Effect of low-frequency KRAS mutations on the response to anti-EGFR therapy in metastatic colorectal cancer.

BACKGROUND: Only patients with wild-type (WT) KRAS tumors benefit from anti-epidermal growth factor receptor (EGFR) monoclonal antibodies (Mabs) in metastatic colorectal cancer (mCRC). Pyrosequencing is now widely used for the determination of KRAS mutation burden and a conservative cut-off point of 10% has been defined. Up until now, the impact of low-frequency KRAS mutations (<10%) on the response to anti-EGFR Mabs has yet to be evaluated. PATIENTS AND METHODS: Tumors from patients receiving anti-EGFR Mabs based on a WT genotype for KRAS, as determined using direct sequencing, have been retrospectively analyzed by pyrosequencing. Patients were categorized as WT (no KRAS mutation) or low-frequency mutation when KRAS mutation was <10% (KRAS low MT). RESULTS: A total of 168 patients treated by anti-EGFR Mabs for mCRC were analyzed. According to pyrosequencing, 138 tumors remained KRAS WT, while 30 tumors were KRAS low MT. In the KRAS low MT and KRAS WT groups, the response rates were 6.7% and 37.0%, respectively, while stabilization amounted to 23.3% versus 32.6% and progression to 70% versus 29% (P < 0.01). Progression-free survival (PFS) was 2.7 ± 0.5 months for KRAS low MT and was 6.0 ± 0.3 months for KRAS WT (P < 0.01). CONCLUSIONS: These results appear to validate consideration of low-frequency KRAS mutation tumors as positive, and justify a large-scale prospective study.

Establishment of a novel anaplastic large-cell lymphoma-cell line (COST) from a 'small-cell variant' of ALCL.

Anaplastic large-cell lymphoma (ALCL) is a distinct biological and cytogenetic entity with a broad spectrum of morphological features (common type, small-cell variant and lymphohistiocytic variant). Few cell lines of ALCL are available and they all originate from primary tumors demonstrating the common type morphology (ie large-sized lymphoma cells). We established a new ALCL cell line (COST) from the peripheral blood of a patient with a small-cell variant of ALCL, at diagnosis. Cells growing in vitro and in SCID mice consisted of two populations, that is, small- and large-sized cells as seen in the patient's tumor. Both large and small malignant cells were positive for CD43/MT1 T-cell associated antigen, perforin, granzyme B and TIA-1, but negative for CD2, CD3, CD5, CD7, CD4 and CD8 antigens. Standard cytogenetic studies as well as multiplex FISH confirmed the presence of the canonical t(2;5)(p23;q35) translocation, but also revealed additional numerical and structural abnormalities. The COST cell line is the first ALCL small-cell variant cell line, and thus provides a potentially useful tool for further functional and molecular studies that should improve our understanding of the small-cell variant of ALCL, which is more frequently complicated by a leukemic phase.

Social network, presence of cardiovascular events and mortality in hypertensive patients.

The aim of this study was to ascertain the relationship between social network and the appearance of mortality (cardiovascular events (CVEs)) in patients with arterial hypertension (AHT). This is a cohort study of 236 patients with a 9-year follow-up. Measurements included age, sex, blood pressure (BP), diabetes, hypercholesterolemia, marital status, social network, social support, stage of family life cycle (FLC), mortality and CVEs. Patients with a low social network registered higher global mortality (hazards ratio (HR) 2.6 (95% confidence interval (CI) 1.3; 5.5)) as did the oldest patients (HR 5.6 (1.9; 16.8)), men (HR 3.5 (95% CI 1.3; 9.3)) and subjects in the last FLC stages (HR 4.3 (95% CI 1.3;14.1)). Patients with low social support registered higher cardiovascular mortality (HR 2.6 (95% CI 1.1; 6.1)) as did the oldest patients (HR 12.4 (95% CI 2.8; 55.2)) and those with diabetes (HR 3.00 (95% CI 1.2; 7.6)). Patients with a low social network registered more CVEs (HR 2.1 (95% CI 1.1; 4.1)) than patients with an adequate network, as did the oldest patients (HR 3.1 (95% CI 1.4; 6.9)), subjects who presented with a higher grade of severity of AHT (HR 2.7 (1.3; 5.5)) and those in the last FLC stages (HR 2.5 (95% CI 1.0; 6.2)). A low social network is associated with mortality and the appearance of CVEs in patients with AHT. Low functional social support is associated with the appearance of cardiovascular mortality.

Increased yield of PCR products by addition of T4 gene 32 protein to the SMART PCR cDNA synthesis system.

Under certain conditions, T4 gene 32 protein is known to increase the efficiency of different enzymes, such as Taq DNA polymerase, reverse transcriptase, and telomerase. In this study, we compared the efficiency of the SMART PCR cDNA synthesis kit with and without the T4 gene 32 protein. The use of this cDNA synthesis procedure, in combination with T4 gene 32 protein, increases the yield of RT-PCR products from approximately 90% to 150%. This effect is even observed for long mRNA templates and low concentrations of total RNA (25 ng). Therefore, we suggest the addition of T4 gene 32 protein in the RT-PCR mixture to increase the efficiency of cDNA synthesis, particularly in cases when low amounts of tissue are used.

[Gene expression profiling by suppression subtractive hybridization (SSH): a example for its application to the study of lymphomas]. / Signature transcriptionnelle par hybridation suppressive soustractive (SSH): un exemple d'application à l'étude des lymphomes.

Suppression subtractive hybridization (SSH) was used to isolate genes that were differentially expressed in anaplastic lymphoma kinase (ALK)-positive and ALK-negative anaplastic large cell lymphoma. In addition, this approach was applied to Hodgkin's disease cases with different clinical outcomes. SSH combines a normalization step that equalizes the abundance of cDNAs within the sequences to be tested and a subtraction step that excludes the common sequences between the target and the control. In a model system, the SSH technique enriches for rare sequences up to 5,000-fold in one round. We have isolated several genes whose expression varied significantly with regard to the tumour subtypes. There were different genes with known or unknown functions. We aim to compare the results of the SSH approach with those obtained with high density filters. In a near future, we would like to design DNA chips specific of each pathology that could be used for clinical purposes (evaluation of prognosis and therapeutic response).

A mesenchymal glioma stem cell profile is related to clinical outcome.

Recent studies have demonstrated a relationship between the expression of stem cell-associated genes and relapses in glioblastoma (GBM), suggesting a key role for tumor stem cells in this process. Although there is increasing interest in this field, glioma stem cells (GSCs) are still poorly characterized, their 'stemness' state and factors maintaining these properties remain largely unknown. We performed an expression profiling analysis of pluripotency in gliomaspheres derived from 11 patients. Comparative analysis between GSCs and H1 and H9 human embryonic stem cells as well as H9-derived neural stem cells indicates major variations in gene expression of pluripotency factors Nanog and OCT4, but a stable pattern for SOX2 suggesting its important function in maintaining pluripotency in GSCs. Our results also showed that all GSC lines have the capacity to commit to neural differentiation and express mesenchymal or endothelial differentiation markers. In addition, hierarchical clustering analysis revealed two groups of GSCs reflecting their heterogeneity and identified COL1A1 and IFITM1 as the most discriminating genes. Similar patterns have been observed in tumors from which gliomaspheres have been established. To determine whether this heterogeneity could be clinically relevant, the expression of both genes was further analyzed in an independent cohort of 30 patients with GBM and revealed strong correlation with overall survival. In vitro silencing of COL1A1 and IFTM1 confirmed the effect of these mesenchymal-associated genes on cell invasion and gliomasphere initiation. Our results indicate that COL1A1 and IFITM1 genes could be considered for use in stratifying patients with GBM into subgroups for risk of recurrence at diagnosis, as well as for prognostic and therapeutic evolution.

[Cultural adaptation and validation of the Medical Outcomes Study Social Support Survey questionnaire (MOS-SSS)]. / Adaptação cultural e validação da versão Portuguesa Questionário Medical Outcomes Study Social Support Survey (MOS-SSS).

The aim of this study was the assesment of psychometric properties of the Portuguese version of the instrument "Medical Outcomes Study - Social Support Survey (MOSSSS)". This questionnaire has been translated and adapted in a Portuguese sample of 101 patients with chronic illness of a rural health centre in Portugal. The average age of patients was 63.4 years, 56.4% female. 29% were illiterate and 2% had completed high school. 78% had arterial hypertension and the 56.4% had diabetes mellitus type 2. The internal consistency was evaluated using Cronbach's alpha. Exploratory and Confirmatory factor analysis were performed in order to confirm reliability and validity of the scale and its multidimensional characteristics. The 2-week test-retest reliability was estimated using weighted kappa for the ordinals variables and intraclass coefficient correlation for the quantitative variables. Cronbach's alphas for the subscales ranged from 0.873 to 0.967 at test, and 0.862 to 0.972 at retest. Exploratory factor analysis revealed the existence of four factors (emotional, tangible, positive interaction and affection support) that explain the 72.71% of the variance. Confirmatory factor analysis supported the existence of four factors that allowed the application of the scale with original items. The goodness-of-fit measures corroborate the initial structure, with chi2/ df=2.01, GFI=0.998, CFI=0.999, AGFI=0.998, TLI=0.999, NFI=0.998, SRMR=0.332, RMSEA=0.76. The 2-weeks test-retest reliability of the Portuguese MOS-SSS as measured by the intraclass correlation coefficient was ranged from 0.941 to 0.966 for the four dimensions and the overall support index. The weighted kappa was ranged from 0.67 to 0.87 for all the items. The MOS-SSS Portuguese version demonstrates good psychometric properties and seems to be useful to measure multidimensional aspects of social support in the Portuguese population.

[Urethral duplication: apropos a case]. / Duplicidad uretral: a propósito de un caso.

We report a case of dorsal blind-ending urethral duplicity in a 37-year-old male who presented dysuria and occasional mucopurulent discharge. The presumptive diagnosis was made on the findings at physical examination and was corroborated by retrograde urethrography. The patient was treated conservatively because the symptoms were minimal.

CT attenuation values and enhancing characteristics of thoracoabdominal lymphomatous adenopathies.

OBJECTIVE: Thoracoabdominal lymphomatous adenopathies have been described as homogeneous and typically showing little enhancement on postcontrast CT. After observing pronounced adenopathic contrast enhancement in one patient with lymphoma, we prospectively investigated the CT enhancing characteristics and attenuation values of pathologic thoracic and abdominal lymph nodes in 25 patients with lymphoma. MATERIALS AND METHODS: Unenhanced CT of the chest, abdomen and pelvis was performed in all cases. The enhancing characteristics and attenuation values of the largest node or group of nodes were evaluated at 1, 2, 5, and 10 min after a 100 ml intravenous bolus injection of contrast medium. RESULTS: On unenhanced CT the nodes were of soft-tissue attenuation (42 +/- 5 HU) in 23 cases and of low attenuation (31 and 28 HU) in 2 cases. The maximum enhancement was achieved at 1 or 2 min postinjection and was low (16 +/- 6 HU) or moderate (31 +/- 6 HU) in 21 cases and pronounced (61 +/- 5 HU) in 4. The patterns of enhancement were classified as homogeneous (no. = 23) inhomogeneous (no. = 1), and peripheral (no. = 1). CONCLUSION: Thoracic and abdominal lymphomatous adenopathies are usually of soft-tissue attenuation on unenhanced CT and can show slight, moderate, or even pronounced enhancement on postcontrast CT.

[Cryotherapy with nitrous oxide in primary care]. / Crioterapia con óxido nitroso en atención primaria.

OBJECTIVE: To study the efficacy, tolerance and safety in primary care of the cryotherapy technique using nitrous oxide. Design. Descriptive, observational and transversal study. SETTING: Mariñamansa-A Cuña Health Centre, Ourense. PARTICIPANTS: All the patients who consulted for cutaneous lesions caused by cryotherapy between March and April 2000 (n=130). MAIN MEASUREMENTS: Age, sex, diagnosis, location, size of lesion, duration of application of the cryogenic agent, previous treatment, side-effects observed, and evolution of the lesion. RESULTS: Mean age was 36.65 (95% CI, 32.57-40.74), of whom 53.8% (95% CI, 44.80-62.50) were women. The most common lesions were common warts (68.5%), actinic Keratosis (12.3%) and plantar warts (7.7%). We found that most lesions were located on an upper limb (50.8%). The mean size of lesions was 8.65 mm2 (95% CI, 4.50-12.80). 69.7% of warts were scraped prior to cryotherapy; and 16.2% received prior keratolysis. We found no side-effects in 32.3% of the patients; the commonest side-effect detected was blisters. Mean number of cryotherapy sessions was 2.82 (95% CI, 2.39-3.29). The technique was tolerated well in 83.8% of cases. We found total cure in 87.7%. Most of the cutaneous lesions that failed were common warts. CONCLUSIONS: Nitrous oxide is effective in most lesions caused by cryotherapy. The technique is tolerated well, can be handled easily and can be used in primary care.

[Influence of social support on patients with essential hypertension]. / Influencia del apoyo social en pacientes con hipertensión arterial esencial.

OBJECTIVE: To determine whether social support affects blood pressure control in patients diagnosed with essential Hypertension. DESIGN: Observational, prospective study, with a year's follow-up. SERRING: Mariñamansa-A Cuña Health Centre, Ourense (Galicia), Spain. PARTICIPANTS: 236 patients diagnosed with essential hypertension in health centre controls. MEASUREMENTS: During the one-year follow-up the following clinical variables were measured: age, sex, blood pressure, severity of hypertension, tobacco consumption, alcohol consumption, presence of diabetes mellitus, hypercholesterolaemia, Body Mass Index and compliance with treatment. Social and family variables were: marital status, cultural level, economic and social status, type of family, stressful vital events and social support. RESULTS: Mean age was 63.51 (62.05-64.96); 66.1% were women. Predominant kind of family was nuclear (64.3%). 30.2% had low social network (0-1 social contacts). We found 22% poor functional social support. Social support remained stable throughout the study. Hypertense patients with poor social networks had, after control for possible confusing variables, their systolic pressure 9.59 mm Hg (2.6716,51) and diastolic pressure 4.29 mm Hg (0.448.15) higher than hypertense patients with broader social networks. CONCLUSION: Hypertense patients with a poor social network had higher blood pressure figures than hypertense patients with wider social networks.

[Environmental stress and cardiovascular reactivity: the effect of stressful life events on hypertense patients]. / Estrés ambiental y reactividad cardiovascular: la influencia de los acontecimientos vitales estresantes en pacientes hipertensos.

OBJECTIVE: To assess the effect of stressful life events (SLE) on the monitoring of patients with essential hypertension. DESIGN: Observational and prospective. SETTING: Primary care. PARTICIPANTS: 236 hypertense patients selected by systematic randomised sampling. MEASUREMENTS: Age, sex, marital status, cultural level, kind of family, presence of SLE, social support, blood pressure, severity of hypertension, tobacco consumption, alcohol, presence of diabetes, hypercholesterolaemia, body mass index, compliance with hypertension treatment. RESULTS. Mean age was 63.51 (62.05; 64.96), with 66.1% being women and 33.9% men. 48.7% of the patients referred to some SLE during the follow-up. These reached a mean of 47.65 (33.16; 62.15) life change units (LCU). Patients with high-impact SLE (>150 LCU) had a significant increase of 10.91 mm Hg (0.43; 21.40) in their systolic blood pressure and of 9.48 beats per minute (3.78; 15.19) in their heart rate, after monitoring for possible factors of confusion. We observed a similar trend for diastolic pressure, but this did not reach statistical significance. CONCLUSION: The presence of SLE has a negative effect on systolic blood pressure in hypertense patients.

[Validation of the chronometer test as a method of screening for hypoacusis in patients aged 65 years and older]. / Validación del test del cronómetro como método de cribaje de hipoacusias en pacientes de más de 65 años.

OBJECTIVE: Evaluation of the chronometry test as a screening procedure for hypoacusia compared with tonal audiometry, in patients over 65 years. DESIGN: Cross-sectional study. SETTING: Mariañamansa health area (Ourense). PATIENTS: The first 49 patients over 65 who came to the medical consultations in June and July 1992 were selected, irrespective of the reasons for their visit; there were 17 males and 32 females, with and average age of 72.4 +/- 4.6 and 71.9 +/- 4.5. Only one patients consulted directly about loss of hearing. MEASUREMENTS AND MAIN RESULTS: A statistically significant association was found between the chronometry test and the audiometry test (gold-standard), significant at p < 0.005 for the left ear and p < 0.05 for the right ear. The test achieved a sensitivity of 0.85 and specificity of 0.76 in the case of the left ear, and 0.84 (sensitivity) and 0.56 (specificity) for the right ear. The kappa rate was 0.55 for the right ear and 0.67 for the left ear, showing agreement that went well beyond chance. CONCLUSIONS: In our view, the chronometry test constitutes an cheap, effective, simple, and replicable procedure for the detection of hypoacusia in patients aged over 65, and one which can usefully be used when no audioscope is available.

[Comparison of family physicians' and gynecologists' use of the intrauterine device (IUD)]. / Comparación en el uso del dispositivo intrauterino (DIU) entre médicos de familia y ginecólogos.

OBJECTIVE: To quantify differences between general practitioners (GPs) and gynaecologists in the technique of insertion and follow-up of the intra-uterine device (IUD). DESIGN: Multicentred, descriptive, longitudinal study. SETTING: Two urban health centres and a family guidance clinic. PARTICIPANTS: Target population (n = 1700) between January 1993 and January 1996. Estimated mean of complications was 25%. Sample size was 247 for alpha = 0.05 and 1-alpha = 0.95. The sample was extended to 300 to allow for possible losses of files, estimated at 20%. MEASUREMENTS AND MAIN RESULTS: The variables age, sex, marital status, educational level, parity, abortions, previous contraception, type of job, type of IUD, post-insertion and follow-up complications, subjective evaluation, removal and average follow-up time, were analysed. 158 (54.9%) of the 288 IUDs finally studied were inserted by GPs, and 130 (45.1%) by gynaecologists. 69.5% were anchor-shaped, and 30.5% T-shaped. In 85.5% no immediate complications were found. Mean follow-up time was 22.67 months (CI 95%, 21.3-24.0), during which time 36.6% had complications detected, which led to removal of the device in 22.3% of complications. We found no statistically significant differences between the two populations for age, marital status, subjective evaluation, number of abortions, parity or previous contraception. Likewise, no differences between G.P.s and gynaecologists were detected for post-insertion or follow-up complications, percentage of IUDs removed, or period of time evaluated. There were differences found for the type of IUD used, with more anchor-shaped IUDs in primary care. There were no differences for the type of IUD or complications requiring its removal. CONCLUSIONS: In the population studied we found no differences in immediate or later complications between IUDs inserted by GPs and by gynaecologists.