Integration of Homeopathy and Complementary Medicine in the Tuscan Public Health System and the Experience of the Homeopathic Clinic of the Lucca Hospital.

INTRODUCTION: The healthcare programs of the Region of Tuscany (Italy) have started the process of integration of some types of complementary medicine (CM), including homeopathy, which began in 1996. The Homeopathic Clinic of Lucca was opened in 1998, followed by the Homeopathic Clinic for Women in 2003, and the Clinic for CM and Diet in Oncology in 2013. METHODS: Observational longitudinal studies conducted on 5,877 patients (3,937 in the general clinic, 1,606 in the women's clinic and 334 in oncology) were consecutively examined from 2003 to 2016. The Outcome in Relation to Impact on Daily Living (ORIDL) was generally used to assess outcomes. RESULTS: Comparing the clinical conditions before and after homeopathic treatment, improvement was observed in 88.8% of general medicine patients with follow-up (45.1%); in particular, 68.1% of the patients had a major improvement in or resolution (ORIDL +2, +3, +4) of their condition. In women, an improvement was obtained in 74.1% cases and a major improvement in 61.2%. In cancer patients with homeopathic and integrative treatment, a significant improvement was observed for all the symptoms during anti-cancer therapy, particularly for hot flashes, nausea, depression, asthenia, and anxiety. CONCLUSIONS: These results suggest that homeopathy can effectively be integrated with allopathic medicine and that the Tuscan experience could provide a useful reference for developing national and European regulations on the use of CM and homeopathy in public healthcare.

Global identification of genes targeted by DNMT3b for epigenetic silencing in lung cancer.

The maintenance cytosine DNA methyltransferase DNMT1 and de novo methyltransferase DNMT3b cooperate to establish aberrant DNA methylation and chromatin complexes to repress gene transcription during cancer development. The expression of DNMT3b was constitutively increased 5-20-fold in hTERT/CDK4-immortalized human bronchial epithelial cells (HBECs) before treatment with low doses of tobacco carcinogens. Overexpression of DNMT3b increased and accelerated carcinogen-induced transformation. Genome-wide profiling of transformed HBECs identified 143 DNMT3b-target genes, many of which were transcriptionally regulated by the polycomb repressive complex 2 (PRC2) complex and silenced through aberrant methylation in non-small-cell lung cancer cell lines. Two genes studied in detail, MAL and OLIG2, were silenced during transformation, initially through enrichment for H3K27me3 and H3K9me2, commonly methylated in lung cancer, and exert tumor suppressor effects in vivo through modulating cancer-related pathways. Re-expression of MAL and OLIG2 to physiological levels dramatically reduced the growth of lung tumor xenografts. Our results identify a key role for DNMT3b in the earliest stages of initiation and provide a comprehensive catalog of genes targeted for silencing by this methyltransferase in non-small-cell lung cancer.

Aerosolised 5-azacytidine suppresses tumour growth and reprogrammes the epigenome in an orthotopic lung cancer model.

BACKGROUND: Epigenetic silencing by promoter methylation and chromatin remodelling affects hundreds of genes and is a causal event for lung cancer. Treatment of patients with low doses of the demethylating agent 5-azacytidine in combination with the histone deacetylase inhibitor entinostat has yielded clinical responses. The subcutaneous dosing route for consecutive days and reduced bioavailability of 5-azacytidine because of inactivation by cytidine deaminase may limit the expansion of epigenetic therapy into Phase III trials. To mitigate these barriers, an aerosol of 5-azacytidine was generated and characterised. METHODS: The effect of aerosol vs systemic delivery of 5-azacytidine on tumour burden and molecular response of engrafted lung tumours in the nude rat was compared. RESULTS: Pharmacokinetics revealed major improvement in the half-life of 5-azacytidine in lung tissue with aerosol delivery. Aerosolised 5-azacytidine significantly reduced lung tumour burden and induced global demethylation of the epigenome at one-third of the comparable effective systemic dose. High commonality for demethylation of genes was seen in tumours sampled throughout lung lobes and across treated animals receiving the aerosolised drug. CONCLUSION: Collectively, these findings show that aerosolised 5-azacytidine targets the lung, effectively reprogrammes the epigenome of tumours, and is a promising approach to combine with other drugs for treating lung cancer.

Erupciones papulosas pruriginosas asociadas a la infección por HIV. Estudio sobre foliculitis eosinofílica y erupción papulosa y pruriginosa / Pruritic papular eruptions in HIV seropositive patients. A study of eosinophilic folliculitis and pruritic papular eruptions

La foliculitis eosinofílica y la erupción papulosa y pruriginosa son patologías que forman parte de las erupciones papulosas y pruriginosas asociadas a la infección por HIV. El objetivo de la presentación fue investigar estas entidades y su relación con el CD4, su correlación con el sexo, la edad y forma de presentación y distribución de las lesiones cutáneas

Solution structure of folic acid. Molecular mechanics and NMR investigation.

The structure of folic acid in solution was investigated by nuclear magnetic resonance (NMR) and theoretical calculations. Dynamical information and geometrical constraints were obtained by carbon-13 relaxation study, homo-nuclear NOESY spectra and hetero-nuclear 1H-13C NOE experiments. This set of experimental data was used for the molecular mechanics and molecular dynamic calculations. The accuracy of the final structure was established by the R(NMR) factor, which was calculated comparing the experimental NOESY cross-peaks intensities and the corresponding values simulated by using the complete relaxation matrix analysis (CORMA) approach.

Dissolution of calcium pyrophosphate crystals by polyphosphates: an in vitro and ex vivo study.

OBJECTIVE: To determine the dissolving ability (DA) of linear pentasodium tripolyphosphate (PSTP), cyclic trisodium metaphosphate (TSMP), polymeric sodium metaphosphate (SMP) on synthetic crystals of calcium pyrophosphate dihydrate (CPPD) and on crystalline aggregates of menisci from patients with chondrocalcinosis (CC). METHODS: Synthetic CPPD crystals were mixed with phosphate buffered saline (PBS), which contained the different polyphosphates, for one hour at 37 degrees C. The calcified menisci were obtained from the knees of four female patients with CPPD disease who underwent total arthroscopic meniscectomy for degenerative meniscal lesions. Meniscal cryosections and fragments were incubated in SMP (15 mg/ml PBS) at 37 degrees C for one hour and 24 hours, respectively. Histological evaluation on meniscal samples after polyphosphate incubation was carried out by ordinary transmitted light microscopy and polarised light microscopy. The dissolution of CPPD crystals by polyphosphates was assessed by atomic absorption spectroscopy, which determined the amount of calcium liberated from synthetic crystals and meniscal fragments. Cytotoxicity of SMP was evaluated by tetrazolium salt assay and by an ultrastructural study on cultured chondrocytes. RESULTS: SMP and PSTP showed higher DA on CPPD crystals than TSMP. Analysis of the DA values at increasing concentrations of SMP showed that a concentration of 15 mg/ml completely dissolved 2.0 mg CPPD crystals. The solution of meniscal CPPD crystals showed a significant increase of calcium concentration after three hours and 24 hours of SMP incubation (p=0.0001; Kruskal-Wallis analysis of variance) compared with fragments incubated in PBS control solution. Macroscopic and microscopic evaluation of meniscal specimens showed a notable reduction of CPPD deposits. A 50% inhibitory dose on cultured chondrocytes was reached at the maximum concentration of SMP used in this work (15 mg/ml); ultrastructural analysis did not show morphological alterations in the treated cells. CONCLUSION: The results of this study indicate that linear polyphosphates are effective in dissolving both synthetic and ex vivo CPPD crystal aggregates. This suggests a potential therapeutic use for these molecules in the treatment of symptomatic CC.

Health care utilization in chronic obstructive pulmonary disease. A case-control study in a health maintenance organization.

BACKGROUND: Information about health care utilization and costs among patients with chronic obstructive pulmonary disease (COPD) is needed to improve care and for appropriate allocation of resources for patients with COPD (COPD patients or cases) in managed care organizations. METHODS: Analysis of all inpatient, outpatient, and pharmacy utilization of 1522 COPD patients continuously enrolled during 1997 in a 172,484-member health maintenance organization. Each COPD case was matched with 3 controls (n = 4566) by age (+/-5 years) and sex. Information on tobacco use and comorbidities was obtained by chart review of 200 patients from each group. RESULTS: Patients with COPD were 2.3 times more likely to be admitted to the hospital at least once during the year, and those admitted had longer average lengths of stay (4.7 vs 3.9 days; P<.001). Mean costs per case and control were $5093 vs $2026 for inpatient services, $5042 vs $3050 for outpatient services, and $1545 vs $739 for outpatient pharmacy services, respectively (P<.001 for all differences). Patients with COPD had a longer smoking history (49.5 vs 34.9 pack-years; P =.002) and a higher prevalence of smoking-related comorbid conditions and were more likely to use cigarettes during the study period (46.0% vs 13.5%; P<.001). CONCLUSIONS: Health care utilization among COPD patients is approximately twice that of age- and sex-matched controls, with much of the difference attributable to smoking-related diseases. In this health maintenance organization, inpatient costs were similar to and outpatient costs were much higher than national averages for COPD patients covered by Medicare.

Utilization in COPD: patient characteristics and diagnostic evaluation.

STUDY OBJECTIVES: Information on current practices of COPD diagnosis and treatment is needed to identify opportunities for improving care. This study describes the clinical characteristics and diagnostic evaluations of COPD patients in a health maintenance organization (HMO) and a university-affiliated county medical center (UMC). DESIGN: Cross-sectional survey performed in a 174,484-member regional HMO and in The University of New Mexico Hospitals and Clinics (UNMH). PATIENTS: Two hundred COPD patients from each system randomly selected from administrative databases based on discharge diagnoses. RESULTS: COPD patients in the UMC, compared to those in the HMO, were younger (mean age, 59.3 vs 66.9 years, respectively), were more likely to be using home oxygen (33% vs 20%, respectively), and had fewer chronic medical conditions (mean number of conditions, 3.1 vs 3.7, respectively) (p < 0.01 for all differences). Approximately half of the COPD patients in both groups continued to smoke cigarettes during the study year. Only 38% of patients in the HMO and 42% in the UNMH system had spirometry results documented in their medical records. CONCLUSIONS: The demographic and clinical characteristics of the COPD patients in these two health-care systems were very different, but smoking status and utilization of diagnostic tests were similar. The diagnosis of COPD in most patients was based only on a history of chronic respiratory symptoms and smoking; spirometry often was not used to confirm the diagnosis. An increased emphasis on smoking cessation and more effective utilization of spirometry are needed to improve the management of COPD in these health-care systems.

Asthma in a Vietnamese refugee population.

Asians and Pacific Islanders comprise a large and growing minority group in the United States, yet data on health status specific to these populations are scant. We conducted an epidemiologic study of asthma in a Vietnamese refugee population to estimate prevalence, evaluate risk factors, and better understand treatments of asthma among Vietnamese individuals. One hundred twenty-four asthma cases were identified from a population of 2,536 new Vietnamese refugees in San Diego (prevalence = 49 per 1,000; 4.9%). Two nonasthmatic control groups of Vietnamese refugees, matched for age and gender with the asthma cases, were recruited for a case-control study, using a questionnaire administered in Vietnamese. Vietnamese asthmatic individuals used both Western and non-Western therapies. Most subjects used traditional health practices, such as coining, cupping, and oil inhalation. As compared with current-refugee controls, the asthmatic subjects used significantly more bleeding (OR: 3.40; 95% CI: 1.06 to 10.80) and herbal ingestion (OR: 1.87; 95% CI: 1.08 to 3.19). As compared with former-refugee controls, the asthmatic subjects used significantly more oil inhalation (OR: 2.58; 95% CI: 1.45 to 4.85), bleeding (OR: 8.64, 95% CI: 1.02 to 73.70), and herbal ingestion (OR: 1.93; 95% CI: 1.02 to 3.67). The presentation and recognition of asthma among the Vietnamese subjects were similar to those in other populations. This information may be helpful in designing culture-specific health-education programs.

1H-NMR studies of postmortem biochemical changes in rat skeletal muscle.

1H-nuclear magnetic resonance (NMR) has been applied to the study of postmortem biochemical changes in perchloric acid extracts of rat skeletal muscle. Several metabolites have been detected and the dependence upon the postmortem time has been considered. The simultaneous quantitative determination of metabolites showing up at very low and very high fields has been suggested to yield a satisfactory delineation of the thanatochronology.

Spin label studies of erythrocytes during storage of blood.

The kinetic behavior of the spin label MAL-6 in the interaction with differently aged human erythrocyte membranes was evaluated by monitoring the rate of disappearance of the room temperature ESR signal due to the MAL-6 spin label added to blood after storage at 4 degrees C or after incubation of red cells at 37 degrees C in a protein-free medium. After 35 days of blood storage or 60 h of erythrocytes incubation at 37 degrees C the decrease of the intensity of the MAL-6 ESR spectra in respect to control samples is markedly enhanced and the correspondent kinetic constants significantly increase. Signal decay of MAL-6 is a further proof that during storage of blood under blood bank conditions or during an artificial ageing of erythrocytes at 37 degrees C, profound modifications occur in the human erythrocyte membrane.

Murine complement component C4 and sex-limited protein: identification of amino acid residues essential for C4 function.

Murine sex-limited protein (Slp) is an isotype of murine complement component C4 that shares 95% sequence identity with C4 as well as the intramolecular thioester necessary for C4 function but has no complement activity. Slp is nonfunctional at least in part because it is not cleaved by the activated form of complement protease C1s (C1s), which proteolytically activates C4 in the classical complement pathway. Slp is also distinct from C4 in that its expression in some mouse strains is under testosterone control. In the present studies, we used site-directed mutagenesis of C4 and expression of the mutant proteins in cultured cells to identify the amino acid substitutions in Slp that are responsible for resistance to C1s cleavage. We focused on sequence changes immediately downstream of the cleavage site in C4 because the arginine at that site is conserved in Slp, but the downstream sequences diverge substantially, with six differences in the first 7 residues followed by a 3-residue deletion in Slp. We found that a C4 mutant carrying only the 3-residue deletion is not cleaved by C1s and has essentially no hemolytic activity, whereas a mutant carrying only the six replacement changes is cleaved by C1s and has normal hemolytic activity. Both mutants have intact thioesters. A third mutant in which two acidic residues in the segment deleted in Slp were replaced by aliphatic residues is also cleaved by C1s, has an intact thioester group, and has normal hemolytic activity. These results indicate that the downstream mutations are responsible for the resistance of Slp to C1s cleavage and suggest that the length rather than the specific sequence of this segment is critical in determining susceptibility to the protease.

Transfected cDNA directs expression of hemolytically active murine C4 in cultured mouse and monkey cells.

Previously cloned and sequenced full-length cDNAs for murine C4 and the closely related sex-limited protein (Slp) have been placed into an eucaryotic expression vector. Transfer of these DNA constructs transiently into monkey COS cells or stably into mouse L cells results in the expression and secretion of hemolytically active mouse C4 and mature Slp. We estimate from hemolytic activities that COS and L cells secrete 0.04 and 3%, respectively, of the C4 level found in mouse plasma. Slp expression is consistently only 10-20% that of C4 although the identical expression system is used for both. Our results show subtle but reproducible cell-type-specific differences in C4 maturation; they also indicate that surprisingly large shifts in electrophoretic mobility on SDS-polyacrylamide gels are induced by a small number of amino acid substitutions. The expression of C4 from cDNAs of known sequence provides a starting point for studies of structure/function relationships in C4 employing site-specific mutagenesis and gene transfer.

Changes in erythrocyte properties during the first hours of life: electron spin resonance of reacting sulfydryl groups.

In an attempt to clarify the mechanism by which the red cells (RBC) of newborn infants are protected against oxidative agents, electron spin resonance (ESR) assays were carried out using the nitroxide radical 4-maleimide-2,2,6,6-tetramethylpiperidinyl-1-oxyl (Mal-6), a sulfydryl-reacting agent. The ESR assays were performed in 24 samples of cord blood, 20 samples of blood from 4-day-old infants, and eight samples of 8-h-old infants. The analyses were carried out on whole blood and washed erythrocytes were resuspended in buffered saline. The same experiments were performed in 10 blood samples from healthy adults as controls. Whole blood, before and after removing the buffy coat, and cell-free plasma were also examined by ESR assay. Cell-free plasma and buffy coats proved not to be appreciably involved in the Mal-6 behavior. The data of the ESR spectroscopy demonstrated a significantly slower reaction rate in the samples of cord blood and in blood of 8-h-old infants, compared to that of 4-day-old infants and adults. No significant differences in Mal-6 behavior could be detected between cord blood and 4-day-old infant blood in the results of ESR assays performed in washed red cells. Chemical determination of RBC-reacting sulfydryl groups and the assays of glutathione also demonstrated the absence of differences between cord blood and blood of 4-day-old infants. The results of our investigation suggest that the RBC-sulfydryl-reacting groups are less involved in the detoxification of oxidative agents during the first hours of life than in the following days.(ABSTRACT TRUNCATED AT 250 WORDS)

Structural and dynamical characterization of piroxicam by 1H- and 13C-NMR relaxation studies.

Carbon spin-lattice relaxation rates of an anti-inflammatory drug, piroxicam, have been measured. These results have been used in determining the reorientational rates of the proton carbon vectors. An analysis of internal motions within the pyridinyl moiety of piroxicam was carried out. Selective proton-carbon nuclear Overhauser effect (NOE) measurements were made in order to determine the solution structure of piroxicam. The effect of indirect NOE arising from exchangeable protons has been analyzed and considered.