Expression of CYP and GST in human normal and colon tumor tissues.

We investigated the immunohistochemical staining characteristics of cytochrome P450 1A1 (CYP1A1), CYPB1, CYP2E1, and glutathione S-transferase P1 (GSTP1), GSTT1, GSTO1, GSTK1 in colon tumor and surrounding normal colon tissues. Tissues were obtained from 47 patients with colon adenocarcinoma and the staining intensity of tumor and control tissues was compared. CYP1A1, CYP1B1, CYP2E1, GSTP1, GSTT1, GSTO1 and GSTK1 expressions in colon cancer cells were significantly greater than those in normal colon epithelial cells. No significant relation was found between the isoenzyme expressions and age, gender, smoking status, tumor grade and tumor stage. The higher expressions of CYP1A1, CYP1B1, CYP2E1, GSTP1, GSTO1, GSTT1 and GSTK1 in tumor than in normal colon tissues may be important for colon cancer progression and development.

Molecular characterization of zeta class glutathione S-transferases from Pinus brutia Ten.

Glutathione transferases (GSTs; EC 2.5.1.18) play important roles in stress tolerance and metabolic detoxification in plants.In higher plants, studies on GSTs have focussed largely on agricultural plants. There is restricted information about molecular characterization of GSTs in gymnosperms. To date, only tau class GST enzymes have been characterized from some pinus species. For the first time, the present study reports cloning and molecular characterization of two zeta class GST genes, namely PbGSTZ1 and PbGSTZ2 from Pinus brutia Ten., which is an economically important pine native to the eastern Mediterranean region and have to cope with several environmental stress conditions. The PbGSTZ1 gene was isolated from cDNA, whereas PbGSTZ2 was isolated from genomic DNA. Sequence analysis of PbGSTZ1 and PbGSTZ2 revealed the presence of an open reading frame of 226 amino acids with typical consensus sequences of the zeta class plant GSTs. Protein and secondary structure prediction analysis of two zeta class PbGSTZs have shared common features of other plant zeta class GSTs. Genomic clone, PbGSTZ2 gene, is unexpectedly intronless. Extensive sequence analysis of PbGSTZ2, with cDNA clone, PbGSTZ1, revealed 87% identity at nucleotide and 81% identity at amino acid levels with 41 amino acids differences suggesting that genomic PbGSTZ2 gene might be an allelic or a paralogue version of PbGSTZ1.

GST isoenzymes in matched normal and neoplastic breast tissue.

The potential to metabolize endogenous and exogenous substances may influence breast cancer development and tumor growth. Therefore we investigated GST activity and the protein expression of glutathione S-transferases (GSTs) isoenzymes known to be involved in the metabolism of endogenous and exogenous carcinogens in breast cancer tissue to obtain new information on their possible role in tumor progression. The interindividual variation in the conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) and of 1,2-epoxy-3-(p-nitrophenoxy) propane (EPNP) with glutathione (GSH) by cytosolic glutathione S-transferases (GSTs) were investigated in human breast matched normal and tumor samples. The GSTA, GSTM, GSTP and GSTT isoenzymes from the crude extracts of matched breast normal and tumor tissues in terms of their immunological properties using western blotting were compared. In most of the samples, the GST activities were higher in the tumor than in the normal cytosolic fractions against both CDNB and EPNP. In the western blotting analysis, it was proved statistically that in normal and tumor epithelial cells, there was difference between GST pi and theta isoenzymes expressions (p0.05). In normal epithelium there was a stronger GST theta expression than in invasive tumor tissues (p=0.013). However, the stronger GST pi expression was observed in tumor epithelium than in normal epithelium in human breast cancers (p=0.000). We found the GSTP protein level and GST activities were higher in the breast tumor than in the normal cytosolic fractions against both CDNB and EPNP, thus implicating a certain biological importance.

CYP and GST polymorphisms and survival in advanced non-small cell lung cancer patients.

UNLABELLED: Several polymorphisms in cytochrome P-450s (CYP)s and Glutathione S-transferases (GST)s have been reported to be associated with survival rates of patients with non-small cell lung cancer (NSCLC) but the studies in this regard are scarce and the results are contradictory. In this study, CYP1A1 (Ile462Val), CYP1B1(Asn453Ser), GST M1, GSTP1 exon 5 (Ile105Val) and exon 6(Ala114Val) and GSTT1 polymorphisms were determined in 138 patients with advanced NSCLC to evaluate their role in survival. Of the studied CYP and GST polymorphisms only GSTP1 exon 6 variant significantly altered (improved) the survival compared to wild type (p=0.036) with median survival of 22.2 months and 16.1 months, respectively. Multivariate analysis also revealed a significant reduction of adjusted hazard ratio of death associated only with the GSTP1 exon 6 variant genotype of 0.45 (95% confidence interval (95% CI), 0.23-0.89, p=0.022). These results show that the GSTP1 exon 6 variant genotype is associated with improved survival in the patients with advanced NSCLC. KEYWORDS: Cytochrome P-450, glutathione S-transferase, non small cell lung cancer, polymorphism, survival.

Metric sex determination from the pelvis in modern Greeks.

The ability to determine sex from unknown skeletal remains is vital, and methods to do this on the various bones of the human skeleton have been researched extensively. Many researchers have emphasized the need for population specific data for methods which are based on measurements, as there are vast differences in body size in various populations. The pelvis is known to be the most sexually dimorphic part of the human body, and no discriminant function formulae for this bone are available for Greek or other Mediterranean groups. The purpose of this study was therefore to develop discriminant functions which can be used for sex determination on measurements of the pelvis of modern Greeks. A sample of 97 male and 95 female pelves in a skeletal collection housed in Heraklion, Crete, was used. Measurements were taken from the articulated pelvis, single os coxae and the sacrum. Discriminant function formulae for all measurements and various combinations were used in order to assess the degree of sexual dimorphism in various parts of the pelvis, and to make the formulae usable on fragmented remains. For the single os coxae, average accuracies of 79.7-95.4% (79.1-93.5% on cross-validation) were found. However, it was found that measurements of the sciatic notch were unreliable and yielded poor results, and it is advisable that this characteristic must only be used as a last resort. Dimensions of the sacrum were not very dimorphic (average accuracy 60.9%), while measurements from the articulated pelvis yielded poorer results than that from single innominate bones. The diameter of the acetabulum was the single most dimorphic characteristic, providing on average 83.9% accuracy when used in isolation.

A study on the antioxidant activities of some new benzazole derivatives.

The in vitro antioxidant properties of some new benzazole derivatives (1-10) such as benzoxazoles, benzimidazoles, and benzothiazoles were determined by their effects on the rat liver microsomal NADPH-dependent lipid peroxidation (LP) level, the scavenging of superoxide anion and the stable radical 2,2-diphenyl-1-picrylhydrazyl (DPPH). Compounds 1, 2, 4 and 6, showed potent scavenging effect on superoxide radical at 10(-3) M. Compound 8, 5-nitro-2-(phenoxymethyl)benzimidazole, strongly inhibited lipid peroxidation at 10(-3) M concentration.

Comparison of NAT1, NAT2 and GSTT2-2 activities in normal and neoplastic human breast tissues.

In this study, arylamine N-acetyltransferases, NATs (E.C.2.3.1.5) and glutathione-S-transferase-T2-2, GSTT2-2 (E.C.2.5.1.18) enzyme activities in the breast tumor and surrounding tumor-free tissues of 22 female breast cancer patients with infiltrating ductal carcinoma were measured. The possible impacts of grade of malignancy, chemotherapy treatment, estrogen receptor status and menopausal status on all enzyme activities were evaluated. The results showed that, both NAT2 and GSTT2-2 display significant differences between tumor and tumor-free breast tissues, while no difference was observed in NAT1. Grade of malignancy seems to be positively associated with NAT1 and negatively associated with GSTT2-2. Though, both NAT2 and GSTT2-2 have increased mean tumor activities, the grade of malignancy, chemotherapy status, menopausal status or estrogen receptor status are not correlated statistically.

Genotoxicity studies on benzimidazole retinoids.

Retinoids consist of a family of naturally occuring compounds including all-trans retinoic acid (ATRA), retinal, retinol (vitamin A), 9-cis retinoic acid, 13-cis retinoic acid as well as a large number of synthetic derivatives. Retinoids are known to elicit diverse pharmacological profiles such as controlling cell differentiation/proliferation and modulating specific premalignant lesions and reducing second primary tumors in patients. Clinical use of retinoids is limited due to their toxicity. Three benzimidazole retinoid derivatives (BITN, BITNm, BITNe) were synthesized and were examined in terms of genotoxicity towards human lymphocyte cultures by sister chromatid exchange (SCE) analysis. It has been found that BITN decreased the number of SCEs 20% at 10(-6) M, but had no effect at 10(-5) M. No significant effect on SCEs was observed for BITNm and BITNe at both concentrations. ATRA increased the SCEs (35%) at 10(-5) M but had no effect at 10(-6) M. The results have shown that benzimidazole retinoids did not induce SCE significantly. Besides this, BITN reduced the SCEs and had a protective effect at low concentration. Since the induction of glutathione S-transferase (GST) is associated with anticancer drug resistance, the effects of BITN, BITNm, BITNe and ATRA on human lymphocyte GSTs were also investigated using CDNB as substrate. BITN and BITNm induced GST activities 54% and 49% respectively at 10(-5) M, but had no effect at 10(-6) M. BITNe induced GST activity 62% at 10(-5) M and 35% at 10(-6) M. ATRA had no effect on GST activity at 10(-5) M.

Acute effect of nebulized budesonide in asthmatic children.

The acute anti-inflammatory effects of inhaled steroids at high doses and their use at home and as emergency treatment of acute asthma attacks in children have been evaluated in many clinical studies. However very little is known about their additional bronchodilator response to systemic steroids plus nebulized salbutamol in the early management in children. Asthmatic patients aged between 5-15 years were investigated in a double-blind, placebo-controlled fashion. Both the study group (Group I) and the control group (Group II) received three consecutive doses of nebulized salbutamol (0.15 mg/kg/dose) and one dose of parenteral methylprednisolone (1 mg/kg/dose, intramuscularly). After this treatment, nebulized budesonide (1 mg/dose) was administered to patients in the study group and placebo (nebulized saline) was administered to patients in the control group. Pulmonary index scoring and peak flow meter was performed to both groups before and after the treatment. There were twelve patients in Group I (mean age: 7.90 +/- 2.34 years) and fourteen patients in Group II (mean age: 9.36 +/- 2.55 years). There was no difference between the two groups with respect to age (p = 0.1421), gender (p = 1.000) and inhaled steroid prophylaxis rate (p = 0.2177). No statistically significant difference was detected between the two groups with respect to the pulmonary index score (p = 0.3528). Yet, there was a statistically significant difference between the two groups with respect to the increase in PEFR (p = 0.0155). The positive acute effect of nebulized budesonide in addition to systemic steroids and nebulized salbutamol in improving the spirometric indices in asthmatic children is an encouraging finding for further investigations of its routine use in the pediatric emergency department.

Antioxidant properties of Ballota species growing in Turkey.

In the present study, the antioxidant properties of ethanol extracts of 16 Ballota species belonging to the Lamiaceae family and growing in Turkey on superoxide anion formation and lipid peroxidation were investigated. All extracts inhibited superoxide anion formation but to different extents. The extracts of Ballota antalyense, Ballota macrodonta, Ballota glandulosissima, Ballota larendana, Ballota pseudodictamnus, Ballota nigra subsp. anatolica, Ballota saxatilis subsp. saxatilis, Ballota rotundifolia, and Ballota saxatilis subsp. brachyodonta exhibited remarkable anti-superoxide anion formation with IC(50) values from 0.50 to 0.87 mg/ml. The extracts of Ballota antalyense, Ballota macrodonta and Ballota glandulosissima have the lowest IC(50) values (0.50, 0.51 and 0.51 mg/ml, respectively) which were rather close to the well-known superoxide anion scavenger alpha-tocopherol (IC(50): 0.22 mg/ml). The extracts of Ballota inaequidens, Ballota glandulosissima, Ballota saxatilis subsp. saxatilis, Ballota macrodonta and Ballota antalyense inhibited lipid peroxidation with IC(50) values from 12 to 20 mg/ml. The extracts of Ballota inaequidens (IC(50): 12 mg/ml) and Ballota glandulosissima (IC(50): 15 mg/ml) exhibited the strongest inhibitory effects on lipid peroxidation compared with alpha-tocopherol (IC(50): 3 mg/ml). The results show that Ballota glandulosissima is the best antioxidant source among these 16 Ballota species.

Estimation of stature from body parts.

Anthropometric technique commonly used by anthropologists and adopted by medical scientists has been employed to estimate body size for over a hundred years. With the increasing frequency of mass disasters, the identification of an isolated lower extremity and the stature of the person it belonged to has created problems for the investigation of the identity of some of the victims. In spite of a need for such a study, there is a lack of systematic studies to identify fragmented and dismembered human remains. The purpose of the paper is to analyze anthropometric relationships between dimensions of the lower extremity and body height. Analysis is based on a sample of middle class male (N=203) and female (N=108) adult Turks residing in Istanbul. The participants are mostly students and staff members of a medical school, and military personnel. Measurements taken are stature, trochanteric height, thigh length, lower leg length, leg length, and foot height, breadth, and length. Of the five variables entered into the regression analysis, all but foot breadth participate in the analysis with leg length as the first and followed by thigh and foot lengths, and finally foot height in males (R(2)). There were also individually calculated formulae for some of these measurements which provided smaller R(2)-values. Student's t-test to assess if there was any intraobserver error in measurements take by individual anthropometrist did not show such any statistically significant difference. In conclusion, the study suggested that estimation of a living height can be made possible using various dimensions of the lower extremity. One must consider differences between populations in order to apply functions as such to others.

Synthesis and antioxidative properties of novel thiazolidinedione/imidazolidinedione compounds as retinoids.

The general term "retinoids" refers to both naturally occurring as well as synthetic compounds which exhibit biological activity similar to vitamin A (retinol). Vitamin A and its two metabolites, retinaldehyde and retinoic acid, are fat-soluble unsaturated isoprenoids necessary for the growth, differentiation and maintenance of epithelial tissues. In this study, we have synthesized thiazolidinedione/imidazolidinedione compounds as retinoids. Their in vitro effects on rat liver microsomal NADPH-dependent lipid peroxidation (LP) levels and superoxide anion formation were determined.

Effects of cigarette smoke with different tar contents on hepatic and pulmonary xenobiotic metabolizing enzymes in rats.

The effects of smoke from cigarettes with two different tar contents (32 mg/cigarette, high tar, and 15 mg/cigarette, low tar) on hepatic and pulmonary monooxygenase (MO) activities (aniline 4-hydroxylase [AH]; aminopyrine N-demethylase [AMND]; 7-ethoxyresorufin O-deethylase [EROD]; p-nitroanisole O-demethylase [p-NAOD]), lipid peroxidation (LP) and reduced glutathione (GSH) levels and glutathione S-transferase (GST) activities toward several substrates (1-chloro-2,4-dinitrobenzene [CDNBI; 1,2-dichloro-4-nitrobenzene [DCNB]; ethacrynic acid [EAA]; 1,2-epoxy-3-(p-nitrophenoxy)-propane [ENPP]) were determined in adult male rats. Adult male rats were exposed to smoke of high- or low-tar cigarettes five times a day, with 1-hour intervals, for 3 days in a chamber where smoke and fresh air lead alternatively and were killed 16 hours after the last treatment. Smoke of both high- and low-tar cigarettes (SHTCC and SLTCC) significantly increased hepatic and pulmonary EROD and p-NAOD activities compared to controls. However, the increase noted by SHTCC on pulmonary EROD activity was higher than that of SLTCC. Hepatic AMND and pulmonary AH activities were significantly increased only by SHTCC. LP level was significantly decreased and increased by SHTCC in liver and lung, respectively, whereas it remained unaltered by SLTCC. Only SHTCC significantly increased GSH level in liver. In the lungs, both SHTCC and SLTCC significantly increased GSH level to the same extent. Hepatic GST activity toward EAA was significantly increased by SHTCC but was significantly decreased by SLTCC. ENPP GST activity was significantly decreased by SHTCC and SLTCC in the livers. In the lungs, all the GST activities examined were significantly depressed by SHTCC whereas only GST activity toward DCNB was reduced significantly by SLTCC. These results reveal that the hepatic and pulmonary MOs and GSTs are differentially influenced by SHTCC and SLTCC in rats.

Plasma carnitine levels in children with Down syndrome.

Carnitine is responsible for several chemical processes, including lipid metabolism, nerve cell conduction, reduction in muscle hypotonia, and limitation in oxidative damage to cells. In patients with Down syndrome (DS), the process of growth is behind that of normal children and neuromuscular control is attained somewhat later. The purpose of this study was to assess variation in levels of carnitine in normal and DS children and the relationship between the amount of carnitine and age. The study involved 30 (15 girls, 15 boys) normal children and 40 (20 girls, 20 boys) DS patients of Turkish ancestry, 6 months to 13 years of age. Carnitine level was determined using Deufel's enzymatic method. Carnitine level was significantly lower in DS patients compared with normal children between 6 months to 5 years of age. Between 5 and 13 years of age, the level of carnitine was about the same in both the normal and DS groups. The results suggest that carnitine level shows a different pattern of age related increase in DS compared to normal children.

Global forensic anthropology in the 21st century.

While it has a long history, the last 30 years have brought considerable advances to the discipline of forensic anthropology worldwide. Every so often it is essential that these advances are noticed and trends assessed. It is also important to identify those research areas that are needed for the forthcoming years. The purpose of this special issue is to examine some of the examples of research that might identify the trends in the 21st century. Of the 14 papers 5 dealt with facial features and identification such as facial profile determination and skull-photo superimposition. Age (fetus and cranial thickness), sex (supranasal region, arm and leg bones) and stature (from the arm bones) estimation were represented by five articles. Others discussed the estimation of time since death, skull color and diabetes, and a case study dealing with a mummy and skeletal analysis in comparison with DNA identification. These papers show that age, sex, and stature are still important issues of the discipline. Research on the human face is moving from hit and miss case studies to a more scientifically sound direction. A lack of studies on trauma and taphonomy is very clear. Anthropologists with other scientists can develop research areas to make the identification process more reliable. Research should include the assessment of animal attacks on human remains, factors affecting decomposition rates, and aging of the human face. Lastly anthropologists should be involved in the education of forensic pathologists about osteological techniques and investigators regarding archaeology of crime scenes.

Skeletal remains of Dr. Eugenio Antonio Berríos Sagredo.

It is often noted that even a well-designed osteological technique may not provide accurate results when applied to single forensic cases. Case studies are ideal to test if this concern is valid, and forensic anthropology is a testing ground for applying a population based standard to individual skeletal remains. Secondly, the increasing role anthropologists have played in forensic sciences has aided the medicolegal disciplines in a number of ways. For example, identification of skeletal remains is now more accurate than ever before. Many of these cases have brought perpetrators to court for justice. The purpose of this paper is to use osteological techniques to analyze skeletal remains and make a positive identification. The victim was found partially buried in the sand near El Pinar, Uruguay in 1995. The analysis indicated that the victim was a 45-year old, white, male who was about 170cm tall. Based on preliminary evidence that the victim might be Dr. Eugenio Antonio Berríos Sagredo, a digital superimposition was made using the victim's photograph and the unknown skull. This examination revealed that the skull corresponded consistently with the individual in the photograph. Results were supported by the fact that personal belongings, such as a medal and wrist watch, also pointed to the same individual. Dental records and radiographs when made available later also indicated the same identity. Dr. Berríos was accused of making nerve gas during the dictatorial regime of former Chilean President General Augusto Pinochet. It was also alleged that he made bombs that killed a Spanish diplomat in his laboratory and a Chilean diplomat in Washington, DC. Many complex techniques are often needed to make a positive identification and such was the case for this study. Because of the nature of anthropology as a holistic discipline, such complexity is an integral part of human biology and behavior and can be used successfully in the forensic sciences and medicolegal investigations.

Sexual diagnosis of the glabellar region.

Determination of sex is a key issue in skeletal identification. Morphological assessment has been one of the oldest approaches by which a particular structure can be evaluated visually. The purpose of this investigation is to evaluate morphological variation in the human glabellar region and its association with sexual dimorphism. The study sample is composed of 90 adult skulls with known sex, race (all Caucasoids), obtained from the Council of Forensic Medicine (Adli Tip Kurumu), Istanbul, Turkey. The glabellar region was cleared of soft tissue and photographs were taken at the closest focused distance to the specimens. The degree of smoothness-roughness of the glabella was classified into four prototypes from 0 (very smooth) to 3 (very rough). Each specimen without knowing its sex was judged as ideal, belonging to one of the prototypes by the senior author. Using the same prototypes, the photographs of the specimens were also rated by ten observers. The results indicated that none of the males were assigned to Prototype 0, and conversely, none of the females were classified as Prototype 3. The judges, however, ranked 8.2% of males as 0 and no females as 3. About 84% of males were in Prototype 2 or higher as judged by the senior author. Males were mostly in Prototype 1 or above, but females in 1 or below. There are various explanations as to why the human skull shows sexual dimorphism, however, the robust nature of the male is the obvious one. The smoothness of the supranasal region has not been associated with any specific well documented anatomic, functional, or sexual aspect of the skull. This study shows that there is an identifiable sexual dimorphism in the supranasal region, but its source has not been well known. It is expected that this results from both the muscles of facial expression and the masticatory function.

The expression of cytochrome P450 enzymes in human breast tumours and normal breast tissue.

Environmental chemicals are one of the risk factors in breast cancer genesis. Cytochrome P450 (CYP) enzymes play a major role in the activation of these chemicals. Using highly specific and sensitive reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. the expression profile of all major xenobiotic metabolizing CYP forms was screened in breast tumour and surrounding tumour free (control) breast tissue in a series of 20 sample pairs obtained from females with infiltrating ductal carcinoma. The levels of CYPIAI mRNA were very low in both tumour and normal tissue. CYP1B1, CYP2B6, CYP2C, CYP2D6, CYP2E1, CYP4B1, and CYP11A1 expressions were positive in both tumours and control tissue. CYP2A6, CYP2A7, CYP2A13, CYP2F1, CYP3A4, CYP3A5. and CYP3A7 mRNAs were expressed neither in tumours nor in control tissue. These results show that several CYPs. responsible for the activation of a quite large number of procarcinogens and genotoxic estrogen metabolites. are expressed in breast tissue with a lack of qualitative differences in CYP expression at mRNA level between breast tumours and surrounding normal breast.