Thiazolidinediones Regulate the Level of ABC Transporters Expression on Lung Cancer Cells.

ATP binding cassette (ABC) transporters related to multidrug resistance (MDR) actively efflux various xenobio-tics from the cells across the cell membrane and decrease a drugs efficiency. Lung cancer is the leading cause of death among all types of cancer in the Czech Republic, and its incidence is still rising. Ciglitazone, rosiglitazone and troglitazone belonging to PPARγ agonist family (formerly used in diabetes mellitus treatment) were selected to investigate their capability to influence expression of ABC transporters on lung cancer cells. Therefore, the effect of PPARγ of agonists on transcription of following ABC transporters was investigated: multidrug resistance protein 1 (MDR1), multidrug resistance-associated protein 1 (MRP1), and breast cancer resistance protein (BCRP). We have investigated if these PPARγ agonists are substrates of ABC transporters using HL60 and HL60 derived cell lines (HL60-MDR1, HL60-MRP1, PLB-BCRP) by cytotoxicity test WST-1. We have mapped the changes in mRNA expression level of those transporters in A549 and HEK293 cells after PPARγ agonists treatment using quantitative reverse transcription real-time PCR (qRT-PCR). All three PPARγ agonists serve as substrates to at least one ABC transporter under study. PPARγ activation correlates with up-regulation of PTEN which may modulate the expression of ABC transporters through PI3K/ Akt signaling pathway. We have shown that rosiglitazone and troglitazone inhibit mRNA expression of MDR1 transporter in both cell lines whereas the expression of MRP1 in HEK293 cell was up-regulated after rosiglitazone treatment and the expression of MDR1 was upregulated after ciglitazone treatment.

Biologic importance and prognostic significance of selected clinicopathological parameters in patients with oral and oropharyngeal squamous cell carcinoma, with emphasis on smoking, protein p16(INK4a) expression, and HPV status.

The aim of this study is to evaluate the biologic importance and prognostic significance of selected clinicopathological parameters in patients with oral (OSCC) and oropharyngeal (OPSCC) squamous cell carcinoma, with emphasis on smoking, protein p16(INK4a) (p16) expression, and human papillomavirus (HPV) status.The study sample consisted of 48 patients with OSCC and 44 patients with OPSCC. Half of the patients were nonsmokers and the other half were gender-, age- and tumor localization-matched smokers. p16 expression was detected in 17/48 (35 %) OSCCs and in 36/44 (82 %) OPSCCs and HPV DNA was present in 7/48 (15 %) OSCCs and in 35/44 (80 %) OPSCCs. The sensitivity and specificity of p16 expression for HPV DNA presence were 0.74 and 0.88, respectively. The OPSCCs were more frequently basaloid (p < 0.001) while the OSCCs were more frequently conventional (p < 0.000001). The OSCCs were more likely to recur locally and to be the cause of death (p = 0.009 in both parameters).The HPV-positive tumors were more frequently localized in oropharynx, were basaloid SCCs and were p16- and HPV-positive (p < 0.000001 in all 4 parameters). The HPV-negative tumors were more frequently localized in oral cavity (p < 0.000001), more frequently asociated with local, regional and locoregional recurence (p = 0.011, p = 0.019 and p = 0.030, respectively) and with tumor-related death (p = 0.003). There was no significant difference with regard to smoking history (p > 0.05). The survival of patients with HPV-positive tumors was significantly longer (median 112 months; 95% CI 54 - 112 months) than that of patients with HPV-negative tumors (median 17 months; 95% CI 12 - 39 months) (p < 0.001). The HPV status of OSCC/OPSCC is an important biological and prognostic parameter and should be examined in all cases, using PCR or immunohistochemical detection of surrogate marker p16. Smoking itself does not seem to be an important prognostic factor.

[Role of human papillomavirus in carcinogenesis of head and neck cancer]. / Uloha lidského papillomaviru v karcinogenezi nádoru hlavy a krku.

Head and neck squamous cell carcinomas develop predominantly in individuals over 40 years of age and more frequently in males. The strongest risk factors for this disease are long-term abuse of tobacco products and alcohol. Recently, several reports of increasing incidence of head and neck cancer in atypical population groups of females or young adults have been published, often in patients with no history of smoking or alcohol abuse. It seems highly probable that at least in a part of these cases, human papillomavirus (HPV) played an important etiological role. Some of the HPV types were proved to cause certain anogenital carcinomas, particularly cervical carcinoma. It seems that in some cases these very HPV types are also involved in head and neck carcinogenesis. Published data on the prevalence of HPV infection in head and neck squamous cell carcinomas vary in different studies. However, it is generally understood that the infection is more commonly present in carcinomas of the oropharynx and palatine tonsils than in oral cavity carcinomas. The hypothesis of sexual transmission of oncogenic HPV types has yet to be confirmed. It is not clear whether current HPV vaccines could possibly decrease the incidence of head and neck squamous carcinomas.

Brain cell RNA: sexual differences in the rat.

Sexual differences in the total content of ribosomal RNA, established by cytophotometric measurements in neurones from selected brain regions, were studied in rats of the Wistar strain. In females of reproductive age, cyclic changes of RNA were synchronized with their oestrous cycle, the values being higher in the oestrous phase than in dioestrus. These changes were observed in pyramidal cells of the hippocampus and of the frontal cortex, in cells of anterior thalamus, of ventromedial and lateral hypothalamus and of tuberculi olfactorii. However, cycling cells were not disclosed in septum and thalamus posterior. A dependence upon the actual level of ovarial hormones was found in ventromedial hypothalamic cells only. In general, the RNA values in males of the same age corresponded to values of dioestrous females. The differences between newborn and 7-day-old pups were not marked enough and did not allow to define the critical period responsible for initiation of this sexual difference. In senescent rats, this difference persisted. During the stable phase of long-lasting dioestrus, the total RNA content in cells of the frontal cortex, hippocampus and hypothalamus was higher in females than in males of the same age which may suggest a faster reduction of this substance in aged males. The prolonged influence of oestrogens in the oestrous phase of the climacteric period (preceding the permanent dioestrus) decreased the RNA values in hippocampal and hypothalamic neurones even below the level established during the permanent dioestrus (and thus reached male values). On the contrary, in frontal cortical neurones, the female values remained higher in the permanent dioestrus as well as during long-lasting oestrus. A discussion concerns the possible participation of genetic determination and of the actual state of ovarial hormones in the manifestation of sexual differences in brain cells of the rat.

Visual evoked potential evidence for magnocellular system deficit in dyslexia.

Some recent studies on dyslexia have suggested a selective abnormality in the magnocellular visual pathway. To verify this hypothesis, we investigated motion-onset visual evoked potentials (VEPs) (predominantly testing the magnocellular system) as well as pattern-reversal VEPs (presumably testing the parvocellular system) in 20 dyslexics and 16 controls (both groups with a mean age of 10.0 years). Although the latencies and amplitudes of the main positive peak of pattern-reversal VEPs did not differ between the dyslexic and control group, the motion specific negative peak of motion-onset VEPs was significantly delayed (p < 0.001) in dyslexics. Our results confirm a selective magnocellular pathway disorder in dyslexics and indicate that the motion-onset VEPs might serve as an objective method for early diagnosis of dyslexia.

Oestrous cycle in septal rat females.

Bilateral lesions of the dorsal part of the septal area were produced in the brain of female rats of Wistar and Long-Evans stocks. The cytogram of their vaginal smears was evaluated repeatedly from the 30th day after surgery and permanent impairment of their oestrous cycle was found. In comparison with intact control females, oestrous phases were detected more frequently whereas dioestrous phases were observed less frequently. The proportions between the individual periods of the oestrous phase (i.e. pro-oestrus, oestrus and metoestrus) were equal in both septal and intact animals. No difference was found between the rat stocks used. Similarly, the previous life history of the rats which had been housed either in a socially impoverished environment (i.e. single cages) or in large communities (colonies) exerted no influence upon the postoperative impairment of the oestrous cycle.

Cathepsin L immunoreactivity in the hypothalamus of normal, streptozotocin-diabetic and vasopressin-deficient Brattleboro rats.

The immunolocalization of cathepsin L in the hypothalamus of normal rats was compared with the distribution of the enzyme in streptozotocin-treated animals and in vasopressin-deficient rats (Brattleboro strain). In rats with a normal metabolic status the neurons of magnocellular nucl. supraopticus and paraventricularis stood out by intense immunostaining for cathepsin L. In rats suffering from an experimentally induced diabetes mellitus and in homozygous Brattleboro rats we observed a strong reduction in enzyme immunoreactivity in these nuclei. Since cathepsin L is capable of splitting certain hypothalamic neuropeptides that are changed in diabetic animals, a role of the enzyme in the metabolism of these peptides is imaginable. Decrease in immunoreactive cathepsin L in vasopressin-deficient rats points to a possible involvement of the enzyme in the control of fluid homeostasis.

Maternal behaviour in septal rat females.

Dorsal lesions of the septum influenced the maternal behaviour of Wistar and Long-Evans females, as well as of their hybrids, in a different way. The phenomenon of infanticide appeared in Wistar females only, whereas mothering was not impaired in others. However, the mouse-killing activity was not enhanced in Wistar females displaying infanticide. On the contrary, they exhibited xenoparental behaviour. If these females had grown up in a species-typical environment, characterized by enriched social stimulation during their critical developmental period, no impairment of maternal behaviour following dorsal septal lesions occurred. The ablation of the whole septum caused a break-down of maternal behaviour with the appearance of infanticide in all females regardless of their stock origin and their individual life history. The specific role of the septum in the control of maternal behaviour is discussed.

Milk fat concentration and growth of rat pups.

The relationship between milk fat concentration and the growth of rat pups from birth to the 20th day was investigated. A total of 36 first-time-lactating Wistar rats and 366 pups from litters of 8-12 were used. The concentration of milk fat was determined by the crematocrit method, on the 2nd, 5th, 10th, 15th, 20th and 25th day of lactation. A wide range of milk fat concentration with a great variability in each individual dam was observed during lactation. To correlate milk fat and pup's growth, the mean value of the milk fat of the individual mothers was calculated for the first 10 days from the values of the 2nd, 5th and 10th day of lactation, and for the second 10 days from the values of the 10th, 15th and 20th days. The results show that pups fed higher fat milk in the first 10 days of life grew faster than pups fed lower fat milk. A significant positive correlation (P < 0.001) was found between the fat concentration of milk and the weight gains of pups in the first 10 days, but not in the older groups. It indicates that the fat concentration of milk is important for the growth of pups mainly in the early postnatal period of life.

Influence of early undernutrition in rats on their body fat and RNA content in VMH neurons.

The RNA content of the ventromedial hypothalamus (VMH) and lateral hypothalamic area (LHA) neurons, and the epididymal, retroperitoneal, and liver fat content of 150-day-old male rats submitted to neonatal undernutrition, were investigated. The neonatal undernutrition was carried out by two different ways. First, by reducing the litters to two pups per nest from birth to weaning. The pups from normal litters (eight pups/nest) served as controls. Secondly, by separating half of the pups (four pups) from normal litters for 8 h daily during the first 5 postnatal days. The remaining nonseparated pups served as their controls. The data show that both groups of early undernourished pups had a significantly increased RNA content in the VMH neurons (RNA content in the LHA neurons was unchanged) and a significantly decreased epididymal, retroperitoneal, and liver fat content in comparison with their controls. The results indicate that early undernutrition leads to a permanent increase in the functional activity of VMH and alteration in fat metabolism in rats.

Influence of litter size on the RNA content of VMH and LHA in preweaning rats.

The nutritional status in the early postnatal period may significantly influence the development and functional activity of food intake regulating mechanisms. Therefore we investigated the weight gains and total RNA content of the ventromedial (VMH) and lateral (LHA) hypothalamic cells of rat pups in normal litters (8 pups/nest) and in reduced litters (2 pups/nest) reared permanently (from birth to weaning) or temporarily (from 1st to 5th day) in reduced nests. A significant decrease of weight gains was found in pups reared in reduced litters during the first 5 days which resulted in significant differences of body weight between the pups from normal and reduced litters on 5th day of life. On the 15th and 30th days, there were no significant differences in body weight between these animals, but the RNA content in VMH cells of pups reared in reduced litters was significantly increased on the 15th and 30th days. The RNA content in the LHA cells was unchanged. The results show that reducing a rat litter to 2 pups caused short-term undernutrition leading to permanent changes of the RNA content in the VMH neurons. The possible consequences of the permanently changed RNA content of this hypothalamic structure are discussed.

[Carotenemia]. / Karotenémie.

The authors demonstrate two infants with a yellow skin coloration which developed as a result of excessive dietary carotenoid intake. The yellow coloration, contrary to jaundice, did not affect the sclerae. Elevated blood carotenoid levels were found and a temporary increase of aminotransferases. In both infants lower IgA levels were recorded. After modification of the diet the yellow coloration disappeared and gradually the aminotransferases reached normal levels.

Ontogeny of ribonucleic acid content in rat cortical neurons influenced by early stimulation.

Developmental changes in the total RNA content were studied in pyramidal cells of the frontal, parietal (somesthetic), temporal (auditory) and occipital (visual) cortical areas in Wistar rats aged 13, 14, 15, 21, 28, 35 and 56 days, and compared with those after complex, visual and acoustic stimulation. In all areas neuronal RNA content reached peak values at the age of 3 weeks, except the parietal cortex, where the values were highest as early as at 15 days (in 21-day pups they were only insignificantly lower). Between the age of 3 and 4 weeks, RNA content of cortical neurons decreased in all three projection areas, whereas in the frontal cortex a significant decline came later--in the 5th week. In the occipital cortex neuronal RNA at 8 weeks reached reliably lower values than in 13-day pups. Neuronal RNA increased at the time of eye-lid opening (13-15 days), highly significantly in the visual cortex. Complex visual and acoustic stimulation raised neuronal RNA content in projection areas of modality involved in stimulation and, moreover, acoustic stimulation did so in the frontal and occipital cortex. On the contrary, complex and visual stimulation decreased RNA contents in the frontal cortex. Developmental dependence, stimulation contingency, and specificity vs. nonspecificity of the observed changes are discussed, and compared with other biochemical, morphological and functional changes that have been found in the period of peaking RNA content, or immediately thereafter.

[The effect of natural and artificial nutrition on vitamin C and E levels in the normal infant]. / Vliv prirozené a umelé výzivy na hladiny vitamínu C a E u fyziologických kojencu.

During the spring months the vitamin C and E serum levels were assessed in 44 normal infants. In addition the serum levels of vitamins C and E were assessed in 17 breastfed infants and their mothers. Low vitamin C levels were recorded in five infants (12.2%). Of these four were bottlefed and one infant had a mixed; mostly formula diet. Two infants with a low vitamin C level had fruit juices and strained fruit. All breastfed infants were well saturated with vitamin C despite the reduced vitamin C level of one nursing mother. Low vitamin E levels were found in two infants (4.5%). Both infants were under six weeks and were bottlefed. After supplementation of the diet with nonmilky products the vitamin E levels rose in breastfed as well as bottlefed children. In breastfed infants older than six weeks the vitamin E levels were significantly higher (p less than 0.001), as compared with bottlefed infants. The results of the investigation confirmed that only breastfeeding or well adapted cows milk formula ensure an adequate vitamin C and E level in normal infants.

[Vitamins C and E in neonates and their mothers]. / Vitamíny C a E u novorozencu a jejich matek.

During the spring months serum levels of vitamin C and E were investigated in 49 women at the end of pregnancy and in their infants at the delivery and on the fifth day after birth. The authors found that 23.3% of the pregnant women (n = 43) who gave birth to mature, normal neonates were not adequately saturated with vitamin C during the spring months. Almost 13% of the infants (12.8%) had lower vitamin C levels on the fifth day after delivery. Two thirds of the mothers (67%) of premature infants (n = 6) had low vitamin C levels. The vitamin E levels in the pregnant women were adequate, 38% of the five-day-old infants had reduced vitamin E levels.

[Follow-up of newly diagnosed patients with diabetes in the Prague population. II. Changes in the group of new diabetics after a 2-year follow-up]. / Sledování nove diagnostikovaných onemocnení cukrovkou v prazské populaci. II. cást: Zmeny v souboru nových diabetiku po dvouletém sledování.

From the total number of 1551 new diabetics and patients with PGT 85 subjects died in the course of two years. Treatment was changed 118 times, failure of PAD treatment was recorded in 11 type 2 diabetics. Three patients originally classified as type 1 were later included in type 2. In three diabetics (of 14) who had sings of diabetic retinopathy already during manifestation of DM marked deterioration of the finding on the fundus occurred. The incidence of macroangiopathic complications did not change.

[Follow-up of newly diagnosed patients with diabetes in the Prague population. I. Incidence of diabetes and disorders of glucose tolerance, methods of detection, complications]. / Sledování nove odhalených nemocných cukrovkou v prazské populaci. I. cást: Incidence cukrovky a poruch glukózové tolerance, zpusob odhalení, komplikace.

The authors followed up the incidence of new cases of diabetes in nine Prague diabetological clinics, the way of manifestation of diabetes and its detection, age dependence, seasonal character and incidence of complications. The incidence of type 2 diabetes was 269, of type 1 2.3 and impaired glucose tolerance 27 per 100,000 population. In 76% diabetes was detected accidentally, most frequently by the health community doctor or factory medical officer; 65% of the diabetes were older than 60 years. 51% suffered from hypertension and in 0.9% of type 2 diabetics retinopathy was found.

Total RNA content of brain cells of inbred mice reared under different conditions.

AB/Jena, DBA/2Jena, C57B1/6 Jena inbred mice and ABD2F1- and B6D2F4-hybrid mice either were reared communally and weaned at day 30 or were reared by isolated mothers and weaned at day 21 (I21). On day 50 the total RNA content of pyramidal brain cells of male offspring was determined by cytophotometry. The studied brain areas were the frontal pole of cerebral cortex (FCC) and the CA1 region of the dorsal hippocampus (DHI). The DBA, C57 and ABD2 mice had a reduced FCC and DHI RNA-content under I21-rearing. Under these conditions the AB strain has the highest RNA-content in both substrates. The results call for the awareness of possible genetic influences when studying the effects of environmental enrichment/impoverishing.