Characterizing and predicting the relationship between translaminar fracture toughness and pull-out length distributions under distinct temperatures.

The translaminar fracture toughness reflects the damage tolerance of a fibre-reinforced composite under longitudinal tension, which often governs the final failure of structures. One of the main energy-dissipation mechanisms that contributes to the translaminar toughness of composites is the fibre pull-out process. The present study aims to quantify and model the statistical distribution of fibre pull-out lengths formed on the translaminar fracture surface of composites, for the first time in the literature; this is done under different temperatures, so that the relationship between pull-out length distributions, micromechanical properties and the translaminar fracture toughness can be established. The fracture surfaces of cross-ply compact tension specimens tested under three different temperatures have been scanned through X-ray computed tomography to quantify the extent of fibre pull-out on the fracture surfaces; the distribution of pull-out lengths showed alarger average and larger variability with an increase in temperature, which also lead to an increase in translaminar fracture toughness. A similar trend has been captured by the proposed analytical model, which predicts the pull-out length distribution based on the analysis of quasi-fractal idealizations of the fracture surface, yielding an overall accuracy of more than 85%. This article is part of the theme issue 'Ageing and durability of composite materials'.

Serum fluctuations of total and free tryptophan levels during the menstrual cycle are related to gonadotrophins and reflect brain serotonin utilization.

BACKGROUND: Serotoninergic (5-HT) neurons are suggested to regulate estrous cycle in animal models. In the present study we evaluated whether a relationship exists between the serotoninergic precursors in the central nervous system and the gonadotrophin-ovarian cyclic function. METHODS: We measured 5-HT precursors [free (FT) and total (TT) tryptophan] and LH, FSH and 17beta-estradiol (E2) levels in the serum of 15 fertile women with normal menstrual cycles during the follicular (cycle days 1-5, 7-11), mid-cycle (cycle days 14-16) and luteal (cycle days 17-19, 22-24) phases. RESULTS: TT and FT were significantly increased in the 7-11 and 17-19 cycle days and were decreased at mid-cycle (P < 0.01), with a cyclic and opposite behaviour when compared to that of FSH and LH. Indeed, correlation analysis through the matrix of mean values showed that LH was negatively correlated to TT (r = -0.636) and FT (r = -0.574), as well as FSH (TT, r = -0.655; FT, r = -0.663), and that TT and FT were positively correlated to each other (r = 0.801; P < 0.001). Furthermore, whilst the two FT peaks reached approximately the same levels in the follicular and luteal phase, TT levels were approximately 30% higher in the luteal than in the follicular phase of the cycle: thus in the first (follicular) phase FT peak was relatively higher than that of TT, whereas the contrary occurred in the second (luteal) phase of the cycle. CONCLUSIONS: Both TT and FT levels have cyclic variations throughout the menstrual cycle, being lowest at mid-cycle (14-16 cycle days), concomitant with the highest LH and FSH concentrations, and higher before and after mid-cycle phase, coinciding with the lowest circulating LH/FSH levels. Since TT and FT levels in the plasma have cyclic changes, our study: (i) suggests that a consumption of serum serotonin precursors takes place concomitant with gonadotrophin release during menstrual cycle; (ii) may represent an in vivo model to investigate this relationship in women in different physiopathological conditions.

Endoscopic selective sampling of human ventricular CSF: a new perspective.

Neuroendoscopy has achieved extensive acceptance among neurosurgeons as a minimally invasive technique for the treatment of patients affected by blocked hydrocephalus. During endoscopic procedures minimal CSF amounts from selected anatomic sites of the ventricles can be withdrawn. Steerable endoscopes are used and their flexibility facilitates the aspiration of CSF during the preliminary inspection through the ventricular cavities, without any interference with the surgical actions or additional risks for the patients. In this preliminary study the concentrations of melatonin and other related metabolites in the lateral ventricles, third ventricle, pineal recess and infundibular recess were examined. The data obtained from a patient affected by blocked hydrocephalus confirmed a constant and significant difference of concentration of these substances, for instance, melatonin levels were found to be much higher in the third ventricle (542 pg/mL in its centre) than in the lateral ventricle (172 pg/mL in the right ventricle). Nevertheless, instead of what we would expect, the highest melatonin concentrations were not found in the pineal recess (438 pg/mL). In the future, neuroendoscopy, beside its evident therapeutic efficacy, could open new perspectives in the study of both CSF biochemistry and physiology, allowing a highly selective approach to the various substances which are released and float in it.

Enzyme activities involved in tryptophan metabolism along the kynurenine pathway in rabbits.

The following enzyme activities of the tryptophan-nicotinic acid pathway were studied in male New Zealand rabbits: liver tryptophan 2,3-dioxygenase, intestine indole 2,3-dioxygenase, liver and kidney kynurenine 3-monooxygenase, kynureninase, kynurenine-oxoglutarate transaminase, 3-hydroxyanthranilate 3,4-dioxygenase, and aminocarboxymuconate-semialdehyde decarboxylase. Intestine superoxide dismutase and serum tryptophan were also determined. Liver tryptophan 2,3-dioxygenase exists only as holoenzyme, but intestine indole 2,3-dioxygenase is very active and can be considered the key enzyme which determines how much tryptophan enters the kynurenine pathway also under physiological conditions. The elevated activity of indole 2,3-dioxygenase in the rabbit intestine could be related to the low activity of superoxide dismutase found in intestine. Kynurenine 3-monooxygenase appeared more active than kynurenine-oxoglutarate transaminase and kynureninase, suggesting that perhaps a major portion of kynurenine available from tryptophan may be metabolized to give 3-hydroxyanthranilic acid, the precursor of nicotinic acid. In fact, 3-hydroxyanthranilate 3,4-dioxygenase is much more active than the other previous enzymes of the kynurenine pathway. In the rabbit liver 3-hydroxyanthranilate 3,4-dioxygenase and aminocarboxymuconate-semialdehyde decarboxylase show similar activities, but in the kidney 3-hydroxyanthranilate 3,4-dioxygenase activity is almost double. These data suggest that in rabbit tryptophan is mainly metabolized along the kynurenine pathway. Therefore, the rabbit can also be a suitable model for studying tryptophan metabolism in pathological conditions.

Melanogenesis by tyrosinase action on 3,4-dihydroxyphenylalanine (DOPA) in the presence of polyethylene glycol: a matrix-assisted laser desorption/ionization mass spectrometric investigation.

The enzymatic reaction between DOPA and tyrosinase, the enzyme considered to be responsible for melanogenesis, was carried out in the presence of polyethylene glycol (PEG). This choice was made in order to increase the solubility of melanins, since these polymers are highly insoluble. The reaction mixtures were sampled at different times, immediately ultrafiltered to remove the enzyme, lyophilized, and analyzed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. The results were very different from those obtained in the absence of PEG. Only a few oligomers of dihydroxyindole (DHI) and dihydroxyindole-2-carboxylic acid (DHICA) were detected in low abundances, whereas new species originating from reaction of PEG with species belonging to the Raper-Mason pattern appeared. The results show that, in the presence of PEG, tyrosinase-catalyzed oligomerization of DOPA exhibits kinetics slower than those observed in the absence of the polymer. However, melanogenesis still takes place in the presence of PEG, as indicated by the formation of black pigments and by the detection of DHI and DHICA oligomers, considered to be the first intermediates in melanin formation.

Effects of ultraviolet irradiation on melanogenesis from tyrosine, Dopa and dopamine: a matrix-assisted laser desorption/ionization mass spectrometric study.

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry experiments were applied to study the influence of ultraviolet (UV) irradiation in melanogenesis. Samples were prepared starting from three different precursors, tyrosine, Dopa and dopamine, in the presence or absence of tyrosinase, the enzyme responsible for the synthesis of melanin. Enzymatic reactions were carried out for 10, 30, 60 and 120 min under UV irradiation at 365 nm, and aliquots were then immediately ultrafiltered and lyophilized. Samples obtained by irradiation of tyrosine solution revealed the formation of 5,6-dihydroxyindole (DHI) oligomers up to pentamers at 120 min; the reaction kinetics were markedly enhanced in the presence of tyrosinase. In the case of Dopa, UV irradiation favored melanogenesis only in the presence of the enzyme; in this case, many reaction pathways were activated, originating various oligomeric species of Dopa, DHI and 5,6-dihydroxyindole-2-carboxylic acid (DHICA). Conversely, when dopamine was used as tyrosinase substrate under UV light, mechanisms of melanogenesis different from those generated by simple enzymatic reaction without irradiation were not activated, as the same oligomeric species were present.

Protein profiles in sera of patients with malignant cutaneous melanoma.

Seventeen samples of sera from patients with malignant cutaneous melanoma at various stages and 14 samples from healthy subjects were analysed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry. Results highlighted the presence of several protein species at molecular weights lower than 30000 Da, presumably originating from proteolysis, in the sera of the patients with melanoma. These species were completely absent in healthy subjects. In particular, the presence and abundance of species with molecular weights in the range 2500-3500 Da exhibit significant variations related to the different clinical stages of the disease.

Tryptophan in human hair: correlation with pigmentation.

The distribution of tryptophan content in human hair of various colours was evaluated, in order to study the accumulation of this amino acid, precursor of serotonin, melatonin and niacin, in hair and the influence on hair pigmentation. Pigmentation is an important factor in determining drug incorporation into hair. Results from 1211 samples of hair from healthy subjects (577 men and 634 women) show that tryptophan levels are significantly higher in males (37.83 +/- 3.45 microg/g dry hair) than in females (26.62 +/- 2.40 microg/g hair). Besides sex, age also influences the distribution of tryptophan in human hair, the highest levels being found in both sexes in the first few years of life, probably due to the influence of milk, and in aging subjects in the groups of 61-80 and > 80 years. In order to investigate the influence of hair colour, hair samples were subdivided according to colour into blond, dark blond, red, light brown, brown, black, grey and white. The hair contents of tryptophan in both sexes was higher in brown and black hair than in blond hair, but in grey and white hair concentrations were the highest, demonstrating that tryptophan accumulates among hair fibres with age. Grouping subjects by age in relation to hair colour, we observed that at ages 1-5 and 6-12 years, colour did not influence tryptophan contents, but at ages 13-19 and 20-40 years tryptophan content increased significantly from blond to brown at 13-19 years and from blond to black at 20-40 years in both sexes. Therefore, variations in tryptophan levels of human hair appear to be correlated with differences in hair colour in both sexes. Tryptophan also accumulates in hair during keratinization, as shown by the presence of high levels of this amino acid in grey and white hair.

Application of matrix-assisted laser desorption/ionization mass spectrometry to the detection of melanins formed from Dopa and dopamine.

Matrix-assisted laser desorption/ionization (MALDI) mass spectrometry was used to study melanogenesis starting from Dopa and dopamine, the latter considered one of the precursors of neuromelanins. These substrates were left to react with the peroxidase - H(2)O(2) system, which is postulated to play an important role in melanin biosynthesis. Samples were prepared by ultrafiltering the substrate - enzyme solution after 30, 60, 120, 240 and 360 min of reaction and aliquots were immediately lyophilized. The reaction of dopamine with peroxidase - H(2)O(2) favoured the formation of dopamine oligomers up to octamers. In contrast, the action of either peroxidase or H(2)O(2) alone, studied for comparison, did not lead to melanin production and only dimeric and trimeric species were observed. Also for Dopa, analogous results were obtained in the presence of either peroxidase or H(2)O(2) alone, without melanin formation. Conversely, Dopa with the peroxidase - H(2)O(2) system led to the formation of a black precipitate after 120 min of reaction, and oligomers of 5,6-dihydroxyindole (DHI), an intermediate of melanogenesis, were detected, together with products of further oxidation. Faster kinetics were observed when Dopa was treated with tyrosinase, the enzyme catalysing the oligomerization of tyrosine to melanins, leading to the formation mainly of DHI oligomers.

Matrix-assisted laser desorption/ionization mass spectrometry in evaluation of protein profiles of infant formulae.

Breast-feeding is the nutritional model in humans, and is continued after birth for variable periods. Milk represents an essential source of growth from both nutritional and functional points of view. When nursing is not possible, indicated or sufficient, artificial feeding becomes essential. Substitutes for mother's milk are usually obtained by modifying the composition of cow's milk either qualitatively or quantitatively. The changes usually involve enzymatic and/or thermal treatment, and for this reason a description of the protein profiles of milk formulae is of great interest. After examination of the results obtained by matrix-assisted laser desorption/ionization (MALDI) in the characterization of protein profiles of cow's milk after different thermal treatments, the application of this analytical technique in the above context appeared interesting. MALDI data for eleven milk formulae, directly acquired from the market, are described and discussed. Results indicate that MALDI mass spectrometry is a particularly powerful method which may be employed either during the production stages of milk formulae or to study the relationships between milk protein content and clinical formula evaluation.

Importance of L-tryptophan metabolism in trypanosomiasis.

African trypanosomiasis or sleeping sickness is caused by extracellular trypanosomes. The presence of seric antibodies directed to a tryptophan-like epitope in trypanosome infected patients and animals led us to investigate the roles of tryptophan in trypanosomiasis. These antibodies are directed against a tryptophan-rich conserved sequence inside the major parasite surface glycoprotein. In vitro, a rapid uptake of tryptophan by trypanosomes is measured. Seric tryptophan levels are decreased during trypanosomiasis. This decrease may be linked with an increase in indoleamine 2,3-dioxygenase (IDO) induced by Interferon-gamma. In vivo inhibition of IDO by norharman provokes a dramatic increase in circulating parasite number. All these data show the essential role of tryptophan in parasite growth. Moreover, antibodies against tryptophan, the decreased concentration of the neurotransmitter serotonin in the brain following infection and the tryptophan metabolites (tryptophol) produced by trypanosomes may participate to the pathophysiological mechanisms provoking sleeping sickness.

Tryptophan catabolism in synovial fluid of various arthropathies and its relationship with inflammatory cytokines.

Synovial fluids (SF) from patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA), gout, and osteoarthritis (OA) were investigated for the levels of interleukin (IL)-1 beta, IL-6 and IL-8, tryptophan (Trp) and indoleamine 2,3-dioxygenase (IDO) activity. Significant differences exist in the levels of IL-1 beta between inflammatory arthritides RA, PsA and gout and non inflammatory arthritis, such as OA. The highest concentration of IL-1 beta was found in RA, that showed high levels also of IL-6 and IL-8. In the same disease we also found the highest IDO activity and the lowest Trp concentration. In addition, IDO activity seems to be related with the decrease in Trp, as demonstrated by the inverse correlation found between these two substances in the SF of all patients.

Liver and kidney kynurenine aminotransferase activity in different strains of rats.

Variations in liver and kidney kynurenine aminotransferase (KAT) activity in Pittsburg-Yoshida, Brown-Norway, albino Wistar, Sprague-Dawley, Long Evans and heterozygous Gunn rats were studied. In liver, values of KAT specific activity, expressed as mumoles of kynurenic acid formed per hour per mg of protein, were different in the groups of Brown-Norway and Pittsburg-Yoshida rats versus Long Evans and Sprague-Dawley rats. The activity expressed as mumoles of kynurenic acid per g of fresh liver showed other differences, being significantly higher in Gunn with respect to other strains of rats and lower in Pittsburg-Yoshida and Brown-Norway rats. In addition, KAT activity was significantly lower in Pittsburg-Yoshida than in Brown-Norway rats. In kidney, the specific activity of kynurenine aminotransferase showed significant differences in the values of Sprague-Dawley and Long Evans rats versus the other strains. The activity expressed per g of fresh tissue was significantly higher in Wistar, Sprague-Dawley, Long Evans and Gunn than in Pittsburg-Yoshida and Brown-Norway rats. No significant differences were found in values between hyperlipidemic Pittsburg-Yoshida and their control Brown-Norway rats. These results high-light the importance of considering various rat strains when inbred animal experimental models are used.

Correlation between tryptophan and hair pigmentation in human hair.

The concentration of tryptophan in human hair of various colours is determined in order to study their correlation with hair pigmentation. The mean levels of this amino acid in hair samples are higher in men than in women. Therefore, sex influences the content of tryptophan in human hair. In addition, age influences the distribution, the highest levels are observed in the 1-5 year age-group and in ageing subjects in the groups up to 61-80 years in both sexes. The hair samples subdivided, according the colour, into blond, dark blond, red, light brown, brown, black, grey, and white demonstrate that in both sexes the concentrations of tryptophan are higher in brown and black hair than in blond hair. However, the tryptophan levels are highest in grey and white hair, showing that tryptophan accumulates among hair fibres with age. Therefore, there is a correlation between tryptophan content and hair pigmentation.

Melanogenesis from 5-hydroxytryptamine, 5,6- and 5,7-dihydroxytryptamines. An in vitro study using MALDI-TOF.

The role of tyrosinase and peroxidase in melanogenesis of 5-hydroxytryptamine, 5,6- and 5,7-dihydroxytryptamines was investigated by matrix-assisted laser desorption/ionization mass spectrometry. Each enzyme was incubated with the tryptamine derivatives and samples were drawn at various times, ultrafiltered and immediately lyophilized. The results indicated that peroxidase promotes oligomerization of 5-HT with fast kinetics but with yields lower than those achieved by tyrosinase. 5,6- and 5,7-DHT formed low molecular mass oligomers in the presence of peroxidase alone. The addition of hydrogen peroxide evidences different reactivity of the two isomers: 5,6-DHT formed immediately a black precipitate while oligomers of the molecule itself and of its oxidation products were detectable for 5,7-DHT.

Seroprevalence to bovine immunodeficiency virus and lack of association with leukocyte counts in Italian dairy cattle.

We report herein on the first serological detection of antibodies to bovine immunodeficiency virus (BIV) in Italy. According to criteria of a stratified-random sampling of dairy cattle reared in the Parma area (a province in the Po Valley, Northern Italy), sera from 3166 cows belonging to 272 herds were collected. In addition, sera of 138 bulls from eight artificial-insemination (AI) centres were sampled. Seventy-eight cows (2.5%) from 16 herds (5.8%) and seven bulls (5.1%) from two AI centres were positive for BIV-R29 antibodies in the IFA-test. IFA-positive sera assayed by Western blot had reaction to different viral proteins: 81 out of 85 sera showed antibody to p26 (considered the BIV major internal core protein); four sera reacted to other viral proteins but not to p26. Peripheral blood leukocytes of 60 seropositive and 60 seronegative animals, belonging to eight BIV-infected herds, were enumerated to assess any effect of BIV infection on white-blood cells. No significant differences were detected between the two groups. These data indicate that BIV infection is present in Italian dairy cattle--but the role of BIV in inducing disease remains unclear.

Determination of copper and zinc levels in human hair: influence of sex, age, and hair pigmentation.

The Cu and Zn levels of both 607 men (1-85 y old) and 649 women (1-92 y old) were determined by atomic absorption spectrometry. Sex does not influence Cu (14.89 + or - 0.89 mu g/g and 15.26 + or - 0.79 mu g/g hair for males and females, respectively) and Zn contents (200.97 + or - 9.68 mu g/g for men and 209.81 + or - 9.49 mu g/g hair for women). Age influences Cu and Zn concentrations, but only significantly in females: Cu levels decrease over 60 y of age; whereas Zn levels increase significantly from age groups 2-5 to 20-40 years. Hair color influences Cu concentrations in both males and females. In males, white hair contains less Cu than black hair; in females, white hair's Cu levels are significantly lower than those of dark blond, red, light brown, and brown hair. There are no significant differences in Zn concentrations with respect to different hair colors, in either males or females.