Occupational asthma induced by fish exposure.

Occupational asthma triggered by inhaling fish-derived aerosols is estimated to affect 2-8% of exposed individuals. This primarily affects workers in the fish processing industry. Fishmongers, rarely experience this issue, as recent research found no significant difference in asthma rates compared to a control group. We report the case of a fishmonger who presented with a 1-year history of rhinoconjunctivitis and asthma. The patient attributed these symptoms to his occupational exposure within the fish market environment, which worsened in the cold storage warehouse. Symptoms improved during holidays. Diagnosis involved skin-prick tests, sIgE (ImmunoCAP-specific IgE) measurements, and bronchial challenge tests, confirming occupational asthma from fish bioaerosol exposure. Parvalbumins, common fish proteins, share structural similarities, leading to cross-reactivity in fish allergy sufferers. In this case, sensitivity to rGad c1 (cod parvalbumin) was identified as the primary trigger for the patient's asthma, and responsible for sensitizations observed across various tested fish species.

Identification and evaluation of inhibitors of the EhGEF1 protein from Entamoeba histolytica.

The Dbl family of guanine nucleotide exchange factors (GEFs) is made up of a vast array of members that participate in the activation of the Rho family of small GTPases. Dbl-family proteins promote the exchange of guanosine diphosphate/guanosine triphosphate (GDP/GTP) in their target molecules, resulting in the activation of a variety of signaling pathways involved in diverse cellular events, such as actin-cytoskeleton remodeling, cellular invasion, cell movement, and other functions. It has been reported that members of the Dbl family have important roles in several cellular events in Entamoeba histolytica. These include activation of the actin cytoskeleton, cytokinesis, capping, uroid formation, cellular proliferation, erythrophagocytosis, cell migration, and chemotaxis. Here, we report the identification and testing of inhibitors of the E. histolytica guanine nucleotide exchange factor 1 (EhGEF1) protein (the research compounds 2BYRF, 2BY05, 2BYT6, 2BYLX, and 2BYPD), which decreased the in vitro ability of the protein to exchange GDP/GTP at its target GTPases, EhRacG and EhRho1, by 14.9-85.2%. Interestingly, the drug 1,1'-(1,2-phenylene)-bis-(1H-pyrrole-2,5-dione), which completely inhibits the GEF activity of the Trio protein in human cells, decreases the GEF activity of the EhGEF1 protein on the EhRacG and EhRho1 GTPases by 55.7% and 3.2%, respectively. The identification and evaluation of such inhibitors opens up the possibility of obtaining a new pharmacological tool to study the function of amoeba GEF proteins, their roles in various Rho GTPase-mediated signaling pathways, and the repercussions of modulating their activities with respect to several mechanisms related to E. histolytica pathogenesis.

Entamoeba histolyticaEhGEF1 structure and mutational analysis: New specific residues critical for function.

This paper reports the EhGEF1-EhRacG and EhGEF1-EhRho1 molecular complexes from Entamoeba histolytica. The not conserved amino acids Gln201,Tyr299, Gln302, Lys312, Asn313, Phe314 and Ile324 were localized, by means of an in silico computational analysis, at the interface of the exposed face from the DH domain of EhGEF1, which are important to establish the contact with its target GTPases EhRacG and EhRho1. Functional studies of nucleotide exchange of Phe314Ala mutant showed a decrement of 80% on EhRacG GTPase; in contrast the Ile324Ala mutant exhibited a reduction of 77%, specifically on EhRho1; meanwhile the Gln302Ala mutant showed a reduction of approximately 50% on the exchange activity for both GTPases. Moreover, the functional studies of the protein EhGEF1 mutants in the conserved residues Thr194Ala, Asn366Ala and Glu367Ala indicated that contrary to what has been reported for other systems, the mutation of these residues did not alter considerably its catalytic activity.

[Quality evaluation of a neuro-development follow-up program of very low birthweight infants]. / Valoración de la calidad de un programa de seguimiento del neurodesarrollo de prematuros de muy bajo peso al nacimiento.

INTRODUCTION: Advances in perinatology have been associated with improved survival of very low birth weight (VLBW;<1500 g) infants. These children have an increased risk of future neurological and development handicaps. A set of quality indicators have been recommended for the neurodevelopment follow-up programs in this group of patients. AIM: To analyze our neurodevelopment follow-up program according to these quality of care indicators in 5 post-discharge follow-up areas: general care; physical health; vision, hearing, speech and language; developmental and behaviour assessment; and psychosocial issues. PATIENTS AND METHODS: Fifty-one VLWB patients born in 2000 who have completed our 6-year follow-up program have been retrospectively studied. The agreement between our program and the standard of quality is evaluated. RESULTS: A copy of a discharge summary of the patient's neonatal course is present in the medical record in 92% of the cases. The inclusion of clinical data and general medical aspects in the medical summary is heterogeneous. Somatometric evaluation for the first two years of age is always present in a filled-in graphic. The international classification of the retinopathy of the preterm (RDP) is used in 100% of the patients. An ophthalmologic examination for RDP is performed in 72% (37/51) of the infants before hospital discharge; of the other 28%, only in 6 patients the date of the outpatient examination is present. Specific evaluations and interventions are performed within the recommended period of time in most of the children with suspected or neurosensory handicaps. More than ninety per cent of the patients are referred to the neurodevelopment specialist and most of them go to the visit (90.2% between 9 and 15 months and 78.4% between 21 and 30 months). Recommendations for a psycho-educational assessment are followed in the extremely premature (BW<1000 g and/or 28 weeks of gestational age) children. The number of patients who follow the program decreases with time. Psychosocial evaluation does not usually appear in the medical records of the patients. CONCLUSION: We have a good adherence to most of the recommended indicators in our follow-up program. The evaluation of these indicators is a useful tool to analyze the quality of a developmental follow-up program and to improve it.

Multi-annual survey of health status disturbance in the Bilbao estuary (Bay of Biscay) based on sediment chemistry and juvenile sole (Solea spp.) histopathology.

The Bilbao estuary (SE Bay of Biscay) is a recovering ecosystem whose sediments are still contaminated. They represent a potential risk for the biota including benthic and demersal species living in direct contact with the sediment. In this context, the present study aims to survey trends of the health status of the Bilbao estuary based on sediment chemistry and sole (Solea spp.) histopathology. Monitoring campaigns were carried out every autumn from 2011 to 2017 along the estuary. Contaminant levels were measured in sediments; liver, gills and gonads of juvenile fish were collected for histopathology. Overall, contaminant levels fluctuated throughout the years, with highest values recorded in the earlier years of the study period. Sole histopathology showed alterations of mild severity. Results permitted to assess the environmental health status of the Bilbao estuary during 7 years, although no clear temporal trend was detected. Longer-term monitoring programmes are necessary to confirm the ecosystem recovery.

Baseline levels of environmental genotoxicity and potential confounding factors using common sole (Solea solea) as sentinel organism.

Micronuclei (MN) and nuclear buds (NB) frequency was assessed monthly over a one-year period in erythrocytes of common soles (Solea solea) from the Basque continental shelf, a minor disturbed area, in order to determine baseline levels and potential biotic and abiotic confounding factors within biomonitoring purposes. Both genotoxic parameters presented seasonal variations, although only NB frequency was positively correlated with environmental variables (i.e. water temperature and salinity). On the contrary, MN and NB were not affected by age and gender. Therefore, samples of combined age and gender can be used for biomonitoring genotoxic effects, although sampling period together with water temperature and salinity should be thoroughly considered for NB. Overall, these findings are the first attempt to establish baseline MN (0.10-0.78‰) and NB (0.13-0.82‰) frequencies in common sole from Basque marine waters, which are useful for upcoming data comparisons and integration within genotoxicity evaluation procedures of similar marine environments under biomonitoring scopes.

Explorations on the ecological role of toxin secretion and delivery in jawless predatory Polychaeta.

Motivated by biotechnological prospects, there is increasing evidence that we may just be scraping the tip of the iceberg of poisonous marine invertebrates, among which the Polychaeta are promising candidates for bioprospecting. Here we show that an inconspicuous phyllodocid uses toxins in its uncanny feeding strategy. The worm, a jawless active predator characterised by its bright green colour, preys on larger invertebrates (including conspecifics) by extracting tissue portions with its powerful proboscis through suction. The animal is even able to penetrate through the valves and plates of live molluscs and barnacles. Observations in situ and a series of experiments demonstrated that the worm compensates its simple anatomy with secretion of a novel toxin, or mixture of toxins, referred to by us as "phyllotoxins". These are carried by mucus and delivered via repeated contact with the tip of the proboscis until the prey is relaxed or immobilised (reversibly). Proteolytic action permeabilises material to toxins and softens tissue to enable extraction by suction. The findings show that toxins are a major ecological trait and therefore play a key role in evolutionary success and diversification of Polychaeta, demonstrating also that understanding adaptative features may become the best showcase for novel animal toxins.

Histopathological baseline levels and confounding factors in common sole (Solea solea) for marine environmental risk assessment.

Liver and gonad histopathology, biometric parameters and hepatic metal bioaccumulation were assessed monthly over a one-year period in common soles from the Basque continental shelf, in order to determine baseline levels and confounding factors within biomonitoring studies. Biometric parameters and hepatic metal bioaccumulation varied according to season and gender. Accordingly, hepatic histopathological traits presented seasonal variations related to the reproductive cycle. However, the hepatic histopathological index showed that seasonality and gender were not significant confounding factors. Conversely, the gonad histopathological index was modulated by season and gender. As for organ comparison, the liver endured more severe histopathological damage than the gonad. In brief, the sampling period and gender may not affect the estimation of hepatic histopathological indices for biomonitoring purposes. Nonetheless, due to different sensitivities to environmental 'noise' variables, the sampling period and gender differentiation should be thoroughly considered for the assessment of gonad histopathology, biometrics and metal bioaccumulation.

Gene promoter hypermethylation in oral rinses of leukoplakia patients--a diagnostic and/or prognostic tool?

Leukoplakia is the most frequent oral precancerous lesion and shows a variable rate of malignant transformation. We hypothesised that the detection of molecular alterations, like the promoter hypermethylation of DNA, in oral cytological samples from patients at risk of developing primary or recurrent tumours could be a valuable diagnostic and prognostic tool in the management of these lesions. Two groups of patients with differing risks of developing oral squamous cell carcinoma (OSCC) were analysed. DNA was extracted from the oral rinse of each patient. The methylation status of the p16, p14 and MGMT gene promoters was determined using a methylation-specific polymerase chain reaction (MSP). Methylation of p16 and MGMT was observed in 44 and 56% of the oral samples, respectively. Only 12% of the cases showed p14 methylation. DNA hypermethylation was more frequent in patients with previous OSCC. DNA promoter hypermethylation is frequent during early oral carcinogenesis and even more so in the later stages. MSP using oral rinses is a non-invasive and highly sensitive technique which could be used to monitor patients with precancerous and cancerous oral lesions.

Neuromodulatory actions of substance P on the muscarinic receptors of the vas deferens of the rat.

The response of post-synaptic neurokinin receptors to SP were not changed by pirenzepine or N-methyl-scopolamine. Atropine led to a slight increase in the EC50 of SP for its post-synaptic neurokinin (NK-A) receptor. In the presence of neostigmine no changes in the Emax and EC50 values of SP for its post- and pre-synaptic receptor site were observed. Only the muscarinic receptor site were observed. Only the muscarinic receptor antagonists, atropine and NMS, elicited statistically significant increases in the Emax of SP at its presynaptic receptor (NK-A). Addition of 7.4-740 nM SP resulted in a decrease in the EC50 and Emax values of ACh for its post-synaptic muscarinic receptor (M1). Conversely, 740 nM SP produced an increase in the EC50 and Emax values of ACh at its pre-synaptic muscarinic receptor (M2). Concentrations of 7.4 and 74 nM SP did not produce statistically significant changes in the Emax of ACh for its pre-synaptic M2 receptor.

Pharmacological actions of substance P in the rat vas deferens.

SP produces two different muscular responses in the isolated vas deferens of the rat. 1) A myotropic, post-synaptic effect that results from stimulation of one subtype of SP receptor, located on the membrane of the smooth muscle (NK-3 receptor). 2) A neurogenic effect at the pre-synaptic level that results from stimulation of another subtype of SP receptor, located on the nerve terminals (NK-1 receptor).

Use of cytological specimens for p53 gene alteration detection in oral squamous cell carcinoma risk patients.

AIMS: Recurrence and multifocal nature are two important characteristics of oral squamous cell carcinoma. Leukoplakia is the most frequent pre-cancerous oral lesion and, in most cases, it is not possible to predict malignant capacity. The objective of this study is to identify p53 alterations in cells taken from the oral cavity of at-risk patients. MATERIALS AND METHODS: The following samples were collected from 34 patients with oral leukoplakia with and without previous carcinoma: oral rinse, a brush swabbed over the lesions and hair roots. Mutational analysis of the p53 gene was performed by single-strand conformation polymorphisms and confirmed by DNA sequencing. RESULTS: We detected 11 mutations in p53 gene in oral cytological specimens. These alterations were observed only in brush cytology samples in patients without previous carcinoma, and in both samples (rinse and brush) in patients with previous carcinoma. Three of these patients had disease recurrence. CONCLUSION: This non-invasive technique may be useful in the follow-up of at-risk patients, and introduces new possibilities to analyse molecular markers before malignant lesions are clinically apparent.

Population frequency distribution of HumF13A01, HumFXIIIB, and HumLIPOL loci in the Basque Country (Northern Spain).

A population study of unrelated individuals from the Basque Country (Northern Spain) was carried out using the GenePrint STR System. The PCR products were separated on denaturing polyacrylamide gels and visualized by silver staining. Three tetrameric loci were evaluated: HumF13A01, HumFXIIIB, and HumLIPOL. All loci fit Hardy-Weinberg expectations, and independence of alleles was found between these STR loci. A comparison with other population groups indicated allele frequencies are well conserved in Caucasians, but differ from other racial groups. The calculated parameters a priori probability of exclusion (Pex) and "power of discrimination" (PD) show how informative these loci are for the determination of identity and relatedness of individuals.

In vitro studies of water-stable cationic carbosilane dendrimers as delivery vehicles for gene therapy against HIV and hepatocarcinoma.

Here we present a synthetic procedure for water-stable carbosilane dendrimers containing ammonium groups at the periphery of type Gn-{[Si(CH2)3N+(Me)(Et)CH2CH2N+Me3]x (CF3SO3 -)y} which have been used as non-viral vectors for transfecting different types of nucleic acids against two different medical problems, HIV and hepatocarcinoma. These systems have shown to be non-toxic in both PBMC and HepG2 cell lines under the experimental conditions and are able to form nanoconjugates with nucleic acids perfectly stable over time and in a wide range of pH values, which leads to the conclusion that the interaction between dendrimer and nucleic acid is very strong. In addition, a high degree of transfection using these nanoconjugates has been observed, ranging from 70-90% depending on the generation and in the particular case of PBMC transfection with anti-HIV oligonucleotides. However, besides of the good properties shown by the dendrimers here prepared as transfecting agents, only moderate effect was observed in functional experiments for hepatocarcinoma, as a result of the strong interaction between dendrimer and nucleic acid. Nevertheless, it is important to mention that an IRS-4 knock-down of 40% in HepG2 achieves an analogous degree of cell sensitization to cancer treatment, which may represent a major advance in the hepatocarcinoma treatment when appropriate dendrimers as transfection agents are used.

EhNCABP166: a nucleocytoplasmic actin-binding protein from Entamoeba histolytica.

The actin cytoskeleton consists of multiple actin binding proteins (ABPs) that participate cooperatively in different cellular functions such as the maintenance of polarity and cell motility as well as the invasion of target cells and regulation of gene expression, among others. Due to the important role of ABPs in the pathogenesis of Entamoeba histolytica, the role of a new nucleocytoplasmic ABP from E. histolytica named EhNCABP166 was investigated. The EhNCABP166 gene encodes a protein with an estimated molecular weight of 166kDa. Structurally, this peptide is composed of two CH domains arranged in tandem at the N-terminus of the protein, followed by an alpha-helical region containing a number of different domains with a low level of homology. Two (Bin1/Amphiphysin/Rvs167) (BAR) domains, one GTPase-binding/formin 3 homology (GBD/FH3) domain, three Bcl2-associated athanogene (BAG) domains, one basic-leucine zipper (bZIP) domain and one poly(A)-binding protein C-terminal (PABC) domain were also present. Molecular and biochemical studies showed that the EhNCABP166 protein is transcribed and translated in trophozoites of E. histolytica. It was also shown that the CH domains are functional and bind to F-actin, whereas the BAR and GBD/FH3 domains interact in vitro and in vivo with different families of GTPases such as Rho and Ras, and with different phosphoinositides. These findings suggest that these domains have the conserved functional properties described in other eukaryotic systems. These domains also interacted with additional GTPase and lipid targets that have not been previously described. Finally, cellular studies showed that EhNCABP166 is localized to the cytoplasm and nucleus of E. histolytica and that it has an important role in phagocytosis, proliferation, and motility of E. histolytica.

Impaired apoptosis in lymphoblasts from Alzheimer's disease patients: cross-talk of Ca2+/calmodulin and ERK1/2 signaling pathways.

We have analyzed the intracellular signals that allow lymphoblasts from Alzheimer's disease (AD) patients to escape from serum deprivation-induced apoptosis. The following observations suggested that modulation of ERK1/2 activity by Ca(2+)/calmodulin (CaM) is involved in preventing apoptosis: (i) ERK1/2 activity seems to support lethality in control cells, as PD98059, the inhibitor of the activating MEK prevented cell death; (ii) control cells show a persistent and higher stimulation of ERK1/2 than that of AD cells in the absence of serum; (iii) CaM antagonists have no effects on control cells, but sensitize AD cells to death induced by serum withdrawal and increased ERK1/2 phosphorylation, and (iv) no apoptotic effects of CaM antagonists were observed in AD cells treated with PD98059. These results suggest the existence of an activation threshold of the ERK1/2 pathway setting by Ca(2+)/CaM-dependent mechanisms, which appears to be the critical factor controlling cell survival or death decision under trophic factor withdrawal.

Validity and reliability issues in published public health and social policy research.

This paper reviews basic measurement concepts associated with objective instruments, argues for the importance of addressing the validity and reliability of instruments in articles reporting the results of empirical research, and discusses common measurement issues through examples observed in public health and social policy journals.

[The Poland anomaly with unilateral peripheral facial paralysis]. / Anomalía de Poland con parálisis facial periférica unilateral.

The authors present the clinical history and physical findings in a 16-month baby girl with the diagnosis of Poland's syndrome associated with another congenital anomaly: unilateral peripheral facial palsy, on the opposite side. The authors believe this is the first report of such an association.

Clinical applications of the diagnosis of p53 alterations in squamous cell carcinoma of the head and neck.

Alteration of the p53 tumor suppressor gene implies an extremely high risk of developing malignancy, and mutation of the gene is one of the most frequent genetic changes found in human cancer. Squamous cell carcinoma of the head and neck (SCCHN) shows a high incidence of p53 tumor suppressor gene alterations; the latter therefore appears to play an important role in the pathogenesis and progression of such neoplasms. The loss of p53 protein activity may be due to many p53 gene mutations or to the action of certain viruses that infect the oral cavity. Local recurrence is the most common cause of mortality after SCCHN surgery; in this sense, p53 gene mutations have been observed in tissue adjacent to the tumor, and constitute a good prognostic marker of tumor recurrence. The analysis of p53 tumor suppressor gene alterations in SCCHN affords important information on the diagnosis, prognosis and treatment of affected patients - such alterations representing an indicator in high risk patients of the convenience of applying more aggressive adjuvant therapies.

Patterns of methylation profiles as diagnostic markers for multiple hepatocellular carcinomas.

BACKGROUND: Hepatocellular carcinoma often displays multiple tumor nodules, thus posing a problem for differential diagnosis between cancers of both multifocal and metastatic origin. Conventionally, pathological criteria have been used for this purpose, but these are largely subjective. In order to facilitate a more objective differential diagnosis of multiple HCCs, we used the patterns of methylation of p16INK4a, p14ARF, and GSTP1 genes as markers for each tumor nodule. METHODS: Sixty-seven nodules from 30 cases of multiple or recurrent HCCs were examined using methylation-specific PCR (MSP) analysis for the detection of methylation profiles. RESULTS: Hypermethylation was detected in 56.7%, 43.3% and 17.9% of the cases for p16INK4a, p14ARF, and GSTP1 genes, respectively. At the genetic level the inter-nodule methylation profiles were heterogeneous in 23 of the cases and homogeneous in another 7, enabling a multifocal origin to be diagnosed in the former and metastatic origin in the latter. CONCLUSIONS: Methylation profiling seems to be useful in differentiating the clonal origins of multiple cancers, as the information yielded by this method is essentially objective.