Effect of Disturbances of Zinc and Copper on the Physical and Mental Health Status of Patients with Alcohol Dependence.

The concentrations of copper and zinc in the tissues of alcohol-addicted people can significantly correlate with the variables describing their mental state. Studies on the homeostasis of zinc in alcohol-dependent patients have often been characterized by low hypozincemia detection. This may be caused by a low content of zinc in blood serum (1%) compared to the average zinc level in the body. Unfortunately, most authors have identified extracellular zinc in their studies. In the available literature, data on the level of copper in patients suffering from alcohol dependence are inconsistent. Our study included 100 alcohol-addicted patients (the study group) and 50 healthy subjects (the control group). Mental state was measured using appropriate psychometric scales. We used inductively coupled plasma mass spectrometry (ICP-MS) to determine copper and zinc content. Our results confirm the purposefulness of the use of zinc concentration in erythrocytes as a diagnostic parameter for low zinc status in alcohol-dependent patients. Alcohol-dependent patients with reduced concentrations of zinc in erythrocytes/copper in blood plasma differed significantly from alcohol-dependent patients with normal concentrations in terms of clinical parameters. With regard to zinc in blood plasma and copper in erythrocytes, this situation has not been found. The clinical symptoms of hypozincemia and copper deficiency in patients addicted to alcohol usually relate to disorders in central nervous system functioning, and they result in a decreased quality of physical and mental life.

The effect of opioid agonists and antagonists on gastrointestinal motility in mice selected for high and low swim stress-induced analgesia.

BACKGROUND: The opioid system in the gastrointestinal (GI) tract plays an important physiological role, but is also responsible for the side effect of opioid drugs, including troublesome constipation in chronic pain treatment. The aim of this study was to characterize and validate a new mouse model to study the effects of opioid agonists and antagonists in the GI tract. METHODS: Six-week-old male Swiss-Webster mice, divergently bred for high (HA) and low (LA) swim stress-induced analgesia (SSIA), were used in the study. To assess the influence of opioid agonists (morphine and loperamide) and antagonists (naloxone hydrochloride, NLX and naloxone methiodide, NLXM) on GI motility, whole GI transit (whole GIT) and upper GIT assays were conducted. To evaluate the expression of opioid receptors in the ileum and colon of HA and LA mice, immune staining was performed. KEY RESULTS: The effect of morphine was more pronounced in HA line, whereas loperamide exerted a stronger effect in LA mice. Furthermore, NLX and NLXM differentially abolished the inhibitory action of the central and peripheral opioid system on whole and upper GIT in HA and LA mice. The differences in GI motility between HA and LA mice coexisted with parallel changes in the expression of opioid receptors in the ileum and colon. CONCLUSIONS & INFERENCES: Differences in the activity of the endogenous opioid system are responsible for the vulnerability to changes in GI motility during treatment with opioids. Our findings validate the HA/LA model for further studies on opioids in the GI tract.

Genetic and environmental predictors of chronic kidney disease in patients with type 2 diabetes and diabetic foot ulcer: a pilot study.

Chronic kidney disease (CKD) is often observed among patients with type 2 diabetes mellitus (T2DM) and diabetic foot (DF) leading to end stage renal disease. The aim of this pilot study was to determine genetic and environmental factors involved in the etiology of CKD among patients with DF. The following polymorphisms were studied: rs1800469, rs759853, rs1553005, rs1799983, rs1801133, rs3134069, rs2073618, rs8192678, rs6330, rs11466112, rs121917832 in terms of alleles distribution in patients with DF and T2DM, with or without CKD. The study includes 101 patients with T2DM and DF. Studied groups were divided into 39 individuals with CKD (cases) and 62 controls, depending on the presence of kidney failure defined as eGFR < 60ml/min/1.73m(2) and coexistence of microalbuminuria > 30 mg/dl in at least 3 urine samples. Cases and controls were matched according to mean age, gender, mean duration of T2DM, mean duration of insulin therapy, mean duration of DF cholesterol levels and smoking frequencies. The study showed that CKD risk factors were the following variables: creatinine level, body weight, hips circumference, ischemic heart disease, hypertension and diabetic retinopathy. Moreover, the results suggest the protective role of the allele C of rs3134069 polymorphism in CKD development in patients with T2DM and DF in the following allelic variants: [AA] vs. [AC] and [AA] vs. [AC + CC]. The allele C was observed to be less frequent than the allele A in patients with T2DM and DF. None of the other following polymorphisms was observed to be a potential risk factor of CKD in T2DM and DF population: rs6330, rs759853, rs1553005, rs1799983, rs1801133, rs1800469, rs8192678, rs11466112, rs121917832. We concluded that the rs3134069 polymorphism seems to be the most likely protective genetic factor in CKD development in patients with T2DM and DF.

Sampling intervals verification in pharmacokinetics studies.

Regulatory agencies do not specify how to plan the sampling intervals in pharmacokinetics (PK) studies. Every interval between each sampling point forms one of the fractions of the area under the curve (AUC). The aim of this study is to propose a method of qualitative evaluation of PK studies, on the basis of the analysis of the partial AUC fields' values. For the pharmacokinetic analysis, average concentrations of high variability drug-itraconazole were used before (BO) and after sampling intervals optimization (AO). PK calculations were performed using Phoenix(TM) WinNonlin 6.3(®) (Certara L.P.) and in house software Biokinetica 4.0. Arithmetic formula and acceptance limit (AL%) was established, below which the mean of partial fields (MAF) value in PK study can be considered optimal. In case of MAF the CV% value before optimization was 125.35 and after the optimization 46.51. In the cases of AUC fractions for several partial fields BO data, the AL% value was exceeded. The values of AUC fractions did not exceed AL% established for AO data. The paper proposes an empirical method of quality assessment, made on the basis of the percentage of the AUC fractions. This method can be used in the quality assessment of PK studies.

Comparison of one-step nucleic acid amplification (OSNA) method and routine histological investigation for intraoperative detection of lymph node metastasis in Polish women with breast cancer.

Sentinel lymph node (SLN) biopsy is a part of the staging procedure in breast cancer patients. Intraoperative molecular analysis for SLN metastases using the one-step nucleic acid amplification (OSNA) method based on reverse-transcription loop-mediated amplification (RT-LAMP) has already been validated in breast cancer. In this study, we compare the intraoperative OSNA method to our routine histological investigation. To evaluate the performance of OSNA in comparison to histology, analysis of 74 SLN from 60 breast cancer patients was conducted with both methods. Of the 22 histologically positive samples, 14 were attributed to macrometastases (++) in the OSNA-CK19 assay and 8 to micrometastases (+). Two samples negative in histopathology were positive in the OSNA method (micrometastases +). Our results show that OSNA is an excellent method for the detection of metastases in lymph nodes and can be applied as an intraoperative diagnostic approach. Intraoperative molecular analysis for SLN metastases using the OSNA method reduces the number of admission days and duration of surgery. To our knowledge this is the first study referring to Polish women.

The association between polymorphisms of the RAD51-G135C, XRCC2-Arg188His and XRCC3-Thr241Met genes and clinico-pathologic features in breast cancer in Poland.

BACKGROUND: XRCC2 and XRCC3 genes are structurally and functionally related to RAD51 which plays an important role in homologous recombination, the process frequently involved in cancer transformation. MATERIAL AND METHODS: In the present work the distribution of genotypes and frequency of alleles of the RAD51 G135C polymorphism, XRCC2 Arg 188His and XRCC3 Thr241Met polymorphism in 790 cases of breast cancer were investigated. The control group consisted of 798 cancer-free blood donors (age +/- 5 years) who were sex and ethnicity-matched. The polymorphisms were determined by PCR-RFLP methods. We also correlated genotypes with the clinical characteristics of breast cancer patients. RESULTS: Our results obtained for the 135G>C polymorphism of the RAD51 gene indicated that both the C/C genotype and the C allele are strongly associated with breast cancer. The Arg/His genotype of XRCC2 (OR = 2.16, 95% CI = 1.48-3.16) and Thr/Met of XRCC3 increased the risk of type I breast cancer occurrence (OR = 2.33, 95% CI = 1.60-3.41). We did not find any association with the RAD51, XRCC2/3 gene polymorphism and estrogen and progesterone receptor status. CONCLUSION: The results support the hypothesis that the polymorphism of RAD51 and XRCC2/3 gene may be associated with the incidence of sporadic breast cancer in Polish women.

Association between polymorphisms of the BRCA2 gene and clinical parameters in breast cancer.

BACKGROUND: A C/T transition - rs4987117 (the Thr1915Met polymorphism) and an A/G transition - rs11571653 (the Met784Val polymorphism) in the BRCA2 gene were linked to breast cancer risk in Polish and Japanese populations, respectively. AIM: To study the association between polymorphisms of the BRCA2 gene and clinical parameters in breast cancer. METHODS: Both polymorphisms were evaluated by RFLP - PCR in blood samples obtained from 117 women with sporadic breast cancer. Patients were stratified by genotype, Bloom - Richardson grade, TNM stage, estrogene and progesterone receptors (PR) status and the linkages of each genotype with each stratum were calculated by logistic regression. RESULTS: Variant genotypes and alleles of both polymorphisms of the BRCA2 gene were inversely related to hormone receptor status for a group of patients with at least one positive receptor status as compared to a group with both receptors negative status (OR 0.27, 95% CI 0.07 - 0.95, p = 0.043 and OR 0.39, 95% CI 0.19 - 0.82, p = 0.013 for Met1915Met homozygote and 1915Met allele, respectively and OR 0.02, 95% CI 0.00 - 0.13, p = 0.0005 and OR 0.43, 95% CI 0.21 - 0.88, p = 0.021, for Val784Val homozygote and the 784Val allele. No association was found between both polymorphisms and Bloom - Richardson grading and TNM staging. CONCLUSIONS: Our results suggest that variant genotypes of the Thr1915Met and Met784Val polymorphisms of the BRCA2 gene may be indicative factors in therapy of ductal breast cancer.

Analysis of RAD51 polymorphism and BRCA1 mutations in Polish women with breast cancer.

AIM: Breast cancer is one of the major killers worldwide. The objectives of this study were to determine the frequency of BRCA1 germ-line mutations and the RAD51 G/C polymorphism in patients with breast cancer. METHODS: 100 breast cancer women provided blood for mutation analysis. Blood samples age matched healthy individuals (n = 106) served as control. The G/C polymorphism and BRCA1 mutations were determined by PCR-RFLP methods. RESULTS: The distribution of the genotypes of the G/C polymorphism RAD51 in both control and patients did not differ significantly from those predicted by the Hardy - Weinberg distribution. There were no significant differences in the genotype distributions and allele frequencies between node-positive and node-negative patients. In present study one Ex20insC mutations of BRCA1 gene was identified in women with breast cancer. CONCLUSION: Our study implies that the G/C polymorphism of the RAD51 gene may not be directly involved in the development and=or progression of breast cancer.

Patients with Paget disease of nipple and with palpable mass in breast have unfavorable prognosis.

Recent data indicate that in breast cancer patients the presence of Paget disease of the nipple may be related to poor prognosis. Therefore, we decided to assess long-term results of the treatment of such patients, and to assess the relationship between the physical and pathological findings and prognosis. The files of 60 patients with Paget disease of the nipple who were treated between 1977 and 2000 were analyzed retrospectively with respect to the results of physical and pathologic examinations, disease recurrence and survival. In 38/60 patients, the cancer was invasive. In 26/60 patients, palpable masses in the breast were diagnosed. The 5-year overall survival probability was 0.68; the probability was 0.82 for patients without palpable masses, 0.51 for those with palpable masses, 0.91 for patients without invasive cancer and 0.58 for patients with associated invasive cancer. In conclusions, patients with Paget disease of the nipple and with palpable mass in the breast had unfavorable diagnosis. The nature of all nipple changes should be explained as early as possible in order to diagnose the disease when no mass is palpable in the breast.

An association between the matrix metalloproteinase 1 promoter gene polymorphism and lymphnode metastasis in breast cancer.

A single guanine insertion (1G/2G polymorphism) in the promoter of the matrix metalloproteinase (MMP-1) gene creates a binding site for the transcription factor and may affect the level of transcription of MMP-1. An elevated level of MMP-1 in cancer cells may facilitate their invasion and contribute to metastasis. To evaluate the contribution of 1G/2G polymorphism in the development and/or progression of breast cancer we genotyped 135 subjects with breast cancer. The 1G/2G polymorphism was determined by the method based on restriction endonuclease digestion. We found that the frequency of the 2G allele was higher in lymphnode-metastasis patients than in the group without metastasis (p < 0.001). We did not find differences between distribution of the genotypes and frequencies of alleles in cancer patients and in healthy subjects served as control. Our results suggest that allele 2G may be associated with lymphnode metastasis in patients with breast cancer and therefore it can be considered as a prognostic marker in this disease.