Local control of nuclear calcium signaling in cardiac myocytes by perinuclear microdomains of sarcolemmal insulin-like growth factor 1 receptors.

RATIONALE: The ability of a cell to independently regulate nuclear and cytosolic Ca(2+) signaling is currently attributed to the differential distribution of inositol 1,4,5-trisphosphate receptor channel isoforms in the nucleoplasmic versus the endoplasmic reticulum. In cardiac myocytes, T-tubules confer the necessary compartmentation of Ca(2+) signals, which allows sarcomere contraction in response to plasma membrane depolarization, but whether there is a similar structure tunneling extracellular stimulation to control nuclear Ca(2+) signals locally has not been explored. OBJECTIVE: To study the role of perinuclear sarcolemma in selective nuclear Ca(2+) signaling. METHODS AND RESULTS: We report here that insulin-like growth factor 1 triggers a fast and independent nuclear Ca(2+) signal in neonatal rat cardiac myocytes, human embryonic cardiac myocytes, and adult rat cardiac myocytes. This fast and localized response is achieved by activation of insulin-like growth factor 1 receptor signaling complexes present in perinuclear invaginations of the plasma membrane. The perinuclear insulin-like growth factor 1 receptor pool connects extracellular stimulation to local activation of nuclear Ca(2+) signaling and transcriptional upregulation through the perinuclear hydrolysis of phosphatidylinositol 4,5-biphosphate inositol 1,4,5-trisphosphate production, nuclear Ca(2+) release, and activation of the transcription factor myocyte-enhancing factor 2C. Genetically engineered Ca(2+) buffers--parvalbumin--with cytosolic or nuclear localization demonstrated that the nuclear Ca(2+) handling system is physically and functionally segregated from the cytosolic Ca(2+) signaling machinery. CONCLUSIONS: These data reveal the existence of an inositol 1,4,5-trisphosphate-dependent nuclear Ca(2+) toolkit located in direct apposition to the cell surface, which allows the local control of rapid and independent activation of nuclear Ca(2+) signaling in response to an extracellular ligand.

Role of heterotrimeric G protein and calcium in cardiomyocyte hypertrophy induced by IGF-1.

In the heart, insulin-like growth factor-1 (IGF-1) is a peptide with pro-hypertrophic and anti-apoptotic actions. The pro-hypertrophic properties of IGF-1 have been attributed to the extracellular regulated kinase (ERK) pathway. Recently, we reported that IGF-1 also increases intracellular Ca(2+) levels through a pertussis toxin (PTX)-sensitive G protein. Here we investigate whether this Ca(2+) signal is involved in IGF-1-induced cardiomyocyte hypertrophy. Our results show that the IGF-1-induced increase in Ca(2+) level is abolished by the IGF-1 receptor tyrosine kinase inhibitor AG538, PTX and the peptide inhibitor of Gßγ signaling, ßARKct. Increases in the activities of Ca(2+) -dependent enzymes calcineurin, calmodulin kinase II (CaMKII), and protein kinase Cα (PKCα) were observed at 5 min after IGF-1 exposure. AG538, PTX, ßARKct, and the dominant negative PKCα prevented the IGF-1-dependent phosphorylation of ERK1/2. Participation of calcineurin and CaMKII in ERK phosphorylation was discounted. IGF-1-induced cardiomyocyte hypertrophy, determined by cell size and ß-myosin heavy chain (ß-MHC), was prevented by AG538, PTX, ßARKct, dominant negative PKCα, and the MEK1/2 inhibitor PD98059. Inhibition of calcineurin with CAIN did not abolish IGF-1-induced cardiac hypertrophy. We conclude that IGF-1 induces hypertrophy in cultured cardiomyocytes by activation of the receptor tyrosine kinase activity/ßγ-subunits of a PTX-sensitive G protein/Ca(2+) /PKCα/ERK pathway without the participation of calcineurin.

Applying the Participatory Slow Design Approach to a mHealth Application for Family Caregivers in Pediatric Ear, Nose, and Throat Surgery.

Pediatric ear, nose, and throat (ENT) surgery is very common, and its outcomes may improve with family education. In this regard, mobile health (mHealth) applications (apps), which are on the rise due to digital transformation, can be beneficial in healthcare. This study outlines the user-centered design and development of a mHealth app (version 5.15.0) to support family caregivers during the perioperative process of pediatric ENT surgery. Conducted over two years in an Italian maternal and child health hospital (January 2020-May 2022), the study employed a participatory design method based on the Information System Research (ISR) framework and guided by the principles of Slow Medicine. Utilizing the Relevance, Rigor, and Design cycles of the ISR framework, the mHealth app's content, functionalities, and technical features were defined and developed. A committee of fifteen experts guided the process with input from 25 family caregivers and 24 healthcare providers enrolled in the study. The mHealth app content was structured around five crucial educational moments characterizing the ENT perioperative period, providing evidence-based information on surgical procedures, strategies for preparing children for hospitalization and surgery, pain management, and post-discharge care. The mHealth app featured a function that sends customized notifications to guide caregivers at specific perioperative stages. The development of mHealth apps by implementing a rigorous, participatory, and Slow design process can foster accessible and family-centered information and care in the field of maternal and child health and beyond.

Venipuncture activates the cerebral cortex in children with intellectual disability.

OBJECTIVE: To evaluate the pattern of cortical activation during a painful procedure, such as a venipuncture, in children with intellectual disability and compare it with that of cognitively healthy children. STUDY DESIGN AND SETTING: A cohort study was conducted and cortical activation was assessed by multichannel cerebral near-infrared spectroscopy to monitor variations in oxyhaemoglobin and deoxyhaemoglobin (Hbb) in children with and without intellectual disability during a venipuncture for blood sampling with topical anaesthesia. Pain and distress were assessed as well using different validated pain scales (visual analogue scale and Non-Communicating Children's Pain Checklist-Postoperative Version for children with intellectual disability), and compared between groups. PARTICIPANTS: 16 children with severe to profound intellectual disability and 20 cognitively healthy peers (age range: 4-17 years). RESULTS: When Hbb was analysed, children with intellectual disability exhibited a bilateral activation of the somatosensory (p<0.006) and right motor cortex (p=0.0045), whereas cognitively healthy peers never showed a cortical activation. Children with intellectual disability also showed more pain than controls (p=0.001). CONCLUSIONS: When subjected to a painful procedure, only children with intellectual disability show an activation of the cerebral cortex, even if topical anaesthesia is applied, and express more pain than cognitively healthy peers. The role of other issues in painful procedures, such as anxiety, fear or physical restraint, deserves further investigation.

Prevalence of SARS-CoV-2 infection in Italian pediatric population: a regional seroepidemiological study.

BACKGROUND: Data on the effective burden of the SARS-CoV-2 pandemic in pediatric population are very limited, mostly because of the higher rate of asymptomatic or paucisymptomatic cases among children. Updated data on COVID-19 prevalence are needed for their relevance in public health and for infection control policies. In this single-centre cross-sectional study we aimed to assess prevalence of SARS-CoV-2 infection through IgG antibodies detection in an Italian pediatric cohort. METHODS: The study was conducted in January 2021 among both inpatients and outpatients referring to Research Institute for Maternal and Child Health "Burlo Garofolo" in Trieste, Friuli Venezia-Giulia, Italy, who needed for blood test for any reason. Collected samples were sent to Italian National Institute of Health for analysis through chemiluminescent immunoassay (CLIA). RESULTS: One hundred sixty-nine patients were included in the study, with a median age of 10.5 ± 4.1 years, an equal distribution for sex (49.7% female patients), and a 55.6% prevalence of comorbidities. Prevalence of anti-SARS-CoV-2 trimeric Spike protein IgG antibodies was 9.5% (n = 16), with a medium titre of 482.3 ± 387.1 BAU/mL. Having an infected cohabitant strongly correlated with IgG positivity (OR 23.83, 95% CI 7.19-78.98, p < 0.0001), while a cohabitant healthcare worker wasn't associated with a higher risk (OR 1.53, 95% CI 0.4-5.86, p 0.46). All of the 5 patients who had previously tested positive to a nasopharyngeal swab belonged to the IgG positive group, with a 3-month interval from the infection at most. CONCLUSION: We assessed a 9.5% SARS-CoV-2 seroprevalence in a pediatric cohort from Friuli Venezia-Giulia region in January 2021, showing a substantial increase after the second peak of the pandemic occurred starting from October 2020, compared to 1% prevalence observed by National Institute of Statistics (ISTAT) in July 2020.

Expression of ECM proteins fibulin-1 and -2 in acute and chronic liver disease and in cultured rat liver cells.

Fibulin-2 has previously been considered as a marker to distinguish rat liver myofibroblasts from hepatic stellate cells. The function of other fibulins in acute or chronic liver damage has not yet been investigated. The aim of this study has been to evaluate the expression of fibulin-1 and -2 in models of rat liver injury and in human liver cirrhosis. Their cellular sources have also been investigated. In normal rat liver, fibulin-1 and -2 were both mainly present in the portal field. Fibulin-1-coding transcripts were detected in total RNA of normal rat liver, whereas fibulin-2 mRNA was only detected by sensitive, real-time quantitative polymerase chain reaction. In acute liver injury, the expression of fibulin-1 was significantly increased (17.23-fold after 48 h), whereas that of fibulin-2 was not modified. The expression of both fibulin-1 and -2 was increased in experimental rat liver cirrhosis (19.16- and 26.47-fold, respectively). At the cellular level, fibulin-1 was detectable in hepatocytes, "activated" hepatic stellate cells, and liver myofibroblasts (2.71-, 122.65-, and 469.48-fold over the expression in normal rat liver), whereas fibulin-2 was restricted to liver myofibroblasts and was regulated by transforming growth factor beta-1 (TGF-beta1) in 2-day-old hepatocyte cultures and in liver myofibroblasts. Thus, fibulin-1 and -2 respond differentially to single and repeated damaging noxae, and their expression is differently present in liver cells. Expression of the fibulin-2 gene is regulated by TGF-beta1 in liver myofibroblasts.

Xerostomia: prevention with Pycnogenol® supplementation: a pilot study.

BACKGROUND: The aim of this pilot study was the evaluation of primary, idiopathic mucosal dryness (xerostomia or dry mouth) in subjects without cancer. METHODS: A group of non-diabetic subjects and a group of diabetics were managed with standard management (SM) or with SM+Pycnogenol® (150 mg/day) for 2 weeks. RESULTS: In total, 48 subjects were included in the study; 24 diabetics and 24 non-diabetics. 12 diabetics and 12 non-diabetics took Pycnogenol® and 12 diabetics and 12 non-diabetics were followed up with standard management only and served as controls. No side effects and no tolerability problems were observed with Pycnogenol®. The registry groups were comparable for characteristics and symptoms at baseline. All otherwise healthy subjects had a BMI<26 kg/m2. In 2 weeks, salivary flow and oxidative stress improved significantly in both groups of subjects (non-diabetics and diabetics) with 150 mg/day Pycnogenol® (P<0.05), while minimal improvements in salivary flow were seen with SM. The subjective score and the number of mucosal breaks and ulcerations, all minimal (<1 mm in length or diameter), were significantly decreased with Pycnogenol® supplementation (P<0.05) with minimal variations in the SM controls. Finally, the mean lysozyme level in parotid saliva samples was significantly increased in the Pycnogenol® group (P<0.05) both in diabetics and non-diabetics. CONCLUSIONS: Based on these preliminary results, Pycnogenol® could be a new, valid option for the treatment of xerostomia.

A nationwide survey of hereditary angioedema due to C1 inhibitor deficiency in Italy.

INTRODUCTION: Hereditary angioedema due to C1-inhibitor deficiency (C1-INH-HAE type I) or dysfunction (C1-INH-HAE type II) is a rare disease characterized by recurrent episodes of edema with an estimated frequency of 1:50,000 in the global population without racial or gender differences. In this study we present the results of a nationwide survey of C1-INH-HAE patients referring to 17 Italian centers, the Italian network for C1-INH-HAE, ITACA. METHODS: Italian patients diagnosed with C1-INH-HAE from 1973 to 2013 were included in the study. Diagnosis of C1-INH-HAE was based on family and/or personal history of recurrent angioedema without urticaria and on antigenic and/or functional C1-INH deficiency. RESULTS: 983 patients (53% female) from 376 unrelated families were included in this survey. Since 1973, 63 (6%) patients diagnosed with C1-INH-HAE died and data from 3 patients were missing when analysis was performed. Accordingly, the minimum prevalence of HAE in Italy in 2013 is 920:59,394,000 inhabitants, equivalent to 1:64,935. Compared to the general population, patients are less represented in the early and late decades of life: men start reducing after the 5(th) decade and women after the 6(th). Median age of patients is 45 (IQ 28-57), median age at diagnosis is 26 years (IQ 13-41). C1-INH-HAE type 1 are 87%, with median age at diagnosis of 25 (13-40); type 2 are 13% with median age at diagnosis of 31 (IQ 16-49). Functional C1INH is ≤50% in 99% of patients. Antigen C1INH is ≤50% in 99% of type 1. C4 is ≤50% in 96% of patients. The chance of having C1-INH-HAE with C4 plasma levels >50% is < 0.05. CONCLUSION: This nationwide survey of C1-INH-HAE provides for Italy a prevalence of 1:64,935. C1-INH-HAE patients listed in our database have a shorter life expectancy than the general population. An increased awareness of the disease is needed to reduce this discrepancy. Measurement of C4 antigen can exclude diagnosis of C1-INH-HAE with an accuracy > 95%. This parameter should be therefore considered for initial screening in differential diagnosis of angioedema.

Prevalence and risk factors for latex-related diseases among healthcare workers in an Italian general hospital.

Latex allergy has become an occupational hazard among healthcare workers. Atopy and degree of exposure have been recognized as predisposing factors for latex sensitization. We investigated the prevalence of latex allergy and the potential risk factors for latex sensitization, by distributing a questionnaire to 284 employees of a general hospital in central Italy. We collected information about occupational history, including specific tasks performed; time of first exposure to latex gloves; number of pairs of gloves; and duration of daily exposure. We also investigated the interval between first exposure and onset of symptoms, as well as the exact circumstances of their appearance. We evaluated pre-existing rhinoconjunctivitis, asthma, atopic and contact dermatitis, and allergies to drugs and foods using prick and patch tests. Latex allergy was established by means of skin-prick test, specific IgE, patch-test, and latex-glove-wearing test. This survey documented a high prevalence of symptoms related to the use of latex (47%) among the hospital staff, demonstrable sensitization to latex was considerably lower (12%), though strongly associated to atopy and duration of occupational exposure. Despite non-specificity, validated questionnaires constitute the most useful means to implement health surveillance and prevention of latex-related diseases among healthcare workers.

Lowered tumor necrosis factor receptors, but not increased insulin sensitivity, with infliximab.

OBJECTIVES: To verify whether Infliximab could modify insulin sensitivity and TNF-alpha and GLUT4 mRNA expression in muscle and adipose tissue of morbidly obese subjects. Soluble TNF receptors I and II (TNFR-I and TNFR-II) were also assayed. RESEARCH METHODS AND PROCEDURES: Six obese subjects were investigated before and 2 weeks after a single intravenous administration of 5 mg/kg Infliximab; insulin sensitivity was evaluated by euglycemic hyperinsulinemic clamp, and TNF-alpha and GLUT4 mRNA expression were assessed by reverse-transcriptase polymerase chain reaction on muscle and adipose tissue. TNF-alpha, TNFR-I, and TNFR-II were determined using the ELISA technique. RESULTS: Infliximab infusion did not affect fasting plasma insulin or fasting plasma glucose levels; whole body glucose uptake did not change significantly. TNF-alpha and GLUT4 mRNA did not show any significant change in muscle or adipose tissue. Serum TNF-alpha was undetectable before and after treatment, whereas TNFR-I and TNFR-II concentrations significantly decreased (p < 0.01). DISCUSSION: An explanation for the absence of effect of Infliximab on insulin resistance in morbidly obese subjects may be the paracrine way of action of this cytokine. Because Infliximab is predominantly distributed within the vascular compartment, its effectiveness in penetrating muscle and adipose tissue is potentially low. The significant decrease of TNFR-I and TNFR-II might be ascribed to a targeted effect of Infliximab on the immune system.

[Genetic composition of the Chilean population. Analysis of mitochondrial DNA polymorphism]. / Composición genética de la población chilena. Distribución de polimorfismos de DNA mitocondrial en grupos originarios y en la población mixta de Santiago.

BACKGROUND: The analysis of mitochondrial DNA restriction site polymorphisms assigns most Latin American aborgines to four haplogroups. These are characterized by determined polymorphic restriction sites and a deletion of 9 base pairs in the intergenic region V. AIM: To study the distribution of mitochondrial DNA haplogroups in Chilean aboriginal groups, as well as in the mixed population of Santiago. MATERIAL AND METHODS: One hundred twenty Aymara subjects and 23 Atacameño subjects from the Northern part of Chile and 162 randomly chosen subjects residing in Santiago were studied. DNA was extracted from peripheral lymphocytes. Mitochondrial DNA was amplified by means of polymerase chain reaction. RESULTS: The frequency of haplogroup B decreases from north to south. Aymaras in the north have the highest frequency (64%) and it is absent among the Yamanas (previously studied) in the extreme South. Haplogroups C and D show an inverse tendency. It is noteworthy that 84% of mitochondrial haplogroups of the mixed population of Santiago are of Amerindian origin whereas the Y-chromosomes are mainly European. CONCLUSIONS: The peculiar distribution of haplotypes indicate that the population of Santiago is the result of an asymmetric mating system in which the females ancestors were mainly Amerindian and the male ancestors mainly European.

Reduced PDK4 expression associates with increased insulin sensitivity in postobese patients.

OBJECTIVE: The aim of this study was to verify whether changes in PDK4 mRNA expression in skeletal muscle in formerly obese subjects who underwent malabsorptive bariatric surgery [bilio-pancreatic diversion (BPD)] might be related to insulin sensitivity improvement, and if these possible modifications might correlate with a reduction of the intramyocytic lipid level. RESEARCH METHODS AND PROCEDURES: Six obese women (body mass index 46.6 +/- 8.2 kg/m(2)) were enrolled in the study. Body composition, euglycemic-hyperinsulinemic clamp and muscle biopsies for skeletal muscle lipid analysis, and semiquantitative reverse transcriptase-polymerase chain reaction were performed before and 3 years after BPD. RESULTS: The average weight loss observed after surgery was approximately 42%. Increased glucose uptake was accompanied by a significant decrease of PDK4 mRNA (R(2) = 0.71, p < 0.001). The amounts of intramyocytic triglycerides correlate directly with PDK4 mRNA (R(2) = 0.87, p = 0.005) and inversely with glucose uptake values (R(2) = 0.75, p < 0.001). DISCUSSION: Our results support the concept that a reduced tissue availability of fatty acids consequent to a massive lipid malabsorption influences glucose metabolism acting through the regulation of PDH complex. In fact, as shown in animals, a higher level of FFA availability is likely to induce overexpression of PDK4 also in humans.