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Dietary supplements are used to implement and balance common dietary habits. The general belief is that natural substances reduce the risk of chronic diseases and amplify sports performance with no harmful side effects. Since sports science students will become professionals of sport activities and may also have a role in suggesting the use of dietary supplements to athletes, the aim of this study was to evaluate if physical activity influences the use of drugs and dietary supplements. A modified version of the International Physical Activity Questionnaire—Short Form (IPAQSF) was administered to perform these evaluations. A total of 1452 students from the University of Messina, Italy, enrolled in sports science courses completed the survey; of these, 1075 (704 male and 371 female students) were physically active in moderate- or high-intensity training. Of those physically active students, 709 (440 male and 269 female) were competitive athletes, identified on the basis of their answer to a specific question also indicating the type of sport they practice. The results suggest that 5.6% of all respondents were regular consumers of pharmaceutical products, compared to just 1.0% of the cohort of competing athletes. In contrast, the consumption of natural supplements was similar (14% vs. 15%) between groups. The most frequently used supplements were vitamins, including vitamin C, vitamin B complex, and multivitamin complex, followed by minerals and amino acids or protein complex. The probability of using dietary supplements was mostly related to the male gender (OR 1.64; 95% CI: 1.17–2.30), having a job (OR 1.45; 95% CI: 1.07–1.96), and, most of all, performing physical activity (OR 3.53; 95% CI: 2.18–5.71). The only factor related to a higher use of drugs was female gender (OR 2.40; 95% CI: 1.52–3.79), and the most used class was antihistaminic, followed by FANS. These results suggest that among the specific population of sports science students, those performing physical activity are less prone to using pharmaceutical products and have healthier habits.

The periaqueductal gray is a mesencephalic structure involved in modulation of responses to stressful stimuli. Structural connections between the periaqueductal gray and the cerebellum have been described in animals and in a few diffusion tensor imaging studies. Nevertheless, these periaqueductal gray–cerebellum connectivity patterns have yet to be fully investigated in humans. The objective of this study was to qualitatively and quantitatively characterize such pathways using high-resolution, multi-shell data of 100 healthy subjects from the open-access Human Connectome Project repository combined with constrained spherical deconvolution probabilistic tractography. Our analysis revealed robust connectivity density profiles between the periaqueductal gray and cerebellar nuclei, especially with the fastigial nucleus, followed by the interposed and dentate nuclei. High-connectivity densities have been observed between vermal (Vermis IX, Vermis VIIIa, Vermis VIIIb, Vermis VI, Vermis X) and hemispheric cerebellar regions (Lobule IX). Our in vivo study provides for the first time insights on the organization of periaqueductal gray–cerebellar pathways thus opening new perspectives on cognitive, visceral and motor responses to threatening stimuli in humans.

Excessively increased training volume and/or intensity and competition can lead to development of overtraining syndrome, causing a performance decrement in athletes. Tracking individual response to exercise intensity is crucial for establishing recovery strategies. We assessed the exercise intensity-dependent variability of stress response biomarkers, namely cortisol (C), testosterone (T), s-IgA, and advanced oxidation protein products (AOPP), in saliva samples of teenage elite water polo players. Saliva was collected on a day of training match (T1) and a day of competitive match (T2), at morning, before and after match. Cortisol/proteins and testosterone/proteins concentrations decreased throughout day T1, whereas increased throughout day T2. The highest values were measured after match on day T2 (2.5 ± 0.5 vs 14.6 ± 6.3 ng/mg; 0.061 ± 0.024 vs 0.371 ± 0.15 ng/mg, respectively). sIgA/proteins and AOPP/proteins concentrations increased throughout both days, and were higher after T2 match than T1 one (respectively, 1073.0 ± 438.2 vs 71.0 ± 17.3 μg/mg; 78.05 ± 24.2 vs 15.98 ± 3.16 nmol/mg, P = .003). Significant differences between concentrations of different biomarkers recorded on T1 and T2 were found only for AOPP, suggesting an increased oxidative stress on day T2. Free testosterone/cortisol ratio on day T2 was lower than that at morning (0.053 ± 0.021 vs 0.107 ± 0.031), indicating an increased catabolic response after competitive match. A highly significant positive correlation was found between Cortisol/Proteins and Testosterone as well as s-IgA/Proteins on day T1, and between Cortisol/Proteins and AOPP on day T2. In conclusion, we found that different types of activities, such a training or competitive session can affect the hormonal response, immunity, and oxidative stress, thereby modulating athletic performance. Our findings also confirm the usefulness of saliva testing as noninvasive way for monitoring the individual response to changes in exercise intensity in teenage elite water polo players.

Background The coexistence of a double superior vena cava (SVC) and a partially left inferior vena cava (PLIVC) with a circumaortic collar, associated with other congenital malformations, was not described previously. Case Description We present a 33-year-old woman in hemodialysis with complete exhaustion of the brachial routes for vascular access, admitted to our Nephrology Unit for a long-term central venous catheter (CVC) implant, usually by us performed under EchoScopic Technique (EST), an echographic venipuncture followed by continuous radioscopic control of guidewire and catheter in all the steps of implant. An intraoperative venography showed a complete stop of right internal jugular vein, a right SVC, a persistent left SVC, a left inferior vena cava in the iliac and subrenal tracts, a circumaortic venous collar in the renal tract, and normal right suprarenal and hepatic tracts. Conclusions The double SVC was related to the persistence of the caudal part of the anterior cardinal veins. As to the PLIVC, the iliac and subrenal parts of the inferior vena cava can be related to the persistent left supracardinal vein, while the circumaortic venous collar to the persistent intersupracardinal and left subsupracardinal anastomoses. All invasive procedures, and particularly those potentially complicated, must be performed under EST, now considered a mandatory tool for CVC implants, owing to the hypothesis of possible venous congenital anomalies.

Background Diffusion tensor imaging (DTI) is a non-invasive MR technique widely employed to study muscle anatomy. DTI parameters have been used to investigate microstructural changes dependent on demographic factors or transient condition such as exercise. The present study is aimed at investigating the diffusion parameters changes of the human calf muscles after a 3-months strength training protocol. Methods Ten young men were trained for improving size and strength of the medial (GCM), lateral gastrocnemius (GCL) and soleus (SL) three times a week, with at least 24 hours between training sessions, for a period of three months. Diffusion weighted magnetic resonance images were acquired at the beginning of the training period (TPRE) and at three months (TPOST) using a 3T scanner. The fractional anisotropy (FA), mean diffusivity (MD) and tensor eigenvalues (λ1, λ2, λ3) were derived from the diffusion weighted imaging data. Results We found a significant increase in λ1, λ2, λ3 and MD values and muscle volumes between TPRE and TPOST in all the examined muscles both for the left and right side. No significant differences were highlighted for FA. Conclusions DTI enables the investigation of muscle microstructure, allowing for the assessment of diffusion parameters variation of the muscle tissue in response to training thus being a useful tool to investigate physiological and pathological changes in skeletal muscle microstructure which could be employed to test the outcomes and the effectiveness of a given training protocol.

The periodontal ligament (PDL) is a highly vascularized connective tissue surrounding the root of a tooth. In particular, the PDL is continuously exposed to mechanical stresses during the phases of mastication, and it provides physical, sensory, and trophic functions. It is known that the application of orthodontic force creates a change in periodontal structures. In fact, these forces generate a pressure on the ligament that closes the vessels. The aim of this study is to observe the modifications of vascular endothelial growth factor (VEGF) in the PDL and extracellular matrix proteins after application of a pre-calibrated and constant orthodontic force at different phases of treatment. We used a 50-g NiTi coiled spring and in vivo samples of PDL of maxillary and mandibular premolars of patients subjected to orthodontic treatment. These teeth were extracted at 1, 7, 14, 21, and 30 days, respectively, by application of force. The extraction of the PDL was effectuated by scarifying the radicular surface on the pressure and tension sides. The mechanical stress induced by the application of force caused an increase in the reactive type of metabolism of extracellular matrix proteins and modulation of neoangiogenesis until restoration.

Aims Oxidative stress increase is a key event for development of sensitivity-related illnesses (SRIs). The aim of this work was to evaluate the influence of a genetic variant in the antioxidant enzyme glutathione peroxidase (GPx1) on oxidative stress development in SRIs. Main methods GPx1 rs1800668 genotype, as well as glutathione, ubiquinone, and DNA damage were assessed in 34 SRI patients and 36 healthy subjects. Key findings Total glutathione, reduced/oxidized glutathione, and ubiquinone were significantly decreased in cases compared with controls, while DNA fragmentation was significantly increased in patients. However, these differences were not associated to GPx1 genetic background. Significance GPx1 rs1800668 polymorphism does not play a major role in SRI-related oxidative stress development.

Methylene-tetrahydrofolate reductase (MTHFR) and paraoxonase 1 (PON1) gene polymorphisms have been associated with hyperhomocysteinemia and oxidative stress increase, that are established cardiovascular risk factors. Given that intense physical activity may increase the susceptibility to adverse cardiovascular outcomes, here we investigated the effects of MTHFR C677T and A1298C as well as PON1 Q192R gene polymorphisms on cardiovascular risk markers in twenty-eight male water polo elite players. The mean plasma levels of homocysteine (Hcy) and advanced oxidation protein products (AOPP) were above reference limits in resting conditions, and increased after competition. Moreover, a positive correlation was found between Hcy and AOPP concentrations, and also between their variations (ratio post-exercise/pre-exercise values) and the variations of lactic dehydrogenase (LDH) and creatine kinase (CK) activities, known as muscle damage markers. The highest Hcy and AOPP values were found in subjects having either MTHFR CT/AC or TT/AA, and PON1 QR192 genotype, respectively. After exercise, Hcy concentrations significantly increased in CT/AC or TT/AA subjects than in athletes having other MTHFR genotypes. A training-induced increase in plasma levels of LDH and CK activities, as well as myoglobin concentrations, was also observed, even if significant differences were found only for CK activity in athletes with MTHFR CT/AC or TT/AA athletes.

The amygdala plays a key role in gathering social cues to context-appropriate responses that require refined motor behavior, involving either direct or indirect connections with sensorimotor-related areas. Although, several studies investigated the structural and functional limbic connectivity of the amygdala both in animals and in humans, less is known about the limbic modulation on sensorimotor-related areas. However, recent evidences suggest the amygdala as a possible cornerstone in the limbic-motor interface. Herein, we used high-resolution diffusion data of the Massachusetts General Hospital-University of Southern California (MGH-USC) Adult Diffusion Dataset, constrained spherical deconvolution-based signal modeling and probabilistic tractography aimed at identifying and reconstructing the connectivity patterns linking the amygdala to the limbic- and sensorimotor-related areas. As regards the limbic network, our results showed that the amygdala has high probability to be connected with the fusiform gyrus and the lateral orbitofrontal cortex. On the other hand, our connectomic analysis revealed a close interplay between the amygdala and the inferior parietal lobule, followed by the postcentral gyrus, the precentral gyrus and the paracentral lobule. The findings of the present study are in line with previous literature and reinforce the idea of the existence of a limbic-motor interface, which is likely to be involved in the emotional modulation of complex functions such as spatial perception and movement computation. Considering that these pathways may play an important role, not on in physiological conditions, but also in pathological context, further studies should be fostered in order to confirm the existence of a limbic-motor interface and its precise functional meaning.

Title: An Immunofluorescence Study About Staining Pattern Variability of Sarcoglycans in Rat’s Cerebral and Cerebellar Cortex. Background: Sarcoglycans are transmembrane glycoproteins which connect extracellular matrix components to cytoskeleton. This protein system has been long studied in muscle but there are few data about its localization in non-muscular tissues. Methods and Findings: In the present report, we have conducted an indirect immunofluorescence study on normal rat’s cerebral and cerebellar cortex. Our results show that in these districts each sarcoglycan is expressed by a “spot-like” staining pattern, with spots of 0.5-2 μm average diameter, extending mainly around the soma of neurons and glial cells. In cerebral cortex, although all sarcoglycans are present, a staining pattern variability for each sarcoglycan, in the different cerebral cortex areas, exists. Instead, the pattern distribution level of sarcoglycans in cerebellar cortex doesn’t change. We also performed a statistical analysis which confirms the immunofluorescence results. Conclusions: Then, the presence of a sarcoglycans variability in cerebral cortex, where it is known the existence of several synaptic network, and the absence of a sarcoglycans variability in cerebellar cortex, where the same synaptic networks are repeated unchanged, suggest that in brain sarcoglycans may be associated with synaptic networks. Moreover, the distribution of sarcoglycans, mainly in post-synaptic regions of the neurons, suggests a role of these proteins in cellular signalling, regulating membrane receptors assembly. We also support that sarcoglycans in glial cells could be associated with the regulation of the mechanism in the brain-blood-barrier.

The post-mortem assessment of sepsis-related death can be carry out by many methods recently suggested as microbiological and biochemical investigations. In these cases, the cause of death is a multiple organ dysfunction due to a dysregulated inflammatory response occurring after the failure of infection control process. It was highlighted also that the heart can be a target organ in sepsis which determines the so-called septic cardiomyopathy characterized by myocardial depression. Several mechanisms to explain the pathophysiology of septic cardiomyopathy were suggested, but very few studies about the structural alterations of cardiac cells responsible for myocardial depression were carried out. The aim of this study was to evaluate whether sarcoglycans (SG) were involved in septic cardiac damage analyzing their expression in sepsis-related deaths and, particularly, if these proteins can be used as markers of septic myocardial dysfunction. Cases of septic-related death confirmed by clinical and autopsy records were investigated and compared to a control group of traumatic deaths. Indirect immunofluorescence analysis was performed to analyze α-SG, β-SG, δ-SG, ζ-SG, ε-SG, and γ-SG. Decrease of fluorescence staining pattern for all tested sarcoglycans was observed in the septic-related deaths compared to normal fluorescence staining pattern of control group. These results provide new findings about the myocytes structural alterations due to sepsis and suggest that these proteins could be used in forensic assessment of septic cardiomyopathy.

Acute exercise is known to induce oxidative stress. Here we assessed the effects of gene polymorphisms SOD2 A16V, CAT −844 G>A, and GPx-1 rs1800668 C>T on oxidative stress markers in 28 elite water polo male players prior to and after a routinely programmed friendly match. The mean plasma concentrations of derivatives of reactive oxygen metabolites (dROMs), as well as lactic dehydrogenase (LDH) activity, creatine kinase (CK) activity, CK-MB, and myoglobin, were significantly increased after exercise, while blood antioxidant potential (BAP) and total free thiols were significantly decreased, compared with those measured before exercise. Advanced oxidation protein products (AOPP) were also increased after exercise but not significantly. We observed that water polo players having either AV16 or VV16 SOD genotype exhibited a significant increase of postexercise AOPP, LDH, CK, and myoglobin plasma levels in comparison with wild-type athletes. Water polo players having either CAT −844 GA or GPx1 CT genotype showed a significant increase of postexercise dROMs plasma levels and, respectively, GPx and CAT enzyme activities in comparison with wild-type subjects. These preliminary results suggest that the screening for gene variants of antioxidant enzymes could be useful to assess individual susceptibility to oxidative stress and muscle damage in water polo players.

The sarcoglycan complex is a trans-membrane system playing a key role in mechano-signaling the connection from the cytoskeleton to the extracellular matrix. While β-, δ-, and e-sarcoglycans are widely distributed, γ- and α-sarcoglycans are expressed exclusively in skeletal and cardiac muscle. Insufficient data are available on the distribution of sarcoglycans in nonmuscular tissue. In the present study, we used immunohistochemical and RT-PCR techniques to study the sarcoglycans also in normal human glandular tissue, a type of tissue never studied in relation to the sarcoglycan complex, with the aim of verifying the real wider distribution of this complex. To understand the role of sarcoglycans, we tested specimens collected from patients affected by benign prostatic hyperplasia and adenocarcinoma. For the first time, our results showed that all sarcoglycans are detectable in normal samples both in epithelial and in myoepithelial cells; in pathological prostate, sarcoglycans appeared severely reduced in number or were absent. These data demonstrated that all sarcoglycans have a wider distribution suggesting a new unknown role for these proteins. The decreased number of sarcoglycans, containing cadherin domain homologs in samples of prostate affected by hyperplasia, and the absence of proteins in prostate biopsies, in cases affected by adenocarcinoma, could be responsible for the loss of adhesion between epithelial cells, which in turn facilitates the progression of benign tumors and the invasive potential of malignant tumors. Anat Rec, 297:327–336, 2014. © 2013 Wiley Periodicals, Inc.

Costameres encircle the myocyte perpendicular to its long axis, and comprise two protein complexes: the dystrophin-glycoprotein complex (DGC) and the vinculin-talin-integrin system. They participate in signaling functions and protect muscle cells from damage induced by workload. The behaviour of those proteins has been a focus of study starting from skeletal and smooth muscle cells to cardiomyocytes, and still represents a topical subject for cardiovascular translational research. This review summarizes the past and present novel approaches of our and other groups of work on this subject of research.

Ureteropelvic junction obstruction is one of the most common causes of hydronephrosis in children. A malfunction of smooth muscle cells is believed to be the underlying mechanism causing obstruction. We investigated the expression of some integrins, talin and β-dystroglycan, considered the main compound of smooth muscle cell cytoskeleton, and active caspase 3 at the level of the ureteropelvic junction obstruction.Specimens were obtained at pyeloplasty in 12 children with ureteropelvic junction obstruction. Six control specimens were obtained during organ explantation. Specimens were divided into renal pelvis, ureteropelvic junction and ureter below the obstruction. Western blot analysis of active caspase 3, and immunofluorescence and polymerase chain reaction analysis were performed for α7A, β1A, α7B and β1D integrins, talin and β-dystroglycan.Talin and β-dystroglycan were slightly impaired in ureteropelvic junction obstruction, while α7B and β1D integrins were severely reduced, and α7A, β1A and active caspase 3 were significantly enhanced compared to controls.We demonstrated activation of apoptosis and a critical alteration of cytoskeleton that might explain the altered function and the increased apoptosis in smooth muscle cells in ureteropelvic junction obstruction. The delayed rearrangement of the cytoskeleton of smooth muscle cells in ureteropelvic junction obstruction might be linked to a postnatal splicing from α7A and β1A to α7B and β1D integrins, respectively. This relationship could explain the common clinical scenario of spontaneous improvement of hydronephrosis in children with suspected ureteropelvic junction obstruction.

In the mature heart, the intercalated disc and costameres provide the cell-cell and cell-matrix junctions respectively. Intercalated disc is situated at the bipolar ends of the cardiomyocytes and the myofibrils are anchored at this structure. The costameres mediate integration with the extracellular matrix that covers individual cardiomyocytes laterally. Costameres are considered as "proteic machinery" that appears to comprise two protein complexes: the dystrophin-glycoprotein complex (DGC) and the vinculin-talin-integrin system. There are structural differences between atrial and ventricular myocytes, but there have been relatively few studies that have analyzed costameres and focal adhesion function in cardiac cells. Our previous study carried out only on atrial myocytes, demonstrated that the DGC and talin-vinculin-integrin complexes had a costameric distribution that, unlike skeletal muscle, it localized only on the I band. We performed a further immunohistochemical analysis extending also the evaluation to the normal human cardiac muscle fibers obtained from ventricle and interventricular septum, in order to define the distribution and the spatial relationship between the proteins of the two complexes also in the other heart districts. Immunoconfocal microscopy of cardiac tissue revealed the costameric distribution of DGC and of vinculin-talin-integrin system, the association of all tested proteins in intercalated disks, in disagreement with other Authors, and in T-tubule with irregular spokelike extensions penetrating toward the center of the cell. Moreover, our data showed that all tested proteins colocalize between each other.

Objectives To investigate the distribution of MTHFR C677T and A1298C as well as PON1 Q192R gene polymorphisms, known to be involved in hyperhomocysteinemia-related cardiovascular risk, in elite athletes. Design and methods Genetic background at MTHFR and PON1 loci and plasma levels of homocysteine, folate, vitamin B 12 and advanced oxidation protein product (AOPP) levels were assessed in thirty-seven 400 m male hurdlers and thirty-four sedentary subjects. Results Homocysteine plasma levels and AOPPs were significantly higher in hurdlers bearing the TT677/AA1298 diplotype than in both hurdlers and sedentary subjects with other diplotypes. A positive dependence between either homocysteine or AOPP plasma levels and MTHFR , but not PON1 genotype, was observed in hurdlers. Conclusions Elite hurdlers, having an unfavorable MTHFR genotype are exposed to increased cardiovascular risk, dependent on alterations of homocysteine and AOPP plasma levels.

Objective: Abdominal aortic aneurysms (AAAs) are characterized by structural remodelling resulting in the gradual weakening and expansion of the aortic wall. Wall stress may furnish a usable indicator to prevent the failure. In order to evaluate this risk, static pressure aging on the cap was used to perform FEA, simulating an aneurysm varying its dimensions from 10 mm to 50 mm. Analyses were carried out by imposing different thickness of the cap, and obtaining correspondent equivalent Von Mises stresses. Understanding how these stresses are distributed and what factors influence stress distributions is critical in evaluating the potential for rupture. Methods: A representative FE model was created in order to simulate the historical evolution of the AAA. CFD analyses were performed to obtain data of the static pressure aging on the model. Patient informed consent and IRB approval were obtained. A linear law was speculated to understand thickness thinning in function of aneurysm’s growth. The obtained pressure maps were used as input to perform elastic linear analyses on the five different FE wrappings. Results: If the bloody pressure is increased, that increases also wall shear stress, and an adaptive increase in arterial luminal size is observed. Results have evidenced peaks of stress varying from 0,004 MPa, for a diameter of 10 mm, to 0,45 MPa, for a diameter of 50 mm. Top and bottom zones of aneurysm result more solicited than the middle ones, as it can be deducted by obtained strain values, ranging from 5,84e-7, for a diameter of 10 mm, to 3,14e-4, for a diameter of 50 mm. In order to evaluate the mechanical behaviour of the cap, related to its thickness at 50 mm of diameter, different FEA were conducted varying thickness, uniformly, from 1 to 0.4 mm. As it is possible to notice, stress increases exponentially while thickness decreases. Conclusions: Results indicate an equivalent Von Mises stress of about 0,45 MPa, close the to failure value, for a critical dimension of 50 mm. This means that failure conditions can depend at least, by two variables: thickness and pressure.

The sarcoglycan complex is a trans-membrane system playing a key role in mechano-signaling the connection from the cytoskeleton to the extracellular matrix. While b-, d-, and e-sarcoglycans are widely distributed, g- and a-sarcoglycans are expressed exclusively in skeletal and cardiac muscle. Insufficient data are available on the distribution of sarcoglycans in nonmuscular tissue. In the present study, we used immunohistochemical and RT-PCR techniques to study the sarcoglycans also in normal human glandular tissue, a type of tissue never studied in relation to the sarcoglycan complex, with the aim of verifying the real wider distribution of this complex. To understand the role of sarcoglycans, we tested specimens collected from patients affected by benign prostatic hyperplasia and adenocarcinoma. For the first time, our results showed that all sarcoglycans are detectable in normal samples both in epithelial and in myoepithelial cells; in pathological prostate, sarcoglycans appeared severely reduced in number or were absent. These data demonstrated that all sarcoglycans have a wider distribution suggesting a new unknown role for these proteins. The decreased number of sarcoglycans, containing cadherin domain homologs in samples of prostate affected by hyperplasia, and the absence of proteins in prostate biopsies, in cases affected by adenocarcinoma, could be responsible for the loss of adhesion between epithelial cells, which in turn facilitates the progression of benign tumors and the invasive potential of malignant tumors.

The muscle nicotinic acetylcholine receptor (AChR) turns over at different rates depending on stage of synaptogenesis and innervation. Tyrosine phosphorylation modulates desensitization, interaction with cytoskeleton and lateral mobility in the membrane of AChR. To determine whether tyrosine phosphorylation also modulates the turnover of AChR, myotubes in vitro were exposed to the tyrosine phosphatase inhibitor pervanadate. Our data indicate that a transient increase of phosphotyrosine levels stabilized a fraction of AChRs. The effects were limited to the non-epsilon subunit-containing AChRs already present in the membrane. Tyrosine phosphorylation of the receptor occurred on the beta subunit, was transient and stable molecules were not selectively tyrosine phosphorylated. The data indicate that modulation of phosphotyrosine levels in muscle cells provides signals to control AChR metabolic stability.

Background The costameres in skeletal muscle fibers were first described by Pardo et al. (1983a) and have been defined as transverse circumferential elements of the cytoskeleton associated to the sarcolemma. Specific immunostaining for vinculin shows that the costameres overlie I bands. However, an exact correlation between the costameres and the Z line is uncertain, although ∼10 proteins so far have been localized in the costameres. To define the exact localization of costameres in human skeletal muscle fibers, we carried out an immunofluorescence study using confocal scanning laser microscopy on the fascia lata muscle of adult males. Methods Samples were fixed in 3% paraformaldehyde; frozen sections were treated with antivinculin, antitalin, antidesmin, and anti-α-actinin, then immunostained with TRITC. For double localization, the TRITC-streptavidin, as a marker for vinculin and FITC-streptavidin a marker for desmin, were used. Results The distance between two subsequent transverse lines of actin-inf indicated that muscle fibers were well stretched. Processing, with different software functions of the images obtained using CLSM, shows that vinculin and talin are only present in the sarcolemmal lattice. Immunostaining for vinculin and double immunostaining for vinculin and desmin demonstrate that costameres superimpose underlying I bands without interruption at the Z line. Immunostaining for talin showed that the protein is located in correspondence with the I band and M line. Conclusions We believe that costameres are “proteic machinery.” The findings of the present study suggest that it is possible to determine the width and the period of each proteic component. In addition, we indicate that costameres are present in correspondence with M line. © 1996 Wiley-Liss, Inc.

Morphological and volumetric variabilities of lateral ventricles are considered indirect indicators of age-and gender-related reductions of white and gray matter. However, no studies have classified lateral ventricles with different morphologies or showed its asymmetric shapes in healthy subjects. We performed an analysis on living subjects, using 3D volume rendering techniques. Eighty-five healthy Caucasian volunteers (49 women and 36 men aged 19–69 years) were scanned by a Philips Achieva 3T R2.6. Three-dimensional reconstruction allowed us to identify three main morphological shapes in living subjects and to show asymmetries between horns. We also assessed the surface deformation of the cerebral ventricles to identify region-specific shape differences in aging healthy adults. Statistical analysis showed significant gender- and age-related volume differences. An increase in lateral ventricle volume appears to be a constant, linear function of age throughout adult life.

The sarcoglycan (SG) complex (SGC) is a subcomplex within the dystrophin-glycoprotein complex (DGC) and is composed of several transmembrane proteins (α, β, δ, γ, ε and ζ). The DGC supplies a transmembranous connection between the subsarcolemmal cytoskeleton networks and the basal lamina in order to protect the lipid bilayer and to provide a scaffold for signaling molecules in all muscle cells. In addition to its role in muscle tissue, dystrophin and some DGC components are expressed in neurons and glia. Very little is known about the SG subunits in the central nervous system (CNS) and some data suggested the presence of ε and ζ subunits only. In fact, mutations in the ε-SG gene cause myoclonus-dystonia, indicating its importance for brain function. To determine the presence and localization of SGC in the human cerebral cortex, we performed an investigation using immunofluorescence, immunoblotting and reverse transcriptase polymerase chain reaction. The results showed that all SG subunits are expressed in the human cerebral cortex, particularly in large neurons but also in astrocytes. These data suggest that the SG subcomplex may be involved in the organization of CNS synapses.

Several studies demonstrated that transglutaminases play a key role in extracellular matrix stabilization needed for cell differentiation. We evaluated transglutaminase expression and activity in the pre-secretory stage of differentiation of the continuously erupting rat incisor. We observed that transglutaminase-mediated incorporation of monodansylcadaverine into protein substrates was specifically located in the apical loop, and along the basement membrane joining mesenchyme and inner dental epithelium in the odontogenic organ. Enzyme activity was associated with mRNAs for transglutaminase 1 and 2. Notably, labelling cells for these isoenzymes were observed in both mesenchymal and epithelial compartments, but not in the basement membrane, in the ameloblast facing pulp anterior region, where ameloblast and odontoblast differentiation begins. These findings demonstrate that transglutaminase 1 and transglutaminase 2 are expressed at a major extent in the pre-secretory stage of regenerating rat incisor, where they probably play complementary roles in cell signalling between mesenchyme and epithelium and extracellular matrix.

During neuromuscular junction maturation, the rapidly degrading receptors (Rr; t1/2 approximately equal to 1 day) are replaced by metabolically stable molecules (Rs; t1/2 approximately equal to 10 days). Rr and Rs do not interconvert, are differently regulated after denervation in adult muscle and are endowed of unique responses to stabilizing agents. In cultured rat myotubes all the epsilon subunit-containing acetylcholine receptors (epsilon-AchRs) are of the Rs type. In the present study we show that Rs exist also in absence of epsilon-AChR and that nonepsilon-(presumably gamma-)AChRs can be included in the Rs pool when epsilon-AChR expression is low. The data indicate that Rs metabolic properties are independent of AChR subunit composition and that epsilon subunit is a signal to efficiently sort AChR molecules to the Rs pool.

Costameres were identified, for the first time, in skeletal and cardiac muscle, as regions associated with the sarcolemma, consisting of densely clustered patches of vinculin; they have many characteristics common to the cell-extracellular matrix-type of adherens junctions. Costameres are considered 'proteic machinery' and they appear to comprise two protein complexes, the dystrophin-glycoprotein complex (DGC) and the vinculin-talin-integrin system. In comparison to skeletal muscle, few studies have focused on cardiac muscle regarding these two complexes, and study is generally relative to dystrophin or to cardiac diseases, such as cardiomyopathies. However, insufficient data are available on these proteins in healthy human cardiomyocytes. For this reason, we performed an immunohistochemical study using human cardiac muscle fibers, in order to define the real distribution and the spatial relationship between the proteins in these two complexes. Our data showed a real costameric distribution of DGC and of the vinculin-talin-integrin system; all tested proteins were present in T-tubule and in intercalated disks. Moreover, our data demonstrated that all tested proteins of DGC colocalized with each other, as all tested components of the vinculin-talin-integrin system, and that all tested proteins of DGC colocalized with all tested proteins of the vinculin-talin-integrin system. Finally, all tested proteins of the two complexes were localized in the region of the sarcolemma over the I band, in 100% of our observations. The present study, for the first time, analyzed the majority of proteins of DGC and of the vinculin-talin-integrin system in cardiac muscle fibers, and it confirmed that DGC and the vinculin-talin-integrin system have a role in the transduction of mechanical force to the extracellular matrix. Finally it attributed a key role in the regulation of action potential duration to cardiac myocytes.