Anti-trypanosomal Lignans Isolated from Salvadoran Peperomia pseudopereskiifolia.

A search for anti-trypanosomal natural compounds from plants collected in El Salvador, a country particularly endemic for Chagas disease, resulted in the isolation of five lignan-type compounds (1-5) from Peperomia pseudopereskiifolia. The lignan derivatives 1, 2, and 4 are new. Their absolute configuration was determined by chemical derivatization. Compounds 1, 5, 6, and 8 exhibited anti-trypanosomal activity against the amastigote form of T. cruzi comparable to that of the existing drug benznidazole.

In Vitro Susceptibility of Kinetoplastids to Celastroloids from Maytenus chiapensis.

Leishmaniasis and Chagas are among the most significant neglected tropical diseases. Due to several drawbacks with the current chemotherapy, developing new antikinetoplastid drugs has become an urgent issue. In the present work, a bioassay-guided investigation of the root bark of Maytenus chiapensis on Leishmania amazonensis and Trypanosoma cruzi led to the identification of two D:A-friedo-nor-oleanane triterpenoids (celastroloids), 20ß-hydroxy-tingenone (celastroloid 5) and 3-O-methyl-6-oxo-tingenol (celastroloid 8), as promising antikinetoplastid leads. They displayed higher potency on L. amazonensis promastigotes (50% inhibitory concentrations [IC50s], 0.44 and 1.12 µM, respectively), intracellular amastigotes (IC50s, 0.83 and 1.91 µM, respectively), and T. cruzi epimastigote stage (IC50s, 2.61 and 3.41 µM, respectively) than reference drugs miltefosine and benznidazole. This potency was coupled with an excellent selectivity index on murine macrophages. Mechanism of action studies, including mitochondrial membrane potential (Δψm) and ATP-level analysis, revealed that celastroloids could induce apoptotic cell death in L. amazonensis triggered by the mitochondria. In addition, the structure-activity relationship is discussed. These findings strongly underline the potential of celastroloids as lead compounds to develop novel antikinetoplastid drugs.

Salvadoran Celastraceae Species as a Source of Antikinetoplastid Quinonemethide Triterpenoids.

Chagas disease and leishmaniasis are among the most widespread neglected tropical diseases, and their current therapies have limited efficacy and several toxic side effects. The present study reports the chemical and antikinetoplastid profiles of extracts from five Salvadoran Celastraceae species against the Trypanosoma cruzi epimastigotes stage and Leishmania amazonensis and Leishmania donovani promastigote forms. The phytochemical profile evinced the presence of flavonoids, tannins, sterols, and triterpenes as the main components in all plant species, whereas quinonemethide triterpenoids (QMTs) were restricted to the root bark of the studied species. Antikinetoplastid evaluation highlights the root bark extracts from Zinowewia integerrima, Maytenus segoviarum, and Quetzalia ilicina as the most promising ones, exhibiting higher potency against T. cruzi (IC50 0.71-1.58 µg/mL) and L. amazonensis (IC50 0.38-2.05 µg/mL) than the reference drugs, benznidazole (IC50 1.81 µg/mL) and miltefosine (IC50 2.64 µg/mL), respectively. This potent activity was connected with an excellent selectivity index on the murine macrophage J774A.1 cell line. These findings reinforce the potential of QMTs as antikinetoplastid agents for the development of innovative phytopharmaceuticals and the plant species under study as a source of these promising lead compounds.

Multistage Detection of Tetrodotoxin Traces in Diodon hystrix Collected in El Salvador.

This study describes a multistage methodology to detect minute amounts of tetrodotoxin in fishes, a plan that may be broadened to include other marine organisms. This methodology was applied to porcupinefish (Diodon hystrix) collected in Punta Chiquirín, El Salvador. A three-stage approach along with post-acquisition processing was employed, to wit: (a) Sample screening by selected reaction monitoring (HPLC-MS/MS-SRM) analyses to quickly identify possible toxin presence via a LC/MS/MS API 3200 system with a triple quadrupole; (b) HPLC-HRFTMS-full scan analyses using an ion trap-Orbitrap spectrometer combined with an MZmine 2-enhanced dereplication-like workflow to collect high-resolution mass spectra; and (c) HPLC-HRMS2 analyses. This is the first time tetrodotoxin has been reported in D. hystrix specimens collected in El Salvador.

Anti-trypanosomal screening of Salvadoran flora.

Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and in Central America, it is considered one of the four most infectious diseases. This study aimed to screen the anti-trypanosomal activity of plant species from Salvadoran flora. Plants were selected through literature search for plants ethnobotanically used for antiparasitic and Chagas disease symptomatology, and reported in Museo de Historia Natural de El Salvador (MUHNES) database. T. cruzi was incubated for 72 h with 2 different concentrations of methanolic extracts of 38 species, among which four species, Piper jacquemontianum, Piper lacunosum, Trichilia havanensis, and Peperomia pseudopereskiifolia, showed the activity (≤ 52.0% viability) at 100 µg/mL. Separation of the methanolic extract of aerial parts from Piper jacquemontianum afforded a new flavanone (4) and four known compounds, 2,2-dimethyl-6-carboxymethoxychroman-4-one (1), 2,2-dimethyl-6-carboxychroman-4-one (2), cardamomin (3), and pinocembrin (5), among which cardamomin exhibited the highest anti-trypanosomal activity (IC50 = 66 µM). Detailed analyses of the spectral data revealed that the new compound 4, named as jaqueflavanone A, was a derivative of pinocembrin having a prenylated benzoate moiety at the 8-position of the A ring.

Bone health and hormonal contraception.

INTRODUCTION: Short-term and long-term steroid contraceptive systems are widely employed in adolescents and premenopausal women; they could induce variation in bone metabolism, but whether these changes increase the overall fracture risk is not yet clear. EVIDENCE ACQUISITION: A systematic search of scientific publications about "hormonal contraceptives" and "bone metabolism" in reproductive age women was conducted. EVIDENCE SYNTHESIS: In adolescent girl, combined oral contraceptives could have a deleterious effect on bone health when their onset is within three years after menarche and when they contain ethinyl estradiol at the dose of 20 mcg. In perimenopausal women, steroid contraceptives seem not influence bone health nor increase osteoporotic fractures risk in menopause. The oral progestogens intake is not related to negative effects on skeletal health. Depot medroxyprogesterone acetate (DMPA) induce a prolonged hypoestrogenism with secondary detrimental effect on healthy bone; the higher bone loss was observed at the DMPA dose of 150 mg intramuscular such as after long-term DMPA-users. Progestin-based implants and intrauterine devices have not negative effect on bone health. CONCLUSIONS: Since sex-steroid drugs induce variations in hormonal circulating concentrations, they may negatively affect bone metabolism. Contraceptive choice should be tailored evaluating any possible effect on bone health. Clinicians should always perform a precontraceptive counselling to identify any coexisting condition that may affect bone health. Further randomized studies are needed to confirm these results.

The role of cholesterol recognition (CARC/CRAC) mirror codes in the allosterism of the human organic cation transporter 2 (OCT2, SLC22A2).

The human organic cation transporter 2 (OCT2) is a multispecific transporter with cholesterol-dependent allosteric features. The present work elucidates the role of evolutionarily conserved cholesterol recognition/interaction amino acid consensus sequences (CRAC and CARC) in the allosteric binding to 1-methyl-4-phenylpyridinium (MPP+) in human embryonic kidney 293 cells stably or transiently expressing OCT2. Molecular blind simulations docked two mirroring cholesterol molecules in the 5th putative transmembrane domain, where a CARC and a CRAC sequence lie. The impact of the conserved amino acids that may constitute the CARC/CRAC mirror code was studied by alanine-scanning mutagenesis. At a saturating extracellular concentration of substrate, at which the impact of cholesterol depletion is maximal, five mutants transported MPP+ at a significantly lower rate than the wild-type OCT2 (WT), resembling the behavior of the WT upon cholesterol depletion. MPP+ influx rate as a function of the extracellular concentration of substrate was measured for the mutants R234A, R235A, L252A and R263A. R234A kinetic behavior was similar to that of the WT, whereas R235A, L252A and R263A activity shifted from allosteric to one-binding site kinetics, very much like the WT upon cholesterol depletion. The impact of cholesterol on protein thermal stability was assessed for WT, R234A and R263A. While the thermal stability of WT and R234A was improved by the supplementation with cholesterol, R263A was not sensitive to the presence of cholesterol. To conclude, the disruption of the CARC/CRAC mirror code in the 5th putative transmembrane domain is sufficient to abolish the allosteric interaction between OCT2 and MPP+.

[Diabetic foot and risk factors]. / Factores de riesgo y pie diabético.

OBJECTIVE: To identify clinic and metabolic risk factors for diabetic neuropathy and peripheral arterial disease in patients evaluated in a diabetic foot unit care. METHOD: From 2000 to 2005 we evaluated the presence of diabetic neuropathy (monofilament, tuning fork and Boulton's clinic scale) and peripheral arterial disease (ankle-brachial index and toe-brachial index) in 304 diabetic patients. We classified patients in four groups: patients without pathology (normal group), with neuropathy (neuropathic group), with peripheral arterial disease (vascular group) and with both pathologies (mixed group) and we compared the characteristics of each group. We analysed other poblational characteristics: age, gender, type of diabetes, duration, microvascular and macrovascular complications, hypertension, smoking habit, antiagregation and mean HbA1c in the last year. RESULTS: Age, frequency of hypertension and coronary disease were significantly higher (p < 0.005) in vascular and mixed group than in normal group (63 +/- 13 and 65 +/- 10 vs. 55 +/- 14; 69.2 and 70.3 vs. 45.5%; 46,2% and 39.2% vs 23.8%, respectively). Frequency of retinopathy, nephropathy and HbA1c were significantly higher (p < 0.05) in neuropathic and mix group than in normal group (62.5 and 66.2 vs. 32.7%; 45.3 and 47.3 vs. 24.8%; 8.1 +/- 1.6 and 8.0 +/- 1.3 vs 7.4 +/- 1.2 respectively). CONCLUSION: This study indicates that the development of diabetic neuropathy is related with worse metabolic control and the presence of other microvascular complications; while age, hypertension and coronary disease are risk factors for peripheral arterial disease.

[Neurally-mediated syncope and occupational accidents: prevention strategies and case report]. / Sincope neuromediata e infortuni sul lavoro: strategie preventive e considerazioni su un caso clinico.

A bus driver came to our observation after an occupational traffic accident due to a syncopal event. The positive result of the tilt testing demonstrated the neurally-mediated nature of the syncope. The accident involved approximately 40 people (all the bus passengers), fortunately without severe injuries or deaths. The described episode indicates the need for a procedural algorithm, commonly approved, applicable in the field of prevention, for those occupational categories with severe accident risk. Indeed, the possibility exists to identify at least a part of the subjects predisposed to neurally-mediated syncope. Fundamental steps for such screening are history taking (looking for previous events, familiarity), the physical examination (useful, for example, to exclude orthostatic hypotension or carotid sinus syncope), and, in particular, the tilt testing, a diagnostic investigation recommended for all the workers who have had a previous syncope and are at high occupational accident risk. Moreover, the reported case recalls the need to strengthen the collaboration between the cardiologist and the occupational health physician.

[A questionnaire to assess school social climate]. / Un cuestionario para evaluar el clima social del centro escolar.

A questionnaire to assess the school social climate (CECSCE) made from items of The California School Climate and Safety Survey is presented. The CECSCE displays a stable factorial Oblimin structure in two social climate factors: 1) relative to the school and 2) relative to the teaching staff. Both factors present a correlation that oscillates between 0.42 and 0.48; and they explain 54,2% and 45,6% of the variance, respectively. Its reliability test-retest is acceptable (r= 0.61) after 9 months. Both factors present gender differences in favour of girls. They also differentiate at the educational level, displaying in 2 masculine course higher means in both factors than in 3 masculine course. The second factor correlates positively with the factors that measure social competence in the Merrell School Social Behaviour Scales and negatively with those that measure antisocial and non adapted behaviour.

[Multicenter study on the prevalence of hyperglycemia among hospitalized patients with parenteral nutrition]. / Estudio multicéntrico de prevalencia de hiperglucemia en pacientes hospitalizados con nutrición parenteral.

OBJECTIVE: The aim of this cross-sectional national multicentric study was to determine the prevalence of hyperglycemia in patients with parenteral nutrition and to assess other clinical factors associated with this complication. METHOD: All Spanish hospital pharmacy services were invited to participate in the study. RESULTS: Twenty eight (28) pharmacy services agreed to participate. The study included 442 patients. The prevalence of hyperglycemia (plasma levels > 200 mg/dL) was 26.7%. Eighty four point two per cent of the patients received less than 3.5 mg/kg/minute of glucose, this infusion rate being considered as the safe threshold. In most patients, follow-up of glycemia was based on capillary blood determination with reactive strips and in 27.6% of the cases in which insulin was prescribed, it was added to the parenteral nutrition bag, in full or in part. No significant correlations were found between glycemia and the clinical factors studied (disorders, fever, medication), except for insulin. CONCLUSIONS: This national multicentric study of the prevalence of hyperglycemia among patients with parenteral nutrition, leaded by hospital pharmacists, was a joint effort aimed to better understand this metabolic complication. Findings are consistent with those reported by other authors and have allowed us to describe the current situation.

Dihydro-ß-agarofuran sesquiterpenes from celastraceae species as anti-tumour-promoting agents: Structure-activity relationship.

Inhibition of tumour promotion in multistage chemical carcinogenesis is considered a promising strategy for cancer chemoprevention. In an ongoing investigation of bioactive secondary metabolites from Celastraceae species, five new dihydro-ß-agarofuran sesquiterpenes (1-5), named Chiapens A-E, and seventeen known ones, were isolated from Maytenus chiapensis. Their structures were elucidated by extensive NMR spectroscopic and mass spectrometric techniques, and their absolute configurations were determined by circular dichroism studies, chemical correlations and biogenic means. The isolated compounds, along with twenty known sesquiterpenes, previously isolated from Zinowiewia costaricensis, have been tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorpol-13-acetate (TPA). Thirty three compounds from this series showed stronger effects than that of ß-carotene, the reference inhibitor. The structure-activity relationship (SAR) analysis revealed that the type of substituent, in particular at the C-1 position of the sesquiterpene scaffold, was able to modulate the anti-tumour promoting activity. Compounds 3, 6, and 33 showed significant effects in an in vivo two-stage mouse-skin carcinogenesis model.

In-bag manual versus uncontained power morcellation for laparoscopic myomectomy: randomized controlled trial.

OBJECTIVE: To evaluate whether manualin-bag morcellation could be efficiently proposed as alternative to the uncontained power technique. DESIGN: Randomized controlled trial. SETTING: Academic hospital. PATIENT(S): One hundred fifty-two premenopausal women eligible for myomectomy were screened, and 104 were randomized. INTERVENTION(S): Patients were randomized into two groups. In the experimental group, "in-bag" protected morcellation was performed. In the control group, patients were treated by uncontained power myoma removal. MAIN OUTCOME MEASURE(S): The primary endpoint was the comparison of morcellation operative time (MOT). The secondary endpoints were the comparisons of total operative time (TOT), simplicity of morcellation (as defined by the surgeon using a visual analogue scale scale), intraoperative blood loss, rate of complications, and postoperative outcomes. RESULT(S): A sample size of 51 per group (n = 102) was planned. Between March 2014 and January 2015, patients were randomized as follows: 53 to the experimental group and 51 to the control group. Most demographic characteristics were similar across groups. MOT was observed to be similar in both study groups (16.18 ± 8.1 vs. 14.35 ± 7.8 minutes, in the experimental and control groups, respectively). Fibroid size was identified as the principal factor influencing morcellation time (Pearson coefficient 0.484 vs. 0.581, in the experimental and control groups, respectively). No significant difference in TOT, simplicity of morcellation, delta Hb, postoperative pain, and postoperative outcomes were observed between groups. CONCLUSION(S): The protected manual in-bag morcellation technique represents a time-efficient and feasible alternative, which does not interfere with surgical outcomes in women undergoing laparoscopic myomectomy. CLINICAL TRIAL REGISTRATION: NCT02086435.

[Benign thyroid nodules in the elderly. Medical treatment]. / I noduli tiroidei benigni nell'anziano. Trattamento medico.

BACKGROUND AND AIM: Benign thyroid nodules are a frequent pathology in the general population and in particular in the aged, but their treatment is still controversial. The percentage by which volume and/or the number of nodules is reduced using L-thyroxine treatment reported in the literature varies widely: from 9 to 68%. The aim of this study was to study the therapeutic efficacy of thyroxine treatment in the aged. METHODS: A retrospective study was performed in 105 elderly out-patients (mean age 68 +/- 6 years) suffering from thyroid nodules forming part of a population of 7452 thyroid patients from central Italy attending the 2nd Medical Clinic at "La Sapienza" University of Rome from 1986 to 1994. Other diseases which might influence treatment were excluded in all subjects. Thyroid hormones, TSH and antithyroid antibodies were assayed using RIA techniques; patients also underwent ultrasonography of the gland, 131I thyroid captation with scintigraphy and cytological analysis using fine needle aspiration. Patients were treated with L-thyroxine (75-150 micrograms/day) and controlled after 6, 12 and 24 months. No collateral effects were reported. RESULTS: The majority of patients (some 85%) remained stationary; just under 10% improved and only a few showed a deterioration of symptoms. CONCLUSIONS: Medical therapy continues, in the authors' opinion, to be the first choice in the treatment of benign thyroid nodules also in elderly patients owing to its easy management, low cost and scarce collateral effects.

[Diabetic ketoacidosis in patients with soft-tissue necrotizing infections]. / Cetoacidosis diabética en pacientes con infecciones necrosantes de tejidos blandos.

Necrotizing soft tissue infection is an uncommon and severe infection of the skin, subcutaneous tissue and superficial fascia which is usually accompanied by severe systemic toxicity. These infections occur more frequently in diabetics and are associated with higher morbidity and mortality rate. The prognosis of necrotizing fasciitis is known to be dependent on early recognition and treatment. Therefore, clinical awareness is important to avoid fatal outcome in patients with diabetes mellffus. We present three patients with undiagnosed type 2 diabetes in whom hyperglycaemia may have facilffated me development of necrotizing tissue infection, which in tum may have precipitated diabetic ketoacidosis in patients who rarely develop this metabolic complication.

[Metabolic syndrome and insulin resistance: prevalence in women with prior gestational diabetes, using two different classifications]. / Síndrome metabólico y resistencia insulínica: prevalencia según distintas clasificaciones en mujeres con diabetes gestacional previa.

OBJECTIVE: To determine the metabolic syndrome prevalence using the ATPIII and the IDF definitions and that of each criterion and to study the differences. To relate the presence of insulin resistance with metabolic syndrome diagnostic. MATERIALS AND METHODS: A total of 1,080 women diagnosed of gestational diabetes who came to follow up were studied. Prevalence of the metabolic syndrome was determined using the ATPIII and the IDF classification and prevalence of each criterion independently. The HOMA insulin resistance index was determined in 575 patients. RESULTS: Metabolic syndrome prevalence was 9.3% according to the ATPIII classification and 17.4% according to the IDF, this coinciding in 94 cases. Only the IDF classification criteria were fulfilled in 94 cases due to abdominal obesity in 13.8%, to fasting glucose in 17% and to both criteria in 69.8%. Six cases only met the ATPIII classification due to absence of abdominal obesity. The HOMA index value was 4.9 (+/- 3.5) and 4.0 (+/- 3) for women diagnosed of metabolic syndrome using the ATPIII or the IDF classification, respectively. Insulin resistance prevalence was 13.7%. The Kappa index of agreement was 0.411 between metabolic syndrome diagnostic by ATPIII and insulin resistance and 0.352 for IDF. CONCLUSIONS: Metabolic syndrome prevalence is almost double when the IDF classification is used than with the ATPIII due to the more strict value of fasting glucose and abdominal obesity. The ATPIII classification better identifies insulin resistance presence than that of IDF.

Toxicology and kinetics of long-term intraventricular infusion of phenytoin and valproic acid in pigs: experimental study.

The effect of continuous intraventricular infusion of phenytoin and valproic acid into the brain of pigs was studied through quantitative measurement of animal behavior, pathological study of animal's brain and measurement of the levels of these drugs in the blood and C.S.F. Two groups of five animals each were treated with increasingly doses of the drugs until the apparition of toxic effects and the dead of animals. Normal behavior was observed with doses up to 3 mg/day of phenytoin and 1.5 mg/day of valproic acid. Toxic effects consisted on severe unsteadiness and muscular rigidity. Pathological study of the brains revealed that there were no damage attributable to the intraventricular infusion of the drugs. The present study suggests that intrathecal or intraventricular infusion of phenytoin and valproic acid could be well tolerated by humans and it leads us to consider subsequent clinical studies in epileptic patients.

[Primary empty sella turcica: clinical, radiological and hormonal evaluation]. / Silla turca vacía primaria: evaluación clínica, radiológica y hormonal.

A retrospective study of 33 patients diagnosed of primary empty sella turcica has been done. Main clinical feature was cephalea, which was present in 51.1% of cases. Diagnosis was done based in Neuroradiologic studies (CAT, Cisternography, Pneumocisternography, Nuclear Magnetic Resonance) finding sellar size being significantly smaller in cases studied after 1980. After Hypophyseal function study, 19 patients did not show any endocrinological disorders, in the other 14 (42.4%), hyperprolactinemia was the most frequent finding (7 patients, 23.3%). 17.8% of cases showed a lowered response of Growth Hormone to insulinic Hypoglycemia. In two cases Panhypopituitarism was found and in case Insipidus diabetes was diagnosed.

[The metabolic control of insulin-dependent diabetes mellitus and diabetic nephropathy]. / Control metabólico de la diabetes mellitus insulinodependiente y nefropatía diabética.

Long-term complications of Insulin Dependent Diabetes Mellitus (IDDM) have been associated with several risk factors, particularly the degree of metabolic control and evolution time of the disease. A study was conducted with 219 randomly selected patients with IDDM at our clinic; evolutive, clinical and analytical parameters were assessed and conventional or multiple insulin therapy was evaluated. The classification of glycated hemoglobin (Hb A1c) in quartiles demonstrated a relatively higher incidence of diabetic retinopathy and nephropathy in the upper quartiles versus the lower quartile (p < 0.05). Likewise, patients with multiple insulin therapy had lower retinopathy (24.5% vs. 50.6, p < 0.001) and nephropathy rates (12.9% vs. 26.6%, p < 0.05) compared with those following a conventional insulin therapy. The multivariate analysis showed a statistically significant regression model (p < 0.001) for microalbuminuria level in patients with no established nephropathy; in these patients, the evolution time of IDDM and their Hb A1c level showed a positive independent association, and the use of multiple insulin therapy was a protective factor. The regression analysis of microalbuminuria levels compared with glycated hemoglobin in patients with no established nephropathy showed a value for Hb A1c of 9% as a break-point; from this point upwards microalbuminuria levels increased more markedly. The multivariate analysis here presented can help identify the presence of microalbuminuria in the pathological range in patients with IDDM followed at a hospital clinic from feasible clinical variables (evolution time, glycated hemoglobin level, program of insulin therapy used) establishing a metabolic objective which helps prevent the development of this complication.