Recent advances in mass spectrometry analysis of phenolic endocrine disruptors and related compounds.

This article reviews recent literature on current methodologies based on chromatography coupled to mass spectrometry to analyze phenolic compounds with endocrine-disrupting capabilities. For this review we chose alkylphenol ethoxylates, bisphenol A, bisphenol F, and their degradation products and halogenated derivatives, which are considered important environmental contaminants. Additionally, some related compounds such as bisphenol diglycidylethers were included. Growing attention has been paid to the mass spectrometric characterization of these compounds and the instrumentation and strategies used for their quantification and confirmation. The current use of gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) methodologies with different mass spectrometers and ionization and monitoring modes is discussed. Practical aspects with regards to the use of these analytical techniques, such as derivatizing reagents in GC-MS, ion suppression in LC-MS, and the most problematic aspects of quantification, are included in the discussion.

Preventing false negatives with high-resolution mass spectrometry: the benzophenone case.

Benzophenone (BP) is one of the many contaminants reported as present in foodstuffs due to its migration from food packaging materials. Liquid chromatography/tandem mass spectrometry (LC/MS/MS) is acknowledged in the literature as the method of choice for this analysis. However, cases have been reported where the use of this methodology was insufficient to unambiguously confirm the presence of a contaminant. In previous work performed by the authors, the unequivocal identification of BP in packaged foods was not possible even when monitoring two m/z transitions (precursor ion - product ion), since ion ratio errors higher than 20% were obtained. In order to overcome this analytical problem a fast, sensitive and selective liquid chromatography/high-resolution mass spectrometry (LC/HRMS) methodology has been developed and applied to the analysis of BP in packaged foods. A direct comparison between LC/HRMS and LC/MS/MS data indicated better selectivity when working with LC/HRMS at a resolving power of 50,000 FWHM (full width at half maximum) than when monitoring two m/z transitions by LC/MS/MS. The resolving power used enabled the detection and identification of Harman as the compound impeding the confirmation of BP by LC-MS/MS. Similar quantitative results were obtained by an Orbitrap mass analyser (Exactive™) and a triple quadrupole mass analyser (TSQ Quantum Ultra AM™).

Multiple-stage mass spectrometry analysis of bisphenol A diglycidyl ether, bisphenol F diglycidyl ether and their derivatives.

The fragmentation of bisphenol A diglycidyl ether (BADGE), bisphenol F diglycidyl ether (BFDGE) and their derivatives was studied by electrospray ionization tandem mass spectrometry. Multiple-stage mass spectrometry and accurate mass measurements were combined to establish the fragmentation pathways. BADGEs and BFDGEs tend to form ammonium adducts under electrospray conditions which fragmented easily. The fragmentation of [M+NH(4)](+) for BADGEs started with the cleavage of the phenyl-alkyl bond, which was followed by the α-cleavage of the ether group to generate the characteristic product ions at m/z 135, [C(9)H(11)O](+), and m/z 107, [C(7)H(7)O](+). The fragmentation of the BFDGE isomer mixtures was studied by on-line reversed-phase liquid chromatography coupled to multiple-stage mass spectrometry (LC/MS(n)). Information obtained from product ion spectra for each BFDGE isomer and its comparison with the fragmentation pathway of BADGE allowed each isomer and the chromatographic elution order to be identified.

Fotocoagulación láser guiada por ultrasonido para el tratamiento del secuestro broncopulmonar fetal. Reporte de dos casos exitosos en Venezuela / Ultrasound-guided laser photocoagulation for the treatment of fetal bronchopulmonary sequestration. Report of two successful cases in Venezuela

El secuestro broncopulmonar es una malformación del aparato respiratorio que consiste en tejido bronquial y pulmonar no funcionante, separado del árbol tráqueo-bronquial y alimentado por un vaso sanguíneo proveniente de la circulación sistémica. La incidencia es de 1 por cada 5000 nacimientos, comprende entre 0,15 % y 6,45 % de las patologías pulmonares. El pronóstico es, generalmente, favorable, reportándose regresión espontánea de la lesión en 50 % a 75 % de los pacientes. Puede ocasionar efecto de masa, comprimiendo el corazón y el pulmón hasta generar cambios hemodinámicos y falla cardíaca. Hay múltiples procedimientos para el tratamiento y manejo, principalmente en casos de gran tamaño y fetos hidrópicos, para mejorar el pronóstico perinatal. Se presentan los dos primeros casos de secuestro broncopulmonar tratados en Venezuela mediante fotocoagulación láser del vaso nutricio y su evolución perinatal, con sobrevida del 100 % y sin ninguna complicación registrada en el periodo perinatal(AU)

Bronchopulmonary sequestration is a malformation of the respiratory system consisting of non-functioning bronchial and pulmonary tissue, separated from the tracheo-bronchial tree and fed by a blood vessel from the systemic circulation. The incidence is 1 in 5000 births, ranging from 0.15% to 6.45% of pulmonary pathologies. The prognosis is generally favorable, with spontaneous regression of the lesion reported in 50% to 75% of patients. It can cause mass effect, compressing the heart and lung to the point of generating hemodynamic changes and heart failure. There are multiple procedures for treatment and management, mainly in large cases and hydropic fetuses, to improve perinatal prognosis. We present the first two cases of bronchopulmonary sequestration treated in Venezuela by laser photocoagulation of the nutrient vessel and their perinatal evolution, with 100% survival and without any complications recorded in the perinatal period(AU)

Liquid chromatography/tandem mass spectrometry (highly selective selected reaction monitoring) for the analysis of isopropylthioxanthone in packaged food.

Isopropylthioxanthone (ITX), usually applied as a mixture of 2- and 4-isomers, is a common photo-initiator in UV inks used in paper- or plastic-based packaging materials. In this work a pentafluorophenylpropyl column (HS F5) has been used to achieve the chromatographic separation of the two isomers. A gradient elution with acetonitrile and a 25mM formic acid-ammonium formate at pH 3.75 are required to provide an Rs of 1.3 between the two compounds. The fragmentation pattern of ITX was studied using two mass analyzers, an ion trap (IT) (multi-stage fragmentation) and a triple quadrupole mass analyzer of hyperbolic rods (accurate mass (AM) measurement). The protonated molecule [M+H](+) observed in the mass spectrometry (MS) spectrum lost an isopropyl group, [M+H-C(3)H(6)](+). Later, this ion fragmented, yielding the radical ion [M+H-C(3)H(6)-CHO](+). The elemental composition of these product ions was confirmed by AM measurement. Electrospray ionization (ESI) was used as an ionization source to couple liquid chromatography (LC) to MS. Instrumental quality parameters of three acquisition modes provided by the triple quadrupole mass analyzer were studied and good run-to-run precision (relative standard deviation, RSD, lower than 10%) and limits of detection (LODs) down to 0.8pg injected in the LC-MS/MS system were obtained. Finally the LC-MS/MS method using H-SRM Q1 acquisition mode was used to analyze 2- and 4-ITX in a range of food samples. The use of highly selective selected reaction monitoring (H-SRM on Q1) resulted in improved selectivity without sensitivity loss.

[Chondroblastoma of the carpal scaphoids]. / Condroblastoma de escafoides carpiano.

A chondroblastoma is a benign tumour of cartilaginous origin which represents less than 1% of all primary osseous tumours. It is typically localised in the epiphysis of the long bones. Some 10% of chondroblastomas affect the bones of the foot and the hand. It is extremely rare for the seat of this lesion to be in the carpal bones. We present a case of chondroblastoma of the carpal scaphoids that was treated through curretage of the lesion and filling with autologous osseous graft from the iliac crest.

Abuse during pregnancy in Mexico City.

OBJECTIVE: Measure the prevalence of physical and sexual abuse during pregnancy, determine the nature and severity of abuse, and assess correlates with abuse. METHOD: A total of 1314 women seeking prenatal care between July 2000 and January 2003 were approached at three public hospitals in Mexico City. An original composite case record form was created to measure physical and sexual abuse before and during pregnancy. RESULT: Forty-one percent of respondents had a history of physical or sexual abuse, with current abuse reported by 11.1%, and abuse during pregnancy by 7.6%. Among abused women, 71% reported an increase in the severity of abuse since becoming pregnant. Logistic regression revealed physically fighting with a partner and a history of abuse best predict violence during pregnancy. CONCLUSION: The severity of abuse among abused women appears to increase during pregnancy. Prenatal care visits in Mexico are an important opportunity for violence screening and intervention.

Massive hand crush: the role of a free muscle flap to obliterate the dead space and to clear deep infection.

Death of tissue and/or deep infection leading to amputation is not an uncommon course of events after massive crush injuries of the central part of the hand. Management of this injury faces the dual problem of having to carry out debridement in the central part of the hand which is radical enough to remove all dead tissue but which, in itself, creates a huge dead space in the depths of the wound. Inadequate debridement and/or leaving a dead space which fills with fluid and detritus behind the flexor tendons leads on to infection, devascularisation of the fingers and amputation. This paper presents the results of very radical debridement of the hand dorsal to the flexor tendons, including the intermetacarpal spaces, and filling the dead space with a well-vascularised free muscle flap in two hands which were referred in a pre-amputation stage, with one already being infected. Both hands were salvaged.

Induction of transforming growth factor-beta 1 production in human cells by herpes simplex virus.

Transforming growth factor-beta (TGF-beta) is a cytokine of particular interest in human retrovirus infections because it can abrogate antigen-specific cellular activation. Although TGF-beta production has been observed in HIV infections, there is no evidence that herpes simplex virus (HSV)-stimulated human cells produce this cytokine. Here we present evidence, for the first time, that in vitro infection of human mononuclear cells with HSV type 1 (HSV-1) induced the release of TGF-beta1 protein. The production of this cytokine was time dependent and was found highly significant (p < 0.001) after 48 h. In addition, we observed that the secretion of TGF-beta1 was dependent on the concentration of human cells. It was found that virus needs to replicate in human cells for the production of TGF-beta1, as UV-inactivated virus did not induce significant production of cytokine protein. Interestingly, increased HSV-1-induced TGF-beta1 production in cultures containing antiinterleukin (IL)-12 or antiinterferon (IFN)-gamma antibodies was observed, whereas an irrelevant antibody had no effect on the production of this cytokine. Taken together, these findings indicate that human cells synthetize TGF-beta1 in response to HSV-1 and at the same time suggest that HSV-1-induced TGF-beta1 production may be one of the mechanisms by which HSV can at least partly evade activation of the host immune system.

Infection of human monocytes with Mycobacterium bovis BCG induces production of CC-chemokines.

DESIGN: CC-chemokines are potent leukocyte activators and chemoattractants, which have an important role in granuloma formation, function critical for the immune responses to mycobacterial infection. This study investigated whether infection of human monocytes with Mycobacterium bovis bacillus Calmette-Guérin (BCG) elicits secretion of RANTES, macrophage inflammatory protein (MIP)-1alpha and MIP-1beta. METHODS: RANTES, MIP-1alpha and MIP-1beta synthesis was measured by the presence of protein secretion in the cell culture supernatant as determined by enzyme-linked immunosorbent assay. To investigate the mechanism of M. bovis BCG stimulation of RANTES, we carried out inhibition assays with antibodies to CD40 and we used an intracellular calcium chelator BAPTA-AM. RESULTS: Infection of human monocytes with M. bovis BCG induced RANTES, MIP-1alpha and MIP-1beta secretion in a dose-dependent manner. This stimulation of CC-chemokines production was not attributed to LPS contamination. M. bovis-induced RANTES secretion was dependent upon bacterial uptake and on tumor necrosis factor (TNF)-alpha. Interestingly, the production of RANTES by M. bovis BCG-infected monocytes occurs through a mechanism that requires intracellular calcium and was significantly inhibited (P<0.05) with antibodies to CD40. CONCLUSIONS: These results suggest that the ability of M. bovis BCG to produce CC-chemokines might lead to protection in the acquired immune response of mycobacterial infection and at the same time indicate that M. bovis BCG-induced RANTES secretion is mediated by CD40 and dependent on the intracellular calcium influx.

Differential induction of TNF-alpha and NOS2 by mitogen-activated protein kinase signaling pathways during Mycobacterium bovis infection.

The role of mitogen-activated protein kinase (MAPK) signaling pathways in the regulation of TNF-alpha and NOS2 production by human monocytes infected with Mycobacterium bovis BCG was examined. Inhibition studies showed that ERK1/2 and p38 MAPK activation were necessary for the monocyte response to M. bovis infection. Analysis of MAPK activation showed rapid phosphorylation of ERK1/2 and p38 in response to M. bovis BCG. Phosphorylation was not due to an autocrine effect of TNF-alpha secretion, since an anti-TNF-alpha antibody had no significant effect on the levels of p38 phosphorylation. The inhibitor PD98059 significantly reduced M. bovis BCG-induced TNF-alpha production and almost completely abrogated phosphorylation of ERK1/2; in addition the potent MEK inhibitor U0126 also abrogated phosphorylation. In contrast, studies using inhibitors selective for ERK1/2 and p38 showed that p38 plays an essential role in the induction of NOS2, whereas the role of ERK1/2 was minor. These results suggest that ERK1/2 and p38 kinases differentially regulate the M. bovis BCG-mediated induction of TNF-alpha and NOS2 in human monocytes.

Abierixin, a new polyether antibiotic. Production, structural determination and biological activities.

A new polyether antibiotic, abierixin, was found in the mycelium of a culture broth of nigericin-producing Streptomyces albus NRRL B-1865. Abierixin was extracted with organic solvents and purified by column chromatography and HPLC. The structure of abierixin was determined by FAB/MS/MS and CI/MS/MS and 1H and 13C NMR spectrometries. Abierixin exhibited weak antimicrobial and ionophorous activities, low toxicity but good anticoccidial activity. Nigericin biosynthesis from abierixin is discussed.

Effect of level of energy intake and influence of breed and sex on the chemical composition of cattle.

Chemical composition of the empty body was determined in 159 animals slaughtered at weights ranging from 121 to 706 kilograms. Holstein and Angus bulls, steers and heifers were fed at two energy levels: ad libitum and 65 to 70% ad libitum. The allometric equation, Y = aXb, was used to determine the effect of energy intake and the influence of breed and sex on the accretion rates of the chemical components relative to the growth of the empty body or fat-free empty body. Group comparisons for chemical composition were made after adjustment by regression to a common empty body weight. The expression of the sex influence on the accretion rates of water, protein and ash relative to the empty body depended on the breed and the energy intake level considered. The accretion rate of chemical fat was not influenced by sex. Genetic differences in the accretion rate relative to the empty body were detected only among animals in the high energy intake group. Regardless of sex, the accretion rates of protein and ash were more rapid (P < .05) in Holsteins than in Angus. However, a breed influence on the accretion rate of chemical fat was detected only among bulls, where Angus had a more rapid accretion rate. Feeding animals at two energy levels resulted in different accretion rates relative to the empty body. In the Angus breed, regardless of sex, the accretion rates of water, protein and ash were more rapid (P < .05) in the low intake group, whereas the accretion rate of chemical fat was slower (P < .05). Among Holsteins, the low energy intake level had a less definite effect; for bulls, the accretion rates of water and chemical fat were more rapid (P < .05) and slower (P < .05), respectively; for steers, and accretion rate of protein was more rapid (P < .05), and for heifers, none of the accretion rates was altered.

[Drug consumption during pregnancy]. / Consumo de fármacos en el embarazo.

The pregnant women receive different types of medications many of whose safety has not been clearly demonstrated. The drug use profiles vary from community to community which justify to make local studies. Two hundred obstetrical patients were interviewed throughout the prenatal control to obtain data on maternal drug consumption. The patients were interrogated with respect to names of drug products, routes of administration, dosage regimens and prenatal consumption or underlying disease states during pregnancy. The interval between administration of a drug and the time of delivery were also noted. In the prenatal period 79% received at least one product. The average number of drug products was 2 with a range from 0 to 6 different drugs. The principal drugs ingested were: vitamins (32%), antibiotics (24%), iron (9%), antiemetics (7%), nonsteroidal anti-inflammatory analgesic (6%) and acetaminophen (5%). The only variable related to drug consumption was the presence of prenatal complications. This study represent an approach for monitoring the medical quality of prenatal care.

[Evaluation of a new suture material, polyglecaprone (25) in gynecology and obstetrics]. / Valoración de un nuevo material de sutura, poliglecaprone (25) en ginecología y obstetricia.

A new synthetic absorbible suture was evaluated (poliglecaprone 25, in the Gynecology-Obstetric practice. 70 patients were studied, 33 obstetrical and 37 gynecological. We practiced trans and postoperative evaluations of the suture, as well as the characteristics of the tensil (stregth or force), memory, adjustment, etc. concluding that it is an excellent suture material and has great advantages over other materials.

[Obstetric characteristics of adolescent pregnancy]. / Características obstétricas de la adolescente embarazada.

Obstetrical characteristics of pregnant adolescents. The aim of the present study was to determine the obstetrical characteristics of pregnant adolescents and to evaluate if the high risk comes from biological factors associate to their age or not. The obstetrical complications and the perinatal results were not different in relation from other groups, with exception of the minors of fifteen years in which the age constitutes an important risk. Nevertheless, the adolescent has other risks to the health such as the risk of acquiring sexual transmission diseases including HIV/AIDS. The child of a teen mother are also more likely to die in infancy, between the first month and the first year of life. Adolescents need information and services to protect their sexual and reproductive health.

[Sexual aggression in adolescents. Epidemiologic study]. / Agresión sexual en adolescentes. Estudio epidemiológico.

Sexual assault is an enormous social and public health problem. Most victims are young women. The aim of this study was to describe the characteristics of this crime in adolescents. The study involved 213 patients separated in two subgroups: adolescents and non adolescents. In adolescents, much of this violence is perpetrated by an acquaintance or relative. Th frequency of sexually transmitted diseases was 20% and rape-related pregnancy was identified in 10% of victims. All victims of sexual assault must be encouraged for both medical and psychological evaluation.

[Association between gestational age, weight, and Apgar score in newborns extracted in pelvic presentation]. / Asociación entre edad gestacional, peso y valor de apgar en recién nacidos extraídos en presentación pélvica.

The analysis for the present study was derived from 343 singleton pregnancies with a breech presentation. The variables associated with Apgar score at 5 minutes were, gestational age and birth weight. This association was observed in all patients and when they were analyzed separately according to route of delivery. The intrapartum and early neonatal mortality was absent in both groups. The perinatal outcome among the vaginally delivered frank breech infants at term was not statistically different compared with the group delivered by caesarean section.

Versatile lipid profiling by liquid chromatography-high resolution mass spectrometry using all ion fragmentation and polarity switching. Preliminary application for serum samples phenotyping related to canine mammary cancer.

Lipids represent an extended class of substances characterized by such high variety and complexity that makes their unified analyses by liquid chromatography coupled to either high resolution or tandem mass spectrometry (LC-HRMS or LC-MS/MS) a real challenge. In the present study, a new versatile methodology associating ultra high performance liquid chromatography coupled to high resolution tandem mass spectrometry (UHPLC-HRMS/MS) have been developed for a comprehensive analysis of lipids. The use of polarity switching and "all ion fragmentation" (AIF) have been two action levels particularly exploited to finally permit the detection and identification of a multi-class and multi-analyte extended range of lipids in a single run. For identification purposes, both higher energy collision dissociation (HCD) and in-source CID (collision induced dissociation) fragmentation were evaluated in order to obtain information about the precursor and product ions in the same spectra. This approach provides both class-specific and lipid-specific fragments, enhancing lipid identification. Finally, the developed method was applied for differential phenotyping of serum samples collected from pet dogs developing spontaneous malignant mammary tumors and health controls. A biological signature associated with the presence of cancer was then successfully revealed from this lipidome analysis, which required to be further investigated and confirmed at larger scale.

The response of Mucor plumbeus to pentachlorophenol: a toxicoproteomics study.

Pentachlorophenol (PCP) represents a critical concern worldwide due to its toxicity and recalcitrance to degradation. The capacity of Mucor plumbeus to transform PCP into several detoxification metabolites, including tetrachlorohydroquinone and several phase II conjugates, was observed by LC-HRMS. The data obtained support the degradation pathway proposed previously. PCP effects in M. plumbeus, an unsequenced species, were investigated using a proteomics approach (bidimensional gel electrophoresis followed by MALDI TOF/TOF analyses). The mycelial proteins identified in the differentially accumulated spots allowed the identification of PCP responsive proteins. The presence of PCP increased the energy demand, altered the cell wall architecture and cytoskeleton and induced a significant stress response. The latter was emphasised by the up-accumulation of protein species associated with defence mechanisms (e.g. HSP70 and cytochrome c peroxidase). Overall the data produced corroborate the capability of PCP to uncouple oxidative-phosphorylation in mitochondria. Importantly, one of the identified mycelial protein species, a NAD- and Zn-dependent ADH, is likely to be involved in PCP degradation. Amongst the fungal secretome analysed, no putative PCP degradative enzymes were detected. This work constitutes the first toxicoproteomic study involving a Zygomycota fungus and the very first concerning the effect of PCP in a fungal proteome.