Immunohistochemically detected high expression of matrix metalloproteinase-2 as predictor of poor prognosis in Duke's B colon cancer

Aim

To demonstrate immunohistochemical expression of matrix metalloproteinase-2 (MMP-2) protein in Duke’s B colon cancer and determine its correlation with age, sex, grade, presence of vascular invasion, and patients’ overall survival.

Method

The study took place from January 1995 to December 1997. We determined the expression of MMP-2 in 152 formalin-fixed, paraffin embedded specimens of Duke’s B colon carcinomas by immunohistochemical analysis using MMP-2 monoclonal antibody. Immunohistochemical expresssion was scored semiquantitatively. Carcinomas were graded as low or high grade. Survival time was analyzed with Kaplan-Meier method, and the log-rank test was used to assess the differences between groups. Cox proportional hazard regression model was used for multivariate survival analysis.

Result

Univariate analysis showed that positive staining for MMP-2, high histological grade, vascular invasion, male sex, and age >60 years were associated with shorter survival in patients with Duke’s B colon cancer (P range from 0.023 to <0.001). Multivariate analysis showed that only MMP-2 overexpression (P < 0.001; hazard ratio [HR] = 3.64) and vascular invasion (P < 0.001; HR = 4.27) were associated with shorter overall survival.

Conclusion

Expression of MMP-2 is an important independent indicator of shorter survival in patients with Duke’s B colon cancer and should be taken into consideration in decision-making on the use of adjuvant systemic therapy in patients with Duke’s B colon cancer.Colorectal carcinoma (CRC) is the third leading cause of cancer-related mortality in developed countries (1). Despite improvements in surgical and adjuvant chemotherapy treatment, mortality from CRC in Western countries remains high, with metastatic spread to the liver occurring in about 50% of patients (2). Although staging remains the most widely used prognostic indicator for CRC, increasing evidence suggests that it is not sufficient for predicting the clinical outcome of these patients (2). This applies especially to patients with intermediate stage diseases (Duke’s B, T3-4N0M0), since clinical management for them has yet to be standardized (2). Clinical staging may be supplemented by the use of biological prognostic markers for invasion and metastasis. They may provide important information needed for the implementation of various novel therapeutic strategies for controlling disease progression and tumor cell dissemination (2).Tumor cell invasion and metastasis are multi-step phenomena, involving the proteolytic degradation of the basement membrane and the extracellular matrix, altered cell adhesion, and physical movement of tumor cells. It was shown that degradation of basement membrane and extracellular matrix play a crucial role in tumor invasion and metastasis (3).Tumor cells secrete proteolytic enzymes or induce host cells to secrete proteases. Extracellular matrix degradation by proteases takes place not only in local invasion, but also in several stages of metastatic cascade, including angiogenesis, intravasation, and extravasation. The proteases involved in extracellular matrix degradation in tumor invasion and metastasis are subdivided into four classes as follows: serine, cysteine, aspartic, and matrix metalloproteinases (MMPs). MMP-2 is responsible for degradation of collagen type IV, which is the major structural protein in the basement membrane. Therefore, activation of MMP-2 is a crucial step in triggering the cascade of tumor invasion and metastasis (4).The prognostic significance of MMP-2 overexpression in humans has been shown in breast cancer (5), head and neck tumors (6), and ovarian carcinomas (7).The aim of this study was to determine the expression of MMP-2 using immunohistochemical methods in a subpopulation of patients with Duke’s B colon cancer and examine its relationship with clinicopathological parameters and patient survival.     

Leptin levels in infertile male patients are correlated with inhibin B, testosterone and SHBG but not with sperm characteristics

In the present study, differences in leptin levels between different groups of male patients presenting with infertility problems and possible correlations between leptin levels and clinical, spermiological, histological and hormonal characteristics were examined. Two hundred and ten male partners from infertile couples were included in the study. Based on clinical examination, spermiogram and testicular histology results, patients were divided into four groups: 42 men with non-obstructive azoospermia, 15 men with obstructive azoospermia, 68 men with oligoasthenoteratozoospermia and 85 men with normozoospermia. Serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B, testosterone, sex hormone binding globulin (SHBG) and leptin were measured. After adjustment for body mass index, there was a negative correlation between serum levels of leptin and inhibin B, total testosterone and SHBG (r = -0.189, p = 0.009, r = -0.250, p = 0.001 and r =-0.221, p = 0.003 respectively) but there was no correlation between leptin and classical sperm characteristics. Our results therefore demonstrate a link between leptin and testicular function, independently of FSH and LH, possibly involving testosterone and SHBG through a regulation of Leydig cell function.     

Personality traits in miners with past occupational elemental mercury exposure

In this study, we evaluated the impact of long-term occupational exposure to elemental mercury vapor (Hg0) on the personality traits of ex-mercury miners. Study groups included 53 ex-miners previously exposed to Hg0 and 53 age-matched controls. Miners and controls completed the self-reporting Eysenck Personality Questionnaire and the Emotional States Questionnaire. The relationship between the indices of past occupational exposure and the observed personality traits was evaluated using Pearson's correlation coefficient and on a subgroup level by machine learning methods (regression trees). The ex-mercury miners were intermittently exposed to Hg0 for a period of 7-31 years. The means of exposure-cycle urine mercury (U-Hg) concentrations ranged from 20 to 120 microg/L. The results obtained indicate that ex-miners tend to be more introverted and sincere, more depressive, more rigid in expressing their emotions and are likely to have more negative self-concepts than controls, but no correlations were found with the indices of past occupational exposure. Despite certain limitations, results obtained by the regression tree suggest that higher alcohol consumption per se and long-term intermittent, moderate exposure to Hg0 (exposure cycle mean U-Hg concentrations > 38.7 < 53.5 microg/L) in interaction with alcohol remain a plausible explanation for the depression associated with negative self-concept found in subgroups of ex-mercury miners. This could be one of the reason for the higher risk of suicide among miners of the Idrija Mercury Mine in the last 45 years.     

Assessment of Attention Deficit/Hyperactivity Disorder among children in families and schools in Mediterranean and continental Croatian towns

Among one of the biggest difficulties in diagnosis and treatment of Attention Deficit/Hyperactivity Disorder (AD/HD) is firstly the involvement of different persons, especially parents and teachers, who, together with experts, make the unavoidable subjects in the process of detecting and treating children with AD/HD. Adding to that the standards of living environment, the process becomes even more complex. The aim of this research is to detect if there is a difference in the assessment of AD/HD, given by the parents and teachers in two urban areas, which have culturally and geographically different characteristics (Middle European and Mediterranean). The assessments of children's behavior were conducted in two randomly selected schools; one in Zadar (Mediterranean town on the Adriatic coast) and the other in Zagreb (Middle European city). The children were evaluated by the experts using DSM IV as a gold standard, and by the parents and teachers using Conners rating scales. Analysis of results showed that there were differences in assessment of AD/HD among experts and parents/teachers, and among parents from two towns. Different results given by parents in Zadar and Zagreb, we reasoned, were obtained because of different sensitivity to the problems of hyperactive/impulsive/attention deficiency children's behavior.     

Prognostic value of S100B protein in carbon monoxide-poisoned rats

OBJECTIVE: To assess the possible role of S100B, a structural protein of astroglial cells, as a biochemical marker in acute carbon monoxide-poisoned rats and to compare its prognostic value with consciousness level, which is one of the major parameters for treatment decision in acute carbon monoxide poisoning. DESIGN: Nonrandomized, controlled interventional trial. SETTING: University laboratory. SUBJECTS: Male Wistar rats weighing 263 +/- 18 g. INTERVENTIONS: The rats were exposed to a mixture of 3000 ppm carbon monoxide in air for 60 mins (group 1) and a mixture of 5000 ppm carbon monoxide in air for 30 mins (group 2). Blood samples were taken from the jugular vein just before and immediately after the carbon monoxide poisoning. The level of consciousness was evaluated at the end of the exposure, and the survival rate was monitored for 7 days. The S100B concentrations were measured with a commercial immunoluminometric assay. MEASUREMENTS AND MAIN RESULTS: In the first group, the unconscious rats after carbon monoxide exposure had significantly higher S100B levels compared with the rats without loss of consciousness. In the second group, the unconscious rats that later died had significantly higher S100B levels compared with the unconscious rats that survived. The S100B levels of all conscious and unconscious surviving rats were not significantly different. The serum level of S100B below 0.44 microg/L predicted survival of carbon monoxide-poisoned rats, with a sensitivity of 100% and a specificity of 86%. CONCLUSIONS: Acute carbon monoxide poisoning is associated with elevated S100B levels. S100B is a better predictor of final outcome than the consciousness level, so it could be used as a prognostic parameter for acute carbon monoxide poisoning in rats.     

Emotional and physical abuse in family: survey among high school adolescents

Aim

To determine the prevalence of different forms of child abuse among high school pupils in Slavonski Brod, Croatia.

Method

The study included 2140 first and fourth-grade pupils aged between 14 and 18 years from all 10 high schools in Slavonski Brod and the area (4 grammar and 6 vocational schools). The pupils were asked to complete an anonymous structured questionnaire during a 45-minute class. The questionnaire, developed for the needs of this study, collected basic demographic data on family life and experience of emotional or physical abuse.

Results

First-grade pupils were more satisfied with their family life than fourth-grade pupils (96.9% vs 91.3%, P<0.001, χ² test). The feeling of being insufficiently loved or unloved was reported by 17.5% of the first-grade and 24.6% of the fourth-grade pupils and a greater percentage of pupils whose parents were divorced or who had a stepparent. Almost 80% of pupils had been verbally or nonverbally punished for disobedience. Emotional abuse was significantly associated with female sex (Nagelkerke R² = 0.87, β = 0.474, P = 0.028), younger age (β = 1.263, P<0.001), and alcoholism in the family (β = 2.037, P<0.001. Physical punishment for disobedience was reported significantly more often by first-grade than fourth-grade pupils (15.6% vs 12.9% P = 0.021, χ² test). Physical abuse was significantly associated (Nagelkerke R² = 0.69) with younger age (β = 0.379, P<0.012), emotional abuse (β = 0.665, P<0.002), alcoholism in the family (β = 1.791, P<0.001) and the lack of parental love (β = -0.645, P<0.001). Possible sexual molestation was admitted by 6.0% boys and 3.3% girls.

Conclusion

Most high school pupils in Slavonski Brod were satisfied with their life at home. Disobedience was usually punished verbally or by aggressive behavior of the parents. Physical punishment was less common and usually did not result in serious injuries.Unfit parents negatively affect the child’s emotional development, which leads to behavioral problems (1-4). Most parents have no intention to hurt their child on purpose, but sometimes they do it out of lack of experience or parenting skills (1). Unfit parents quite often acquired dysfunctional behavior patterns in their childhood (1).Main forms of child abuse include physical, emotional, and sexual harassment, and neglect (5,6). The signs of physical abuse are usually visible. Emotional abuse may take many forms, such as ignoring, belittling, or intimidation. It negatively affects the development, distorts the positive self-image of the child, leaves long-term consequences, and may lead to risky or aggressive behavior in adolescence or adulthood (5). Abuse is defined as a permanent type of behavior that “erodes and corrodes” the child’s integrity (7). In the strict sense, the term does not include occasional spanking, or “losing patience” with the child. Fit mothers react inadequately in 10% of total time spent with the child, as opposed to unfit mothers who do so in 80%-90% of the time (7).Long-term negative family conditions are predictive of aggressive behavior, anxiety, depression, delinquency, and social problems in adolescence or adult life (8), for example adolescent pregnancy (9,10), risky sexual behavior and drug addiction (11-13), emotional vulnerability (14), lower verbal intelligence quotient (15), alcoholism, suicidal behavior, and job abandonment (11,16-18). According to the US Department of Health and Human Services, the prevalence of abuse in the USA for the year 2000 was about 12.3/1000 children (19). More than 60% of children were neglected, 20% were physically, 10% sexually, and 7% emotionally abused. The highest percentage of victims were infants and children up to 3 years of age, more often girls than boys, and more than 80% of abusers were parents (19).According to a retrospective study from the United States that involved 1000 adults, about 30% of women and more than 40% men reported to have experienced some form of abuse in childhood; 5% reported to have been sexually and as many as 19% physically abused (20). In Thailand, as many as 77% of sixth graders in the elementary school were physically punished by whipping or lashing with a belt, and 95% were emotionally harassed by being yelled at or humiliated in various ways (21). In Croatia, the number of abused children has increased by 74% over the last 10 years, and the prevalence of sexual child abuse has tripled (22). According to the Croatian Ministry of Internal Affairs, 13 698 cases of child abuse were recorded in the period between 1991 and 1995, and only 10% of the abused children were physically examined and treated in health care institutions (22).A study among Croatian elementary school children in 2004 showed that 35% were victims of various forms of abuse and school violence, 19% were passive victims, 8% provocative victims, and 8% were bullies (23). In Sisačko-moslavačka County, 10%-20% of high school pupils experienced some form of harassment in childhood (24).According to the 2001 census, Slavonski Brod and its surroundings had 124 349 inhabitants and was the sixth largest town in Croatia. Before the war 1991-1995, it was a prosperous industrial and agro-industrial center with developed social services, but during the war it suffered considerable infrastructural damage and hosted many refugees from adjacent Bosnia, part of which took permanent residence. Post-war recession took its toll and nowadays Slavonski Brod has become one of the most impoverished towns in Croatia (25). Such socio-economic situation opens the possibility for increased violence and child abuse. The aim of this study was to determine the frequency of adolescent abuse in Slavonski Brod.     

Apocrine adenoma of the breast coexistent with invasive carcinoma

We describe a case of apocrine adenoma with simultaneous occurrence of invasive ductal carcinoma in the breast of a 53-year-old woman. Apocrine adenoma affecting the breast is very rare. The lesion is composed of back-to-back ducts and papillary fronds covered with apocrine cells, and it is sharply demarcated from the surrounding breast tissue. The patient presented with a palpable nodule with skin retraction in her right breast, where ultrasound examination identified a 5-mm hypoechogenic nodule. The ultrasound also revealed in the surrounding breast parenchyma an additional abnormal finding suggestive of carcinoma. Histologic examination of the excised specimen showed that the hypoechogenic nodule represented an apocrine adenoma in proximity to the invasive ductal breast carcinoma. This is the first report that describes the simultaneous occurrence of these two lesions in the same breast.     

Neopterin--an early marker of surgical stress and hypoxic reperfusion damage during liver surgery.

Neopterin is elevated in infections, autoimmune diseases and post-transplant. Recently neopterin elevation was linked to stress response and malignancy. To determine early changes of serum neopterin caused by surgical stress and to investigate their association with other inflammatory markers and with malignancy, we measured neopterin, C-reactive protein (CRP) and procalcitonin (PCT) levels at four predefined time-points within 24 hours in 27 patients admitted for liver resection. Our results show that neopterin increased during operation and the increase was not related to preoperative neopterin levels. On the first day after surgery neopterin level was not significantly different from postoperative levels. In patients with malignant disease neopterin concentration before operation was higher than in patients with non-malignant disease, however, the increase in neopterin concentration during operation was not different between both patient groups. During surgery CRP and PCT did not increase significantly. On the first postoperative day CRP and PCT were elevated and their levels correlated with neopterin (Pearson's correlation coefficient r=0.51 and r=0.76, respectively). We conclude that neopterin elevation during liver resection contributes major part to the increased levels observed on the first postoperative day. Perioperative neopterin release can/may be related to stress response and hypoxia produced during operation. Using this marker, hypoxic reperfusion damage could be detected earlier and more accurately.