A complete chitinolytic system in the atherinopsid pike silverside Chirostoma estor: gene expression and activities.

The expression and digestive activity of pike silverside Chirostoma estor endogenous chitinases were analysed in samples from four life stages: whole eggs; larvae; juvenile intestine and hepatopancreas and adult intestine and hepatopancreas. A chitinase cDNA was cloned and partially sequenced (GenBank accession number: FJ785521). It was highly homologous to non-acidic chitinase sequences from other fish species, suggesting that it is a chitotriosidase. Quantitative PCR showed that this chitinase was expressed throughout the life span of C. estor, with maximum expression in the hepatopancreas of juveniles. Chitotriosidase and chitobiosidase activities were found at all life stages, along with a very high level of N-acetyl glucosaminidase (NAGase). The chitotriosidase activity could be encoded by the cloned complementary (c)DNA, although additional chitinase genes may be present. The chitotriosidase activity appeared to be transcriptionally regulated only at the juvenile stage. The expression and activity of chitinases tended to increase from the early to juvenile stages, suggesting that these variables are stimulated by chitin-rich live food. Nevertheless, the feeding of juvenile and adult fish with both live food and a balanced commercial diet seemed to provoke significant reductions in pancreatic NAGase secretion and/or synthesis in the gut. Moreover, all chitinase activities were lower in adults, probably reflecting a higher intake and use of the balanced diet. The observation of chitotriosidase and chitobiosidase activities together with a very high NAGase activity suggest the presence of a complete and compensatory chitinolytic chitinase system that enables this stomachless short-gut fish species to use chitin as an energy substrate. These novel findings suggest that dietary inclusions of chitin-rich ingredients or by-products might reduce the farming costs of C. estor without impairing performance.

Ghrelin stimulates myogenic differentiation in a mouse muscle satellite cell line and in primary cultures of bovine myoblasts.

Ghrelin is an acylated hormone that influences food intake, energy metabolism and reproduction, among others. Ghrelin may also stimulate proliferating myoblast cell differentiation and multinucleated myotube fusion. The aim of this work was to assess the effect of human ghrelin (hGHRL) and human ghrelin fragment 1-18 (hGHRL1-18) on myoblast differentiation by means of mRNA expression and protein level. Two types of cells were tested, the cell line i28 obtained from mouse skeletal muscle and primary cultures of bovine myoblasts. Both ghrelin and its N-terminal fragment hGHRL1-18 were used at concentrations of 0, 0.01, 0.1, 1, 10 and 100 nm. Treatments were applied to pre-confluent cultures and were maintained for 4 days. We determined that between 0.1 and 100 nm, hGHRL and hGRHL1-18 had similar effects on myogenic differentiation of i28 cells (p < 0.01). On the other hand, only the higher concentrations (10 and 100 nm) of hGHRL stimulated bovine myoblast differentiation. These results could be attributed to the presence, in both i28 cells and in bovine myoblasts, of the mRNA for GHS-R1a and CD36 receptors. The use of ghrelin in livestock production is still questionable because of the limited effects shown in this study, and additional research is needed in this field.

Beta-carotene is incorporated or mobilized along with triglycerides in bovine adipose tissue in response to insulin or epinephrine.

Pasture fed cattle ingest substantial amounts of beta-carotene (beta-C). Not all of the carotenoid compound is transformed into vitamin A, but the surplus is deposited in adipose tissue (AT). The mechanisms of beta-C incorporation and mobilization are unknown. Two experiments were conducted using explants from bovine AT cultured in vitro. First, beta-C incorporation by explants from three animals was examined with different beta-C concentrations (0, 1, 5 and 20 microm) and different times of incubation (every 5 h up to 25 h). The data showed a significant increase of beta-C concentration in explants only for 20 microm beta-C. Secondly, effects of insulin and epinephrine on beta-C and triglyceride (TG) contents of explants were studied. Explants from six animals were incubated with either hormone and 0 or 20 microm beta-C for 20 h. Both TG and beta-C contents were affected positively by insulin and negatively by epinephrine. Interestingly, changes in ratios of beta-C/TG (hormone vs. control) were similar (1.7 x 10(-3) and 1.8 x 10(-3)), respectively, for insulin and epinephrine, indicating that beta-C level is directly related to TG content. We also report the presence of mRNA for beta-C 15, 15' oxygenase in bovine AT. The in vitro culture system using explants from bovine AT is a promising model to investigate factors that might affect the accumulation and metabolism of beta-C.

Tricomicosis palmelina causa poco frecuente de afectación del tallo piloso / Trichomycosis palmelina: rare cause of hair shaft affection

Resumen La tricomicosis o tricobacteriosis palmelina es un motivo de consulta poco frecuente en los servicios de dermatología, es importante reconocerla y diferenciarla de otras patologías ya que puede ser difícil de diagnosticar. La tricomicosis o tricobacteriosis es una condición infecciosa superficial asintomática, rara, causada en la mayoría de los casos por Corynebacterium flavescens (también llamada Corynebacterium tenuis), perteneciente a la familia corinebacteriaceae, las cuales, son bacterias gram positivas, pleomórficas, inmóviles, no encapsuladas, aerobias o anaerobias facultativas. Afecta pelos de la región axilar, zona púbica, escrotal e interglutea; se debe al aumento de la población bacteriana de la microbiota normal, adherida a la cutícula del pelo. Esta afectación es propia de climas húmedos, afectando predominantemente a adultos jóvenes, de sexo masculino, deportistas y homosexuales en un entorno de pobreza y hacinamiento. El objetivo del presente reporte es comunicar un caso de tricomicosis palmelina flava en región púbica en un adulto joven y describir las características clínicas de esta patología relativamente poco conocida.

Abstract Trichomycosis or palmelin trichobacteriosis it is an infrequent reason for consultation in dermatology services, important to recognize and differentiate it from other pathologies since it can be difficult to diagnose. Trichomycosis or trichobacteriosis is a rare, asymptomatic superficial infectious condition, caused in most cases by Corynebacterium flavescens (also called Corynebacterium tenuis), belonging to the Corynebacteriaceae family, which are gram-positive, pleomorphic, immobile, non-encapsulated bacteria, aerobic or facultative anaerobic. It affects hairs in the axillary region, pubic, scrotal and intergluteal area; It is due to the increase in the bacterial population of the normal microbiota, attached to the hair cuticle. This affectation is typical of humid climates, predominantly affecting young adults, males, athletes and homosexuals in an environment of poverty and overcrowding. The objective of this article is to report a case of trichomycosis palmellina flava in the pubic region present in a young adult and to present the clinical characteristics of this relatively unrecognized condition.

Effect of resveratrol and lipoic acid on sirtuin-regulated expression of metabolic genes in bovine liver and muscle slice cultures.

Sirtuins (Sirt) are NAD-dependent deacetylases that are activated by the antioxidants resveratrol (RSV) and lipoic acid (LA). The objective of this study was to determine in bovine liver and muscle slice cultures the effect of RSV and LA treatment on the expresssion of Sirt1, Sirt3, peroxisome proliferator-activated receptor γ coactivator 1α (PPARGC1A), and the forkhead box O transcription factors FoxO1 and FoxO3 as well as other factors involved in glucose and lipid metabolism and related to Sirt activity. Tissue slices from crossbred bulls were treated during 60 min with 40 or 80 µ RSV and 30, 100, 300, or 1,000 µ LA under restricted conditions (Krebs-Ringer buffer without nutrients) and fed conditions (2.5 m propionate in combination with 1 n glucagon) for liver slices or with 0.01 µ epinephrine for muscle slices. Quantitative real-time PCR was used to analyze the expression of the mRNA for the genes studied and western blot analysis for the expression of the protein for Sirt1. Our results show that the expression of the mRNA for Sirt1 was enhanced by RSV in liver under restriction ( ≤ 0.0112) and by LA in muscle, more under restriction ( ≤ 0.0121) than after epinephrine administration ( < 0.0001). Sirt3 is affected in a dose-dependent manner by both compounds in both tissues and under both metabolic conditions ( ≤ 0.0452). The expression of the protein for Sirt1 was increased by LA in both tissues under restricted conditions ( = 0.0026 and = 0.0201, respectively) but in liver also in fed conditions ( = 0.0016). Genes involved in the antioxidant response were upregulated in both tissues. These results indicate that bovine Sirt respond differently to RSV and LA stimulation than monogastric Sirt do and that gluconeogenesis in ruminants is not related to Sirt to the same degree as in monogastric species. However, these results provide information about the possible role of Sirt in ruminant metabolism.

Phase I and pharmacological studies of the cryptophycin analogue LY355703 administered on a single intermittent or weekly schedule.

LY355703 is a synthetic derivative of the marine cryptophycins, cytotoxic agents which induce mitotic arrest by binding at the microtubule vinca binding domain. Promising preclinical features of LY355703 were the 40-400 greater potency than paclitaxel or vinca alkaloids, the broad spectrum of antitumor activity in xenografts and the antitumour activity in multidrug resistant (MDR)-expressing murine tumours. Aims of this study were to define the maximum tolerated dose (MTD) and the dose recommended for Phase II, the pattern of toxicity, the pharmacokinetic profile and to document hints of antitumour activity of LY355703 given as 2-h infusion on day 1 every 3 weeks (Study 1) or, later on, on days 1, 8 and 15 every 4 weeks (Study 2). The latter weekly regimen was selected because of the acute dose-related toxicity reported in Study 1. The dose was escalated using a modified Continual Reassessment Method. Pharmacokinetic studies were performed on day 1 of cycle 1 in both studies; LY355703 plasma concentrations were assessed by liquid chromatography with tandem mass spectrometry. A total of 35 adult patients with solid tumours entered Study 1; the dose was escalated from 0.1 to 1.92 mg/m(2); at this dose 2 of 5 patients presented grade 3 neuropathy and myalgias; 1.48 mg/m(2) was then recommended for Phase II study. A total of 8 patients were treated in Study 2 at 1 mg/m(2); cumulative long-lasting neuroconstipation and neurosensory toxicity precluded the completion of the cycle in 9 out of 15 cycles; the clinical development of the weekly regimen was then discontinued. Other toxicities included cardiac dysrhythmia and mild alopecia. Pharmacokinetics of LY355703 appeared to be linear over the dose range studied. The administration of LY355703 on a 3-week schedule is associated with an acute dose-dependent peripheral neuropathy and myalgia of high interpatient variability for which possible risk factors and pharmacokinetic correlates could not be identified.

Phase I and pharmacokinetic studies of PNU-159548, a novel alkycycline, administered intravenously to patients with advanced solid tumours.

PNU-159548 (4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyl-daunorubicin) is the lead compound of a novel class of cytotoxic agents (alkycyclines) with a unique mechanism of action combining DNA intercalation with alkylation of guanines in the DNA major groove. The objectives of two phase I studies were to assess the dose-limiting toxicities (DLTs), to determine the maximum tolerated dose (MTD) and to study the pharmacokinetics (PKs) of PNU-159548 and its active metabolite PNU-169884 when administered intravenously (i.v.) over 10 or 60 min to patients with advanced solid tumours. Patients were treated with escalating doses of PNU-159548, courses repeated every 21 days at doses ranging from 1.0 to 16 mg/m(2). For pharmacokinetic analysis, plasma sampling was performed during the first course and assayed using a validated high-performance liquid chromatographic assay with mass spectrometric detection. 69 patients received a total of 161 courses. The MTD was reached at 14 and 16 mg/m(2) in heavily (HP) and minimally pretreated/non-pretreated (MP) patients, respectively, with thrombocytopenia as the DLT. A hypersensitivity reaction was observed in 8 patients across all dose levels, characterised by fever with chills, erythema, facial oedema and dyspnoea. The PKs of PNU-159548 and PNU-169884 were linear over the dose range studied. A significant correlation was observed between the percentage decrease in platelet count and the AUC of PNU-159548. In these studies, the DLT of PNU-159548 was thrombocytopenia. The recommended dose for phase II studies of PNU-159548 is 12 and 14 mg/m(2) administered i.v. over 10 min, once every 21 days, in HP and MP patients, respectively.

Response to first-line chemotherapy and long-term survival in patients with multiple myeloma: results of the MM87 prospective randomised protocol.

In this study we evaluated whether a good response to conventional chemotherapy, i.e. a significant tumour reduction, is a prerequisite for improved survival in multiple myeloma (MM). Between January 1987 and March 1990, 341 consecutive previously untreated patients with MM received chemotherapy within the prospective, multicentre, randomised Protocol MM87. Of these, 258 patients were evaluable for both response and long-term survival and 244 (94.6%) have died. The median survival of all patients was 40 months (6-162 months). The median survival did not differ between patients who had complete response (CR) (50 months (9-162 months)), partial response (PR) (46 months (8-147 months)) or stable disease (SD) (41 months (7-135 months)). The median survival was shorter (13.6 months (6-135 months)) (P<0.0001) in patients whose disease progressed while they were receiving first induction chemotherapy. Causes of death were more frequently (P=0.04) related to MM in patients who had progressive disease (PD) than in patients who had a CR or PR or SD. The main clinical and laboratory characteristics were similar in the four groups. These data indicate that patients who maintain SD during first-line chemotherapy have a prognosis similar to that of patients who attain a response. Only patients whose disease progresses have a distinctly worse outcome.

A prospective, controlled, nonrandomized study on prophylactic parenteral dichloromethylene bisphosphonate (clodronate) in multiple-myeloma.

Bone resorption by osteoclasts causes neoplastic bone disease, which is a significant cause of death in multiple myeloma (MM). Counteracting bone resorption with prophylactic bisphosphonates has delayed bane disease, and this is expected to improve survival. Between January, 1987 and March, 1990, 341 evaluable previously untreated, consecutive patients with MM entered a prospective, multicenter study in which cytostatic therapy was randomized. The first 148 patients recruited were not planned for prophylaxis and the following 193 were scheduled to receive parenteral, prophylactic clodronate. Clodronate was administered at a dose of 600-1000 mg/4-6 weeks and was started at diagnosis and continued throughout survival time. Data on clodronate prophylaxis were evaluated on both an intention-to-treat and a compliance analysis basis. The rate of response and the duration of response were independent of clodronate prophylaxis. Progression of skeletal disease occurred less often in patients who received the drug than in those who were not given prophylaxis (50.5 vs 34.8%; p<.02 by compliance analysis). Survival was longer for patients on clodronate prophylaxis than for those who were not planned for (p<.02 by intention to-treat-analysis) or for those who did not receive clodronate prophylaxis (p<.009 by compliance analysis). Local pain associated with i.m. administration was the only significant side effect of clodronate. Parenteral clodronate prophylaxis prolongs survival in MM, probably because it allows better control of bone disease and reduces deaths related to it.

The use of peripheral-blood hematopoietic progenitors mobilized with standard-dose chemotherapy plus granulocyte-colony-stimulating factor to support multicyclic dose-intensive chemotherapy for advanced breast-cancer.

This study was aimed at determining: (a) the degree of mobilization of peripheral blood hematopoietic progenitors (PBSC) induced by a single course of standard-dose chemotherapy (CT) followed by G-CSF and the feasibility and safety of the administration of multiple courses of intensified CT with repeated PBSC reinfusions; (b) the relationship between the number of mononuclear cells (MC) in S-phase of the cell cycle (as evaluated by DNA flow cytometry, FCM), the CRT-GM and the CD34(+) cells in the leukapheresis product. Six patients with metastatic breast cancer received a course of standard FEC (5-FU 600 mg/m(2), epirubicin 75 mg/m(2), cyclophosphamide, CTX, 600 mg/m(2), day 1) followed by G-CSF (5 mu g/kg twice a day, from day 3 until leukapheresis), which served as both initial treatment for their disease as well as the PBSC mobilization technique. Collected PBSC were fractionated and reinfused, without G-CSF, following each of further 5 subsequent intensified FEC (HD-FEC: 5-FU 750 mg/m(2), epirubicin 100 mg/m(2), CTX 1,000 mg/m(2)) courses planned at 21-day intervals. The individual hematopoietic reconstitution curves showed superimposable profiles for all patients, and the leukaphereses were performed between days 7 and 10 after the first CT course. A median of 18.8x10(9) (10.4-35.6) MC, 9.3 (2.6-23.3) CD34(+) cells x 10(6)/kg body weight and 9.8 (1.6-27.3) CFU-GM x 10(4)/kg body weight were collected from each patient (with 1 or 2 phereses). All patients received the planned 5 courses of HD-FEC followed by PBSC reinfusion, without experiencing haematological cumulative toxicity >WHO grade 3 for WBC and >grade 2 for PLT. No >grade 3 non-hematological toxicity was recorded. There were no treatment-related delays in CT administration so that the delivered average relative dose-intensity (ARDI) was 1.65. A good correlation was seen between the percentage of MC in S-phase and the number of CFU-GM (R(2)=0.566, p<0.0065) or the number of CD34(+) cells (R(2)=0.625, p<0.0031) in the leukapheresis product. A single course of standard FEC+G-CSF is effective in mobilizing sufficient amounts of PBSC to support 5 additional courses of HD-FEC, which could represent an alternative to single, myelo-suppressive CT programs. DNA analysis by FCM should be further investigated as a rapid method for PBSC quantification, since proliferating MC and CFU-GM were closely related.

Advanced breast-cancer - a randomized study of different doses of epirubicin associated with fixed doses of cyclophosphamide and 5-Fluorouracil.

A moderate increase in the dose of anthracycline could be feasible and of clinical benefit in advanced breast cancer. Between April 1991 and April 1994, 69 consecutive patients with recurrent or metastatic breast cancer were randomly treated with two regimens, including different dosages of epirubicin (75 versus 100 mg/m(2)) associated with the same dosage (600 mg/m(2)) of cyclophosphamide and 5-fluoruracil (75-FEC vs 100-FEC). Patients were planned to receive 6 courses at 21 day-intervals. Thirty-six patients received the 75-FEC regimen and 33 received the 100-FEC regimen. The two groups were comparable for age, menopausal status, disease-free interval, previous therapy, performance status and sites of disease: Over the whole study, the 100-FEC regimen has allowed a 18% actual increase in the epirubicin dosage over the 75-FEC regimen. Overall response rate was 56% for the 100-FEC and 51% for the 75-FEC, with the 100-FEC inducing some more complete responses than the 75-FEC (38% vs 23%). Survival (but not time to progression) tended to be longer with the 100- than with the 75-FEC (median: 20 vs 13 mos, p<0.09). Nonhematologic side effects Were similar. Hematologic toxicity was slightly higher with 100- than with 75-FEC, with granulocyte colony stimulating factors used to recycle in the scheduled time in the 15 and 4% of courses, respectively.

Multicyclic dose-intensive chemotherapy with circulating progenitor cell support for high-risk primary breast cancer.

The feasibility and safety of the administration of multiple cycles of dose-intensive chemotherapy (CT) supported with repeated reinfusions of circulating progenitor cells (CPCs) were evaluated in a prospective study of adjuvant initial therapy of poor-prognosis breast cancer. Eighteen patients with resectable breast cancer involving >/= 10 axillary nodes or >/= 5 axillary nodes and negativity of the estrogen receptor status received a cycle of standard FEC regimen (5-FU 600 mg/m2, epirubicin 60 mg/m2, CTX 600 mg/m2, i.v. on day 1) followed by G-CSF as CPC mobilization technique. Collected CPCs were fractionated and reinfused, with G-CSF, after each of the 4 subsequent cycles of high-dose FEC (HD-FEC) (5-FU 750 mg/m2, epirubicin 120 mg/m2, CTX 3 g/m2, i.v.) planned at 21 day intervals. The median numbers of CD34+ cells and CFU-GM collected (with one or two leukaphereses per patient) were 9.7x10(6)/kg (range: 2.5-22.9) and 9.9x10(4)/kg (range: 1.9-27.3), respectively, and day 9 was the median first day of procedure (range: 8-12) after FEC. All patients received the 4 courses of HD-FEC (for a total of 72 cycles). Hemopoietic recovery was rapid after each cycle and there was no treatment-related delays in CT administration. Mucositis was the major non-hematological toxicity. There were 2, 3, 7 and 9 episodes of WHO grade 3/4 mucositis in cycles 1, 2, 3 and 4, respectively. These severe episodes lasted a median of 4 days (range: 2-6) but no patient required parenteral nutrition. The mean +/- SD total hospital stay lasted 10 +/- 2 days. The delivery of 4 cycles of dose-intensive FEC CT supported by CPCs (mobilized with a single course of standard-dose FEC + G-CSF) is feasible and safe. It could represent an effective alternative strategy to other more aggressive programs for the adjuvant therapy of high-risk early breast cancer.

The pheromonal restoration of cyclic activity in young estrogenized persistent estrus female rats is a vomeronasal effect.

Eight sham-operated (SO) and six vomeronasalectomized (VMNX) young adult female rats were used to demonstrate the participation of the vomeronasal organ (VNO) in the pheromonal restoration of cyclic activity (PRCA). All rats were normal four-day cycling before and after surgery and they received a single subcutaneous injection of 2 mg of estradiol benzoate (EB) to induce an anovulatory persistent vaginal estrus. Fifteen days after EB injection, they were treated by spraying in the nostrils, every 10 min for one hour on only one day, 50 muL of urine from young fertile males. SO rats showed PRCA in the following days, while VMNX did not. These results support the VNO as the receptor of the male urinary pheromone (or pheromones) responsible for PRCA, demonstrating that it is possible to improve the function of aging ovaries acting with an exteroceptive stimulus.

Involvement of the corticoadrenal hormones in the pheromonal restoration of reproductive activity in aging rats.

The involvement of the adrenal progesterone and corticosterone in the early gonadotropin secretion associated with the pheromonal restoration of ovarian cyclic activity (PRCA) in aging female rats is studied. PRCA is induced by male urinary pheromones and is preceded by an alpha-adrenergic-mediated release of the hypothalamic decapeptide luteinizing hormone-releasing hormone and plasma increases of estradiol, progesterone and the gonadotropins luteinizing hormone and follicle stimulating hormone. Aging reproductive Wistar female rats were used to study the effects of bilateral adrenalectomy and of a subcutaneous injection of the antisteroid RU486 on plasma levels of corticosterone, progesterone and gonadotropins in rats stimulated with nasal spraying of male urine (MU) or saline. The results demonstrate that progesterone and corticosterone released by MU are from adrenal origin, and that these adrenal secretory products are critical for MU-induced increase of gonadotropins. This suggests that olfactory stimulation of ACTH release stimulates adrenal release of progesterone and corticosterone, and both trigger the events that initiate the activation of the hypothalamus-pituitary-ovarian axis that leads to PRCA.

Effect of nasal instillation of female urine or vaginal exudate on testosterone secretion in isolated and anesthetized male rhesus monkeys (Macaca mulatta).

Two male adult rhesus monkeys were individually placed in cages with a pulling device in order to immobilize the animals for anesthesia. The room was temperature-controlled having a light/dark period of 12/12 hours. The animals were rapidly immobilized and immediately anesthetized with ketamine i. m. (10 mg/kg of body weight). They were bled four times at 15, 30, 45, and 60 mins after the ketamine injection, twice a week during 6 weeks. When necessary, maintenance doses of ketamine were administered. The levels of serum testosterone in experimental conditions (nasal instillation of female urine or a suspension of vaginal exudate) showed significant lower values with respect to those in control conditions (saline instillation). The control levels of testosterone tend to increase up to 60 mins. The testosterone from samples obtained in experimental conditions did not show such an increase, remaining similar during the sampling and similar to the 15 min control levels that could be considered as basal. These results seem to point out some chemical information from females capable of modifying the pattern of secretion of testosterone of the males in the above mentioned experimental conditions.

Effect of nasal instillation of female urine or vaginal exudate on serum cortisol and prolactin levels in isolated and anesthetized male rhesus monkeys (Macaca mulatta).

Two male adult rhesus monkeys were used and caged individually. The room was temperature-controlled having a light-dark period of 12/12 hours. The animals were rapidly immobilized and immediately anesthetized with ketamine i. m. (10 mg/kg of body weight). They were bled four times at 15, 30, 45 and 60 mins after the ketamine injection, twice a week for 6 weeks. When necessary, maintenance doses of ketamine were administered. The serum levels of cortisol and prolactin after nasal instillation of a suspension of vaginal exudate showed lower values than in control conditions (nasal instillation of saline). The control levels of cortisol tended to increase up to 60 mins, whilst in experimental conditions (nasal instillation of female urine or vaginal exudate) did not show such an increase. Cortisol remained similar during the sampling and similar to the 15 mins control levels, but the difference is statistically significant only after instillation of vaginal exudate as compared with control. The levels of prolactin did not show significant variations during sampling either in control or after female urine instillation. However, the instillation of vaginal exudate decreased the prolactin levels at 15 mins which tended to recover the control levels up to 60 mins. These results support the hypothesis, discussed in a previous paper, that some chemical information from females suppresses or mitigates the effect of acute stress resulting from handling the animals before anesthesia.

CD-44: a marker of metastases of head and neck melanomas.

Specimens of 38 melanomas of the oral cavity and facial skin and 23 lymph nodes with melanoma metastases were stained immunohistochemically for the presence of variants 5, 6, 7 and 8 of CD-44 antigen. A positive reaction was observed only for variant 7-8 in primary melanomas as well as in lymph nodes with metastases from melanoma. The CD-44 score (percentage of CD-44 positive cells chi intensity of reaction) correlated with metastatic behaviour of tumours and was greater in metastatic lymph nodes than in primary tumours.

DNA content and cell kinetics in colorectal carcinoma: flow cytometric analysis of primary tumor and liver metastases.

AIMS AND BACKGROUND: Both flow cytometric DNA ploidy and proliferative activity have been indicated as potential prognostic indicators in colorectal cancer. Due to tumor biological heterogeneity, these parameters are best assessed with multiple sampling. METHODS: We undertook a prospective study on 52 patients with Duke's D colorectal tumors looking at multiple samples of the primary tumors and liver metastases. DNA ploidy and tumor proliferative activity (derived from proliferating cell nuclear antigen, PCNA FCM expression) were evaluated. RESULTS: Of primary tumors, 36/52 (69.2%) were aneuploid in at least 1 sample, with a median value of the DNA index of the aneuploid peak of 1.58. On liver metastases, 42/52 (80.7%) patients were aneuploid in at least 1 sample with a median DNA index of the aneuploid peak of 1.64. Identical or nearly identical histograms from different tumor samples were observed in only 18/52 (34.6%) of the primary tumors and in 15/52 (28.8%) of the liver metastases. The PCNA values for primary tumors ranged from 5 to 28% (median value = 16.5%). In the liver metastases, PCNA values ranged from 12 to 38% (median value = 19.8%). Proliferative activity was lower for diploid than for aneuploid tumors. DNA ploidy and PCNA expression of the deep specimen of primary tumors were similar to those of the liver metastasis of the same patient while this concordance was not complete in the case of superficial biopsy specimens. CONCLUSIONS: If correctly performed, FCM techniques allow an accurate analysis of DNA ploidy and proliferative activity and both these measurements can offer considerable potential for a more comprehensive approach to colorectal cancer.

Histological development of the vomeronasal complex in the pre- and postnatal rat.

The vomeronasal complex of the rat was studied by means of different staining techniques and light microscopy between the 13th day of gestation and 42 days of postnatal life. The anlage of the vomeronasal organ in the 13-day-old embryo consisted of a cluster of cells proliferating from the olfactory placode towards the medial line. The vomeronasal organ was well developed by the end of gestation, showing the same ratio between receptor and receptor-free epithelium as in the adult animal. However, full development of the epithelia did not occur until the end of the second postnatal week. Glandular rudiments were observed within the vomeronasal area in 17-day-old fetuses, but the gland showed no sign of activity until the 7th day of postnatal life, when a slight PAS-positive reaction was observed in their cells' cytoplasm. This PAS-positive reaction intensified considerably during the second week and corresponded to the adult picture by the end of this time. The content of the vomeronasal glands contrasted with that of the glands flowing into the nasal fossa, which did not show any PAS-positive reaction but which stained positive for alcian blue. We also studied the evolution of the blood vessels such as the capsule of the complex. This capsule showed complete ossification by the beginning of the 3rd postnatal week, with the exception of certain regions which remained cartilaginous in the adult. Some capillaries were observed in the complex in 17-day-old fetuses, and by 20 days of gestation some of them could be observed penetrating the receptor epithelium. Subsequently, the vascularization of the complex became so profuse that one could consider the vomeronasal complex to consist of erectile tissue.

Low cleavage activity of 15,15'dioxygenase to convert beta-carotene to retinal in cattle compared with goats, is associated with the yellow pigmentation of adipose tissue.

Two experiments (one with twelve heifers and the other with 14 goats) were designed to evaluate the effect of increasing dietary beta-carotene concentration on the activity of the cleaving enzyme 15,15' beta-carotene dioxygenase and the concentration of beta-carotene and retinol in selected tissues. The experiments lasted 120 days. During the first 90 days animals were offered a beta-carotene unsupplemented diet. In the following 30 days, they were distributed to one of three treatments: 0, 5.5 or 352 mg of beta-carotene/kg of dry matter intake. All animals were slaughtered at the end. In heifers the concentration of beta-carotene in plasma, reflected the level of beta-carotene fed. Goats had detectable levels of beta-carotene only on day 10 of supplementation. In the liver, beta-carotene concentrations were highest with the 352 dose in both species. Heifers had the highest concentration of beta-carotene in the adipose tissue. In bovines, no interaction between beta-carotene treatment and intestinal sampling site was found (P > 0.2) for the activity of 15,15 dioxygenase. Across beta-carotene levels, results of the enzyme assay were: 0.19, 0.32 and 0.45 nmol retinal/(mg S-9 protein/h) (P < 0.01) for 0, 5.5 and 352 mg beta-carotene.kg dry matter intake -1.d-1, respectively; across intestinal sampling sites results of the enzyme assay were 0.45, 0.43 and 0.08 nmol retinal/(mg S-9 protein/h) (P < 0.01) for duodenum, jejunum and ileum, respectively. Caprine data showed an interaction between beta-carotene treatment and intestinal sampling site (P < 0.05) for the activity of 15,15 dioxygenase. The results for treatment 0 were: 1.4, 1.4 and 0; for treatment 5.5: 1.41, 1.42 and 0.13; and for treatment 352: 1.46, 1.99 and 0.48 nmol retinal/mg S-9 protein/h for duodenum, jejunum and ileum, respectively. The lower levels of duodenal and jejunal 15,15'dioxygenase activity in cattle compared with goats, may explain the greater pigmentation of adipose tissue in the former ruminant specie.