaining their initial LAg levels. The kinetics of expression loss was rapid for CD8, CD56, and CD28, and more than 80% of initial expression was lost in the early stages of apoptosis but was slower for CD3, CD5, and CD4. For CD3 and CD5, expression was dependent on the apoptotic stimulus used. It is interesting that loss of antigen expression was independent of cell size. This phenomenon was also found in nonmanipulated, highly pure CD19 B lymphocytes of peripheral blood mononuclear cells from B chronic lymphocytic leukemia patients. Loss of LAg expression appeared to be a common feature of apoptotic lymphocytes under all the conditions assayed. The different kinetic patterns of LAg loss suggest apoptotic cells might actively regulate this process.

Antigens, CD↗

Epithelioid malignant peripheral nerve sheath tumour in a dog.

An unusual morphological variant of malignant peripheral nerve sheath tumour, in which the cells exhibited a predominantly epithelial morphology, was diagnosed in a dog. The animal had a history of progressive ataxia. Post-mortem examination revealed an infiltrative growth in the sublumbar region, encroaching on the L1 and L2 vertebral bodies; additional masses were observed in the kidneys, liver and lungs. Histologically, the subcutaneous mass was composed of a dense population of spindle-shaped cells arranged in fascicles. The internal masses were composed of epithelioid cells arranged in a nodular pattern. Immunohistochemically, the spindle-shaped cells of the subcutaneous mass were positive for S-100, and the epithelioid cells of the internal growths exhibited weak S-100 immunoreactivity.

Animals↗

Expression of alphav, alpha4, alpha5 and beta3 integrin subunits, fibronectin and vitronectin in goat peri-implantation.

Integrins are glycoprotein heterodimers located in the cell membranes that stimulate intercellular adhesion and act as extracellular matrix (ECM) protein receptors. Although integrins have been detected in the implantation sites of various species, little is known about their participation in ruminant non-invasive placentation. The objective of this study was the detection of alphav, alpha4, alpha5, beta1 and beta3 integrin subunits and of two of their ligands, fibronectin and vitronectin, to determine their participation in the caprine peri-implantation process. On Day 21 post-coitum (pc), endometrial epithelium and trophoblastic cells showed an intense alphav and beta3 integrin subunits expression and moderate staining for alpha4 and alpha5. On Day 23 pc, integrin expression decreased noticeably and only a weak staining of alpha4 and beta3 integrin subunits were observed. No beta1 integrin subunit expression was detected on either of the days studied. Fibronectin (FN) expression in trophectodermic and endometrial epithelium was weak or moderate on the days studied while vitronectin (VN) expression in the same tissues was moderate or strong on Day 21 pc but decreased on Day 23 pc. These results suggest that alphavbeta3 integrin, alpha4 and alpha5 subunits, VN and FN are expressed in caprine endometrium and blastocyst and may play a role in the cascade of the implantation process.

Animals↗

Genetic organization of the catabolic plasmid pJP4 from Ralstonia eutropha JMP134 (pJP4) reveals mechanisms of adaptation to chloroaromatic pollutants and evolution of specialized chloroaromatic degradation pathways.

Ralstonia eutropha JMP134 (pJP4) is a useful model for the study of bacterial degradation of substituted aromatic pollutants. Several key degrading capabilities, encoded by tfd genes, are located in the 88 kb, self-transmissible, IncP-1 beta plasmid pJP4. The complete sequence of the 87,688 nucleotides of pJP4, encoding 83 open reading frames (ORFs), is reported. Most of the coding sequence corresponds to a well-conserved IncP-1 beta backbone and the previously reported tfd genes. In addition, we found hypothetical proteins putatively involved in the transport of aromatic compounds and short-chain fatty acid oxidation. ORFs related to mobile elements, including the Tn501-encoded mercury resistance determinants, an IS1071-based composite transposon and a cryptic class II transposon, are also present in pJP4. These mobile elements are inefficient in transposition and are located in two regions of pJP4 that are rich in remnants of lateral gene transfer events. pJP4 plasmid was able to capture chromosomal genes and form hybrid plasmids with the IncP-1 alpha plasmid RP4. These observations are integrated into a model for the evolution of pJP4, which reveals mechanisms of bacterial adaptation to degrade pollutants.

Adaptation, Physiological↗

A previously unexposed forest soil microbial community degrades high levels of the pollutant 2,4,6-trichlorophenol.

2,4,6-trichlorophenol (2,4,6-TCP) is a hazardous pollutant that is efficiently degraded by some aerobic soil bacterial isolates under laboratory conditions. The degradation of this pollutant in soils and its effect on the soil microbial community are poorly understood. We report here the ability of a previously unexposed forest soil microbiota to degrade high levels of 2,4,6-TCP and describe the changes in the soil microbial community found by terminal restriction fragment length polymorphism (T-RFLP) analysis. After 30 days of incubation, about 50% degradation of this pollutant was observed in soils amended with 50 to 5,000 ppm of 2,4,6-TCP. The T-RFLP analysis showed that the soil bacterial community was essentially unchanged after exposure to up to 500 ppm of 2,4,6-TCP. However, a significant decrease in richness was found with 2,000 and 5,000 ppm of 2,4,6-TCP, even though the removal of this pollutant remained high. The introduction of Ralstonia eutropha JMP134 or R. eutropha MS1, two efficient 2,4,6-TCP degraders, to this soil did not improve degradation of this pollutant, supporting the significant bioremediation potential of this previously unexposed, endogenous forest soil microbial community.

Bacteria↗

Voriconazole in the treatment of invasive mold infections in transplant recipients.

Mortality due to invasive mold infections in solid organ transplant recipients is very high despite therapy with amphotericin B, including lipid formulations. Voriconazole is a triazole with a good activity against molds, including Aspergillus spp. and Scedosporium spp. Experience with voriconazole is limited, but preliminary results in patients with these infections are promising. Reported here is the experience with voriconazole administered on a compassionate-use basis to five patients with invasive mold infections: four solid organ recipients and one patient with an autoimmune disorder. Four patients had invasive Aspergillus fumigatus infection (3 lung infections, 1 abdominal infection) and one had invasive ocular Scedosporium apiospermum infection. The MIC of voriconazole was < or =1 microg/ml for all isolates (NCCLS performance standards for microdilution assay, proposed standard M38-P). Voriconazole was administered as primary therapy in a patient with Scedosporium infection and, in patients with Aspergillus infections, after persistence of positive culture despite a cumulative dose of 3 g of a lipid formulation of amphotericin B. Voriconazole was administered for a median time of 80 days (range, 60-90 days). No visual disturbances were observed, but one patient presented a moderate increase in liver enzymes. An increase in the levels of immunosuppressive drugs (tacrolimus or cyclosporine) was detected in all patients during coadministration with voriconazole. A clinical response was observed in all patients (complete response, n=3; partial response, n=2), and a microbiological response was observed in all but one patient. Furthermore, a good relationship between the MIC of voriconazole and outcome was observed. Voriconazole is an effective and safe therapy for treatment of invasive mold infections in solid organ recipients. To avoid toxicity with this drug, however, the dosing of immunosuppressive drugs must be reduced.

Adult↗

[Evaluation of exercise tolerance in patients treated with praziquantel for chronic schistosomiasis with no signs of cardiopulmonary impairment].

OBJECTIVE: The purpose of this study was to evaluate whether a progressive cycle ergometer test (PCET) can detect minimal functional abnormalities that might indicate compromise of the pulmonary vascular bed in patients treated with praziquantel for chronic schistosomiasis who have no clinical signs of cardiopulmonary impairment. PATIENTS AND METHOD: We studied 9 patients (whose mean (SD) age was 38 (18) years and 10 control subjects aged 32 (14) years. Both groups were evaluated by spirometry and a PCET to determine maximum oxygen consumption, heart rate, minute ventilation and gas exchange. Echocardiograms were used to rule out the presence of associated heart disease. RESULTS: Lung function was similar among patients and controls. Slight decreases in maximum oxygen consumption and heart rate reserve were observed with effort during the PCET, with normal respiratory response. No echocardiographic abnormalities that might indicate the presence of pulmonary vascular occlusive disease were observed. CONCLUSIONS: The results showed that the resting lung function is normal in these patients. However, there was a slight decrease in exercise tolerance, probably related to physical detraining. The data obtained during exercise indicated that pulmonary vascular occlusive disease is unlikely in these patients. Abnormalities may appear in later stages of the disease or in patients who do not receive early medical treatment.

Adult↗

Efficient degradation of 2,4,6-Trichlorophenol requires a set of catabolic genes related to tcp genes from Ralstonia eutropha JMP134(pJP4).

2,4,6-Trichlorophenol (2,4,6-TCP) is a hazardous pollutant. Several aerobic bacteria are known to degrade this compound. One of these, Ralstonia eutropha JMP134(pJP4), a well-known, versatile chloroaromatic compound degrader, is able to grow in 2,4,6-TCP by converting it to 2,6-dichlorohydroquinone, 6-chlorohydroxyquinol, 2-chloromaleylacetate, maleylacetate, and beta-ketoadipate. Three enzyme activities encoded by tcp genes, 2,4,6-TCP monooxygenase (tcpA), 6-chlorohydroxyquinol 1,2-dioxygenase (tcpC), and maleylacetate reductase (tcpD), are involved in this catabolic pathway. Here we provide evidence that all these tcp genes are clustered in the R. eutropha JMP134(pJP4) chromosome, forming the putative catabolic operon tcpRXABCYD. We studied the presence of tcp-like gene sequences in several other 2,4,6-TCP-degrading bacterial strains and found two types of strains. One type includes strains belonging to the Ralstonia genus and possessing a set of tcp-like genes, which efficiently degrade 2,4,6-TCP and therefore grow in liquid cultures containing this chlorophenol as a sole carbon source. The other type includes strains belonging to the genera Pseudomonas, Sphingomonas, or Sphingopixis, which do not have tcp-like gene sequences and degrade this pollutant less efficiently and which therefore grow only as small colonies on plates with 2,4,6-TCP. Other than strain JMP134, none of the bacterial strains whose genomes have been sequenced possesses a full set of tcp-like gene sequences.

Bacterial Proteins↗

Immunohistochemical expression of p53, fibroblast growth factor-b, and transforming growth factor-alpha in feline vaccine-associated sarcomas.

Fifty feline sarcomas associated with vaccine-site injection were evaluated to determine the immunohistochemical expression of p53 protein, basic fibroblast growth factor (FGF-b), and transforming growth factor-alpha (TGF-alpha). Forty-one tumors (82%) were fibrosarcomas (FS), eight (16%) were malignant fibrous histiocytomas (MFH), and one (2%) was a chondrosarcoma (CS). Overexpression of p53 protein was observed in the nuclei of tumor cells in 28 (56%) sarcomas; FGF-b expression was found in the cytoplasm of tumor cells in 40 (80%) feline sarcomas, but the staining was more intense in the spindle-shaped cells of FS than in polygonal or round cells of MFH. The single CS faintly expressed FGF-b. The majority of feline vaccine-associated sarcomas (43 of 50, 86%) expressed moderate or intense staining for TGF-alpha in the cytoplasm of tumor cells. Heterogeneous immunolabeling for p53, FGF-b, and TGF-alpha was present in neoplastic, multinucleated giant cells. Intense expression of FGF-b was statistically associated with younger cats (P < 0.01) and with tumors with nodular growth patterns (P = 0.02). In addition, sarcomas negative for p53 protein expressed FGF-b more frequently than did p53-positive tumors (P = 0.04). The frequency of FGF-b immunostaining was significantly higher in sarcomas with intense expression of TGF-alpha (P = 0.05). Immunohistochemical detection of p53 protein, FGF-b, and TGF-alpha suggests that these growth-regulating proteins may play different roles in the development of sarcomas associated with vaccine sites.

Animals↗

Fibroblastic growth factor receptor (FGF-R) expression during uterine involution in goat.

To investigate the possible participation of fibroblastic growth factors (FGFs) in endometrial involution, 20 multiparous goats, slaughtered on days 0, 1, 4, 10, 16 and 22 postpartum (pp), were used. Samples of different parts of the previous pregnant horns were taken and processed using streptoavidin-biotin-peroxidase complex method to analyse FGF receptor (FGF-R) expression. The percentage of positive cells in luminal epithelium, superficial and deep glands and stroma was evaluated. Epithelial, glandular and stromal cells exhibited FGF-R immunoreactivity. No differences between caruncular and inter-caruncular epithelium were observed and staining was most evident in the superficial glands. The greatest degree of FGF-R expression was seen on days 10 and 16 pp, coinciding with epithelial and stromal cellular regeneration. These results suggest that caprine uterine involution is associated with variations in the expression of FGF-R.

Animals↗

Early amniocentesis with the filtration technique: neonatal outcome in 123 singleton pregnancies.

OBJECTIVE: To evaluate results of a prospective study of pregnancies in which early amniocentesis with the filtration technique was performed at 10-13 weeks' gestation (mean 12.3 weeks' gestation). METHODS: 123 singleton pregnancies in which early amniocentesis with the modified filtration technique was performed at 10-13 weeks' gestation (mean 12.3 weeks' gestation). The amniotic fluid was aspirated into the syringe and reinjected through the filter. RESULTS: All the procedures were performed successfully by a single needle insertion. Neither dry taps nor filtration failures occurred. The mean time of amniocentesis was 4.02 min (95% confidence interval, 3 min and 36 s to 4 min and 18 s). The karyotyping success rate was 99.2%. Temporary amniotic fluid leakage occurred in three women (2.4%). There were 110 (89.4%) live births. Two cases of stillbirth occurred at week 38 and week 40. Two unintended losses occurred within three weeks after sampling (1.62%). Another additional unintended fetal death was notified at the 20-week screening ultrasonography. The total fetal-loss rate was 10.6%. In one case, talipes equinovarus was detected at the 20-week screening ultrasound study. CONCLUSIONS: Further studies are needed to determine the risk of amniotic leakage and its relation to duration of the procedure in patients undergoing early amnifiltration.

Amniocentesis↗

Clinical pattern and therapeutic results achieved in 1490 patients with germ-cell tumours of the testis: the experience of the Spanish Germ-Cell Cancer Group (GG).

OBJECTIVE: To describe the clinical characteristics and treatment results obtained with the application of a homogeneous treatment protocol in 1490 patients with germ-cell tumours (GCT) registered in the 55 hospitals belonging to the Spanish Germ-Cell Cancer Group (GG) during the period between January 1994 and April 2001. METHODS: In general, surveillance was the common policy for stage I patients without local poor prognosis factors, whereas they received adjuvant chemotherapy in case those factor were present. Chemotherapy schedules used in advanced cases were cisplatin and etoposide (EP) for seminoma and BEP or BOMP-EPI in non-seminoma, according to whether the patient was in the good or poor prognosis IGCCCG (International Germ-Cell Cancer Collaborative Group) group. Excision of residual masses was mandatory in non-seminomatous germ-cell tumour (NSGCT). RESULTS: Initial local symptomatology was increased testis size in 90% of cases. Sonography was an excellent diagnostic tool to suggest tumour. Non-seminoma (64.2%) was more frequent than seminoma (35.8%). Approximately 10% had the antecedent of cryptorchidism. Non-seminoma patients were 7 years younger than seminoma. Right testis was involved predominantly. Pre-orchidectomy tumour markers were elevated in 21% of seminoma (betaHGC) and 79% in non-seminoma (alphaFP and/or betaHGC). Scrotum violation occurred in only 1.8%. There were significant differences among stage I and the IGCCCG prognosis groups related to a longer interval between the first symptom and orchiectomy. Eighteen percent of non-seminomatous germ-cell tumour belonged to the poor prognosis IGCCCG group. With a median follow-up to 33 months, this series has achieved a 3 year overall survival of 98% for seminoma and 94% for non-seminoma. Only 10% of excised residual masses present after chemotherapy contained malignant cells. CONCLUSION: Spanish GCT have a similar clinical pattern to that described in the other occidental countries except for a slight increased proportion of non-seminoma upon seminoma. Co-operative groups as GG are unique structures to obtain quick and wide experience on the treatment of testis tumours, contributing to achieve a high cure rate.

Adolescent↗

[Foam cell nephropathy in heterozygous female with Fabry's disease].

We describe the clinical and pathological characteristics of a 49 year-old heterozygous female carrier of Anderson-Fabry's disease. Light microscopy and ultrastructural study of a renal biopsy showed the presence of foam cell nephropathy and galactosylceramide deposits affecting podocytes, the parietal epithelium of Bowman's capsule and the distal tubular cells, endothelial cells and medullary interstitial elements. Retrospectively, the presence of storage disease was confirmed in a hysterectomy specimen obtained two years previously. Our observation shows that heterozygotes for this disorder can not only carry and transmit the disease but may also develop pathological deposits in various organs. A renal biopsy from these carriers allows precise identification of the disease, facilitates adequate genetic counseling and gives the option of enzyme replacement therapy in patients who have pathological deposits.

Fabry Disease↗

Microwave irradiation for rapid epidermis-dermis separation and improved epidermal cell immunodetection.

We explored the effects of microwave irradiation on epidermal-dermal separation and subsequent immunostaining of epidermal cells. Epidermal sheets were obtained after incubation in 0.02 M EDTA in PBS and microwave irradiation with 4 pulses of 420 watts for 5 sec, with a total incubation period of 4 min. The control epidermal sheets were immunostained for Langerhans cells and dendritic epidermal T cells using a conventional immunoperoxidase method. The experimental immunodetection of these cells was assisted by incubating the primary antibodies for 10 min at 70 watts. We showed a simple and rapid method for separation of the epidermal-dermal junction and immunostaining of epidermal cells with optimal morphological preservation.

Animals↗