Clock gene per 2 daily rhythm: Correlation with the serum level of uncoupling protein 1 (UCP1) in goat and horse.

Recent studies evidenced that the circadian rhythm of Per2 is involved in adaptive thermogenesis by the modulating transcription of uncoupling protein 1 (UCP1). For this purpose, we investigated the linkage between the daily rhythm of Per2 and UCP1 in ruminant and non-ruminant mammalian species. Five clinically healthy, not pregnant, and not lactating Maltese female goats and five clinically healthy, not pregnant, and not lactating Italian Saddle horses were enrolled in the study. All animals were housed under natural photoperiod (sunrise 05:05, sunset 20:55) and environmental temperature and humidity. Goats were kept individually in 3.0 × 2.0 m box, horses were housed individually in 3.5 × 3.5 m box; all boxes were equipped with an opening window. On each subject, blood samples were collected every 4 h for a 48-h period. The Per2 gene expression was determined on blood samples collected in PAX gene Blood RNA Tube, whereas UCP1 concentration was assessed on serum. Per2 and UCP1 levels were statistically influenced by the species (p < 0.0001) and the time of data collection (p < 0.0001), but not by the day of monitoring. Per2 showed daily rhythmicity, statistically different in mesor and amplitude between the two species, diurnal in goats, nocturnal in horses; with the same robustness. UCP1 did not show daily rhythmicity. During the experimental period the two parameters showed a negative correlation in horses. According to the findings herein obtained, we can claim that the role of Per2 in the thermogenesis induced by the beige adipocytes throughout UCP1 activation did not reflect what found in other mammal species, but further studies are required to establish their correlation in equids.

Thermographic ocular temperature correlated with rectal temperature in cats.

Body temperature reflects the animal health and/or disease conditions. During clinical examination, temperature measurement is a basic step in veterinary medicine. The conventional method used is the rectal thermometry, particularly stressful in some subjects, especially for cats. A less stressful alternative method, such as infrared thermal imaging camera, is used in various fields of medicine for diagnosis, prognosis and correct therapeutic approaches. To evaluate the usefulness of infrared thermal imaging for the assessment of ocular temperature, twenty cats of different breeds (European, Siamese and Persian, 4-6 years old, mean body weight 4.3 ± 0.30 Kg) were enrolled in the study. In order to evaluate the applicability of the ocular temperature assessment through thermal imaging as a tool for measuring the animal's body temperature, the obtained values were compared with the rectal temperature values recorded in each cat by means of a digital thermo-camera. There were no differences between left and right eye; and a difference of about 1.19 °C between the ocular and rectal temperature value was recorded (p < 0.0001). Rectal and ocular temperatures were positively correlated (p < 0.0001; r = 0.93). In conclusion, we show that ocular temperature is an alternative method for body temperature measurement that can be used in clinical evaluation of cats, especially in cases where rectal temperature recording is not possible.

Leukocyte modifications during the first month after foaling in mares and their newborn foals.

During early post-partum period both neonatal foals and peripartum mares are most susceptible to diseases. The aim of this study was to establish physiologic modifications of leukogram during the first month after foaling in mares and their newborn foals. To this end blood samples were collected from nine mares and nine foals (T0-T10), every three days from the 1st day until the 30th day after foaling. Samples were analysed for white blood cell (WBC) count and differential leucocyte counts. Two-way repeated measure analysis of variance (ANOVA) showed, in postpartum mares WBC showed significant higher values at T0 (9.02±0.76) in respect to other time points, and at T2 (8.08±0.53) and T3 (7.92±0.59) compared to T1 (6.98±0.43), whereas in foals lower WBC values at T0 (6.11±0.49) compared to other experimental periods except T1 (6.90±0.94), and at T1 compared to T8 (7.95±0.61) and T10 (7.90±0.36) were observed. The differential leucocyte counts showed significant modifications in the percentage of neutrophils (π<0.001) and lymphocytes (p<0.001) both in postpartum mares and in foals during the experimental period. Furthermore ANOVA showed significant differences between postpartum mares and foals (P<0.01) in all studied parameters, and between postpartum mares and control mares in WBC and neutrophils values. The obtained results provide suitable information about the influence of foaling on leukogram of periparturient mares and reveal WBC dynamics in newborn foals during the first month post-partum.

Neurodevelopmental trajectories of letter and speech sound processing from preschool to the end of elementary school.

Learning to read alphabetic languages starts with learning letter-speech-sound associations. How this process changes brain function during development is still largely unknown. We followed 102 children with varying reading skills in a mixed-longitudinal/cross-sectional design from the prereading stage to the end of elementary school over five time points (n = 46 with two and more time points, of which n = 16 fully-longitudinal) to investigate the neural trajectories of letter and speech sound processing using fMRI. Children were presented with letters and speech sounds visually, auditorily, and audiovisually in kindergarten (6.7yo), at the middle (7.3yo) and end of first grade (7.6yo), and in second (8.4yo) and fifth grades (11.5yo). Activation of the ventral occipitotemporal cortex for visual and audiovisual processing followed a complex trajectory, with two peaks in first and fifth grades. The superior temporal gyrus (STG) showed an inverted U-shaped trajectory for audiovisual letter processing, a development that in poor readers was attenuated in middle STG and absent in posterior STG. Finally, the trajectories for letter-speech-sound integration were modulated by reading skills and showed differing directionality in the congruency effect depending on the time point. This unprecedented study captures the development of letter processing across elementary school and its neural trajectories in children with varying reading skills.

Association of TCF7L2 gene variants with low GAD autoantibody titre in LADA subjects (NIRAD Study 5).

AIMS: We previously demonstrated the presence of two different populations among adult-onset autoimmune diabetes (latent autoimmume diabetes of adults; LADA) having high or low titre of antibodies to glutamic acid decarboxylase (GADA). The transcription factor 7-like 2 (TCF7L2) gene has been recognized as the major gene associated with Type 2 diabetes. The aim of the present study was to evaluate whether the phenotypic heterogeneity of LADA based on GADA titre is associated with TCF7L2 polymorphisms. METHODS: Two hundred and fifty patients identified as LADA, divided into two subgroups with low (< or = 32 arbitrary units) or high (> 32 units) GADA titre, 620 subjects with Type 2 diabetes [from the Non-Insulin Requiring Autoimmune Diabetes (NIRAD) study cohort of 5330 subjects] in addition to 551 consecutive cases of Type 1 diabetes and 545 normoglycaemic subjects were analysed for the rs12255372 and rs7903146 polymorphisms of the TCF7L2 gene using Taqman. RESULTS: The genotype and allele distributions of the two polymorphisms revealed similar frequencies in subjects with low GADA titre and Type 2 diabetes. High GADA titre, Type 1 diabetes and controls also showed comparable frequencies. A significant increase of GT/TT genotypes of the rs12255372 single-nucleotide polymorphism (SNP) and CT/TT genotypes of the rs7903146 SNP was observed in low GADA titre and Type 2 diabetes compared with high GADA titre, Type 1 diabetes and controls (P < or = 0.04 for both comparisons). The risk alleles of both variants were increased in low GADA titre and Type 2 diabetes compared with high GADA titre, Type 1 diabetes and control subjects (P < 0.02 for all comparisons). CONCLUSIONS: TCF7L2 common genetic variants of susceptibility are associated only with low GADA antibody titre in LADA patients.

Cardinal directions for visual optic flow.

As we move through our environment, the flow of deforming images on the retinae provides a rich source of information about the three-dimensional structure of the external world and how to navigate through it. Recent evidence from psychophysical [1] [2] [3] [4], electrophysiological [5] [6] [7] [8] [9] and imaging [10] [11] studies suggests that there are neurons in the primate visual system - in the medial superior temporal cortex - that are specialised to respond to this type of complex 'optic flow' motion. In principle, optic flow could be encoded by a small number of neural mechanisms tuned to 'cardinal directions', including radial and circular motion [12] [13]. There is little support for this idea at present, however, from either physiological [6] [7] or psychophysical [14] research. We have measured the sensitivity of human subjects for detection of motion and for discrimination of motion direction over a wide and densely sampled range of complex motions. Average sensitivity was higher for inward and outward radial movement and for both directions of rotation, consistent with the existence of detectors tuned to these four types of motion. Principle component analysis revealed two clear components, one for radial stimuli (outward and inward) and the other for circular stimuli (clockwise and counter-clock-wise). The results imply that the mechanisms that analyse optic flow in humans tend to be tuned to the cardinal axes of radial and rotational motion.

Benign breast disease and consumption of beverages containing methylxanthines.

The relationship between methylxanthine (Mx) consumption and benign breast disease was evaluated in a case-control study of 288 women with histologically confirmed benign breast lumps (203 dysplastic lesions and 85 benign tumors) and 2 groups of control women--285 patients in the hospital for acute conditions apparently unrelated to the consumption of Mx-containing beverages and 291 outpatients. The relative risk estimates of dysplastic breast lesions (fibrocystic disease), with allowance for all identified potential distorting factors, for women who drank 1-2 or 3 or more cups of coffee per day were 4.1 and 6.4, respectively, when the hospital controls were the comparison group and 2.0 and 3.7, respectively, when the outpatient controls were the comparison group. The relationship was even stronger when the total consumption of Mx-containing beverages (coffee plus tea) was considered and increased with increasing duration of use. The association was not explained by any of the major risk factors for fibrocystic breast diseases or by differences in general characteristics or other lifestyle habits between cases and controls. Mx consumption was not related to the risk of benign breast tumors (fibroadenomas). These findings support the hypothesis that Mx consumption is related to the risk of dysplastic lesions of the breast.

High testosterone and low progesterone circulating levels in premenopausal patients with hyperplasia and cancer of the breast.

Serum testosterone, progesterone, luteinizing hormone, prolactin, and sex hormone-binding globulin levels were measured in 55 normal controls, in 31 patients with hyperplastic alterations of breast epithelium, and in 23 patients with breast cancer. All patients and controls were premenopausal, and they were comparable for age, weight, and body surface. In the controls, the mean level of testosterone [0.47 +/- 0.16 (S.D.) ng/ml] was lower and the mean level of progesterone (17.63 +/- 8.11 ng/ml) was higher than in breast cancer patients [testosterone level, 0.62 +/- 0.22 ng/ml (p less than 0.005); progesterone level, 11.4 +/- 8.0 ng/ml (p less than 0.005)] and in patients with breast epithelial hyperplasia [testosterone level, 0.55 +/- 0.2 ng/ml (p less than or equal to 0.05); progesterone level, 13.9 +/- 8.6 ng/ml (p less than 0.05)]. No difference was found in the mean circulating levels of luteinizing hormone, prolactin, or sex hormone-binding globulin between controls and patients. These results confirm previous findings of increased urinary testosterone excretion in women with anovulatory menstrual cycles and epithelial hyperplasia or cancer of the breast and strongly support the hypothesis that androgens play an important role in both induction and development of breast cancer.

Increased androgenic activity and breast cancer risk in premenopausal women.

Blood and urine specimens from 27 premenopausal breast cancer patients and 62 healthy controls have been compared with respect to concentration of testosterone and progesterone in blood and of testosterone and androstanediol in urine, measured in the luteal phase of the menstrual cycle. There was a strong positive association between the concentration of the two androgens, either in blood or urine, and breast cancer risk. A strong association was also observed with decreasing levels of progesterone. The association was statistically significant (p for trend less than 0.01) for each hormone; the rate ratios were 10.2 for serum testosterone (highest category), 5.6 for serum progesterone (lowest category), 8.4 for urinary testosterone (highest category), and 5.2 for androstanediol (highest category). The rate ratio for women presenting both high serum testosterone and low progesterone was 21.8 (4.1 to 116.1). Considering the exposure to at least one of three androgens at the highest level and low progesterone, the rate ratio was as high as 90.2 (8.2 to 989.7). This study provides evidence for the hypothesis that increased androgenic activity is an important risk indicator for breast cancer, particularly when associated with anovulation, as indicated by low serum progesterone level.

Serum and urinary androgens and risk of breast cancer in postmenopausal women.

Serum levels of testosterone, dihydrotestosterone, androstenedione, dehydroepiandrosterone sulfate, and sex hormone-binding globulin and urinary levels of testosterone and androstanediol were compared in 75 women with breast carcinoma and 150 age-matched healthy controls. Odds ratios for quartiles of hormones, adjusted for known potential confounders, were computed using conditional logistic regression. Risk of breast cancer was positively associated with levels of all androgens in serum and urine but appeared stronger for testosterone (for trend, P = 0.03) and dehydroepiandrosterone sulfate (for trend, P = 0.06) in serum and for testosterone (for trend, P = 0.001) and androstanediol (for trend, P = 0.04) in urine. The adjusted odd ratios for high versus low quartiles were 2.7 (95% confidence interval, 1.1-6.5) for serum testosterone, 2.8 (1.1-7.4) for dehydroepiandrosterone sulfate, 4.7 (1.8-12.1) for urinary testosterone, and 3.4 (1.4-8.7) for urinary androstanediol. These observations suggest that endogenous androgenic hormones may play an important role in the epidemiology of postmenopausal breast cancer in women.

Androgens and breast cancer in premenopausal women.

We investigated the role of androgens in premenopausal breast cancer by comparing serum testosterone, dihydrotestosterone, androstenedione, dehydroepiandrosterone sulfate, progesterone, sex-hormone-binding globulin-binding capacity, and urinary testosterone and androstanediol in 63 women with breast adenocarcinoma and 70 healthy controls of similar age. With variables dichotomized at the 75th percentile, the age-adjusted relative risk was 3.4 (95% confidence interval, 1.6-7.3) for high versus low levels of serum testosterone, 2.1 (0.9-4.8) for urinary testosterone, and 2.5 (1.1-5.9) for serum dihydrotestosterone. We observed no differences in other hormones. The strength of the associations changed markedly with increasing time to the onset of the next menses. The risk for testosterone and dihydrotestosterone, which was negligible in women with onset within 5 days of sampling, increased progressively to nearly 10-fold higher than in unstratified data in women with onset 10 days or more after sampling. This study provides arguments in favor of a role for increased androgenic activity in premenopausal breast cancer. It also suggests that unknown factors related to cycle length may be important in modulating the strength of the association with testosterone. The results are discussed also in reference to possible biases and inadequacies in study design.

Isolation, characterization and binding properties of two rat liver fatty acid-binding protein isoforms.

Mammalian liver has only one fatty acid-binding protein (L-FABP) while the liver of non-mammalian vertebrates expresses a liver basic FABP (Lb-FABP) in addition to other members of the FABP family. We explore the possibility that L-FABP isoforms accomplish, in the liver of mammals, the metabolic functions corresponding to the different FABPs present in the liver of non-mammalian vertebrates. We have isolated isoforms I and II which have a different residue 105, Asn in the former and Asp in the latter. We made a conformational comparison of the apo-isoforms by intrinsic fluorescence emission and fourth-derivative spectroscopy, native-state proteolysis and unfolding curves. Ligand affinity was studied by measuring cis-parinaric acid displacement by different ligands. They have differences in their molecular conformation, including the environment of the binding site. Isoform II has probably a more open conformation than isoform I, thus allowing the binding of a greater variety of ligands. The affinity of isoform II for lysophospholipids, prostaglandins, retinoids, bilirubin and bile salts is greater than that of isoform I. These characteristics of rat L-FABP isoforms I and II suggest that they may accomplish different functions as happens with those of the different FABP types in non-mammalian species.

Binding properties of Echinococcus granulosus fatty acid binding protein.

EgFABP1 is a developmentally regulated intracellular fatty acid binding protein characterized in the larval stage of parasitic platyhelminth Echinococcus granulosus. It is structurally related to the heart group of fatty acid binding proteins (H-FABPs). Binding properties and ligand affinity of recombinant EgFABP1 were determined by fluorescence spectroscopy using cis- and trans-parinaric acid. Two binding sites for cis- and trans-parinaric acid were found (K(d(1)) 24+/-4 nM, K(d(2)) 510+/-60 nM for cis-parinaric acid and K(d(1)) 32+/-4 nM, K(d(2)) 364+/-75 nM for trans-parinaric). A putative third site for both fatty acids is discussed. Binding preferences were determined using displacement assays. Arachidonic and oleic acids presented the highest displacement percentages for EgFABP1. The Echinococcus FABP is the unique member of the H-FABP group able to bind two long chain fatty acid molecules with high affinity. Structure-function relationships and putative roles for EgFABP1 in E. granulosus metabolism are discussed.

Clinical nutrition in gerontology: chronic renal disorders of the dog and cat.

The recent advances in the nutrition of companion animals has resulted in a longer possible life-span for dogs and cats and an improvement in their quality of life. Numerous studies about geriatric animals show that an aging dog or cat requires a specific nutritional formulation that considers the metabolic changes associated with age. A correct diet plays an important role in the treatment of some chronic pathologies in aging animals, particularly those for which the aging process modifies the organ function. A correct diet can provide therapeutic support to the administration of drugs that can sometimes compromise organ function. In the present study, we identify key aspects of the clinical nutrition during chronic renal disorders of dogs and cats, diseases with an elevated incidence and a major cause of mortality in geriatric animals. The aim of nutritional treatment for dogs and cats affected by chronic renal disorders is to improve the quality and length of life, assuring an adequate amount of energy and slowing the progression of renal failure. To improve treatment efficacy it is necessary to prepare different dietary rations during the various stages of disease, on the basis of clinical signs and laboratory data.

Critical evaluation of the use of thermography in the investigation of scintigraphically cold thyroid nodules.

One hundred and fifty-five cases of scintigraphically "cold" thyroid nodules were examined telethermographically to determine their thermal gradient. All nodules hyperthermic greater than or equal to 1.5 degrees C were successively removed, as were those (regardless of the thermal gradient) present in men, those in women younger than 21 years and older than 45 years of age, and those clinically suspect, the latter regardless of age or sex of the patient. Of 95 operated cases, 79 proved to be benign or nontumoral nodules, and 16 were carcinomas at histology. Hyperthermia greater than or equal to 1.5 degrees C had been present in 43% of the benign cases and in only 25% of the carcinomas. Out of ten cases were hyperthermia greater than or equal to 2 degrees C, none was a carcinoma. A malignancy was found in 26% of 23 normothermic cases, 17.6% of 34 slightly hyperthermic cases, and only 10.5% of 38 markedly hyperthermic (greater than or equal to 1.5 degrees C) cases. There was no evident correlation between the thermal gradient and the clinical diameter of the thyroid nodule. Our study affirms that thermography is not reliable when used to select cold thyroid nodules for surgical removal.

Fine needle aspiration cytology in superficial lymph nodes: an analysis of 266 cases.

The results of fine needle aspiration cytology of superficial lymph nodes in 275 patients are discussed. Nine cases were lost to follow-up. Of the remaining 266 aspirates, 152 were classified as cytologically malignant, 79 as benign, three as suspicious for malignancy, and 32 as unsatisfactory owing to scanty cellularity. The sensitivity of cytology for metastatic cancer was 96.5%. The results of aspiration biopsy from lymphomas were less accurate (67.5% sensitivity). There were no false positives and nine (11.3%) false negatives. Open biopsy of three suspect lymph nodes proved them to be malignant. The aspiration procedure is easy, safe and reliable. The diagnosis of benignity cannot, however, be determined without an open biopsy, particularly for lymphomas.

Localized mastalgia as presenting symptom in breast cancer.

The importance of breast pain as a presenting symptom of breast cancer was assessed in 200 women with localized mastalgia but negative physical examination and in 478 women with operable breast cancer. In the first group, mammography detected five cases of subclinical breast cancer at the site of pain. In the second group, 86 patients (18%) reported localized pain as presenting symptom, followed, at different intervals, by the detection of a breast lump. Localized pain can be considered a presenting symptom of breast cancer thus requiring a careful physical and mammographic examination, especially when risk factors are associated.

Diagnostic efficacy of the clinical-radiological-cytological triad in solid breast lumps: results of a second prospective study on 631 patients.

The clinical-radiological-cytological triad was used for diagnostic evaluation in 631 women over 30 years old with a solid breast lump, excluding clinically obvious cancers. All the lumps were subsequently removed surgically, except for 105 cases which spontaneously regressed within 2 months. 285 of the 526 nodules removed were cancers, and 162 of 285 (57%) did not exceed 20 mm in size. The sensitivity of the individual tests (physical examination, mammography and fine-needle aspiration cytology) of the malignant lumps was respectively 0.83, 0.73 and 0.60, and it increased to 0.95 when they were associated. Of the remaining 346 benign lumps the specificity of the three tests was respectively 0.60, 0.78 and 0.98. The predictive value of a positive response (certain or probable) to the three tests was 0.63, 0.74 and 0.94. The certain positive responses of cytology (131 of 285 cases) reached a predictive value of 1.00. The predictive value of a negative response for the three tests was respectively 0.81, 0.80 and 0.82; in the absence of positive responses, the predictive value for benignancy of the triad was 0.93 (177 of 190 cases). The systematic use of the diagnostic triad and the organizational platform allowed the clinicians to select malignant cases and plan inpatient/outpatient surgical treatment.

Purification and structural characterization of a fatty acid-binding protein from the liver of the catfish Rhamdia sapo.

We report here the isolation of a fatty acid-binding protein (FABP) from the liver of the catfish Rhamdia sapo. The purification procedure involves gel filtration, anion-exchange chromatography and reverse-phase high-performance liquid chromatography. The purified protein is basic (pI > 8.7) and migrates on sodium dodecyl sulfate-gel electrophoresis as a single entity of about 15 kDa. Its amino acid composition resembles those of FABPs isolated from other animals. Unlike mammalian liver FABPs, catfish liver FABP contains at least one tryptophan residue per molecule. No significant cross-reactivity was observed between the purified protein and polyclonal antibodies against either rat liver FABP or rat heart FABP. Amino acid sequencing of peptides obtained by digestion with Lys-C revealed that the catfish protein is structurally more similar to chicken liver FABP (69% identity in a 67-residue overlap) than to human liver FABPs (36%), nurse shark (Ginglymostoma cirratum) liver FABP (30%) and human heart FABP (31%). Taken together, these results suggest that catfish liver FABP is far more closely related to chicken liver FABP than to the FABPs isolated from the liver of mammals or elasmobranchs.

[Drug resistance and hormone dependence of a case of breast cancer]. / Chemioresistenza ed ormonodipendenza di un caso di cancro mammario

A case of breast cancer was treated with mastectomy followed by cyclical combination chemotherapy with CMF for a year. In the course of chemotherapy cancer developed in the contralateral breast. Mastectomy was followed within two months by lung metastases. After ovariectomy the lung metastases cleared almost completely. The case shown how the hormone dependence of a breast cancer can remain unimpaired even after prolonged chemotherapy and notwithstanding its failure and secondly that amenorrhea due to chemotherapy need not mean endocrine failure of the ovary.