Chlamydia pneumoniae and Mycoplasma pneumoniae in young children from China with community-acquired pneumonia

Eighty-five cases community-acquired pneumonia (CAP) in children 5 years or younger, confirmed by chest X-ray, and 185 age-matched control patients with diarrhea or dermatitis from the Outpatient Department at Beijing Children's Hospital were enrolled into this study. Nasopharyngeal swab specimens were obtained from all subjects. Real-time PCR-based fluorescence assays were performed for Chlamydia pneumoniae and Mycoplasma pneumoniae. A nested PCR was also run for C. pneumoniae for comparison of assays. C. pneumoniae was found in 3 (3.5%) of CAP cases and in 4 (2.1%) of controls (P = 0.51). M. pneumoniae was found in 6 (7.1%) of CAP cases and in none of the controls (P = 0.001). The agreement rate of the 2 applied PCR methods used for C. pneumoniae detection was 98.5%. Our study demonstrates that M. pneumoniae may play a significant role in CAP affecting children up to 5 years in China, whereas C. pneumoniae in nasopharyngeal specimens was not associated with CAP in this age group.     

Study of the diurnal variation of human lymphocyte subsets

The diurnal variation of peripheral white cells and in particular of different lymphocyte subsets has been investigated in 11 normal subjects. Monoclonal antibodies (Mab) phenotyping total T lymphocytes (UCHT1), cytotoxic/suppressor (UCHT4), helper T cells (Leu 3a), B lymphocytes (231) and K/NK cells (H25) have been used. In addition to the traditional parametric statistics, the chronobiological approach of cosinor was adopted in order to inferentially obtain the waveform covering the 24-hr period. Cosinor analysis validated the existence of a significant rhythm for leucocytes (p = 0.04, lymphocytes (p = 0.03), and Leu 3a positive cells (p = 0.04). The best fitting period was equal to 28 hr. No circadian rhythm was documented for lymphocyte subsets positive for Mab UCHT4, 231, H25, by testing the variability with a frequency ranging from 1 cycle every 12hr to 1 cycle every 28 hr. By the non-inferential analysis of temporal curves, a significant peak of Leu 3a positive cells at 20.00 hr (p less than 0.001) was observed. The acrophase has been recorded by cosinor analysis at 20.36 hr. We conclude that the circadian variability of Leu 3a positive cell profile must be taken into account when the helper/suppressor ratio is calculated. The time-dependence of this lymphocyte subpopulation suggests that when therapy with corticosteroids is considered in autoimmunity, it should be planned by keeping the peak of Leu 3a positive cells as a marker time.     

Ethyl glucuronide and ethyl sulphate determination in serum by liquid chromatography-electrospray tandem mass spectrometry

BACKGROUND: Ethyl glucuronide (EtG) and ethyl sulphate (EtS) are two ethanol metabolites that can be detected in serum up to 8 h after ethanol elimination. Their presence is therefore indicative of recent ethanol consumption in case of delayed sampling after an event (e.g. car crash). METHODS: A LC-ESI-MS-MS method for the determination of EtG and EtS in serum was developed and validated. Two product ions together with the parent ions were monitored for identification. Pentadeuterated-EtG was used as internal standard. RESULTS: Excellent linearity for EtG (from 0.22 to 45 micromol/l) and EtS (from 0.40 to 80 micromol/l) was observed (r(2)>or=0.9998). LOD and LLOQ were 0.04 and 0.20 micromol/l for EtG and 0.08 and 0.40 micromol/l for EtS, respectively. Accuracy (bias) and precision (relative standard deviation), studied at four different quality control levels, were always better than 7%. Matrix effects were found to be negligible. The method was applied to several samples obtained from known alcoholics and social drinkers. CONCLUSIONS: A sensitive and specific determination of EtG and EtS in serum samples was achieved despite a simple and fast sample preparation. To our knowledge, this is the first fully validated method for the simultaneous determination of the two alcohol metabolites in serum.     

A cytostatic drug improves control of HIV-1 replication during structured treatment interruptions: a randomized study

OBJECTIVE: To study the effect of highly active antiretroviral therapy (HAART) with and without hydroxyurea (HU) on changes in plasma viral load (VL) set-point, and on HIV-1-specific responses, after five cycles of structured treatment interruptions (STI). METHODS: A group of 20 patients taking HAART for chronic HIV infection with VL < 20 copies/ml were randomized to continue HAART or HAART plus HU for 24 weeks followed by five STI cycles. HU was also stopped in cycles 1-3 but continued in cycles 4 and 5. The number of individuals maintaining a VL set-point < 5000 copies/ml during the fifth interruption were determined. RESULTS: VL remained < 5000 copies/ml in eight out of nine patients in the HU group and in four out of ten patients in the HAART group after a median 48 weeks of follow-up after the fifth interruption ( P=0.039). By STI cycle 5, there was a significant increase in the neutralizing activity (NA), in both magnitude and breadth of the total cytotoxic T lymphocyte (CTL) response and in lymphoproliferative response (LPR) from baseline. No significant differences were observed between HAART and HU groups in NA, CTL and LPR at any time-point. There were no differences in the NA titers at any time-point between responder and non-responder patients. There was a trend for higher CTL and LPR levels in responder patients (P= 0.10). CONCLUSIONS: In this randomized, controlled study of STI with cycles of HAART or HAART plus HU, a lower peak VL rebound and a lower VL set-point was achieved in patients continuing HU while other drugs were discontinued. HU did not blunt anti-HIV-1-specific responses; however, control of VL did not correlate with anti-HIV-1-specific cellular immune responses.     

Autologous tandem transplantation in patients with primary progressive or relapsed/refractory lymphoma

Patients with primary progressive or refractory Hodgkins disease (HD) or aggressive non-Hodgkins lymphoma (NHL) have a particularly poor prognosis. Here we report the results of autologous tandem transplantation in these patients. Patients aged 18–55 years with primary progressive or refractory relapsed HD and aggressive NHL were included. Patients received high-dose etoposide (2000 mg/m²) followed by peripheral blood stem cell harvest (PBSC). The first high-dose chemotherapy (TMC) consisted of thiotepa (750 mg/m²), mitoxantrone (40 mg/m²), and carboplatin (990 mg/m²). Patients with no change (NC), partial remission (PR), or complete remission (CR) after TMC then received BEAM with carmustine (300 mg/m²), etoposide (1200 mg/m²), cytarabine (1600 mg/m²), and melphalan (140 mg/m²). Patients with bulky disease (>5 cm) or residual lymphoma received involved field radiotherapy. Twenty-five patients were included (HD=10, NHL=15, median age 34 years). Two patients with HD achieved a CR and five patients a PR [response rate (RR) 70%]. Three patients (30%) experienced treatment failure including two deaths due to peritransplant complications. Five patients with aggressive NHL were in CR and two patients in PR (RR 46%). Of the eight patients (56%) with treatment failure, three had progressive disease and five died from peritransplant complications. Freedom from treatment failure (FFTF) and overall survival (OS) for all patients after 12 months was 28% and 40%, respectively. Tandem HDCT followed by autologous stem cell transplantation (ASCT) offers a chance of cure in these poor prognostic patients, but is associated with risks.J.-P. Glossmann and J. O. Staak have contributed equally to this work.