The utility of a standardised breath sampler in school age children within a real-world prospective study.

Clinical assessment of children with asthma is problematic, and non-invasive biomarkers are needed urgently. Monitoring exhaled volatile organic compounds (VOCs) is an attractive alternative to invasive tests (blood and sputum) and may be used as frequently as required. Standardised reproducible breath-sampling is essential for exhaled-VOC analysis, and although the ReCIVA (Owlstone Medical Limited) breath-sampler was designed to satisfy this requirement, paediatric use was not in the original design brief. The efficacy of the ReCIVA at sampling breath from children has been studied, and 90 breath-samples from 64 children (5-15 years) with, and without asthma (controls), were collected with two different ReCIVA units. Seventy samples (77.8%) contained the specified 1 l of sampled-breath. Median sampling times were longer in children with acute asthma (770.2 s, range: 532.2-900.1 s) compared to stable asthma (690.6 s, range: 477.5-900.1 s;p= 0.01). The ReCIVA successfully detected operational faults, in 21 samples. A leak, caused by a poor fit of the face mask seal was the most common (15); the others were USB communication-faults (5); and, a single instance of a file-creation error. Paediatric breath-profiles were reliably monitored, however synchronisation of sampling to breathing-phases was sometimes lost, causing some breaths not to be sampled, and some to be sampled continuously. This occurred in 60 (66.7%) of the samples and was a source of variability. Importantly, multi-variate modelling of untargeted VOC analysis indicated the absence of significant batch effects for eight operational variables. The ReCIVA appears suitable for paediatric breath-sampling. Post-processing of breath-sample meta-data is recommended to assess the quality of sample-acquisition. Further, future studies should explore the effect of pump-synchronisation faults on recovered VOC profiles, and mask sizes to fit all ages will reduce the potential for leaks and importantly, provide higher levels of comfort to children with asthma.

µ-Opioid Activity in Chronic TMD Pain Is Associated with COMT Polymorphism.

Clinicians have the dilemma of prescribing opioid or nonopioid analgesics to chronic pain patients; however, the impact of pain on our endogenous µ-opioid system and how our genetic profile (specifically catechol-O-methyltransferase [COMT] polymorphisms) impacts its activation are currently unknown. Twelve chronic temporomandibular disorder (TMD) patients and 12 healthy controls (HCs) were scanned using positron emission tomography (PET) with [11C]carfentanil, a selective radioligand for µ-opioid receptors (µORs). The first 45 min of each PET measured the µOR nondisplaceable binding potential (BPND) at resting state, and the last 45 min consisted of a 20-min masseteric pain challenge with an injection of 5% hypertonic saline. Participants were also genotyped for different COMT alleles. There were no group differences in µOR BPND at resting state (early phase). However, during the masseteric pain challenge (late phase), TMD patients exhibited significant reductions in µOR BPND (decreased [11C]carfentanil binding) in the contralateral parahippocampus (P = 0.002) compared to HCs. The µOR BPND was also significantly lower in TMD patients with longer pain chronicity (P < 0.001). When considering COMT genotype and chronic pain suffering, TMD patients with the COMT158Met substitution had higher pain sensitivity and longer pain chronicity with a 5-y threshold for µOR BPND changes to occur in the parahippocampus. Together, the TMD diagnosis, COMT158Met substitution, and pain chronicity explained 52% of µOR BPND variance in the parahippocampus (cumulative R2 = 52%, P < 0.003, and HC vs. TMD Cohen's effect size d = 1.33 SD). There is strong evidence of dysregulation of our main analgesic and limbic systems in chronic TMD pain. The data also support precision medicine by helping identify TMD patients who may be more susceptible to chronic pain sensitivity and opioid dysfunction based on their genetic profile.

HPLC & NMR-based forced degradation studies of ifosfamide: The potential of NMR in stability studies.

PURPOSE: The aim of this study is to conduct a forced degradation study on ifosfamide under several stress conditions to investigate the robustness of the developed HPLC method. It also aims to provide further insight into the stability of ifosfamide and its degradation profile using both HPLC and NMR. METHODS: Ifosfamide solutions (20mg/mL; n=15, 20mL) were stressed in triplicate by heating (70°C), under acidic (pH 1 & 4) and alkaline (pH 10 & 12) conditions. Samples were analysed periodically using HPLC and FT-NMR. RESULTS AND DISCUSSION: Ifosfamide was most stable under weakly acidic conditions (pH 4). NMR results suggested that the mechanism of ifosfamide degradation involves the cleavage of the PN bond. For all stress conditions, HPLC was not able to detect ifosfamide degradation products that were detected by NMR. CONCLUSION: These results suggest that the developed HPLC method for ifosfamide did not detect the degradation products shown by NMR. It is possible that degradation products co-elute with ifosfamide, do not elute altogether or are not amenable to the detection method employed. Therefore, investigation of ifosfamide stability requires additional techniques that do not suffer from the aforementioned shortcomings.

HCV infection in Egyptian patients with acute hepatitis.

The diagnostics of community-acquired acute HCV hepatitis in an endemic area was studied in 110 Egyptian patients with acute jaundice. In the first week of the jaundiced period 30 of 110 patients (27.3%) had anti-HCV antibodies. The majority already showed high levels of anti-HCV IgG (25/30), associated with anti-HCV IgM in nine of them. Five patients showed only an HCV IgM reactivity. Seven had also anti-HEV and/or anti-HBV: their jaundice could then be related to an acute infection caused by those viruses. All patients were infected with genotype 4a, in three associated with the 3a. During the follow-up five patients seroconverted for IgG, while their anti-HCV IgM did not show a uniform pattern of reactivity. Patients with positive serology suspected of an acute HCV infection were older than the patients with other acute hepatitis and showed a lower peak of ALT level. Seroconversion during acute hepatitis strongly indicated HCV as the etiologic agent. However, the detection of anti-HCV IgG antibodies in the jaundiced period showed that the majority of patients had already seroconverted to anti-HCV antibodies; in most of them it is possible to hypothesize a reactivation of a chronic HCV infection.

Hepatitis C virus-specific B cell activation: IgG and IgM detection in acute and chronic hepatitis C.

BACKGROUND/AIMS/METHODS: Immunological responses to hepatitis C virus infection have not been fully studied. In an attempt to clarify some immunopathogenetic aspects of B cell activation during acute and chronic hepatitis C virus infection and to identify markers of chronicity or of recovery, the humoral response in hepatitis C virus-infected patients was studied. RESULTS: In children with acute jaundice, with negative markers of acute hepatitis A, B and E, six of 87 (6.9%) had detectable anti-HCV IgM, and only one (1.1%) had detectable anti-HCV IgG. In adults with acute jaundice, with negative markers of acute hepatitis A, B and E, ten of 23 (43.5%) had detectable anti-HCV IgM associated in eight patients with detectable anti-HCV IgG. In chronic hepatitis C virus-infected adult patients, all anti-HCV IgG seropositive, four of 14 (28%) patients had detectable HCV-IgM in serum. In vitro specific antibody production was inducible in a minority of patients. In acute and chronic hepatitis C virus-infection, IgM-HCV serology did not correlate with viremia as detected by polymerase chain reaction. CONCLUSIONS: Therefore, the polymerase chain reaction remains at the moment the only direct marker to demonstrate hepatitis C virus viral replication in patients with acute and chronic hepatitis while anti-HCV IgM analysis alone has only a limited diagnostic value in hepatitis C virus-infection.

HCV infection in patients with primary defects of immunoglobulin production.

We tested for infection with hepatitis C virus (HCV) in 58 patients affected by humoral immunodeficiencies: 43 common variable immunodeficiency (CVI), two hyper IgM syndrome (HIM), two IgG subclass deficiency, four ataxia-telangiectasia (AT), and seven X-linked agammaglobulinaemia (XLA). While the assessment of serum specific HCV antibodies in some of these patients was not informative because of the impairment in specific antibody production, the reverse transcriptase polymerase chain reaction (RT-PCR) assay used to detect serum HCV RNA was a useful method for diagnosing infection. We found that 38% of late onset hypogammaglobulinaemic patients (CVI, HIM or IgG subclass deficiency) had evidence of HCV infection. HCV infection was not detectable in patients with XLA or AT. The majority of our patients had persistent viraemia, and those who underwent liver biopsy showed histological findings of chronic hepatitis. Moreover, we could demonstrate in vitro that eight of 18 HCV-infected patients were actively producing anti-HCV antibodies, despite their impaired antibody production. The high rate of HCV infection in hypogammaglobulinaemic patients could be related to several nosocomial routes of transmission, including intravenous immune globulin administration. Despite the persistent viremia only two patients had cirrhosis and none had hepatocarcinoma.

Submandibular space abscess due to Actinobacillus actinomycetemcomitans.

A submandibular space abscess is reported in which a pure culture of Actinobacillus actinomycetemcomitans was identified. The bacterium may often be overlooked as a pathogen due to its slow growth and its requirement for carbon dioxide for primary isolation. As A. actinomycetemcomitans is often resistant to commonly used antibiotics, proper management is based on careful utilization of microbiologic tests and clinical judgement. In this case prompt surgical drainage and appropriate antibiotic therapy resolved the abscess.