Effects of heating and recovery media pH on the heat resistance of Alicyclobacillus acidoterrestris Ad 746 spores.

AIMS: Alicyclobacillus acidoterrestris is a sporulating, acidophilic bacterial species which spoils acidic beverages such as fruit juices. This work aims to quantify the heat resistance of A. acidoterrestris spores and their recovery potential as a function of heating and recovery media pH. METHODS AND RESULTS: The heat treatments were carried out with the strain of A. acidoterrestris Ad 746 in Bacillus acidoterrestris thermophilic medium. The pH of the heating medium from pH 7 to pH 2 nonsignificantly reduced the heat resistance. However, the pH levels of the recovery media strongly affected the apparent heat resistance of this strain. The maximum heat resistance was found when the pH was 4·70 and decreased when the pH decreased to pH 2·8, close to the minimum growth pH and when the recovery medium pH increased to pH 5·3. CONCLUSION: The heating medium pH has a slight effect on the spore heat resistances of this acidophilic species. However, the pH of the recovery media strongly affected the apparent heat resistance of this strain. SIGNIFICANCE AND IMPACT OF THE STUDY: The obtained parameters quantifying the heat resistance of A. acidoterrestris spores are tools to optimize the heat treatments and to control its development.

A subset of walnut allergic adults is sensitized to walnut 11S globulin Jug r 4.

BACKGROUND: The role of sensitization to commercially available allergens of English walnut (Juglans regia) Jug r 1, 2 and 3 in walnut allergy has been previously investigated in walnut allergic adults and was unable to explain all cases of walnut allergy. OBJECTIVES: Identify recognized walnut allergens, other than the ones previously investigated (Jug r 1-3), in walnut allergic adults and determine the sensitization frequency and diagnostic value. METHODS: Three different in-house walnut extracts were prepared and analysed on SDS-PAGE blots to identify allergenic walnut proteins. Immunoblots and immunoprecipitation, followed by LC-MS analysis, were performed to screen for, and confirm, IgE binding to walnut allergens in selected walnut allergic adults. In a cohort of 55 walnut challenged adults, including 33 allergic and 22 tolerant, sensitization to native and recombinant walnut allergen Jug r 4 was assessed using immunoblotting and immuno-line blot (EUROLINE), respectively. RESULTS: Screening of sera of 8 walnut allergic adults identified Jug r 4 as an allergen in our population. In the total cohort of 55 subjects, 5 were positive for Jug r 4 on immunoblot and 10 on EUROLINE. All but one EUROLINE positive subject had a positive food challenge (sensitivity 27%, specificity 95%, PPV 90%, NPV 47%). All 5 subjects positive on immunoblot were also positive on EUROLINE. LC-MS analysis showed a lack of Jug r 4 in the ImmunoCAP extract. Co-sensitization to other 11S albumins (eg hazelnut Cor a 9) was common in Jug r 4 sensitized subjects, potentially due to cross-reactivity. CONCLUSIONS: Walnut 11S globulin Jug r 4 is a relevant minor allergen, recognized by 27% of walnut allergic adults. It has a high positive predictive value of 90% for walnut allergy. Specific IgE against Jug r 4 occurred mostly with concomitant sensitization to other walnut components, mainly Jug r 1.

Myocarditis and inflammatory bowel diseases: A single-center experience and a systematic literature review.

BACKGROUND: Myocarditis and inflammatory bowel diseases (IBD) are rare conditions, but may coexist. Myocarditis in IBD may be infective, immune-mediated, or due to mesalamine toxicity. A gap of knowledge exists on the clinical features of patients that present myocarditis in association with IBD, especially for endomyocardial biopsy-proven cases. Our aims are: 1) to describe the clinical characteristics of patients with an associated diagnosis of myocarditis and IBD in a single-center hospital, 2) to perform a systematic review of the literature of analogous cases. METHODS: We retrospectively analyzed data of patients followed up at the outpatient Cardio-immunology and Gastroenterology Clinic of Padua University Hospital, to identify those with an associated diagnosis of myocarditis and IBD. In addition, a systematic review of the literature was conducted. We performed a qualitative analysis of the overall study population. RESULTS: The study included 104 patients (21 from our single center cohort, 83 from the literature review). Myocarditis in IBD more frequently affects young (median age 31 years) males (72%), predominantly with infarct-like presentation (58%), within an acute phase of the IBD (67%) and with an overall benign clinical course (87%). Nevertheless, a not negligible quote of patients may present giant cell myocarditis, deserve immunosuppression and have a chronic, or even fatal course. Histological evidence of mesalamine hypersensitivity is scarce and its incidence may be overestimated. CONCLUSIONS: Our study shows that myocarditis in association with IBD, if correctly managed, may have a spontaneous benign course, but predictors of worse prognosis must be promptly recognized.

[Minimally invasive treatment of chronic subdural hematoma in adults. Results in 116 patients]. / Tratamiento mínimamente invasivo del hematoma subdural crónico del adulto. Resultados en 116 pacientes.

INTRODUCTION: Chronic subdural hematoma in adults (CSDH) has a global crude incidence of 14.1/100,000 per year in our institution captive population. There is no single treatment protocol. In our hospital we choose a minimal invasive technique (trans-marrow puncture) without general anaesthesia due to the age of the population. A descriptive study of patients with CSDH and treatment results, including a laterality analysis, is presented. MATERIAL AND METHODS: We retrospectively searched patients with (CSDH) between January 1998 and May 2009. The diagnosis was made by neuroimaging techniques in all patients. The preferred treatment was trans-marrow puncture; exceptionally some patients were treated by burr holes or craniotomy. RESULTS: We found 127 patients. Age, gender, midline displacement, hospitalisation days, and number of procedures, were not a predictive factor of mortality in the first month. A slight majority (55%) of CSDH were on the left side, with no statistically significant difference. There were 6 (4.7%) deaths during hospitalisation. In our series cumulative mortality at six months was 11.8%. Markwalder scale at admission was not a predictive factor of statistically significant mortality. An 80% of the patients received trans-marrow puncture as single procedure was performed on 80% of the patients. CONCLUSIONS: The results of our study suggest that trans-marrow puncture is an acceptable procedure, with low mortality, and less hospitalisation days and complications. Mortality, associated mechanisms, age, gender, midline displacement are no different than in others previous publications. We found a higher frequency of hematomas to the left, as in other series. Meta-analysis studied need to be performed to determine more accurately the frequency of this dominance.

A method of obtaining, processing, and analyzing human intestinal secretions for antibody content.

Human intestinal secretions can be readily obtained using a commercially available intestinal lavage solution. Although such secretions contained abundant protease activity, significant loss of immunoglobulins was prevented by the addition of a mixture of protease inhibitors. The total content of IgA, IgM, and IgG antibody in secretions was measured using sandwich ELISA. In the secretions of ten normal volunteers IgA was most abundant (197 micrograms/ml +/- 103 SD) followed by IgM (12.5 micrograms/ml +/- 6.8 SD) and IgG (0.24 micrograms/ml +/- 0.04 SD). The IgA in secretions was predominantly secretory IgA as shown by sucrose density centrifugation. The effect of intestinal secretions on the sensitivity of the antigen-specific ELISA was tested by adding murine myeloma IgA anti-TNP added to samples of human secretions. IgA anti-TNP activity could be detected as low as 1 ng/ml, and there was no evidence of interference with the ELISA by other constituents in the secretions. Using these methods an antigen-specific secretory IgA anti-cholera toxin B subunit response in the secretions of volunteers given an oral B subunit vaccine was readily demonstrated.

Dose ranging pharmacokinetic trial of high-dose alicaforsen (intercellular adhesion molecule-1 antisense oligodeoxynucleotide) (ISIS 2302) in active Crohn's disease.

BACKGROUND AND AIMS: To evaluate the safety, pharmacokinetics and clinical efficacy of the intercellular adhesion molecule-1 antisense phosphorothioate oligonucleotide alicaforsen (ISIS 2302) at 250-350 mg in Crohn's disease. METHODS: : Patients (> 50 kg) with active Crohn's disease (Crohn's disease activity index > or = 220) were assigned by gender, randomly, to two alicaforsen treatment groups: 300 or 350 mg, infused intravenously three times a week for 4 weeks. All patients weighing 36-50 kg received 250 mg of alicaforsen. Background aminosalicylates, antibiotics, immunosuppressives and corticosteroids were permitted, but tumour necrosis factor-alphainhibitors were prohibited. The primary end-point was clinical remission (Crohn's disease activity index < or = 150). RESULTS: Twenty-two patients were enrolled with a mean baseline Crohn's disease activity index of 304. Steroids were used by 27%, 5-aminosalicylic acid by 68% and immunosuppressives by 27%; 23% had previously received infliximab. Five subjects withdrew after one to three infusions for infusion-related symptoms. Nine patients (41%) experienced clinical remission. Fifty-three per cent of the evaluable subjects receiving more than three infusions experienced remission (18% at week 8; 29% at week 12). The overall response, using a minimum decrease of 70 in the Crohn's disease activity index, was 41-47% for the evaluable group, at weeks 8 and 12. The median duration of remission was 14 weeks. Plasma pharmacokinetic results showed overlapping levels (Cmax, AUC) for the three doses. The infusion-related reaction profile consisted of fever, chills, headache, nausea, emesis or arthralgias, typically occurring 2-4 h after completion of the first infusion. Reactions were less frequent in patients receiving background corticosteroids. The 2-4-h transient post-infusion partial thromboplastin time prolongation values, a class effect of phosphorothioate oligonucleotides, were 18, 21 and 23 s for 250, 300 and 350 mg, respectively. CONCLUSIONS: Alicaforsen (ISIS 2302), at fixed doses of 300 and 350 mg, achieved the desired drug exposure and may be an effective therapy for Crohn's disease. Infusion-related reactions were observed less frequently in patients on corticosteroids, and with decreasing frequency with continued treatment.

Adrenal insufficiency in high-risk surgical ICU patients.

STUDY OBJECTIVES: To examine the incidence and response to treatment of adrenal insufficiency (AI) in high-risk postoperative patients. DESIGN: Prospective observational case series. SETTING: Large urban tertiary-care surgical ICU (SICU). PARTICIPANTS: Adults > 55 years of age who required vasopressor therapy after adequate volume resuscitation in the immediate postoperative period. INTERVENTIONS: Each patient underwent a cosyntropin (ACTH) stimulation test; at the discretion of the clinical team, some patients were empirically given hydrocortisone (100 mg IV q8h for three doses) before serum cortisol values became available. MEASUREMENTS: Adrenal dysfunction (AD), defined as serum cortisol < 20 microg/dL at all time points, with Delta cortisol (60 min post-ACTH minus baseline) of < or = 9 microg/dL; functional hypoadrenalism (FH), defined as serum cortisol < 30 microg/dL at all time points or Delta cortisol (60 min post-ACTH minus baseline) < or = 9 microg/dL; and AI, as the presence of either AD or FH. RESULTS: One hundred four patients were enrolled with a mean age (SD) of 65.2 +/- 16.9 years. AI (AD plus FH) was found in 34 of 104 patients (32.7%): AD was found in 9 patients (8.7%), FH in 25 patients (24%), and normal adrenal function in 70 patients (67.3%). The absolute eosinophil count was significantly higher in the combined AD and FH groups compared with the group with normal adrenal function (p < 0.05). Forty-six of 104 patients (44.2%) received hydrocortisone; 29 (63%) could be weaned from treatment with vasopressors within 24 h. This beneficial effect of hydrocortisone reached statistical significance in the FH group when compared with untreated patients (p < 0.031); a similar trend was seen in the AD group (p = 0.083). Mortality was also lower in the hydrocortisone-treated AI patients (5 of 23 [21%] vs 5 of 11 [45%] in those not receiving hydrocortisone; p < 0.01). CONCLUSION: There is a high incidence of AI among SICU patients > 55 years of age with postoperative hypotension requiring vasopressors. There is also a significant association between hydrocortisone replacement therapy, resolution of vasopressor requirements, and improved survival.

[Idiopathic hypophysitis in elderly patients. Report of 2 cases and review of literature]. / Hipofisitis idiopática en pacientes de edad avanzada. Presentación de dos casos y revisión de la literatura.

INTRODUCTION: Hypophysis inflammatory tumors are a non frequent cause for hypopituitarism. The motive of outpatient visit is headache. It is more frequent in pregnancy and immediate post-labor women. The pathology shows a lymphocytic inflammatory infiltrated or granulomas with giant cells (these can be accompanied with an infectious or autoimmune systemic disease associated). CLINICAL CASES: We present two elderly female patients, without infectious or autoimmune pathology associated, with suggestive abnormalities of hypophysial adenoma by MRI. Both presented a suspecting clinical manifestation of intracranial expansive lesion (progressive intensive headaches, with partial compromise of the III left pair in the second one), noticing by laboratory a subclinical hormonal dysfunction. Surgery of tumoral exeresis was underwent in both patients, showing a granulomatous hypophysitis in one and a lymphocytic hypophysitis in the other. CONCLUSION: Hypophysis inflammatory tumors must be considered as a preoperative differential diagnosis of every hypophysial tumor without important hormonal dysfunction associated, mainly in women and what is more in pregnancy or lying-in women, taking into account that surgery of tumoral resection can worsen a subclinic hypophysial dysfunction.

[Chronic and subacute subdural haematoma. An epidemiological study in a captive population]. / Hematoma subdural crónico y subagudo. Estudio epidemiológico en una población cautiva.

INTRODUCTION: Although chronic and subacute subdural haematomas (CSSH) are amongst the commonest neurosurgical conditions, there are few studies on their incidence in the general population. OBJECTIVES: To determine the overall annual rate, the specific rates according to age and sex based on the Official Argentinian National Census of 1991 (OANC 91) for CSSH. PATIENTS AND METHODS: The Hospital Privado de Comunidad de Mar del Plata attends a captive population of 89,500 persons from the Instituto Nacional de Servicios Sociales de Jubilados y Pensionados (INSSJP) and the Prepaid Medical Schemes (PMP) of our institution. We studied the patients of INSSJP and PMP who had CSSH between 1992 and 1996. We determined the annual overall rate and the specific rates according to age and sex, and fitted to the OANC 91. RESULTS: 1. Annual overall rate: 14.1 CSSH/100,000 persons/year. 2. Specific rate for women: 11.6 CSSH/100,000 persons/year. 3. Specific rate for men: 18.1 CSSH/100,000 persons/year. 4. Specific rate 71-80 years old: 18.8 CSSH/100,000 persons/year. 5. Rate fitted to OANC9: 3.1 CSSH/100,000 persons/year. CONCLUSIONS: Our overall rate is higher, and the specific rate for the age group 71-80 years is intermediate, with regard to the rates found in other studies. Neuroepidemiological investigation should be stimulated so that more clinical studies are made regarding the results and costs based on the population.

[First episode non-traumatic subarachnoid haemorrhage. An epidemiological study of incidence and of arterial hypertension as a risk factor]. / Primer episodio de hemorragia subaracnoidea no traumática. Estudio epidemiológico de su incidencia y de la hipertensión arterial como un factor de riesgo.

AIMS: Our aim was to conduct an epidemiological study of the incidence of non-traumatic subarachnoid haemorrhage (NTSAH) and to evaluate a history of arterial hypertension (AHT) as a risk factor (RF). We also sought to determine the gross annual rate (GAR) and specific rates, by sex and age, of first episode non-traumatic subarachnoid haemorrhage (FENTSAH) adjusted to the Censo Nacional de Población de la Argentina (CNPA), and to carry out a population-based case-control study about the history of AHT as a RF in an Argentine community. PATIENTS AND METHODS: Our institution, the Hospital Privado de Comunidad de Mar del Plata, Argentina, attends a population of about 89,500 individuals exclusively and completely. There is a single filing system of case histories with all the diagnoses in a database. Patients who suffered a FENTSAH between the years 1992 and 1996 (n=62) were investigated to determine the specific GAR by sex and age and adjusted to the 1991 CNPA. The population was compared at random with two controls per case for the study of a history of AHT as a RF, and was then stratified according to sex and age. RESULTS: 1. GAR: 13.6/100,000 inhab./year; 2. Specific rate for males: 10.5/100,000 inhab./year; 3. Specific rate for females: 15.9/100,000 inhab./year; 4. CNPA adjusted rate: 7.3/100,000 inhab./year; 5. A significantly important number of patients with FENTSAH presented a history of hypertension as compared to controls subjects (odds ratio: 4.73; CI 95%: 2.39 9.34). CONCLUSIONS: Our rates are not substantially different from the rates found in other western countries and our findings indicate that a history of AHT is a RF for FENTSAH.

Biomarker discovery by plasma proteomics in familial Brugada Syndrome.

Brugada Syndrome (BS) is a polygenic inherited cardiac disease characterized by life-threatening arrhythmias and high incidence of sudden death. In this study, two-dimensional gel electrophoresis (2D-PAGE) coupled to mass spectrometry (LC-MS/MS) was used to investigate specific changes in the plasma proteome of BS patients and family members sharing the same gene mutation (SCN5AQ1118X), with the aim to identify novel disease biomarkers. Our data demonstrate that the levels of several proteins were significantly altered in BS patients compared with controls. In particular, apolipoprotein E, prothrombin, vitronectin, complement-factor H, vitamin-D-binding protein, voltage-dependent anion-selective channel protein 3 and clusterin were considerably increased in plasma sample of BS patients, whereas alpha-1-antitrypsin, fibrinogen and angiotensinogen were considerably decreased; moreover, post-translational modifications of antithrombin-III were detected in all affected individuals. On the light of these results, we hypothesize that these proteins might be considered as potential markers for the identification of disease status in BS.

Transthoracic echocardiographic guidance of patent ductus arteriosus occlusion with an Amplatz® canine duct occluder.

Placement of an Amplatz canine duct occluder (ACDO) is usually performed by fluoroscopy (Nguyenba and Tobias, 2007). The latter technical approach presents limitations, mostly due to radiation exposure, making this practice dangerous for the patient and operators. In this study, we describe the successful placement of an Amplatz Canine Duct Occluder device by using transthoracic echocardiographic (TTE) guidance, performed on an 11-month-old female mongrel dog with a grade VI/VI continuous heart murmur diagnosed with patent ductus arteriosus (PDA). The TTE is useful in eliminating exposure to radiation and is more versatile than fluoroscopy in conveying real-time detailed information concerning the position of the ACDO.

Proteomic analysis in canine leishmaniasis.

The aim of this study was to perform a proteomic analysis on serum of dogs naturally infected with Leishmania parasite. Sera from 24 dogs, n. 8 with high IFAT titre of anti-Leishmania antibodies (>or= 1:640), n. 8 with uncertain titre (= 1:40), and n. 8 with IFAT negative were used. Sera of each group were pooled together to form three pools: P (high titre); U (uncertain titre); and N (negative). The P pool was analyzed, using a mass spectrometry-based approach to search for Leishmania proteins (qualitative analysis). In a second experiment, protein signal intensities of U and P pools were compared with the signal intensities of N pool by a quantitative mass spectrometry method based on isotopic dilution. The quantitative analysis detected a total of 70 proteins, of which 17 and 5 resulted over- and under-represented in sample P, respectively.

In-line coupling of low-pressure short capillary electrochromatography columns to electrospray ionisation mass spectrometry.

A new in-house designed and constructed injection valve for capillary electrochromatography (CEC) based on a rotating injection part with compartments for the eluent as well as for the sample has been coupled to a mass spectrometer via a sheath flow electrospray ionisation (ESI) interface, using short capillary columns of 15 cm length. The CEC columns were packed with 3 microm C(18) bonded silica particles, and a mixture of peptides was analysed using an ammonium acetate/acetonitrile eluent. A significant increase in the signal-to-noise ratio was obtained when the peptides were dissolved in water with the same content of organic modifier as in the eluent with an addition of 0.5% (v/v) acetic acid. When the CEC analysis was performed without any additional pressure, the separation current sometimes dropped tremendously due to bubble formation, caused by different permeability in the first and packed part of the column causing an extremely low electroosmotic flow. The separation current was restored to its original value by applying only 7 bar at the inlet of the CEC column, and the separation performance for the test peptides was recovered. A comparison of the CEC performance of peptides in pure CEC mode and in low-pressure CEC mode is reported.

Capillary electrochromatography/nanoelectrospray mass spectrometry for attomole characterization of peptides.

The successful coupling of capillary electrochromatography (CEC) to an ion trap mass spectrometer via a nanoelectrospray interface (nESI) is described. Using a conductively coated tip butted to the end of a CEC column, it was possible to obtain a stable spray without any sheath liquid being employed. Selected small peptides were separated with CEC columns (100 microm i.d./25 cm long) packed with 3 microm Hypersil C8 or C18 bonded silica particles with an eluent composed of ammonium acetate/acetonitrile. Peptide mixtures of desmopressin, peptide A, oxytocin, carbetocin and [Met(5)]-enkephalin were detected in the mid-attomole range, which is the lowest amount analyzed using CEC combined with MS detection. It was also observed that sensitivity can be compromised at higher separation voltages. We demonstrate that CEC/nESI-MS, at the current stage of development, represents one of the most sensitive systems for peptide analysis.

Tratamiento mínimamente invasivo del hematoma subdural crónico del adulto. Resultados en 116 pacientes / Minimally invasive treatment of chronic subdural hematoma in adults. Results in 116 patients

Introducción: El hematoma subdural crónico del adulto (HSDCA) tiene una incidencia bruta global en la población cautiva de nuestra institución (pacientes capitados y planes) de 14,1 casos por 100.000 habitantes/año. No existe un protocolo único de tratamiento. En nuestro hospital, dada la edad avanzada de nuestros pacientes, optamos por una terapia mínimamente invasiva (punción transósea [PTO]) sin anestesia general. El objetivo es realizar un estudio descriptivo de pacientes con HSDCA, analizar la lateralidad y los resultados del tratamiento efectuado. Material y métodos: Se detectaron en forma retrospectiva pacientes con HSDCA internados entre enero de 1998 y mayo de 2009 inclusive. El diagnóstico se basó en neuroimágenes. El tratamiento de elección fue la PTO, realizándose por excepción trepanación o craneotomía. Resultados: Se identificó a 127 pacientes. La edad, el sexo, el desplazamiento de línea media, los días de internación y el número de procedimientos no fueron factores predictivos de letalidad a los 30 días del diagnóstico. Un 80% de los pacientes recibió PTO como único procedimiento. La mediana de internación estos pacientes fue de 4 días. El 55% de los hematomas fueron izquierdos, sin diferencia estadísticamente significativa. Se registraron 6 defunciones en internación (4,7%.). La letalidad acumulada a los seis meses fue del 11,8%. La escala de Markwalder al ingreso no fue predictiva de letalidad estadísticamente significativa. Conclusiones: Los resultados de nuestro estudio sugieren que la PTO es un procedimiento aceptable, de baja letalidad, con escasos días de internación y complicaciones. La letalidad, los mecanismos asociados, la edad, el sexo y el desplazamiento de la línea media no difieren de lo publicado en las diferentes series. Encontramos una mayor frecuencia de hematomas hacia la izquierda, al igual que en las diferentes series, debería realizarse un estudio meta-analítico para determinar con mayor precisión la frecuencia de este predominio (AU)

Introduction: Chronic subdural hematoma in adults (CSDH) has a global crude incidence of 14.1/100,000 per year in our institution captive population. There is no single treatment protocol. In our hospital we choose a minimal invasive technique (trans-marrow puncture) without general anaesthesia due to the age of the population. A descriptive study of patients with CSDH and treatment results, including a laterality analysis, is presented. Material and methods: We retrospectively searched patients with (CSDH) between January 1998 and May 2009. The diagnosis was made by neuroimaging techniques in all patients. The preferred treatment was trans-marrow puncture; exceptionally some patients were treated by burr holes or craniotomy. Results: We found 127 patients. Age, gender, midline displacement, hospitalisation days, and number of procedures, were not a predictive factor of mortality in the first month. A slight majority (55%) of CSDH were on the left side, with no statistically significant difference. There were 6 (4.7%) deaths during hospitalisation. In our series cumulative mortality at six months was 11.8%. Markwalder scale at admission was not a predictive factor of statistically significant mortality. An 80% of the patients received trans-marrow puncture as single procedure was performed on 80% of the patients. Conclusions: The results of our study suggest that trans-marrow puncture is an acceptable procedure, with low mortality, and less hospitalisation days and complications. Mortality, associated mechanisms, age, gender, midline displacement are no different than in others previous publications. We found a higher frequency of hematomas to the left, as in other series. Metaanalysis studied need to be performed to determine more accurately the frequency of this dominance (AU)

Tratamiento mínimamente invasivo del hematoma subdural crónico del adulto. Resultados en 116 pacientes. Respuesta de los autores / Minimally invasive treatment of chronic subdural haematoma in adults. Results in 116 patients. Response by the authors

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