Breast cancer chemotherapy treatment monitoring based on serum sample Raman spectroscopy.

In this paper, breast cancer patients were monitored throughout their chemotherapy treatments (CHT), with blood serum sample Raman spectroscopy and multivariate analysis, approximately for a year. First of all, we discriminate between healthy and clinically diagnosed breast cancer patients. Breast cancer detection in terms of sensitivity and specificity were 87.14% and 90.55% respectively. Although no shifts of peaks in mean spectrum of samples from breast cancer patients were found with respect to the mean spectrum from control patients, some peaks did show clear differences in intensity, the greatest disparities found at 509, 545, 1063, 1103, 1338, 1556, 1083 and 1449 cm- 1 are associated with amino acids and phospholipid, 1246 and 1654 cm- 1, corresponding to amide III and I, respectively. Other peaks of interest encountered at 450, 661, 890, 917 and 1405 cm- 1 are associated to glutathione. Then, 6 breast cancer patients were monitored during their chemotherapy treatments, the results were in complete correspondence with their medical records, enabling a detailed study of the evolution of each patient's cancer. A special interest arose in the possible correlation between the intensity of Raman peak, 450 cm- 1, corresponding to glutathione and evolution of cancer throughout CHT, i.e., glutathione appears to be a good candidate as breast cancer biomarker. The results confirmed that Raman spectroscopy and PCA are, not only a good support to current breast cancer detection techniques, but could also be excellent techniques to monitor more efficiently breast cancer patients undergoing CHT, using blood serum samples which are a lot less invasive than other methods.

Urinary neutrophil gelatinase-associated lipocalin and monocyte chemoattractant protein 1 as biomarkers for lupus nephritis in Colombian SLE patients.

Background Information regarding urinary biomarkers in Mestizo and Afro-Latin-American patients is very limited. We investigated whether levels of urinary neutrophil gelatinase-associated lipocalin (NGAL), and monocyte chemoattractant protein 1 (MCP-1) are good biomarkers to differentiate patients with lupus nephritis among Latin-American systemic lupus erythematosus (SLE) patients. Methods SLE patients meeting the revised American College of Rheumatology classification criteria for SLE were recruited. Urinary levels of NGAL and MCP-1 were measured using a commercial ELISA kit. Serum anti-C1q antibodies were measured by ELISA. SLE activity was measured with the systemic lupus erythematosus disease activity index (SLEDAI). Mann-Whitney tests were used to compare data and Spearman's rank correlations were used to examine associations between continuous variables. In addition, receiver operating characteristic curves were performed. Results One hundred and twenty SLE patients were recruited (87% women) with a median age of 32.8 ± 12.1 years and median disease duration of 7.3 ± 6.9 years. Afro-Latin-Americans had a significantly higher prevalence of lupus nephritis and higher SLEDAI scores than Mestizos. The three biomarkers were significantly higher in patients with lupus nephritis than in patients without lupus nephritis. In addition, urinary NGAL and MCP-1 were significantly higher in patients with active lupus nephritis than in inactive lupus nephritis. Urinary NGAL levels were significantly higher in Afro-Latin-American patients. A receiver operating characteristic curve for urinary biomarkers for lupus nephritis in all SLE patients showed a good level of sensitivity and specificity. Conclusion In our cohort of SLE patients, we found that urinary NGAL and MCP-1 in addition to anti-C1q antibodies were useful biomarkers for the identification of renal involvement and discrimination of active lupus nephritis among patients with renal disease.

Expression and localization of angiopoietin family in buffalo ovarian follicles during different stages of development and modulatory role of angiopoietins on steroidogenesis and survival of cultured buffalo granulosa cells.

The present study investigated the expression and localization of angiopoietin (ANPT) family members in buffalo ovarian follicles of different size. It also looked at the role of ANPTs in estradiol secretion and mRNA expression of phosphoinositide-3-kinase-protein kinase B signaling pathway cellular proliferation (phosphoinositide-dependant kinase and protein kinase B [AKT]) and proapoptotic (BAD) factors with caspase 3 in cultured buffalo granulosa cells (GCs). The mRNA and protein expression of ANPT-1 was greatest (P < 0.05), whereas ANPT-2 was reduced (P < 0.05) in preovulatory follicles as compared to F1 follicle. Tyrosine kinase with immunoglobulin-like and EGF-like domains 1 transcripts and protein expression did not change in all follicular groups, whereas tyrosine kinase with immunoglobulin-like and EGF-like domains 2 mRNA was highest (P < 0.05) in theca interna but not GC layer of preovulatory follicle. All members of ANPT family were localized in GC and theca interna showing a stage specific immunoreactivity. Cultured GCs were treated with ANPT-1 and ANPT-2 separately at doses of 1, 10, and 100 ng/mL and in combination at 100 ng/mL for three incubation periods (24, 48, and 72 hours). Estradiol secretion was highest (P < 0.05) at 100 ng/mL at 72 hours of incubation when GCs were treated with either protein alone. The mRNA expression of phosphoinositide-dependant kinase and AKT was highest (P < 0.05), and BAD with caspase 3 was lowest (P < 0.05) at 100 ng/mL at 72 hours of incubation, when cultured GCs were treated separately with each protein or in combination. The immuoreactivity of AKT, pAKT, and pBAD were maximal, whereas BAD was minimal with 100 ng/mL at 72 hours when cultured GCs treated with either protein alone. The findings indicate that ANPTs are expressed in a regulated manner in buffalo ovarian follicle during different stages of development where they may promote steroidogenesis and GC survival through autocrine and paracrine actions.

Loss of a clinical attachment level in a child: differential diagnosis with aggressive periodontitis. Case report.

A nine-year-old boy presented with a serious periodontal lesion caused by a rubber band that had been used to close a medial diastema. Surgery was required to remove it. The impaction of a foreign body in the periodontal space should be suspected when a clinical attachment level is lost during childhood.

[The relationship between human papillomavirus and cyclin D1 gene amplification in squamous cell carcinomas of the head and neck]. / Relación entre el virus del papiloma humano y la amplificación del gen de la ciclina D1 en los carcinomas epidermoides de cabeza y cuello.

INTRODUCTION: infection by oncogenic subtypes of human papillomavirus (HPV) and cyclin D1 gene (CCND1) amplification are frequent events in head and neck squamous cell carcinomas. The objective of this paper is to establish the relationship between the presence of human papillomavirus (HPV) gene sequences and the development of CCND1 gene amplification in these tumours. MATERIALS AND METHODS: 59 squamous cell carcinomas of the head and neck were studied for HPV types 6b and 16 and CCND1 gene amplification by polymerase chain reaction. RESULTS: HPV DNA was detected in 14 tumors (24%). Ten of them were positive for the HPV type 6b and 4 for the HPV type 16. CCND1 gene amplification was found in 15 cases (25%). Although we have found a higher frequency of CCND1 amplification in the HPV-positive cases (36%, versus 22% in the HPV-negative cases), these differences were not statistically significant (P= 0,32). CONCLUSIONS: The presence of HPV gene sequences does not seem to be related to a significative higher incidence of CCND1 gene amplification in the squamous cell carcinomas of the head and neck.

[Metastases in both internal auditive meatus of nasosinusal melanoma]. / Metastasis en ambos conductos auditivos internos de melanoma nasosinusal. A propósito de un caso.

Primary malignant melanoma of the nasal and paranasal sinusses is rare and represent 3% of malignant nasosinusal tumors. The prognosis of this tumor is poor. The evolution is unpredictable; there is very commonly a series of local recurrences which may be well controlled, followed by remote metastases remaining long quiescent. There are no statistical difference in local control or survival between patients receiving surgery alone and those receiving surgery and radiotherapy. Chemotherapy is used for metastatic forms. We present a case of a patient with a primary malignant melanoma of the nasal mucosa. Tree years after diagnosis we found intracranial metastases in left and right auditive internal meatus.

[Lateral sinus thrombosis due to cholesteatoma. Report of a case and literature review]. / Trombosis del seno lateral secundaria a colesteatoma. A propósito de un caso y revisión de la literatura.

After introduction of antibiotics, endocranial complications in otitis media are less common. Lateral sinus thrombophlebitis (LST) incidence has markedly decreased but mortality is still high. This complication should be considered in patients with ear discharge, fever and neurological symptoms. CT-scan and MRI enables early diagnosis and have a role in detecting addition intracranial complications. Surgical intervention should be aggressive and anticoagulation should be considered. We present a case of a woman with othorrea, fever and neurological symptoms. The CT-scan shows us a cholesteatoma that grows intracranial and a LST as complication of it.

[DNA extraction using Chelex resin for oncogenic amplification analysis in head and neck tumours]. / Extracción de ADN con resina Chelex en el análisis de la amplificación oncogénica en carcinomas de cabeza y cuello.

DNA extraction from tissues can be the most laborious and complex step in amplifying DNA by PCR when phenol-choroform procedure is used. We compare this lengthy, slow and expensive extraction method with other two based in the use of Chelex-100 resin. This chelating resin has been applied for extracting DNA from different tissues to use with the PCR. These procedures are simple, rapid and do not require multiple steps. In this study we compared DNA extraction from 30 head and neck squamous cell carcinomas (HNSCC) using organic solvent precipitation, Chelex 100 resin with and without proteinase K pretreatment. The results show that proteinase K-Chelex 100 procedure is as efficient as the phenol-chloroform one.

[Inactivation of p53 and amplification of cyclin D1 in squamous cell carcinomas of the head and neck]. / Inactivación de P53 y amplificación de la ciclina D1 en los carcinomas epidermoides de cabeza y cuello.

P53 and CCND1 (cyclin D1) genes play a critical role in the cell cycle regulation. Abnormalities of these genes are frequent in different types of cancers, including those of the head and neck. The aim of this work is to investigate whether P53 inactivation (determined by loss of heterozygosity analysis) is related to CCND1 gene amplification (determined by differential PCR analysis), and if these alterations are correlated with clinical outcome in a series of 56 patients with squamous cell carcinoma of the head and neck. Loss of heterozygosity of the P53 gene was found in 39 cases (70%) and CCND1 amplification in 17 cases (30%). Both abnormalities together were found in 11 cases (20%), without a significant association between them (P = 0.83). No relationship was found between P53 inactivation, the clinico-pathological parameters analyzed and the clinical outcome. CCND1 amplification was associated with advanced T-stages (P = 0.02), nodal metastases (P = 0.01) and a decreased survival (P = 0.002). The combination of both abnormalities shows a pattern that seems to be additive, since it was associated with an increase in tumor recurrences and a decrease in survival that was higher than for either of them individually. In conclusion, P53 and CCND1 abnormalities are frequent in squamous cell carcinomas of the head and neck. The combined analysis of these abnormalities seems to be more informative than either of them individually and may have a prognostic value in these carcinomas.

[CCND1 oncogene amplification and cellular DNA content in squamous cell carcinomas of the head and neck]. / Amplificación del oncogén CCND1 y contenido celular de ADN en los carcinomas epidermoides de cabeza y cuello.

Cyclin D1 protein (encoded by the CCND1 gene) contributes to the progression of the cell cycle in the G1/S checkpoint. Cyclin D1 overexpression (for instance as a consequence of CCND1 amplification) might result in loss of control over genetic damage at this point and in an accumulation of chromosomal aberrations. In this work we analyze whether CCND1 amplification is associated with a higher incidence of alterations in cellular DNA content. 31 squamous cell carcinomas of the head and neck were studied. CCND1 amplification was determined by polymerase chain reaction. Cellular DNA content was determined by flow cytometry. CCND1 amplification was found in 6 (19%) cases. Thirteen (42%) cases were diploid and 18 (58%) were aneuploid. Two (33%) of the 6 cases with CCND1 amplification were aneuploid compared with 16 (64%) of the cases without CCND1 amplification (P = 0.36). We conclude that CCND1 amplification is not associated to a higher incidence of chromosomal aberrations in squamous cell carcinomas of the head and neck.

[Molecular changes in epidermoid carcinoma of the oropharynx]. / Alteraciones moleculares en los carcinomas epidermoides de la orofaringe.

In most of the studies about molecular alterations in squamous cell carcinomas of the head and neck there is not distinction between the different subsites of this area. The objective of this study is to describe the molecular alterations in squamous cell carcinomas of the oropharynx. Twenty-nine oropharyngeal carcinomas, with a minimum follow-up of 36 months, were studied. The molecular alterations analyzed were: the amplification of 11q13 region (in the 29 cases), and the MYC and ERBB1 oncogenes (in 22 cases); the integration of Human Papillomavirus (HPV) types 6b and 16 (in 22 cases); the loss of heterozygosity (LOH) of p53 and N-acetyltransferase-2 (NAT2) gene (in 12 and 13 informative cases, respectively); and the cellular DNA content (in 13 cases). The most frequent alterations found were the LOH at p53 (67%), and NAT2 (54%) locus, followed by 11q13 amplification (49%). ERBB1 amplification was found in 14% of the cases, and MYC amplification only in one (5%). Integration of the HPV was found in 23% of the cases. Nine (69%) of the 13 analyzed cases were aneuploid. The only alteration with a prognostic significance was 11q13 amplification that showed a tendency to be associated with a higher frequency of nodal metastases and tumor recurrence.

Differences in stroke subtypes among natives and caucasians in Boston and Buenos Aires.

BACKGROUND AND PURPOSE: Several issues regarding ethnic-cultural factors, sex-related variation, and risk factors for stroke have been described in the literature. However, there have been no prospective studies comparing ethnic differences and stroke subtypes between populations from South America and North America. It has been suggested that natives from Buenos Aires, Argentina, may have higher frequency of hemorrhagic strokes and penetrating artery disease than North American subjects. The aim of this study was to validate this hypothesis. METHODS: We studied the database of all consecutive acute stroke patients admitted to the Ramos Mejia Hospital (RMH) in Buenos Aires and to the Beth Israel Deaconess Medical Center (BIMC) in Boston, Massachusetts, from July 1997 to March 1999. Stroke subtypes were classified according to the Trial of Org 10172 in Acute Stroke Treatment (TOAST) criteria. All information on patients (demographic, clinical, and radiographic) was recorded prospective to the assessment of the stroke subtype. RESULTS: Three hundred sixty-one and 479 stroke patients were included at RMH and BIMC stroke data banks, respectively. Coronary artery disease was significantly more frequent in BIMC (P:<0.001), whereas tobacco and alcohol intake were significantly more frequent in RMH (P:<0.001). Intracerebral hemorrhage (P:<0.001) and penetrating artery disease (P:<0.001) were significantly more frequent in the RMH registry, whereas large-artery disease (P:<0.02) and cardioembolism (P:<0.001) were more common in the BIMC data bank. CONCLUSIONS: Penetrating artery disease and intracerebral hemorrhage were the most frequent stroke subtypes in natives from Buenos Aires. Lacunar strokes and intracerebral hemorrhage were more frequent among Caucasians from Buenos Aires than Caucasians from Boston. Poor risk factor control and dietary habits could explain these differences.

Relationship between red cell folate and cervical intraepithelial neoplasia and cervical human papillomavirus infection.

OBJECTIVE: The aim of this study was to determine the relationship between red cell folate levels and the presence of cervical intraepithelial neoplasia (CIN) or cervical human papillomavirus infection (HPV) (or both). For that purpose, we designed a case-control study. MATERIALS AND METHODS: One hundred three asymptomatic women who were between the ages of 20 and 76 and attending the gynecological clinic of the Military Hospital in Bogota, Colombia, were selected. Their mean age was 37.9 years. Inclusion criteria combined a colposcopic examination and a cervicovaginal Papanicolaou smear; accordingly, patients were divided into women without CIN or HPV (55 women, the control group) and women with CIN or HPV (or both) (48 women, the study group). Red cell folate levels were determined by radioimmunoassay. RESULTS: Statistically significant differences in folate levels were found between cases [2.55 ng/ml; standard deviation, 0.75; 95% confidence interval, 2.34-2.73] and controls (2.96 ng/ml; standard deviation, 0.77; 95% confidence interval, 2.75-3.14; p < .008). The odds ratio between the folate levels and CIN or HPV (or both) was 0.49 (p = .01). Red cell folate levels of the individuals participating in this study were not found to be associated with parity, the use of oral contraceptives, cigarette smoking, or age. CONCLUSIONS: High red cell folate levels appear to provide a protective effect against the development of CIN or HPV (or both).

[Maximum oxygen consumption and serum lipids in diabetics, controls, and athletes]. / Consumo máximo de oxígeno y los lípidos séricos en diabéticos, controles y atletas.

Although it has been pointed out that the main cardiovascular risk factors in diabetic are dyslipidemias, hypertension, and cigarette smoking, very few studies have analyzed other risk factors, such as sedentarism and maximal aerobic capacity (VO2 max). The purpose of this study was to evaluate VO2 max and blood lipids (total cholesterol = TC, high density lipoproteins cholesterol = HDL; low density lipoproteins cholesterol = LDL; triglycerides = TG), in 19 diabetics subjects, 19 sedentary and 19 long distance runners. The diabetics of 5-10 years of evolution were non obese (body fat < 28%), and in regular metabolic control (Hb A1 < 12%). The athletes had more than five years of uninterrupted training. The diabetics had significantly lower VO2 max, than the other groups. The mean +/- SEM (mL/kg/min) in diabetics was 30.5 +/- 1.6 versus 41.3 +/- 2.3 in the controls and 54.2 +/- 2.5 in the athletes. There were significant group differences in TG (218 +/- 44 in diabetics vs 106 +/- 16 in controls and 94 +/- 10 mg/dL in athletes) and in HDL (27.3 +/- 4.3 in diabetes versus 34.1 +/- 4.3 and 43.9 +/- 6.3 mg/dL). These data suggest that sedentarism and low VO2 max may be important risk factors in diabetics. An aerobic program for them could perhaps change their blood lipids favorably as their VO2 max was significantly correlated with TG (r = -0.45), HDL (r = 0.52) and the TC/HDL ratio (r = -0.57).

Otomicosis: diagnóstico por el laboratorio / Otomycoses, laboratory diagnosis

Entre 1982 y 1991 se analizaron en el Laboratorio de Micología del Hospital Universitario del Valle (H.U.V.), en Cali, Colombia, un total de 377 pacientes con diagnostico de otitis de etiología a aclarar; se seleccionaron 113 casos de los cuales 107 mostraron examen directo y cultivo positivo para hongos, mientras que en 6 el examen directo reveló la presencia de estructuras micoticas pero los cultivos fueron negativos. Se analiza la frecuencia de los agentes causales, la distribución de la micosis de acuerdo con el sexo, los factores predisponentes y los métodos de diagnostico en el laboratorio. El hongo Aspergillus flavus fue el agente causal mas aislado (33.3 por ciento) siguiendole en orden de frecuencia Candida sp (26.3 por ciento). El sexo femenino se vio afectado en mayor proporción (53.1 por ciento). Factores como el calor, la humedad excesiva, el cambio de PH, algunos elementos nutricionales, la manipulación con materiales variados y el uso de esteroides favorecen el desarrollo de la infección. La combinación del examen directo y/o cultivo (seriado) permiten confirmar la etiología del proceso, para brindar un tratamiento adecuado y mejorar el pronostico de la micosis

Laboratory evaluation of stain-length passive dosimeters for monitoring of vinyl chloride and ethylene oxide.

The effects of vapor concentration in the range of 1.2 to 5.1 ppm (vinyl chloride) and 8.3 to 29.1 ppm (ethylene oxide) on the response of new stain-length passive dosimeters were evaluated separately in a dynamic exposure chamber. The vinyl chloride dosimeter was prepared with a permanganate impregnated blend of Chromosorb W and silica gel, while a silica gel-coated plastic strip (TLC plate) impregnated with dichromate was used to detect ethylene oxide. The use of a TLC plate as the inert support allowed us to reduce the amount of reagent loaded per length of tube, thus significantly enhancing in the sensitivity of the unit, which was necessary for accurately detecting ethylene oxide at these low concentrations. At the vinyl chloride exposure of 8 ppm-hrs the length of stain was 0.76 cm and the 95% confidence interval about this point was +/- 1.4 ppm-hrs (18%). For the ethylene oxide dosimeter the length of stain at the exposure of 80 ppm-hrs was 0.90 cm and the 95% confidence interval about this point was +/- 16 ppm-hrs (20%). Although some shortening of the stain was noted at low relative humidity (26%) in the vinyl chloride device, no effect on the dosimeter response was observed over the range of relative humidity of 35 to 96%. The ethylene oxide dosimeter response was not affected by relative humidity in the range of 28 to 90%. The use of a TLC plate as the inert support of the colorimetric reagent has proven to be an excellent means of improving the sensitivity of these stain-length passive dosimeters.(ABSTRACT TRUNCATED AT 250 WORDS)