Colonization with Helicobacter is concomitant with modified gut microbiota and drastic failure of the immune control of Mycobacterium tuberculosis.

Epidemiological and experimental observations suggest that chronic microbial colonization can impact the immune control of other unrelated pathogens contracted in a concomitant or sequential manner. Possible interactions between Mycobacterium tuberculosis infection and persistence of other bacteria have scarcely been investigated. Here we demonstrated that natural colonization of the digestive tract with Helicobacter hepaticus in mice is concomitant with modification of the gut microbiota, subclinical inflammation, and drastic impairment of immune control of the growth of subsequently administered M. tuberculosis, which results in severe lung tissue injury. Our results provided insights upon the fact that this prior H. hepaticus colonization leads to failures in the mechanisms that could prevent the otherwise balanced cross-talk between M. tuberculosis and the immune system. Such disequilibrium ultimately leads to the inhibition of control of mycobacterial growth, outbreak of inflammation, and lung pathology. Among the dysregulated immune signatures, we noticed a correlation between the detrimental lung injury and the accumulation of activated T-lymphocytes. Our findings suggest that the impact of prior Helicobacter spp. colonization and subsequent M. tuberculosis parasitism might be greater than previously thought, which is a key point given that both species are among the most frequent invasive bacteria in human populations.

Gut microbiota trajectory in pediatric patients undergoing hematopoietic SCT.

Acute GvHD (aGvHD) is the main complication of hematopoietic SCT (HSCT) during the treatment of hematological disorders. We carried out the first longitudinal study to follow the gut microbiota trajectory, from both the phylogenetic and functional points of view, in pediatric patients undergoing HSCT. Gut microbiota trajectories and short-chain fatty acid production profiles were followed starting from before HSCT and through the 3-4 months after transplant in children developing and not developing aGvHD. According to our findings, HSCT procedures temporarily cause a structural and functional disruption of the gut microbial ecosystem, describing a trajectory of recovery during the following 100 days. The onset of aGvHD is associated with specific gut microbiota signatures both along the course of gut microbiota reconstruction immediately after transplant and, most interestingly, prior to HSCT. Indeed, in pre-HSCT samples, non-aGvHD patients showed higher abundances of propionate-producing Bacteroidetes, highly adaptable microbiome mutualists that showed to persist during the HSCT-induced ecosystem disruption. Our data indicate that structure and temporal dynamics of the gut microbial ecosystem can be a relevant factor for the success of HSCT and opens the perspective to the manipulation of the pre-HSCT gut microbiota configuration to favor mutualistic persisters with immunomodulatory properties in the gut.

Multiplex polymerase chain reaction and ligation detection reaction/universal array technology for the traceability of genetically modified organisms in foods.

A multiplex polymerase chain reaction (PCR) system was developed for the simultaneous detection of target sequences in genetically modified soybean (Roundup Ready) and maize (MON810, Bt176, Bt11, and GA21). Primer pairs were designed to amplify the junction regions of the transgenic constructs analyzed and the endogenous genes of soybean (lectin) and maize (zein) were included as internal control targets to assess the efficiency of all reactions. This multiplex PCR has constituted the basis for an efficient platform for genetically modified organism traceability based on microarray technology. In particular, the ligation detection reaction combined to a universal array approach, using the multiplex PCR as target, was applied. High specificity and sensitivity were obtained.

Development of a peptide nucleic acid polymerase chain reaction clamping assay for semiquantitative evaluation of genetically modified organism content in food.

In the present study a peptide nucleic acid (PNA)-mediated polymerase chain reaction (PCR) clamping method was developed and applied to the detection of genetically modified organisms (GMO), to test PCR products for band identity and to obtain a semiquantitative evaluation of GMO content. The minimal concentration of PNA necessary to block the PCR was determined by comparing PCRs containing a constant amount of DNA in the presence of increasing concentration of target-specific PNA. The lowest PNA concentration at which specific inhibition took place, by the inhibition of primer extension and/or steric hindrance, was the most efficient condition. Optimization of PCR clamping by PNA was observed by testing five different PNAs with a minimum of 13 bp to a maximum of 15 bp, designed on the target sequence of Roundup Ready soybean. The results obtained on the DNA extracted from Roundup Ready soybean standard flour were verified also on DNA extracted from standard flours of maize GA21, Bt176, Bt11, and MON810. A correlation between the PNA concentration necessary for inducing PCR clamping and the percentage of the GMO target sequence in the sample was found.

The self selected speed of running in recreational long distance runners.

The aim of this study was to test the hypothesis that the self selected speed in running (vss) is dependent upon the same factors that determine maximal speed in endurance events (e. g. the anaerobic threshold). Experiments were carried out on 8 recreational long distance runners (42.1 +/- 8.6 years of age, 70.1 +/- 10.6 kg of body mass, 1.74 +/- 0.06 m of body height) while they were participating in a 14 day relay race. During the "race" the subjects were not requested to perform maximally but only to cover their running turn (1 hour per day) at their preferred pace. The relationships between heart rate (HR), perceived exertion (RPE), blood lactate concentration ([La]b) and speed (v) were determined in each subject, before the race, during an incremental running test. From these relationships the speed corresponding to a 4 mM concentration of lactate in blood (v4mM) was calculated and found to be 14.3 +/- 1.8 km x h(-1) (n = 8). At this speed the RPE and HR values were 13.6 +/- 1.4 and 156.4 +/- 12.8 bpm, respectively. The average values of speed (vss, 13.4 +/- 0.6 km x h(-1)), RPE (13.5 +/- 1.4) and HR (154.4 +/- 7.6 bpm) measured during the race (n = 47) were not significantly different from those measured at the lactate threshold (v4mM, RPE4mM and v4mM). However, vss and the average HR during the race showed significantly lower variances than v4mM and HR4mM suggesting that, besides the need of avoiding lactate accumulation in blood, other factors must be involved in the choice of speed in running.

Seizures, ventricular tachycardia, and rhabdomyolysis as a result of ingestion of venlafaxine and lamotrigine.

Few cases of overdoses have been described involving venlafaxine, lamotrigine, or a combination of the two agents. We describe a combined venlafaxine and lamotrigine ingestion in a patient presenting with a seizure, ventricular tachycardia, and rhabdomyolysis. We conclude that patients with overdoses that involve venlafaxine can exhibit severe cardiac effects in addition to seizures, especially if venlafaxine is combined with other agents.

Cardiovascular responses to blockade of GABA synthesis in the hypothalamus of the spontaneously hypertensive rat.

Previous studies have suggested that a decreased inhibitory input onto neurons within the posterior hypothalamus (PH), a known pressor area, may contribute to hypertension in the spontaneously hypertensive rat (SHR). Recent experiments from this laboratory have shown that neurons in the PH of the SHR have an altered and elevated discharge frequency compared to those in the normotensive rat. In addition, biochemical studies have reported that there is a decreased concentration of the inhibitory neurotransmitter, GABA, in the hypothalamus of the SHR. The objective of the present study was to assess any variations in GABAergic modulation of cardiovascular activity in SHRs compared to normotensive Wistar-Kyoto (WKY) rats and Sprague-Dawley (SD) rats. Arterial pressure and heart rate responses to microinjections of the GABA synthesis inhibitor 3-mercaptopropionic acid (3-MP) into the posterior hypothalamic area of anesthetized young (6-8 weeks) and mature (11-16 weeks) hypertensive and normotensive rats were recorded. Microinjection of 3-MP elicited increases in arterial pressure of 17.4 +/- 3.9 mmHg, 18.1 +/- 7.8 mmHg, 16.9 +/- 6.4 mmHg, and 10.4 +/- 3.5 mmHg in the mature WKY, mature SD, young WKY, and young SHR, respectively. In addition, heart rate was elevated by 33.2 +/- 21.9 beats/min, 70.0 +/- 25.3 beats/min, 56.3 +/- 15.0 beats/min and, 45.9 +/- 10 beats/min in the mature WKY, adult SD, young WKY, and young SHR groups, respectively. In contrast, microinjection of 3-MP into the posterior hypothalamus of adult SHRs produced no significant change in arterial pressure (-5.0 +/- 1.8 mmHg) or heart rate (+5.3 +/- 6.1 beats/min).(ABSTRACT TRUNCATED AT 250 WORDS)

Hypothalamic GABAergic mechanism involved in respiratory response to hypercapnia.

Previous studies have demonstrated that suprapontine areas of the brain modulate the respiratory responses to hypoxia and hypercapnia. The purpose of the present study was to determine if neurons in the posterior hypothalamus are responsible for this modulation. The respiratory (monitored from diaphragmatic activity) and cardiovascular responses to hypoxia and to hypercapnia were examined in anesthetized rats before and after microinjection of a GABA synthesis inhibitor (3-mercaptopropionic acid, 3-MP) into the posterior hypothalamus. Unilateral micro-injection of 3-MP into the posterior hypothalamus elicited an increase in minute diaphragmatic activity (+54.9 +/- 15.8%), arterial pressure (10.5 +/- 3.2 mmHg) and heart rate (26.8 +/- 10.7 min-1) after a delay of 15-20 minutes. The respiratory responses to hypercapnia but not the cardiovascular responses were greatly accentuated after hypothalamic microinjections of 3-mercaptopropionic acid. In contrast, none of the responses (increases in diaphragmatic activity and heart rate; fall in arterial pressure) elicited by hypoxia were altered after microinjections of the GABA synthesis inhibitor into the posterior hypothalamus. These findings indicate that a GABAergic inhibition of posterior hypothalamic neurons modulates the respiratory response to hypercapnia.