Population differentiation in the context of Holocene climate change for a migratory marine species, the southern elephant seal.

Understanding observed patterns of connectivity requires an understanding of the evolutionary processes that determine genetic structure among populations, with the most common models being associated with isolation by distance, allopatry or vicariance. Pinnipeds are annual breeders with the capacity for extensive range overlap during seasonal migrations, establishing the potential for the evolution of isolation by distance. Here, we assess the pattern of differentiation among six breeding colonies of the southern elephant seal, Mirounga leonina, based on mtDNA and 15 neutral microsatellite DNA markers, and consider measures of their demography and connectivity. We show that all breeding colonies are genetically divergent and that connectivity in this highly mobile pinniped is not strongly associated with geographic distance, but more likely linked to Holocene climate change and demographic processes. Estimates of divergence times between populations were all after the last glacial maximum, and there was evidence for directional migration in a clockwise pattern (with the prevailing current) around the Antarctic. We discuss the mechanisms by which climate change may have contributed to the contemporary genetic structure of southern elephant seal populations and the broader implications.

Microbiological polyphasic approach for soil health evaluation in an Italian polluted site.

The use of microorganisms as bioindicators of soil health is quite a new feature, rarely considered for the soil health evaluation in chronically-polluted industrial sites, and still suffering of the bias related to the technique applied. In this work we applied a microbiological polyphasic approach, relying on soil indigenous microorganisms as bioindicators and combining culture-dependent and -independent methods, in order to evaluate soil health of four sites (1a, 1b, 2a and 2b) inside a chemical factory with a centenary activity. Functional as well as structural aspects were comprehensively considered. Results were related to the kind of pollutants found in each site. Heavy metal pollution was recorded in sites 1b and 2b, while both organic and inorganic substances were detected in sites 1a and 2a. Based on the chemical and physical properties of the four soils, site 1b and 2b grouped together, while 1a and 2a were separated from the others. The density of the culturable bacteria was very low in site 2a, where only gram-positive were found. According to the identification of culturable bacteria, site 2a showed the lowest similarity with the other sites. Microbial activity was detected only in sites 1b and 2b. PCR-DGGE (Denaturing Gradient Gel Electrophoresis), was performed on the culturable, total and active microbial communities. Consistently with the identification of culturable bacterial strains, the molecular profile of the culturable fraction of site 2a, was clearly separated from the molecular profiles of other sites in cluster analysis. Molecular fingerprintings of the whole and active bacterial communities differed among the sites, but clustered according to the pollutants present in each site. The presence of possible key species in each site has been discussed according to the whole and active species. Since the results obtained by microbiological analysis are consistent with the chemical data, we suggest that the use of this microbiological polyphasic approach and of microorganisms as intrinsic bioindicators, can be suitable for the evaluation of soil health.

Comparison of two quantitation methods in HPLC: standardless versus calibration with external standards. Application to the analysis of amino acids in fruit juices.

A distinctive method of high-performance liquid calibration peak-area quantitation, called the standardless or absolute method, allows the absolute number of moles of the main amino acids found in twenty-six commercial fruit juices to be determined. The theory of the standardless model is briefly illustrated. The results found with this alternative method are compared with the reference procedure (calibration with external standards) by using difference plots and regression analysis. The presence of both systematic, constant, and proportional errors between the two methods is disclosed and discussed.

Impact of Bt corn on rhizospheric and soil eubacterial communities and on beneficial mycorrhizal symbiosis in experimental microcosms.

A polyphasic approach has been developed to gain knowledge of suitable key indicators for the evaluation of environmental impact of genetically modified Bt 11 and Bt 176 corn lines on soil ecosystems. We assessed the effects of Bt corn (which constitutively expresses the insecticidal toxin from Bacillus thuringiensis, encoded by the truncated Cry1Ab gene) and non-Bt corn plants and their residues on rhizospheric and bulk soil eubacterial communities by means of denaturing gradient gel electrophoresis analyses of 16S rRNA genes, on the nontarget mycorrhizal symbiont Glomus mosseae, and on soil respiration. Microcosm experiments showed differences in rhizospheric eubacterial communities associated with the three corn lines and a significantly lower level of mycorrhizal colonization in Bt 176 corn roots. In greenhouse experiments, differences between Bt and non-Bt corn plants were detected in rhizospheric eubacterial communities (both total and active), in culturable rhizospheric heterotrophic bacteria, and in mycorrhizal colonization. Plant residues of transgenic plants, plowed under at harvest and kept mixed with soil for up to 4 months, affected soil respiration, bacterial communities, and mycorrhizal establishment by indigenous endophytes. The multimodal approach utilized in our work may be applied in long-term field studies aimed at monitoring the real hazard of genetically modified crops and their residues on nontarget soil microbial communities.

High-performance liquid chromatographic analysis of free amino acids in fruit juices using derivatization with 9-fluorenylmethyl-chloroformate.

A simple, rapid, and reliable reversed-phase high-performance liquid chromatographic method for the analysis of 16 amino acids of main interest in commercial fruit juices (pear, orange, grapefruit, pineapple, peach, and apricot) is described. No sample cleanup is required. The pH of the fruit juices is adjusted to alkaline value (8.5) using 200 mM borate buffer, then amino acid is converted to stable derivatives using 9-fluorenylmethyl-chloroformate. The excess of derivatization reagent is removed by a hydrophobic amine, 1-amino-adamantane hydrochloride. The derivatization procedure is simple, fast, and described in detail. Amino acids are detected at 263 nm and eluted within 35 min. The calibration, precision (< or = 6.1%), and recovery (102% +/- 4%) of the method are reported. The conditions of separation are optimized; however, serine partially overlapped with aspartic acid. The amino acid profile of fruit juices is consistent with data from the literature.

Removal of ochratoxin A in red wines by means of adsorption treatments with commercial fining agents.

The presence in wine of the fungal metabolite, ochratoxin A (OTA), represents a serious risk for consumer health. A variety of fining agents, including activated carbon, silica gel, potassium caseinate, egg albumin, and gelatin, was evaluated in relation to their abilities to remove OTA in fortified wines. Freundlich adsorption isotherms were used to model the adsorption behavior between ochratoxin A and the fining agent. Potassium caseinate and activated carbon were found to be the best fining agents that could be used to remove OTA in wine. Potassium caseinate removed up to 82% of OTA when used at 150 g/hL, whereas activated carbon showed the highest specific adsorption capacity due to a high surface area per mass and low adsorption of total polyphenols.

Comparison of different immunoaffinity clean-up procedures for high-performance liquid chromatographic analysis of ochratoxin A in wines.

Three immunoaffinity clean-up procedures to analyse ochratoxin A (OTA) in wines were compared. The direct wine clean-up with Ochraprep and OchraTest columns gave equivalent results in terms of recovery and precision if compared with the reference procedure involving a preliminary extraction of OTA with chloroform. OTA quantification limit in wine ranged from 0.020 to 0.045 microg/l. The 'on-flow' OTA emission spectrum (excitation 333 nm) showed a maximum at 460 nm and could be used to confirm the quantitative results. The analysis of 11 red and white wines gave no significant quantitative differences between the three clean-up techniques.

[Importance of dual isotope myocardial tomoscintigraphy in the detection of coronary disease in the graft among 96 heart transplant recipients]. / Intérêt de la tomoscintigraphie myocardique double isotope dans la détection de la maladie coronaire du greffon chez 96 transplantés cardiaques.

With a survival rate of 70% at 3 years, cardiac transplantation is the best treatment for end-stage heart disease. However, progressive development of graft atherosclerosis is frequent. Diagnosis of transplant coronary disease remains difficult and non-invasive tests have proved relatively insensitive. Therefore, coronary angiography performed annually is still the gold-standard test for the detection of heart transplant vasculopathy. We analyzed the records of 96 patients (82 men and 14 women) who were transplanted from 1986 to 1996. Mean age was 53 +/- 2.7 and time elapsed from transplantation was mean 5.3 +/- 10 years. All patients had rest myocardial TI 201 perfusion SPECT, followed by MIBI gated SPECT after exercise. MIBI gated SPECT allows simultaneous evaluation of perfusion, regional LV function and global ejection fraction. Angiocoronarography, performed in all patients during the six months following radionuclide investigation, showed the presence of coronary heart vasculopathy in nine (9.3%). Seven of these patients had abnormal dual isotope imaging and 2 of them had normal perfusion but altered LV regional function. Sensitivity of dual isotope scintigraphy was 77% and specificity was 97.7%. Dual isotope scintigraphy is helpful to detect coronary vasculopathy in heart transplant recipients and may reduce indications of angiocoronarography.

Central alveolar hypoventilation (Ondine's curse) treated with negative pressure ventilation.

A 22 year old woman with central alveolar hypoventilation (CAH) underwent repeated nocturnal polysomnographic recordings to assess the efficacy of negative pressure ventilation (NPV) administered by pneumowrap. The NPV successfully prevented hypoxia during most of the recording time; however, a sudden severe hypoxic episode not related to an apnoea occurred during a recording night with NPV. This made home use advisable together with an ear oximeter fitted with an alarm.

Pancreatic serous cystadenomas. Report of 8 cases with a mean follow up of 7 years.

Serous cystadenoma of the pancreas are rare tumors and have little or no malignant potential. We report our experience in the management of eight cases of these tumors in the last 22 years. All the patients were women with a mean age of 59 years. All the cysts caused symptoms. Ultrasound and CT-scan were useful in the diagnosis of the pancreatic cystic tumor out not in determining the nature of these lesions clear. FNA-biopsy was performed in 6 cases but in only case was the diagnosis confirmed. All tumors were resected. Four radical pancreatoduodenectomies, two distal pancreatectomies and two cystectomies were performed. Mean followup was 83.5 months. All patients are alive and with no signs of recurrence. Complications include an external pancreatic fistula, an acute cholangitis and a case of delayed gastric emptying. In all cases the histological diagnosis was serous cystadenoma of the pancreas. We conclude that resection of these tumors is mandatory although they are supposed to be benign, in order to avoid complications and because malignant transformations has been related to nonresective treatment.

Thrombotic thrombocytopenic purpura associated with primary tuberculosis.

Thrombotic thrombocytopenic purpura (TTP) during infectious diseases is a known, but rare event. In this paper a case of TTP associated with primary infection by Mycobacterium tuberculosis is described. Various therapeutic approaches were used with the patient: fresh frozen plasma infusions and plasma exchange, specific anti-tuberculous therapy, anti-platelet drugs and steroids. A complete remission occurred 3 months after the onset of the acute disease. A hypothesis on the pathogenesis of TTP might be an increased pro-coagulant activity of interleukin 1 (IL-1) on endothelial cells.

External negative pressure ventilation techniques.

External ventilation was introduced at the beginning of the century to treat acute and chronic respiratory failure. Long-term negative pressure ventilation (NPV) has proved useful in patients with respiratory failure, secondary to a restrictive impairment of neuromusculoskeletal origin. Although NPV may be successfully used in acute-on-chronic respiratory failure in patients with chronic airflow obstruction (CAO), its use in the long-term management of this type of patient seems much less promising. The various types of ventilators, iron lung, pneumo-wrap and cuirass are discussed. All these prostheses, except for the pneumo-wrap, are connected to a pump able to generate negative and positive pressure. NPV must be a controlled ventilation, because an effective trigger is not yet available. The most important feature of the ventilator must be the ability to vary the ventilatory pattern to adapt the machine drive to the patient drive. The goal of NPV is not only to normalize gas exchange, but also to restore an effective spontaneous ventilation. This kind of ventilation has no negative effects on haemodynamics, because it does not vary physiological gradients of transthoracic pressure. However, due to lack of coordination between the respirator and the pharyngeal muscles of the patient, obstructive apnoeas may occur during NPV in sleeping patients.

Lymphangioleiomyomatosis: clinical course.

Pulmonary lymphangioleiomyomatosis (LAM) is a rare disease of smooth muscle proliferation that generally leads to death, due to respiratory failure, within 10 yrs. The disease almost exclusively affects women of child-bearing age and rarely in postmenopausal years. High resolution computed tomographic (CT) patterns are characteristic and almost pathognomonic for LAM. Attempts to treat this disease with hormonal therapy have shown that a number of hormonal manipulations may not be helpful, but that surgical or radiant oophorectomy alone is the most effective treatment, even if the disease is so rare as to preclude randomized trials. Castration, as a means of early therapy, has been associated with a stable respiratory picture in our patients.

Breathing disorders during sleep in chest wall diseases.

Chest wall deformity due to kyphoscoliosis may present with two altered breathing patterns during sleep; 1) progressive O2 desaturation particularly during REM sleep and 2) central apnoea also during REM sleep but rarely accompanied by O2 desaturation. No typical breathing pattern abnormality is seen in the rigid spine syndrome.

Intermittent negative pressure ventilation in patients with restrictive respiratory failure.

Thirty one patients in stable respiratory failure (arterial oxygen tension (PaO2) 67 +/- 20 mmHg (8.9 +/- 2.7 kPa) and arterial carbon dioxide tension (PaCO2) 59 +/- 10 mmHg 7.9 +/- 1.3 kPa)), secondary to non-obstructive ventilatory impairment, were treated by intermittent negative pressure ventilation (INPV), using a pneumowrap or poncho during the night. Daytime arterial blood gas measurements, taken before and after the ventilation, revealed a substantial improvement in PaO2, PaCO2 and maximal inspiratory pressure (Pimax). This improvement persisted during the six month follow-up of home nocturnal ventilation. The polysomnographic, recording during mechanical ventilation by poncho, in five patients, showed a general improvement in the quality and structure of sleep. All patients returned to normal arterial oxygen saturation (SaO2) values (> 90%), except for one. In three patients, the appearance of obstructive events was noted but the desaturation that they caused was, remarkably, lower than that caused by central events in baseline recording. We conclude that non-invasive treatment by INPV, in patients with neuromuscular and chest wall disease, is the method of choice as an alternative to intermittent positive pressure ventilation and to tracheostomy.

Suppression of ventilatory muscle activity in healthy subjects and COPD patients with negative pressure ventilation.

We evaluated the ability of NPV to suppress EMGd and EMGint in seven patients with severe COPD and five normal subjects. Subjects were studied either without (A) or with mouthpiece and nose clip (B). Electromyographic suppression was assessed comparing EMG activity during NPV with the control activity without a mouthpiece and prior to the initiation of the NPV run. In normal subjects, in A, NPV resulted in a partial suppression of EMGd; in B, prior to NPV, EMGd rose compared with A prior to NPV. In patients, in A, NPV resulted in a suppression of both EMGd and EMGint. In B, prior to NPV, both EMGd and EMGint rose compared with A prior to NPV. Thus, it seems that NPV is able to produce a consistent reduction in inspiratory muscle EMG activity. This variable NPV ability would have to be assessed for better selection criteria for patient candidates in a rehabilitation program.