Effect of intercellular adhesion molecule 1 deficiency on the development of apical periodontitis.

AIM: To evaluate the specific role of ICAM-1 in host responses against endodontic infection. METHODS: Apical periodontitis was experimentally induced in the mandibular first molars of ICAM-1 knockout and wild-type (WT) mice by pulp exposure to the oral environment. At 7, 21 and 42 days following pulp infection, the animals were euthanized and the jaws were prepared for analysis under conventional and fluorescence microscopy (histopathologic and morphometric analysis), immunohistochemistry (polymorphonuclear leucocytes), enzyme histochemistry (osteoclasts and cementoclasts) and RT-PCR (IL-1 α, TNF-α, INF-γ, IL-10, RANK, RANKL and OPG). A generalized linear model with GLIMMIX procedure with Satterthwaite approximation method of degrees of freedom, Tukey-Kramer, pseudo-ranking nonparametric, Bonferroni-Holm multiple testing adjustment, analysis of variance (ANOVA) and the Tukey's multiple comparisons tests were used to evaluate the statistical differences between the groups using SAS 9.4 and the GraphPad Prism 5.0 software (α = 0.05). RESULTS: Compared to WT mice, ICAM-1 knockout mice had significantly greater bone resorption (P < 0.05), reduced recruitment of neutrophils to periapical inflammatory tissues (P < 0.05) and an increased number of fibroblasts (P < 0.05) at all experimental periods. The osteoclast number was significantly higher in ICAM-1 KO than that of WT animals at all times (P < 0.05), while there was no significant difference between the groups regarding cementoclasts. At day 21, the level of IL-1α, RANK, RANKL and IL-10 had increased significantly in tissues from ICAM-1 KO versus WT mice (P < 0.05), while no significant difference was observed in TNF-α and OPG levels (P > 0.05). Tissue levels of INF-γ were significantly lower in ICAM-1 KO than those in WT mice (P < 0.05). CONCLUSION: ICAM-1 deficiency impaired the host response against endodontic infection, resulting in increased tissue destruction.

Masticatory muscle activity evaluation by electromyography in subjects with zygomatic implants.

BACKGROUND: Zygomatic implants are an alternative treatment in the rehabilitation of atrophic maxilla to promote stability in the stomatognathic system. The aim of this study was to compare the electromyographic (EMG) activity of masseter and temporalis muscles in controls and in individuals with complete implant-supported dentures anchored in the zygomatic bone. MATERIALS AND METHODS: Fifty-four volunteers of both genders (mean age 52.5 years) were selected and distributed into two groups: Individuals with zygomatic implant (ZIG; n=27) and fully dentate patients (CG; n=27). MyoSystem-BR1 was used to assess masseter and temporalis muscles EMG activity in different mandibular movements: protrusion, clenching, maximal voluntary contraction (MVC) with Parafilm M®, right and left laterality and chewing (peanuts and raisins). Data was processed, normalized (MVC) and analyzed using the SPSS 21.0. Student t-test (P ≤ 0.05) was used for group comparison. RESULTS: The results were statistically significant (P ≤ 0.05) for protrusion, clenching, right and left laterality and raisin chewing. For the mandibular posture conditions, the ZIG obtained higher EMG activity patterns when compared to CG. For the masticatory performance during chewing of peanuts and raisins, the ZIG showed higher EMG mean values when compared to CG. CONCLUSIONS: The zygomatic implant promoted an active response of the muscle fibers (hyperactivity) during both mandibular posture and chewing conditions, probably due to the absence of periodontal receptors, which play a significant role for preparing a bolus for swallowing.

The type II "receptor" as a decoy target for interleukin 1 in polymorphonuclear leukocytes: characterization of induction by dexamethasone and ligand binding properties of the released decoy receptor.

Whereas the signaling function of the interleukin 1 (IL-1) receptor type I (IL-1R I) has been well documented, the type II "receptor" has been suggested to act as a decoy target for this cytokine. Since IL-1 may represent a key target of the immunomodulatory and antiinflammatory properties of glucocorticoids (GC), the aim of this study was to investigate the effects of dexamethasone (Dex) on IL-1R expression in human polymorphonuclear leukocytes (PMN), which express predominantly the type II molecule (IL-1R II). We found that Dex augments the levels of steady state transcripts encoding the IL-1R I and, most prominently, those of IL-1R II. Dex induced both transcripts via transcription-dependent mechanisms and by prolongation of the mRNAs half-lives. Inhibition of protein synthesis superinduced basal and Dex-augmented IL-1R II mRNA, whereas it completely inhibited the induction by Dex of IL-1R I transcripts. Induction of IL-1R II mRNA by Dex was associated with augmented membrane expression and release of the type II IL-1 binding molecule. This effect was mediated by the GC receptor. Other steroids (17 beta-estradiol, progesterone, and testosterone) were ineffective. The concentrations of IL-1 alpha and IL-1 receptor antagonist required to displace the binding of IL-1 beta to the soluble form of the decoy molecule induced by Dex from PMN were, respectively, 100 and 2 times higher compared with IL-1 beta. The induction by Dex of the type II receptor, a decoy molecule for IL-1, may contribute to the immunosuppressive and antiinflammatory activities of Dex.

Intracellular motor-driven transport of rodlike smooth organelles along microtubules.

Molecular motors are fascinating proteins that use the energy of ATP hydrolysis to drive vesicles and organelles along cytoskeleton filaments toward their final destination within the cell. Several copies of these proteins bind to the cargo and take turns transporting the cargo attaching to and detaching from the track stochastically. Despite the relevance of molecular motors to cell physiology, key aspects of their collective functioning are still unknown. In this work we propose a one-dimensional model for the transport of extensive and smooth organelles driven by molecular motors. We ran numerical simulations to study the behavior of the cargo for different motor configurations, focusing on the transport properties observable in the experiments, e.g., average speed of the organelle and variations in length. We found that active motors drive the cargo using two different mechanisms: Either they locate in front of the cargo and pull the organelle or they situate at the cargo lagging edge and push. Variations in the organelle length is in close relation with the fraction of motors in each configuration, which depends on the resisting load. The results of this model were contrasted with experimental data obtained from the tracking of rodlike mitochondria during active transport in Xenopus laevis melanophores.

Dielectric properties of healthy and diabetic alloxan-induced lenses in rabbits.

The dielectric properties of the eye lens were studied for healthy and alloxane-induced diabetic rabbits in the frequency range from 500 Hz to 100 kHz electric field and temperatures from 25 to 50 °C. In the full temperature range, the average relative permittivity and dielectric loss values for a healthy lens are lower than those recorded for diabetic tissue. Dielectric relaxation of polar amino acids on the alpha-crystallin surface with a characteristic frequency of 7 kHz in the range of 25-50 °C for healthy and diabetic samples is accompanied by the activation energy of proton conductivity with an average values of 33 and 39 kJ mol-1, respectively. The permittivity decrement, which characterizes the size of the dielectric dispersion with a central relaxation time of 0.023 ms for a diabetic sample, is more than twice as high as for a healthy sample. Measurements on the rabbit eye lens were carried out at ambient temperature above and below the physiological range, since these conditions provide an appropriate pattern of dielectric behavior for the diagnosis of clinical dysfunction of the human lens.

Changes in integrin and E-cadherin expression in neoplastic versus normal thyroid tissue.

UNLABELLED: BACK: The functional organization of polarized epithelia depends mostly on adhesion molecules belonging to the integrin and cadherin families. These molecules either recognize basement membrane components, such as laminins, or form intercellular junctions via homotypic interactions. Such tissue organization is often disrupted upon neoplastic transformation, and the resulting loss of functional polarization and cell cohesion might be a prerequisite for the invasive and metastatic behavior of carcinomas. PURPOSE: We studied modifications on thyroid adhesive mechanisms at various stages of neoplastic progression in terms of adhesion molecule expression, topography, and functional regulation by tyrosine kinases. Starting from this working hypothesis, we sought to identify one or more biological markers that would be suggestive of malignant transformation and poorer prognosis and that could be developed as a reliable indicator(s) in early diagnostic steps. METHODS: The study was carried out on both surgical samples and the corresponding fine-needle aspiration biopsy smears (numbers of specimens collected: 19 adenomas, seven follicular carcinomas, 13 papilary carcinomas, and 39 normal tissues). Immunohistochemistry of tissue sections and smears and immuno-precipitation and western blot analysis of protein extracts were done with a battery of monoclonal and polyclonal antibodies. Northern blotting was performed on RNA extracts from frozen tissue samples and use of an integrin subunit beta4 complementary DNA probe. RESULTS: Our findings can be summarized as follows: 1) In normal thyroid cells, the cooperative role of integrin alpha6beta4 and laminin 5/kalinin in hemidesmosome-mediated adhesion adhesion is missing, and recognition of the basal lamina occurs via integrin alpha3beta1 and laminin 1 and/or 2 (this pattern being maintained in adenomas but altered in carcinomas regardless of their histotype or differentiation grade); 2) only in carcinomas with clinical and/or histologic aggressiveness do neoexpression of integrin subunit beta4 and loss of laminin 2/merosin occur, indicating de novo assembly of integrin alpha6beta4; 3) pericellular redistribution and cytoskeletal disconnection of the E-cadherin-catenin complex occur; and 4) basal E-cadherin tyrosine phosphorylation decreases in carcinomas as compared with that in normal and adenomatous tissue. CONCLUSION: The malignant progression of thyroid tumors involves marked rearrangement of cell-basement membrane and cell-cell adhesion molecules and changes in their cytoskeleton linkage. These rearrangements are also easily and reproducibly detected on fine-needle aspiration biopsy smears. IMPLICATIONS: Immunodetection of adhesion molecules in sections and/or fine-needle smears may complement the toolbox of thyroid surgical pathologists; it may expand the possibilities of achieving a correct early diagnosis of thyroid tumors and of gaining some prognostic information on thyroid tumors.

[Exposure to crystalline silica in the air during renovation of residential buildings]. / Esposizione a silice libera cristallina nelle attività di ristrutturazione in edilizia abitativa.

Exposure to respirable cristalline silica dust during construction activities can cause silicosis and, in accordance with International Agency for Research on Cancer (IARC), even lung cancer. Among the different facets of construction, residential buildings are of particular interest because the lack of exposure data for workers employed. In this sector a dry sand-blasting of brick ceiling and renewal activities has been investigated. The survey has been carried out in a civil dwelling and 58 samples have been collected using the SKC aluminium cyclone and Door-Oliver plastic cyclone at flow-rate 2.2-1.7 l/min. The respirable dust samples were analysed by x-ray diffraction using NIOSH Method 7500. Even if the results of this study should be considered preliminary, they showed SLC respirable concentrations even many time above TLV-TWA during a sand-blasting activities and some times also during the other activities. Awaiting the correct classification from European Community, it would be desiderable that the workers would had appropriate information and prevention on the specific risks associated to the dust in the construction activities.

Charge selectivity of proteinuria in type I diabetes explored by Ig subclass clearance.

To investigate the role of protein charge in early diabetic proteinuria, the clearance of proteins differing in charge and/or size (anionic and cationic Igs, albumin) was evaluated in 98 insulin-dependent (type I) diabetic patients selected as a representative sample of the 418 patients attending our clinics. Of the patients, 12.9% were microalbuminuric and 4.8% were macroalbuminuric. Anionic and total IgG clearances were significantly increased in 30.6 and 12.2% of patients and were correlated with duration of disease. Anionic IgG4 clearances were increased in patients (9.2%) with normal IgG excretion, suggesting that charge-selectivity impairment is responsible for protein loss. Anionic Ig clearances were also higher in some patients (14.3%) with normal albumin clearance, probably as a result of different glomerular filtration and/or tubular reabsorption. The anionic-cationic IgG clearance ratio tended to increase in parallel with albumin clearance, but once above macroalbuminuric levels, it tended to fall again, indicating the concomitant presence of size-selectivity loss. The anionic IgG clearance and the anionic-cationic IgG ratio, in addition to albumin excretion, may be valuable in assessing early kidney protein charge-selectivity impairment and better characterizing normoalbuminuric patients and those in the preclinical stage of diabetic nephropathy.

Lifelong physical exercise delays age-associated skeletal muscle decline.

Aging is usually accompanied by a significant reduction in muscle mass and force. To determine the relative contribution of inactivity and aging per se to this decay, we compared muscle function and structure in (a) male participants belonging to a group of well-trained seniors (average of 70 years) who exercised regularly in their previous 30 years and (b) age-matched healthy sedentary seniors with (c) active young men (average of 27 years). The results collected show that relative to their sedentary cohorts, muscle from senior sportsmen have: (a) greater maximal isometric force and function, (b) better preserved fiber morphology and ultrastructure of intracellular organelles involved in Ca(2+) handling and ATP production, (c) preserved muscle fibers size resulting from fiber rescue by reinnervation, and (d) lowered expression of genes related to autophagy and reactive oxygen species detoxification. All together, our results indicate that: (a) skeletal muscle of senior sportsmen is actually more similar to that of adults than to that of age-matched sedentaries and (b) signaling pathways controlling muscle mass and metabolism are differently modulated in senior sportsmen to guarantee maintenance of skeletal muscle structure, function, bioenergetic characteristics, and phenotype. Thus, regular physical activity is a good strategy to attenuate age-related general decay of muscle structure and function (ClinicalTrials.gov: NCT01679977).

Dopamine-sensitive cAMP generating system in rat extracerebral arteries.

The effect of dopamine (DA) on the 3',5'-cyclic adenosine monophosphate (cAMP) generating system in rat extracerebral arteries was studied in order to identify DA receptors. The carotis interna and the basilar artery as well as the arteries of the circle of Willis were homogenized and centrifuged. This extracellular arterial preparation was used for studying the effect of DA, norepinephrine and of drugs interfering with alpha- and beta-adrenergic as well as with dopaminergic receptors on cAMP production. DA increased the concentration of cAMP. The DA-elicited increase of cAMP levels was inhibited by dopamine receptor blocking agents (phenothiazine derivatives and butyrophenones) but unaffected by alpha- or beta-adrenoceptor blocking drugs or by the ergot-like DA agonist bromocriptine, or by sulpiride. The findings appear to be indicative of the existence of only D1 receptors within the blood vessels studied.

Characterization of [3H]5-hydroxytryptamine uptake within rat cerebrovascular tree.

The in vitro uptake of tritiated serotonin ([3H]5HT) was studied in a preparation of rat extracerebral arteries. The uptake of [3H]5HT was time- and temperature-dependent and of high affinity; linear regression analysis gave a Km value of 6.48 X 10(-7) M for the specific uptake. Bilateral superior cervical ganglionectomy was without effect on [3H]5HT uptake while it significantly reduced the uptake of tritiated norepinephrine by the preparation of rat extracerebral arteries. The serotonergic neurotoxin 5,7-dihydroxytryptamine and lesions to both the medial and the dorsal raphe nuclei caused a marked loss of [3H]5HT uptake but did not change the uptake of tritiated norepinephrine. Competition studies with norepinephrine, desimipramine (a noradrenergic uptake blocker), nomifensine (a dopaminergic uptake blocker) and fluoxetine (a 5HT uptake blocker) confirmed the specificity of the [3H]5HT uptake mechanism. Histoautoradiographic studies showed the highest density of silver grains at the level of the adventitial-medial border of the basilar artery. Fluoxetine inhibited the accumulation of silver grains within the adventitial-medial border in the blood vessel studied. The present data further support the view that a neuronal serotonergic system may play a role in the control of blood flow in the cerebrovascular tree.

Dopamine-sensitive adenylate cyclase in rabbit renal artery.

The effect of dopamine on 3',5'-cyclic adenosine monophosphate (cAMP) generation in the rabbit renal artery was studied. Dopamine added to homogenates of rabbit renal artery increased the concentration of cAMP. The stimulation of cAMP levels elicited by dopamine was remarkably reduced by the dopamine receptor antagonists fluphenazine and haloperidol, but it was unaffected by the ergot-like dopaminergic agonist bromocriptine. These findings appear to indicate that the effects of dopamine on rabbit renal artery are mediated through a D1 receptor.

Effects of prolonged picotamide therapy on platelet activity in patients with peripheral arterial disease.

An open study was carried out in 14 patients with peripheral arterial disease to investigate the effects of prolonged therapy with picotamide on platelet activity. Patients received daily oral doses of 900 mg picotamide for 1 month, 600 mg per day during the second month and 300 mg per day from the third to the sixth month of the study. Measurements were made before and during therapy of blood coagulation parameters and factors influencing platelet function, i.e. plasma beta-thromboglobulin and serum thromboxane B2. The results showed that there were no significant variations in platelet count, prothrombin time, partially activated thromboplastin time, presence and amount of fibrinogen in blood, and antithrombin III. Examination for fibrinogen degradation products was constantly negative and unaltered during therapy. Although plasma beta-thromboglobulin values did not vary significantly, there was a significant and progressive reduction throughout treatment in serum levels of thromboxane B2.

Beta-adrenoceptors in the rat kidney. Immunohistochemical study.

The direct histochemical detection of beta-blocker binding sites was studied in sections of rat kidney using an immunohistochemical technique developed in our laboratory. Frozen sections of rat kidney were incubated in a solution of (-)alprenolol, washed, exposed to fluorescent (-)alprenolol antibodies (FAA) and then observed at a fluorescence microscope. Strong fluorescence was found within the wall of renal artery and vein, but primarly in the artery. At the level of blood vessels (-)alprenolol binding sites were located chiefly in the media and in the intima. The renal glomerulus, the loop of Henle and collecting tubules appear to be free of any fluorescence. Consequently they do not have beta-adrenoceptors. On the contrary, the glomerular afferent and afferent arterioles, the cellular elements of the juxtaglomerular apparatus, proximal and distal convoluted tubules, are rich in (-)alprenolol binding sites. At higher magnifications the immunoreactivity appears to be located in the basal membrane of cellular elements which indicates that (-)alprenolol binding sites are membrane receptors. The direct immunohistochemical detection of beta-blocker binding sites in the kidney may offer useful information concerning the site of action of beta-blockers at the level of an important target organ for this class of drugs.

Beta adrenergic receptors in the parathyroid glands.

The location of beta-adrenoceptors in human parathyroid gland was studied using an immunohistochemical method. Frozen sections of human parathyroid glands, taken from surgical samples, were treated with (-)-alprenolol, washed and exposed to (-)-alprenolol antibodies conjugated with fluorescent dyes. The (-)-alprenolol was bound to the parathyroid principal cells and to the main blood vessels. On the contrary, adrenergic nerve fibres, demonstrated with formaldehyde fluorescence technique, were only located within the walls of main blood vessels. The findings are discussed.

Role of advanced glycation end-products (AGE) in late diabetic complications.

To evaluate accumulation of advanced glycation end-products (AGE) in diabetes and its possible correlation with late diabetic complications, AGE levels were measured by spectrofluorimetry in eye lens and sciatic nerve proteins and isolated tail tendon collagen of rats with experimental diabetes of 3- and 6-month duration. The values obtained were compared to those from age-matched control rats and correlated with cataract presence and somatosensory evoked potential (SEP) alterations. Diabetic animals had increased AGE levels in all tissues at both times; cataract developed in 29% of diabetic rats at 3 months and in 57% at 6 months; SEP conduction velocity was reduced in diabetic animals both at 3 (54.5 +/- 1.8 S.E.M. m/s vs. 73.9 +/- 1.0, P < 0.0001) and 6 months (59.5 +/- 1.4 vs. 71.5 +/- 1.6, P < 0.0001) from diabetes induction. No eye lens AGE level differences were observed when cataract presence was considered. Interestingly, in diabetic rats, increased sciatic nerve AGE levels were associated with reduced SEP. These data show that: (1) AGE levels are increased as early as 3 months from development of hyperglycemia; (2) other factors, in addition to an enhanced rate of fluorescent AGE formation, might play important roles in the pathogenesis of diabetic cataract; (3) increased peripheral nerve AGE levels are associated with SEP alterations.

A charge selectivity impairment in protein permselectivity is present in type 2 diabetes.

A possible loss in kidney charge permselectivity of proteins before any manifestation of nephropathy has been sought in type 2 (non-insulin-dependent) diabetes by assessing the clearances of proteins differing in charge and/or size (anionic and cationic immunoglobulins, albumin). Eighty-five consecutive outpatients with type 2 diabetes were studied and compared with 101 normal subjects. Of the patients, 14.1% were microalbuminuric and 2.3% macroalbuminuric. A significant increase in protein clearances was observed in diabetic patients in comparison with normal subjects: the median of albumin clearance was 0.09 ml/min, interquartile range (IR) 0.04-0.31 (P < 0.01 vs normals); that of anionic immunoglobulins (IgG4) 0.02 ml/min, IR 0.04-0.05 (P < 0.005 vs normals); and that of neutral/cationic immunoglobulins (IgG) 0.13 ml/min, IR 0.07-0.19 (P < 0.01 vs normals). The anionic/cationic immunoglobulin ratio median was 0.22, IR 0.11-0.43, and exceeded the upper limit of normal values in 29.4% of all patients. IgG4 clearance was positively correlated with albumin clearance (r = 0.72) and with IgG clearance (r = 0.98). Nevertheless anionic immunoglobulin clearance was increased in a number of patients (17.3%) with normal IgG excretion and even in patients (15.1%) with normal albumin clearance. Clearances of IgG4 and IgG, but not that of albumin, were correlated with the duration of diabetes. Thus, an increased anionic/cationic IgG ratio in type 2 diabetes highlights a charge selectivity defect in protein permselectivity; this selective proteinuria may reflect more accurately than does microalbuminuria an early kidney abnormality in this form of diabetes.

The cholinergic innervation of human pancreatic islets.

The cholinergic innervation of pancreatic islets was investigated in the human using operatory samples. In order to analyze the nature of stained cholinergic nerve fibers some specimens were incubated in a solution containing 6-hydroxydopamine (6-HDA) to obtain a selective degeneration of adrenergic nerves. Cholinergic nerve fibers are present in human pancreatic islets, and appear to be organized in an external peri-insular plexus. Some nerve fibers from the peri-insular plexus enter the islets and seem to innervate directly various types of endocrine insular elements. The 6-HDA treatment does not alter the distribution pattern of cholinergic nerve fibers within pancreatic islets.

Effect of long-term treatment with acetyl-L-carnitine on structural changes of ageing rat brain.

The effects of long-term treatment (11 months) with acetyl-L-carnitine (75 mg/kg daily) on the morphology of brain and optic nerve was studied in 16 senescent (22-month-old) Wistar rats (nine untreated, seven treated). Five young rats (aged 3 months) were used for comparison. Senescence was found to cause a structural disorganization of cerebral cortex, hippocampus and cerebellar cortex, and a decrease in the volume densities of the pyramidal neurons of layers 2 and 5 of the prefrontal cortex. An impaired myelination of the pyramidal tract and of the optic nerve was also observed. Besides improving the structural organization of the cerebral areas under study, treatment with acetyl-L-carnitine increased the volume densities of pyramidal neurons of the prefrontal cortex layers under observation. It must be added that myelination of the pyramidal tract and optic nerve was found to be less impaired after acetyl-L-carnitine administration.

Enhanced nonenzymatic glycation of eye lens proteins in experimental diabetes mellitus: an approach for the study of protein alterations as mediators of normal aging phenomena.

The levels of advanced nonenzymatic glycation endproducts (ACE) were investigated by spectrofluorimetry in eye lens proteins obtained from rats with experimental diabetes of 3 and 6 months duration and from normal age-matched control rats. Diabetic animals showed higher AGE levels at both times studied. However the older control animals showed protein ACE levels comparable to those of the experimental 3 months diabetic group. These data suggest that a pathological phenomenon such as enhanced nonenzymatic glycation, associated to diabetic hyperglycemia, can be considered as a process leading to an accelerated aging of proteins. Thus experimental diabetes mellitus may be used as a model to investigate physiological protein senescence.