Diagnosis of biofilm infections: current methods used, challenges and perspectives for the future.

The diagnosis of biofilms continues to be a challenge, and there is no standardized protocol for such a diagnosis in clinical practice. In addition, some proposed methodologies are expensive to require significant amounts of time and a high number of trained staff, making them impracticable for clinical practice. In recent years, mass spectrophotometry/matrix-assisted laser desorption ionization time of flight (MALDI-TOF) has been applied it in biofilm studies. However, due to several problems and limitations of the technique, MALDI-TOF is far from being the gold standard for identifying biofilm formation. The omics analysis may prove to be a promising strategy for the diagnosis of biofilms in clinical laboratories since it allows the identification of pathogens in less time than needed for conventional techniques and in a more specific manner. However, omic tools are expensive and require qualified technical expertise, and an analysis of the data obtained needs to be careful not to neglect subpopulations in the biofilm. More studies must therefore be developed for creating a protocol that guarantees rapid biofilm identification, ensuring greater chances of success in infection control. This review discusses the current methods of microbial biofilm detection and future perspectives for its diagnosis in clinical practice.

Sub-ångstrom experimental validation of molecular dynamics for predictive modeling of extended defect structures in Si.

In this Letter we present the detailed, quantitative comparison between experimentally and theoretically derived structures of the extended {311} defect in silicon. Agreement between experimental and theoretical column positions of better than ±0.05 nm has been achieved for all 100 atomic columns in the defect structure. This represents a calculated density of 5.5×10(14) silicon interstitials per cm(2) on {311} planes, in agreement with previous work [S. Takeda, Jpn. J. Appl. Phys., Part 2, 30, L639 (1991)]. We show that although the {311} defect is made up of five-, six-, seven-, and eight-member rings, the shape of these rings varies as a function of position along the defect, and these variations can be determined experimentally with high precision and accuracy. The excellent agreement between the calculated and experimentally derived structure, including the position of atomic columns and the shape of the distinct structural units of the defect, provides strong evidence for the quality and robustness of the molecular dynamics simulation approach for structural studies of defects. The experimental approach is straightforward, without the need for complicated image processing methods, and is therefore widely applicable.

Apoptotic and cell cycle response to homoharringtonine and harringtonine in wild and mutant p53 hepatocarcinoma cells.

This study aimed to evaluate the modes of action of harringtonine (HT) and homoharringtonine (HHT) alkaloids in cell with wild (HepG2/C3A) and mutant p53 (HuH-7.5). We performed assays for cytotoxicity, genotoxicity, induction of apoptosis, cell cycle phase, and membrane integrity. Obtained data were compared with the relative expression of mRNA of genes related to proliferation, apoptosis, cell cycle control, metabolism of xenobiotics, and reticulum endoplasmic stress. The relative expression of the genes showed an increase in apoptosis-inducing mRNAs, such as TNF and BBC3, as well as a reduction in BCL2 and BAK. The mRNAs of CYP2E1 and CYP2C19 xenobiotic metabolism genes increased in both lineages, while CYP3A4 increased only in the HuH-7.5 lineage. The mRNA expression of endoplasmic reticulum (ER) stress genes (ERN1 and EIF2AK3) was shown to increase in HHT and HT treatments. A similar increase was recorded in the mRNA expression of the TRAF2 gene. The changes observed in this study support the hypothesis that ER stress was more strongly associated with TNF induction, causing cell death by apoptosis in p53 mutant cells. This result with wild and mutant p53 cells may have clinical implications in the use of these compounds.

[Evaluation of depressive symptoms and resilience in patients on pharmacotherapeutic follow-up]. / Evaluación de síntomas depresivos y resiliencia en pacientes bajo seguimiento farmacoterapéutico.

This study aimed to carry out the pharmacotherapeutic follow-up of patients with depression and to assess its impact on the resilience of the patients. Patients were followed-up for 8 months. The pharmacist evaluated depressive symptoms, resilience, and the need for pharmaceutical intervention. The measurement tools used were the Dader method, PHQ-9, and a resilience scale. Data were analysed using BioStat 5.0 software and the performing of the Wilcoxon and Pearson correlation tests. There was a reduction in the rate of depressive symptoms from 12.9 to 5.2 (P<.0001), and an increase in the resilience score from 112.4 to 149.0 (P<.0001). Pharmaceutical interventions were made to resolve the drug related problems in the form of oral communication between pharmacist-patient or pharmacist-patient-doctor. The pharmaceutical care was effective in decreasing depression and contributed to the increased resilience of patients.

Independent prognostic value of laminin receptor expression in breast cancer survival.

We studied the possible prognostic role of laminin, estrogen (ER), and progesterone (PR) receptors and other pathological factors in relation to the disease-free interval and overall survival of female breast carcinoma patients. Multivariate analyses of clinical and pathological data with respect to the above survival time variables were performed by Cox regression. The statistical dependence of prognosis on ER, PR, and tumor size was based on the discriminant cutoff value that could best distinguish between survival curves. Axillary nodal status was the most significant independent factor in the prediction of both disease-free interval and overall survival of these patients. Use of the information on laminin receptor expression, PR concentration, tumor size, lymphocytic infiltrate, and tumor necrosis improved significantly the prediction of the risk of recurrence. Patients with tumors expressing laminin receptors had 40% less risk of recurrence (P = 0.0209) than those with no expression. On the other hand, four covariates were independently predictive of the risk of death: axillary nodal status, lymphocytic infiltrate, PR and ER concentration. There was a marginally significant (10% level) interaction between tumor size and lymphocytic infiltrate with respect to the prediction of the risk of recurrence. The above sets of variables were used to classify patients into risk groups for the prediction of recurrence and death.

A prospective randomized evaluation of three schedules of mesna administration in patients receiving an ifosfamide-containing chemotherapy regimen: sustained efficiency and simplified administration.

Chemotherapy with oxazaphosphorines, such as ifosfamide, is often limited by unacceptable urotoxicity. Without uroprotection hemorrhagic cystitis becomes dose-limiting. Mesna, a thiol compound, is a drug able to bind the toxic metabolites, forming nontoxic compounds in the urine. A total of 122 patients were enrolled in this study and 228 chemotherapy cycles with an ifosfamide-containing regimen were performed (225 evaluable). Mesna was given at the same total dose as the ifosfamide in all arms. On arm A, mesna was given i. v. in equal doses 15 min before and 4 h and 8 h following the ifosfamide dose. On arm B, mesna was given in three equivalent doses 15 min before (i.v.) and 4 h (i.v.) and 8 h (p.o., double dose) following ifosfamide. On arm C, mesna was given i.v. in two equal doses given 15 min before and 4 h following. The incidence of urotoxicity was very low (lower than 15%) in the three arms, 0% in A, 1.36% in B and 2.70% in C. All three arms were equally efficient. Schedule C was considered superior to the others, since it was equally effective, simpler and more convenient.

Tumor and serum beta-2-microglobulin expression in women with breast cancer.

To investigate whether the tumor expression of beta-2-microglobulin (beta 2-M) could serve as a marker of tumor biologic behavior, the authors studied specimens of breast carcinomas from 60 consecutive female patients. Presence of beta 2-M was analyzed by immunohistochemistry. No significant correlations were found between tumor beta 2-M expression and several histologic attributes such as type, histologic and nuclear grades, mitotic index, necrosis, vascular invasion, and lymphocytic infiltration. Likewise, beta 2-M was not associated with markers of disease extension such as TNM, (UICC, classification of malignant tumors) staging and axillary lymph node involvement or with estrogen, progesterone, and glucocorticoid receptor levels. However, there was a significantly positive association between tumor beta 2-M expression and the degree of lymphocytic infiltration in the tumor tissue. Beta 2-M serum levels were determined by an enzyme-linked immunosorbent assay in samples from 22 of the above women. Although some of the highest values had been obtained in women with larger (T4) primary tumors, the authors failed to detect any statistical relationship between beta 2-M expression in the tumor with serum levels or between serum beta 2-M and the above histologic, laboratory, and clinical factors.

Identification of the external branch of the superior laryngeal nerve during thyroidectomy.

Seventy-six patients underwent preoperative vocal evaluation and were randomized into 3 groups: (1) those with the superior thyroid pole dissected by the first author, with the external branch of the superior laryngeal nerve (EBSLN) identified by means of a nerve stimulator; (2) those patients whose dissection was executed by a resident, with no nerve search; and (3) those whose dissection was undertaken by the first author, without any nerve search. Postoperative analysis consisted of voice evaluation and electromyography of the cricothyroid muscle. No lesion occurred in patients in group 1. Twenty-eight percent of patients in group 2 and 12% in group 3 experienced a complete lesion of the EBSLN (p = 0.0123). When the patients in group 1 were compared with the patients with 62 nerves corresponding to nonoperated thyroid lobes, patients in group 1 exhibited no increased risk, whereas a significantly increased hazard was evident in both groups 2 (p = 0.0002776) and 3 (p = 0.0346393). In this study, effective prevention of iatrogenic EBSLN lesions during thyroidectomies was achieved only by the intraoperative identification of the nerve with the nerve stimulator.

Clinical and laboratory signs of reperfusion: are they reliable?

We studied 101 patients (88 men and 13 women, mean age 54.5 +/- 10 years) who arrived at the hospital during the first 6 hours of acute myocardial infarction evolution. Our objective was to assess the reliability of clinical and laboratory signs of recanalization using intravenous streptokinase as a thrombolytic agent. The mean time between the beginning of infusion and coronary arteriography was 53.83 +/- 43 hours. The positive predictive values for pain, arrhythmia, ST segment and enzymes were 97.9%, 94.2%, 91.8% and 90.8%, respectively; the negative predictive values were 46.8%, 40.8%, 37.2%, and 50% in the same order. Sensitivity was 65.7%, 62.8%, 58.4% and 77.6% and specificity 95.6%, 86.9%, 82.6% and 73.9%, respectively. The positive predictive value, calculated on the basis of the presence of each variable alone or in association showed a probability of recanalization of 76.9% for one sign, 84% for two, 96.3% for three and 100% for all four. When we compared the positive predictive values of each variable according to the interval between the beginning of pain and admission to the hospital (during the first 3 hours or between 3 and 6 hours) our results were 100%/94% for pain (P = NS), 97%/88% for arrhythmia (P = NS), 100%/75% for ST segment (P = 0.004), and 97%/80% for enzymes (P = 0.019). The same analysis applied to negative predictive values showing 22%/62% (P = 0.007), 17%/55% (P = 0.008), 21%/47% (P = NS), 27%/61% (P = NS) for pain, arrhythmia, ST segment and enzymes, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

[Effects of denervation of the soleus muscle of rats]. / Efeitos da denervação em músculo soleus de ratos.

Histological aspects and changes in wet weight were studied on rat soleus muscles after denervation by cutting the sciatic nerve. The muscles were removed after a variable period of time from the third to the fiftieth day. There was a progressive fall off in wet weight and after the third week the muscles underwent 50% atrophy. Histological changes were most striking on sarcolemmal nuclei; they became vesicular, enlarged with prominent nucleolus and there was an apparent increase in their number. The muscle fibres changed their shape, becoming rounded and there was a 50% reduction in muscle fibre diameter after 50 days. Connective tissue abnormalities were minimal after 50 days and the striations remained intact.

Pharmaceutical care to patients treated with antidepressants.

Depression is considered one of the ten most disabling conditions on the planet, causing physical, personal and social limitations. Antidepressants are the most commonly used drug treatment for eliminating or controlling the symptoms of depression. A variety of factors can contribute to treatment abandonment, particularly severe side effects. The present article is aimed to provide information and guidelines on the development of a pharmacotherapy follow-up program for patients treated with antidepressant medication. According to this practice, the pharmacist evaluates whether the three following requirements of a proper pharmacotherapy are being observed: need, effectiveness and safety. If one of these requirements is not met, the pharmacist seeks to solve the problem, by intervening in the pharmacotherapy, within the scope of his/her skills. Pharmaceutical care is aimed to improve adherence to treatment and minimize side effects, as well as the occurrence of drug interactions. Thus, the present article presents and discusses the main strategies of pharmaceutical care to achieve the proposed objectives.

A influência do acompanhamento farmacoterapêutico na adesão à terapia anti-hipertensiva e no grau de satisfação do paciente / Influence of pharmacotherapeutic follow-up on patient compliance and satisfaction with antihypertensive therapy

O objetivo desta pesquisa foi proporcionar o serviço de Acompanhamento Farmacoterapêutico (AFT) a pacientes hipertensos usuários da Farmácia Popular de Alfenas (MG), além de avaliar a adesão do usuário aos medicamentos antes e após o AFT e o grau de satisfação com o serviço prestado. O trabalho foi realizado com 27 pacientes de ambos os sexos e com faixa etária entre 40 e 70 anos ou mais. Os pacientes foram divididos em grupo controle e teste, ambos responderam ao questionário de adesão no início do estudo e, ao final, apenas os pacientes do grupo teste o fizeram. Os pacientes do grupo teste receberam Acompanhamento Farmacoterapêutico durante dez meses (2008) e, ao final desse período, responderam o questionário de satisfação pelo serviço. Um dos motivos mais importantes para a não adesão foi o esquecimento de administrar o medicamento (67% para ambos os grupos). Outros fatores, como a falta de informação sobre a doença e a não compreensão sobre como utilizar os medicamentos, foram sanados com a intervenção farmacêutica. Dos pacientes participantes desta pesquisa, 80% achavam importante o trabalho conjunto do farmacêutico com o médico e 100% disseram que continuariam a utilizar o serviço prestado e o indicariam a amigos e parentes. Os dados sugerem que as intervenções farmacêuticas foram efetivas no sentido de aumentar a adesão no grupo que recebeu AFt e que os pacientes ficaram satisfeitos com o serviço prestado.

This research was designed to provide a Pharmacotherapeutic Follow-up (PF) service to hypertensive patients who attend the Popular Pharmacy in Alfenas (Minas Gerais, Brazil) and to assess user adherence to medication before and after the PF and the degree of satisfaction with the service rendered. This study was carried out with 27 patients of both sexes, aged 40 years and above. The patients were divided into two groups, control and test; all patients answered the adherence questionnaire at the beginning of the study and, at the end, only those in the test group. The patients in the test group received PF for ten months (2008) and, at the end of this period, they were asked to complete a questionnaire regarding their satisfaction with the service. One of the commonest reasons given for non-adherence to medication was forgetfulness (67% for both groups). Other problems, such as lack of information on the disease and poor understanding of how to take the medicines, were solved by pharmaceutical intervention. According to 80% of the patients involved, this teamwork between pharmacist and physician is important, and 100% reported they would continue to use the service and would recommend it to friends and relatives. The data suggest that pharmaceutical interventions were effective in improving adherence to medication among patients in the group that received PF and that the patients were satisfied with the service given.

Soft-tissue sarcomas of the head and neck in adults and children: experience at a single institution with a review of literature.

Ninety-four soft-tissue sarcomas of head and neck occurring among 10,700 malignancies, seen during the period 1953-1980 in the head-and-neck surgical department, were studied with reference to age, sex, ethnic origin, site, histological type and survival. The objective was to review our own experience with sarcomas in these sites seen in the Department of Pathology, A.C. Camargo Hospital. The predominant site was the head. Fibrosarcoma was the most frequent type in adults and rhabdomyosarcoma, located exclusively in the head, was the most frequent in children.

Genetic alterations in c-erbB-2 protooncogene as prognostic markers in human primary breast tumors.

c-erbB-2 DNA amplification and mRNA expression were analyzed by dot and Southern blots in 65 human primary breast tumors. Gene amplification was observed in 21 of 65 (32.3%) and elevated levels of c-erbB-2 transcript in 14 of 60 (23.3%) of the tumors analyzed. Only 55% of the tumors with c-erbB-2 gene amplification presented gene overexpression, showing an incomplete correlation between gene amplification and overexpression. No statistically significant correlation was observed between c-erbB-2 genetic alterations and other prognostic factors in breast cancer. However, patients with tumors presenting c-erbB-2 gene amplification and/or overexpression appeared to have a shorter disease-free interval than patients without c-erbB-2 genetic alterations. High levels of c-erbB-2 gene amplification were more powerful predictors of risk of recurrence than was overexpression of the gene. Cox univariate-bivariate analyses suggested that gene amplification was independent of nodal status to predict recurrence in breast cancer.

Estrogen and progesterone receptor mRNA levels in primary breast cancer: association with patient survival and other clinical and tumor features.

The relative expression of estrogen receptor (ER) and progesterone receptor (PR) mRNA transcripts was measured in 71 primary breast-cancer biopsies. ER and PR binding activity were estimated in parallel by the dextran-coated-charcoal method. There was a close correlation between the amount of ER mRNA and estradiol binding activity. Tumors from post-menopausal patients contained higher levels of ER mRNA than those from pre-menopausal patients. Northern-blot analysis indicated the presence of a major band of 6.3 kb in all ER mRNA-positive tumors. Some tumors showed, in addition, 3.7- and 2.4-kb transcripts. PR binding activity and overall PR mRNA levels correlated moderately. PR mRNA and ER mRNA were associated. Four PR mRNA species with estimated sizes of 11.4, 4.5, 3.7 and 2.5 kb were detected in 14% of the PR mRNA-positive tumors. The 3.7-kb transcript was detected to varying degrees in all PR mRNA-positive biopsies, accompanied in some tumors by the 2.5-kb species. ER and PR mRNA levels > or = 50 pg/5 micrograms total RNA correlated with prolonged survival of the patients. In addition, high ER mRNA levels were associated with absence or minimal necrosis and vascular invasion together with absence or minimal level of tumor lymphocytic infiltration, but not with age, clinical stage, tumor size or overexpression of c-myc or c-erbB-2 mRNA. PR mRNA was not statistically associated with any of the above clinicopathological features. A bivariate analysis showed that both ER and PR mRNA levels were able to predict overall survival independently of the lymph-node status.

Epidemiological and clinical correlations with genetic characteristics of Wilms' tumor: results of the Brazilian Wilms' Tumor Study Group.

The epidemiological and clinical features of Wilms' tumor (WT) were analyzed in 176 patients admitted to the Brazilian WT Study Group. The occurrence of congenital anomalies (9.1%) and the M:F ratio (0.83) were comparable to those observed by the US National WT Study and the International Society of Pediatric Oncology trials. Bilateral cases were younger on average than unilateral cases (37.2 vs. 45.0 months). Ethnic group and gender were also associated with age, with non-white children being generally older (46.1 months) than whites (39.5 months), and boys being younger than girls (37.0 vs. 46.1 months). However, the most important factor in association with age at diagnosis was disease stage. No early disease patients were diagnosed after 8 years of age (mean: 37.5 months), whereas 10% of those with advanced disease were diagnosed between the ages of 8 and 10 years (mean: 56.3 months). There were no clear distinctions in age distributions on the basis of the presence of tumor multicentricity, intra- or perilobar nephroblastomatosis, and of a combination of putative genetic determinants. The mean age at diagnosis for cases with congenital defects was higher than that for the remaining patients. Some of the study results are in support of the recessive oncogene model for WT. However, the possible confounding effect of disease stage and the strong interdependence of the remaining factors may be masking important relations in regard to mutational events occurring during embryogenesis.

Plasminogen activator expression and steroid hormone receptors in female breast cancer: a multifactorial study.

In order to evaluate the relationship of plasminogen activator (PA) activity to the expression of estrogen (ER) and progesterone (PR) receptors, we assayed primary tumor specimens from 121 cases of female breast cancer. Other clinical and histopathological variables were also investigated with respect to their possible association with PA activity in the samples. Statistical correlations were examined by stratified analysis techniques and by multiple regression methods. PA activity was higher in tumors which were ER+PR+ than in those exhibiting other subsets of joint receptor expression, a finding which was particularly predominant in tumors from post-menopausal women. Of the other variables examined, only disease stage, nodal status and vascular infiltration presented marginal positive statistical associations with PA activity, although this was not confirmed by multivariate analysis. The latter technique showed that the presence of PR was sufficient to explain the variation in PA levels. However, ER emerged as the sole significantly explanatory factor when ER+PR- patients were removed from analysis. Both PR and age (negatively) were independent contributory factors for PA activity in the analysis of post-menopausal women. None of the variables examined emerged as being significantly associated with PA when data from pre-menopausal patients were used. These findings indicate that PA activity in breast tumor samples is statistically associated with the expression of functional estradiol receptors, although to a lesser extent than PR.

Surgical treatment of colon and rectum adenocarcinoma in elderly patients.

The results are presented of disease-free interval and overall survival in 53 elderly patients with colon or rectum adenocarcinoma treated with curative surgery. The average age was 75.3 years (median = 75.0); 21 patients were male and 32 were female. Tumor location was as follows: rectum 18 (34%), sigmoid 17 (32.1%), right colon 14 (26.4%), and transverse colon 4 (7.5%) All patients were surgically treated following the classical patterns for tumor resection. After pathological examination, which included the histological grade differentiation, the disease stage was reevaluated following the pTNM system. Overall and disease-free survival at 5 years, for all patients, independent of histological grade differentiation and disease stage, were 75.3% and 55.5%, respectively. Overall survival at 5 years for patients with grades I and II histological differentiation was 74.1% and 85.0%, respectively. None of the grade III patients (2 cases) survived more than 1 year. The 5 years disease-free survival for patients with histological grade differentiation I and II was 56.8% and 60%, respectively. There was no statistically significant difference in overall survival for patients with stages SI, SII, and SIII, but the disease-free survival at 5 years by stages was found to be significant with rates of 100%, 67.6%, and 22.6%. Postoperative mortality was 4 (7.5%). The postoperative mortality and survival rates obtained in this group of patients encourage us not to consider age as a limiting factor for curative surgical treatment.