Dietary glycemic index, glycemic load and metabolic profile in children with phenylketonuria.

BACKGROUND AND AIMS: No data exist in the current literature on the glycemic index (GI) and glycemic load (GL) of the diet of phenylketonuric (PKU) children. The aims of this study were to examine the dietary GI and GL in PKU children on a low-phenylalanine (Phe)-diet and to evaluate whether an association may exist between the carbohydrate quality and the metabolic profile. METHODS: Twenty-one PKU children (age 5-11 years) and 21 healthy children, gender and age matched, were enrolled. Dietary (including GI and GL) and blood biochemical assessments were performed. RESULTS: No difference was observed for daily energy intake between PKU and healthy children. Compared to healthy controls, PKU children consumed less protein (p = 0.001) and fat (p = 0.028), and more carbohydrate (% of total energy, p = 0.004) and fiber (p = 0.009). PKU children had higher daily GI than healthy children (mean difference (95% confidence interval), 13.7 (9.3-18.3)) and higher GL (31.7 (10.1-53.2)). PKU children exhibited lower blood total and low density lipoprotein cholesterol (LDL) levels (p < 0.01) and higher triglyceride level (p = 0.014) than healthy children, while glucose and insulin concentrations did not differ. In PKU children the dietary GL was associated with triglyceride glucose index (Spearman's correlation coefficient = 0.515, p = 0.034). CONCLUSION: In PKU children a relationship of the dietary treatment with GI and GL, blood triglycerides and triglyceride glucose index may exist. Improvement towards an optimal diet for PKU children could include additional attention to the management of dietary carbohydrate quality.

Diet in children with phenylketonuria and risk of cardiovascular disease: A narrative overview.

AIMS: The aim of this paper is to review the possible relationship of restricted phenylalanine (Phe) diet, a diet primarily comprising low-protein foods and Phe-free protein substitutes, with major cardiovascular risk factors (overweight/obesity, blood lipid profile, plasma levels of homocysteine, adiponectin and free asymmetric dimethylarginine (ADMA), oxidative stress and blood pressure) in PKU children. DATA SYNTHESIS: In PKU children compliant with diet, blood total cholesterol, low-density lipoprotein cholesterol (LDL-C), plasma ADMA levels and diastolic pressure were reported to be lower and plasma adiponectin levels to be higher compared to healthy controls. No difference was observed in overweight prevalence and in high-density lipoprotein cholesterol (HDL-C) levels. Inconsistent results were found for plasma homocysteine levels and antioxidant status. CONCLUSIONS: PKU children compliant with diet seem to display non-different cardiovascular risks compared with the healthy population. Well-designed longitudinal studies are required to clarify the potential underlying mechanisms associated with PKU and cardiovascular risk factors.

Short and long term health effects of parental tobacco smoking during pregnancy and lactation: a descriptive review.

A great deal of attention has been focused on adverse effects of tobacco smoking on conception, pregnancy, fetal, and child health. The aim of this paper is to discuss the current evidence regarding short and long-term health effects on child health of parental smoking during pregnancy and lactation and the potential underlying mechanisms. Studies were searched on MEDLINE(®) and Cochrane database inserting, individually and using the Boolean ANDs and ORs, 'pregnancy', 'human lactation', 'fetal growth', 'metabolic outcomes', 'obesity', 'cardiovascular outcomes', 'blood pressure', 'brain development', 'respiratory outcomes', 'maternal or paternal or parental tobacco smoking', 'nicotine'. Publications coming from the reference list of studies were also considered from MEDLINE. All sources were retrieved between 2015-01-03 and 2015-31-05. There is overall consistency in literature about negative effects of fetal and postnatal exposure to parental tobacco smoking on several outcomes: preterm birth, fetal growth restriction, low birth weight, sudden infant death syndrome, neurodevelopmental and behavioral problems, obesity, hypertension, type 2 diabetes, impaired lung function, asthma and wheezing. While maternal smoking during pregnancy plays a major role on adverse postnatal outcomes, it may also cumulate negatively with smoking during lactation and with second-hand smoking exposure. Although this review was not strictly designed as a systematic review and the PRISMA Statement was not fully applied it may benefit the reader with a promptly and friendly readable update of the matter. This review strengthens the need to plan population health policies aimed to implement educational programs to hopefully minimize tobacco smoke exposure during pregnancy and lactation.

Effect of genotype, gender and feed restriction on growth, meat quality and the occurrence of white striping and wooden breast in broiler chickens.

Due to their importance for the control of meat quality in broiler chickens, the present study aimed at identifying the factors associated with the occurrence of myopathies and characterizing the meat properties when affected by myopathies. To this aim, a total of 768 broiler chickens were reared until slaughter (46 d) to evaluate the effect of genotype, gender, and feeding regime (ad libitum vs. restricted rate, 80% from 13 to 21 d of age) on performance and meat quality. Standard broilers were heavier (3,270 vs. 3,139 g; P<0.001) and showed lower feed conversion (1.56 vs. 1.61; P<0.001) than the high-yield broilers. Males showed higher final live weight (3,492 vs. 2,845 g) and lower feed conversion (1.54 vs. 1.63) than females (P<0.001). Feed restriction decreased final live weight (3,194 vs. 3,142 g; P<0.01) and feed conversion (1.60 vs. 1.57; P<0.01) compared to ad libitum feeding. At gross examination, feed restriction tended to increase white-striped breasts (69.5 vs. 79.5%; P<0.10), whereas females showed less wooden breasts than males (8.0 vs. 16.3%; P<0.05). White-striped fillets had higher pHu (5.87 vs. 5.83), and lower a* (-0.81 vs. -0.59) and b* color indexes (13.7 vs. 14.5) (P<0.05), whereas wooden breast fillets exhibited higher cooking losses (25.6 vs. 22.1%) and AK-shear force (4.23 vs. 2.84 kg/g) compared with normal fillets (P<0.001). At histological examination, 3.1% of pectoralis major were normal, 26.6% mildly degenerated, 45.3% moderately degenerated, and 25.0% severely degenerated. In conclusion, genotype had a moderate effect on growth without modifying myopathy occurrence. In contrast, gender and feed restriction affected performance, meat quality, and breast abnormalities.

Memory changes in patients with hippocampal sclerosis submitted to surgery to treat mesial temporal lobe epilepsy.

PURPOSE: This study was performed with the purpose of analysing the relationship between epileptological and surgical variables and post-operative memory performance, following surgery for refractory mesial temporal lobe epilepsy (MTLE) due to hippocampal sclerosis (HS). METHODS: Logical memory (LM) and visual memory (VM) scores for immediate and late follow-up of 201 patients operated for MTLE/HS were reviewed. Scores were standardized with a control group of 54 healthy individuals matched for age and education. The Reliable Change Index (RCI) was calculated to verify individual memory changes for late LM and VM scores. A multiple linear regression analysis was carried out with the RCI, using LM and VM scores as well as the clinical variables. RESULTS: A total of 112 (56%) patients had right HS. The RCI of the right HS group demonstrated that 6 (7%) patients showed improvement while 5 (6%) patients showed decreased scores in late LM; for late VM, 7 (8%) patients presented improvement, and 2 (3%) patients showed poorer scores. RCI of the left HS group showed that 3 (3%) individuals showed improved scores, while scores of 5 (4%) patients worsened for late LM; for late VM, 3 (3%) patients presented higher scores and 6 (5%) showed lower scores. Left HS and advanced age at onset of the first epileptic seizure were predictors of late LM loss (p<.05). CONCLUSION: Left MTLE/HS and seizure onset at advanced ages were predictive factors for the worsening of late LM. We observed poorer baseline LM function in the left HS group and improvement of LM in some patients who had resection of the right MTL. Patients in the right HS group showed a higher percentage of reliable post-operative improvement for both VM and LM scores.

Margin involvement and clinical pattern of basal cell carcinoma with mixed histology.

BACKGROUND: Mixed basal cell carcinoma (BCC) has not been sufficiently and specifically studied. OBJECTIVE: The aim of this study was to estimate in adults the prevalence of mixed cases observed among primary BCCs and to compare clinical and anatomical features of mixed vs. single BCCs, with focus on the incomplete excision. PATIENTS AND METHODS: A total of 3636 histologically confirmed primary BCCs were examined. Data on gender, age, histological subtype, anatomical location and margin involvement were collected. Mixed type was defined as a combination of two or more single subtypes. RESULTS: Prevalence of single and mixed BCCs was 82.2% and 17.8% respectively. Prevalence of BCCs on the upper limbs was higher in mixed than single cases (8.8% vs. 4.0%; P<0.001) while prevalence on the back was lower (16.9% vs. 23.7%; P<0.001). Tumour was aggressive in 59.1% of mixed vs. 16.0% of single BCCs (P<0.001). Margin involvement was more prevalent in mixed than in single BCCs (16.7% vs. 9.6%; P<0.0001). At multivariate analysis being mixed vs. single BCC was associated with aggressiveness of tumour (OR=8.5, 95% CI, 6.9-10.4), lateral margin involvement (OR=1.98, 95% CI, 1.42-2.76) and subject being man (OR=1.31, 95% CI, 1.10-1.60) but not with deep involvement of margin or anatomical location. CONCLUSION: Among BCCs, the mixed type may be observed in adults with relatively high rate and may represent a complex and individual subset of BCCs with potential aggressive behaviour.

Seasonal effects on hematological and innate immune parameters in sea bass Dicentrarchus labrax.

The temperate aquatic environment is affected by two primary components of season, temperature and photoperiod, during the annual cycle. Many organisms respond to seasonal change physiologically, behaviorally or both. The aim of this study was to investigate the effect of seasonality on cortisol, hematological and innate immune parameters in European sea bass reared under traditional semi-intensive aquaculture. Sea bass (Dicentrarchus labrax) were reared in an outdoor pond and serum cortisol, hematocrit, leucocrit, serum lysozyme activity and total glutathione were bimonthly monitored over a 14-months period. The effect of seasonality was observed for all parameters carried out, with generally higher values in summer and lower in winter. These results could improve the understanding of the influence of seasonal cues on the immune system and hematological parameters in fish in order to optimize the husbandry practices.

Spindle and/or epithelioid cells nevi in pediatric age: are they different from those of adults? A clinical review of 187 cases.

AIM: Spindle and/or epithelioid cells nevi represent the spectrum of a clinico-pathologic entity with different characteristics. Aim of the study is to provide information about the differences in characteristics of these nevi for different groups of age. METHODS: Two different groups are considered: younger than 15 years and older than 15 years. An analysis of 187 spindle and/or epithelioid cells nevi was performed. Forty-five pediatric patients (24 males and 21 females) and 142 adult patients (44 males and 98 females) were examined. Age, sex, type of nevus, location, clinical characteristics were evaluated. RESULTS: Spindle and epithelioid cells nevi were observed in 53% of the pediatric and in 45% of adult patients. Female more frequently presented with spindle nevus cell both in pediatric (56%) and in adult (70%) cases. In pediatric patients, the anatomical distribution was prevalent in the areas of the head and neck. Pigmentation was not a distinctive feature of pediatric cases and only interested the spindle and epithelioid cells nevi. The regularity of borders was not a distinctive character for neither of the groups of patients. Uniformity in color occurred more frequently in pediatric patients. CONCLUSION: Spindle and/or epithelioid nevi belong to the same spectrum of pathologies, they behave differently in the different groups of age thus permitting a certain degree of clinical distinction in different age groups.

Hospital admissions for respiratory conditions in children and outdoor air pollution in Southwest Milan, Italy.

AIM: To evaluate whether a relationship exists in hospital admissions for respiratory conditions in children with outdoor pollution in the Southwest Milan, Italy. METHODS: Daily count of hospital admissions for asthma, upper or lower respiratory diseases (LRD) occurred among children aged < or =14 years at the San Paolo Hospital during 2007-2008 was recorded. Pollutants included the particular matter of <10 microm diameter, ozone, carbon monoxide (CO) and nitrogen dioxide (NO(2)), reported on the same day of admission and 1 up to 5 days before admission. The moving weekly average (MWA) was also considered. RESULTS: Total respiratory admissions were associated with the same day level of CO (30.6% increase per 1 mg/m(3) increase, 95% confidence interval, 3.9-69.9%) and with the MWA of NO(2) (9.0% increase per 10 microg/m(3) increase, 1.2-16.8%). The effect of CO was stronger on upper respiratory diseases (URD) (lag 0, 21.3% increase, 6.4-38.3%). The effect of NO(2) was stronger on LRD (MWA, 5.3% increase, 0.9-10.2%). Multipollutant models confirmed the role of CO on URD and that of NO(2) on LRD. CONCLUSION: Carbon monoxide and nitrogen dioxide may be associated with emergency hospital admissions for respiratory conditions among children in Southwest Milan.

Precision-based exercise as a new therapeutic option for children and adolescents with haematological malignancies.

Children and adolescents with haematological malignancies (PedHM) are characterized by a severe loss of exercise ability during cancer treatment, lasting throughout their lives once healed and impacting their social inclusion prospects. The investigation of the effect of a precision-based exercise program on the connections between systems of the body in PedHM patients is the new frontier in clinical exercise physiology. This study is aimed at evaluating the effects of 11 weeks (3 times weekly) of combined training (cardiorespiratory, resistance, balance and flexibility) on the exercise intolerance in PedHM patients. Two-hundred twenty-six PedHM patients were recruited (47% F). High or medium frequency participation (HAd and MAd) was considered when a participant joined; > 65% or between 30% and < 64% of training sessions, respectively. The "up and down stairs'' test (TUDS), "6 min walking" test (6MWT), the "5 Repetition Maximum strength" leg extension and arm lateral raise test (5RM-LE and 5RM-ALR), flexibility (stand and reach), and balance (stabilometry), were performed and evaluated before and after training. The TUDS, the 5RM-LE and 5RM-ALR, and the flexibility exercises showed an increase in HAd and MAd groups (P < 0.05), while the 6MWT and balance tests showed improvement only in HAd group (P < 0.0001). These results support the ever-growing theory that, in the case of the treatment of PedHM, 'exercise is medicine' and it has the potential to increase the patient's chances of social inclusion.

Effect of dietary supplementation with yeast cell wall extracts on performance and gut response in broiler chickens.

BACKGROUND: The dietary supplementation of yeast cell wall extracts (YCW) has been found to reduce pathogenic bacteria load, promote immunoglobulin production, prevent diseases by pro-inflammatory responses, and alter gut microbiota composition. This study evaluated growth and slaughter results, health, gut morphology, immune status and gut transcriptome of 576 male chickens fed two diets, i.e. C (control) or Y (with 250-500 g/t of YCW fractions according to the growth period). At 21 and 42 d the jejunum of 12 chickens per diet were sampled and stained with hematoxylin/eosin for morphometric evaluation, with Alcian-PAS for goblet cells, and antibodies against CD3+ intraepithelial T-cells and CD45+ intraepithelial leukocytes. The jejunum sampled at 42 d were also used for whole-transcriptome profiling. RESULTS: Dietary YCW supplementation did not affect final live weight, whereas it decreased feed intake (114 to 111 g/d; P ≤ 0.10) and improved feed conversion (1.74 to 1.70; P ≤ 0.01). Regarding the gut, YCW supplementation tended to increase villi height (P = 0.07); it also increased the number of goblet cells and reduced the density of CD45+ cells compared to diet C (P < 0.001). In the gut transcriptome, four genes were expressed more in broilers fed diet Y compared to diet C, i.e. cytochrome P450, family 2, subfamily C, polypeptide 23b (CYP2C23B), tetratricopeptide repeat domain 9 (TTC9), basic helix-loop-helix family member e41 (BHLHE41), and the metalloreductase STEAP4. Only one gene set (HES_PATHWAY) was significantly enriched among the transcripts more expressed in broilers fed diet Y. However, a total of 41 gene sets were significantly over-represented among genes up-regulated in control broilers. Notably, several enriched gene sets are implicated in immune functions and related to NF-κB signaling, apoptosis, and interferon signals. CONCLUSIONS: The dietary YCW supplementation improved broiler growth performance, increased gut glycoconjugate secretion and reduced the inflammatory status together with differences in the gut transcriptome, which can be considered useful to improve animal welfare and health under the challenging conditions of intensive rearing systems in broiler chickens.

Effect of feed restriction timing on live performance, breast myopathy occurrence, and muscle fiber degeneration in 2 broiler chicken genetic lines.

During recent years, research on meat quality in poultry has aimed to evaluate the presence and consequences of breast myopathies as well as the factors which can affect their occurrence by modifying the growth rate. A total of 900 broiler chickens were reared until slaughter (48 D) to evaluate the effect of 2 genetic lines (A vs. B) and feeding plans (ad libitum [AL], early restricted [ER], from 13 to 23 D of age, and late restricted [LR], from 27 to 37 D of age; restriction rate: 80%) on performance, meat quality, and breast muscle myopathies. Calsequestrin and vascular endothelial growth factor (VEGF) expressions, and muscle fiber degeneration (MFD) were recorded at 22, 36, and 48 D. Chickens in the AL treatment had greater final live (P < 0.01) and carcass weights and proportion of pectoralis major muscle (P = 0.04) compared to chickens in the LR treatment, whereas chickens in the ER treatment had intermediate final live (3,454 g) and carcass weights, and proportion of pectoralis major muscle (25.6%). Chickens of line A were heavier than chickens of line B (P < 0.001), and had a greater feed conversion rate. Chickens of line A also had a greater dressing out percentage (P < 0.001), but a lower proportion of pectoralis major muscle (P = 0.04), as well as a greater meat pH (P < 0.001), meat cooking losses (P < 0.01), and shear force of the pectoralis major muscle (P = 0.03). Calsequestrin and VEGF mRNA were significantly lower in ER and LR chickens compared to AL chickens after feed restriction and during refeeding (P < 0.05). MFD scores increased with chicken age (P < 0.001) and differed between genetic lines (P < 0.001). Neither feeding plan nor genetic line affected the occurrence of white striping or wooden breast condition.

Height of the cerebellar vermis and gestational age at birth.

OBJECTIVES: The objective of this study was to construct reference ranges of the neonatal cerebellar vermis height with respect to the gestational age at birth. METHODS: This observational study assessed 434 neonates born at 25-42 weeks' gestation. The neonates were singleton and appropriate in size for gestational age, and did not exhibit any abnormalities or neonatal disease. Gestational age was based on the date of the last menstrual period and confirmed by ultrasound examination performed within the 12(th) week of pregnancy. Sonographic measurements of the height of the cerebellar vermis in the mid-sagittal plane were performed within 24 h of birth by the same neonatal sonographer. Reference ranges (5(th), 50(th) and 95(th) centiles) were estimated by a mean and SD model based on least-squares polynomial regression. RESULTS: Neonatal sonographic measurements were obtained in all cases. Mean (SD) maternal age was 30.2 (4.3) years. Mean cerebellar vermis height adjusted for gestational age did not differ between males and females, the mean adjusted difference being 0.012 (95% CI, - 0.009 to 0.033) cm. Mean cerebellar vermis height (cm) against gestational age (weeks) was suitably modeled by a linear-cubic polynomial as - 1.784 + 0.137 x GA - 0.000019 x GA(3) (SD = - 0.147 + 0.008 x GA), where GA = gestational age. CONCLUSIONS: Reference ranges for the height of the cerebellar vermis at birth with respect to gestational age at birth have been constructed in appropriate-for-gestational-age neonates.

Immunohistochemical localization of constitutive and inducible heat shock protein 70 in carp (Cyprinus carpio) and trout (Oncorhynchus mykiss) exposed to transport stress.

In the present work we investigated by immunohistochemistry the cellular localization of constitutive as well as inducible heat shock protein 70 in several tissues of common carp (Cyprinus carpio) and rainbow trout (Oncorhynchus mykiss) exposed to transport stress.In carp, the constitutive form (HSC70) was detected only in red skeletal muscle of both control and stressed animals. In the same species, the inducible form (HSP70) was evident in the epithelia of renal tubules,gills and skin of stressed animals, whereas in controls only red skeletal muscle exhibited an immunopositivity to HSP70 antibody. In trout, immunostaining to HSC70 antibody was found mainly in the epithelia of intestine, gills and skin of both control and stressed animals although the reactivity was generally higher in animals exposed to transport stress. In the same species immunostaining to HSP70 antibody was observed only in red skeletal muscle and epidermis of control animals.

Usefulness of chemical-shift MRI in discriminating increased liver echogenicity in glycogenosis.

BACKGROUND: Glycogen storage diseases are inherited defects which cause accumulation of glycogen in the tissues. Hepatic steatosis is defined as accumulation of fat within hepatocytes. On sonography, liver shows increased echogenicity both in glycogen storage diseases and steatosis. Liver hyperechogenicity in glycogen storage diseases may depend on accumulation of glycogen and/or fat. Chemical-shift magnetic resonance imaging can discriminate tissues only containing water from those containing both fat and water. AIM: The primary aim of the present study was to evaluate the usefulness of liver chemical-shift magnetic resonance imaging for detecting liver steatosis in patients with metabolic impairment due to glycogen storage diseases. SUBJECTS: Twelve patients with type I (n=8) or type III (n=4) glycogen storage diseases were studied and compared to 12 obese-overweight subjects with known liver steatosis. As control group 12 lean normal voluntary subjects were recruited. METHODS: Liver was evaluated by sonography and chemical-shift magnetic resonance imaging to calculate hepatic fat fraction. RESULTS: A significant difference in echogenicity between patients with glycogen storage diseases and normal subjects was observed (p<0.05), while this difference was not present between overweight-obese and glycogen storage diseases patients. On the contrary, fat fraction was similar between glycogen storage diseases patients and normal subjects and different between glycogen storage diseases patients and overweight-obese (p<0.05). CONCLUSION: The present data suggest that chemical-shift magnetic resonance imaging may exclude fat deposition as a cause of liver hyperechogenicity in subjects with glycogen storage diseases.

Dietary macronutrient intake during the first 10 years of life in a cohort of Italian children.

OBJECTIVE: To describe the longitudinal pattern of dietary energy and macronutrient intakes during the first 10 years of life in a sample of healthy Italian children. PATIENTS AND METHODS: Sixty-one healthy children were followed from birth to 10 years of age. At 1, 5, 8, and 10 years of age, anthropometric measurements were taken and nutritional habits evaluated using a standardized food frequency questionnaire. RESULTS: Compared to the Italian recommended dietary allowances, the children's diet was high in protein at any age, high in lipids from age 5 onward, and high in calories at 5 years of age. At 1 year, mean (SD) protein, carbohydrate, and lipid intake, expressed as percentage of total energy, was 20% (3%), 48% (7%), and 34% (5%), respectively. Afterward, the corresponding mean value ranged around 14% to 15%, 57% to 60%, and 29% to 32%. The mean glycemic load and overall glycemic index ranged around 143 to 150 and 58 to 59, respectively. A difference between boys and girls was found in the longitudinal pattern for the intake of energy (P < 0.0001) and glycemic load (P < 0.0001). The individual longitudinal dietary pattern varied over time for the intake of total energy (P < 0.0001), any macronutrient (P < 0.0001), and overall glycemic index (P = 0.05). CONCLUSIONS: Although the amount of energy intake was in accordance with the Italian recommended dietary allowances, children consumed a high-protein and high-lipid diet. Early and continuous dietary education of children and parents may provide support for adequate nutritional habits throughout their childhood.

Quantitative RT-PCR analysis and immunohistochemical localization of HSP70 in sea bass Dicentrarchus labrax exposed to transport stress.

In aquaculture, fish are exposed to stressful conditions, which cause an increased synthesis of heat shock proteins (HSPs) at the cellular level. In this work we considered the expression of the constitutive and inducible forms of HSP70 as an indicator of stress caused by transport, during development of the sea bass (Dicentrarchus labrax), a teleost fish of high value for aquaculture. Qualitative RT-PCR analysis revealed expression of inducible HSP70 gene in larvae and fry (25, 40 and 80 days) as well as in adult tissues (liver, brain, muscle, gills, kidney, gonads, heart, spleen and skin) of both control and stressed animals. Expression of inducible HSP70 mRNA examined in different adult tissues by Real-Time PCR, was significantly higher in skin and skeletal muscle of stressed animals than in controls. Immunolocalization of inducible and constitutive forms of heat shock protein 70 (HSP70 and HSC70), reported here for the first time, demonstrated an ubiquitous distribution of HSC70 protein in several tissues of both stressed and control animals (at all stages), while inducible HSP70 protein was found only in skeletal muscle of stressed animals. In all stressed animals, regardless of their developmental stage, cortisol levels were higher than in control animals.

Effect of age on the occurrence of muscle fiber degeneration associated with myopathies in broiler chickens submitted to feed restriction.

To evaluate muscle fiber degeneration (MFD) associated with white striping and wooden breast, pectoralis major of 192 broilers differing for genotype (standard vs. high breast yield), gender, and feeding regime (ad libitum vs. restricted rate 80% from 13 to 21 d of age) were sampled at 14, 21, 28, 35, and 46 d of age for histological analyses by hematoxylin and eosin (H&E) staining to evaluate tissue morphology, Masson's trichrome to identify collagen presence, and Oil red and Nile blue for lipid presence. Microvessels (diameter ≤15 µm), nuclei positive to anti-cleaved lamin A and monoclonal proliferating cell nuclear antigen (PCNA) antisera were counted to assess apoptotic and regenerative processes, respectively. Significant differences were found according to feeding system, age, and their interactions. The frequency of chickens with MFD was higher with ad libitum than restricted feeding (75.0% vs. 62.5%; P = 0.01) and increased with age (18.8%, 28.1%, 75.1%, 96.9%, and 96.9% at 14, 21, 28, 35, and 46 d). However, at 14 d a similar frequency (18.8%) was found in all broilers; at 21 d, MFD occurred more in broilers fed ad libitum than in those under restriction (50.0% vs. 6.3%; P < 0.01); at 28 d differences were reduced (87.5% vs. 62.5%; P  = 0.10) to disappear by 35 (100% and 93.8%) and 46 d (96.9% and 96.9%). The number of microvessels decreased with age (20.7 to 9.46; P < 0.001) and the number of nuclei positive to the anti-cleaved lamin A antibody increased. At histology, MFD at 46 d corresponded to loss of typical cross striations, massive necrotic process, degenerating fibers surrounded by inflammatory cells, scattered fibers in an abundant collagen-rich connective tissue, numerous adipose cells; necrotic fibers showed a high percentage of apoptotic nuclei, and regenerating fibers appeared positive to anti-PCNA antibody. In conclusion, MFD soon occurred after 2 wk of growth and increased dramatically within 28 d. Early feed restriction reduced MFD as long as animals were restricted, but no residual effect was recorded after re-alimentation.

Long-term culture of muscle explants from Sparus aurata.

Although there are mammalian myoblast cell lines, no fish myoblast cell line has been developed so far. The aim of this study was to develop a culture system of muscle explants for fish, as explants provide an approximation of the in vivo conditions for cell proliferation and differentiation, and enable a close comparison with events in muscle regenerating in vivo. Here we describe the main features of a long-term in vitro culture system for muscle explants from Sparus aurata fry. At the time of sampling, the original fibres were damaged and subsequently degenerated as shown by the loss of parvalbumin (PV) and presence of apoptotic nuclei. This mechanical damage provoked a myogenic response by activation of myogenic precursor cells. After a few days, new mononucleate cells aligned with the original fibres were seen in the explants, some with proliferating cell nuclear antigen (PCNA-) and Myf-5-positive nuclei, indicating proliferation and their myogenic fate. By 1 week, multinucleate cells with desmin immunoreactivity but PCNA- and Myf5-negative nuclei were present, equivalent to differentiated, postmitotic myotubes. Some of these myotubes were also immunoreactive for PV and insulin-like growth factors (IGFs). By 11 days, many of the myotubes were also immunoreactive for myostatin (MSTN). By 23 days, many of the myotubes had increased in diameter, were packed with myofibrils, and were strongly PV-positive and immunoreactive for MSTN, IGF-I and IGF-I receptor. This study shows that a proliferative process occurs in the explants despite the death of the original muscle fibres, and new muscle fibres expressing growth regulators are formed by regeneration from myogenic precursors present in the explants at the time of sampling.

Dietary glycemic load and macronutrient intake in healthy Italian children.

The objective is of this study is to examine the relationship of dietary glycemic load (GL) and overall glycemic index (OGI) with macronutrients intake, body mass index (BMI) and insulin sensitivity in healthy children. The subjects comprised of 105 healthy non-obese eight-years old children, 60 boys and 45 girls. A Food Frequency Questionnaire (FFQ) evaluating dietary habits, GL and OGI. Insulin sensitivity was evaluated by the homeostatic model assessment (HOMA). GL was positively associated with dietary total (correlation coefficient, r=0.57) and starch (r=0.67) carbohydrates, daily consumption of pasta and white bread, cooked potatoes, bakery products and cookies, and negatively with dietary fats (r=-0.52). OGI was positively associated with daily consumption of white bread and cookies, and negatively associated with soluble carbohydrates (r=-0.35), and consumption of fibres, proteins, fruit, legumes and carrots. No significant association was found of GL or OGI with BMI or insulin sensitivity. In healthy children, GL and OGI may represent a useful indicator of quality of diet.