Subtelomeric deletions detected in patients with idiopathic mental retardation using multiplex ligation-dependent probe amplification (MLPA)

Subtelomeric rearrangements are responsible for 5% to 10% of cases of unexplained mental retardation. Despite their clinical relevance, methods to screen for these cytogenetically invisible abnormalities on a routine base are scarce. We screened patients with idiopathic mental retardation for subtelomeric aberrations using multiplex ligation-dependent probe amplification (MLPA). This recently developed technique is based on PCR amplification of ligated probes hybridized to chromosome ends. Currently, 41 telomeres can be screened in just two multiplex reactions. Four subtelomeric rearrangements (5.3%) were detected in a group of 75 patients with mild to severe mental retardation in combination with dysmorphic features and/or a familial history of mental retardation: two terminal 1p deletions, a terminal 1q deletion, and a terminal 3p deletion. Deletions could be verified by FISH and marker analysis. In one case the MLPA indicated a terminal 21q deletion due to a 3-bp deletion at the site of the probe, giving a false-positive rate of 1.3%. This study demonstrates that MLPA is a fast and reliable screening method, potentially suitable for use in routine diagnostics.     

Unexplained chronic fatigue and core conflictual relationship themes: A study in a chronically fatigued population

Objective. Unexplained fatigue syndromes are multidimensional phenomena that involve a constellation of symptoms. This paper explores whether typical relationship patterns are associated with self‐reported and clinically rated fatigue symptoms in chronically fatigued patients. Method. Relationship patterns were assessed by means of the core conflictual relationship theme (CCRT) method. This method examines transference patterns, and was applied to interview data collected from chronically fatigued patients (N=30). Chronic fatigue was assessed by means of a self‐report questionnaire and was also rated clinically. Results. Both self‐reported and clinically rated fatigue correlated with relationship themes. The intensity of fatigue related to the perception of others as not respecting and as negatively interfering. The typical reaction of the self to relationships consists of feeling disrespected, anger, passivity, and reduced feelings of self‐consistency. Conclusion. Patients' perception of interpersonal relationships as distressing may be pivotal in understanding these results. Implications for clinical intervention and future research are indicated.     

Dexamethasone decreases the pro- to anti-inflammatory cytokine ratio during cardiac surgery

Background. Cytokines regulate inflammation associated withcardiopulmonary bypass (CPB). Pro-inflammatory cytokines maycause myocardial dysfunction and haemodynamic instability afterCPB, but the release of anti-inflammatory cytokines is potentiallyprotective. We studied the effects of dexamethasone on pro-and anti-inflammatory cytokine responses during coronary arterybypass grafting surgery. Methods. Seventeen patients were studied: nine patients receiveddexamethasone 100 mg before induction of anaesthesia (group1) and eight patients acted as controls (group 2). Plasma levelsof tumour necrosis factor (TNF)-     

Does sucralfate reduce early side effects of pelvic radiation? A double-blind randomized trial.

STUDY AND METHODS: A double-blind placebo-controlled study randomized 108 patients to investigate the effect of sucralfate on gastrointestinal side effects of pelvic radiation. RESULTS: Overall, pelvic radiation with the administered doses and fields and performed according to nowadays technical standards, was well tolerated. Comparison of the mean scores and the peak reactions for radiotherapy discomfort, diarrhoea and number of stools per day in the 80 evaluable patients showed no statistically significant difference between sucralfate and placebo. CONCLUSION: Based on these results, the use of sucralfate can not be recommended as standard practice.     

Isolation and characterization of tumor-infiltrating lymphocytes from cervical carcinoma

The evidence that virus-induced tumors generally elicit T-cell responses prompts the notion that HPV-related cervical carcinoma would be amenable to treatment by T-cell-mediated adoptive therapy. Therefore, we cultured and cloned tumorinfiltrating lymphocytes (TIL) from a patient with cervical carcinoma and studied the in vitro characteristics of these TIL by using the established autologous tumor-cell line. After stimulation of bulk TIL cultures with 1,000 Units/ml recombinant interleukin 2 (rIL-2), followed by limiting dilution, T-cell clones were generated in the presence of 20 U/ml rIL-2 and irradiated autologous tumor cells, PBLs and EBV-transformed B-cell lines. Phenotypically, all clones were CD3/CD8-positive with a heterogeneous CD56 expression. All expressed preferential cytolytic activity against autologous tumor cells, did not lyse autologous lymphoblasts, and were cytotoxic against the NK-sensitive cell line K562. A minor lytic capacity was detectable on allogeneic cervical tumor-cell lines or tumor-cell lines of other histologic types. Cytotoxicity against the autologous tumor could be inhibited by anti-CD3, anti-CD8 and anti-ICAMI but not by anti-HLA class-1 (W6/32, B9.12.I), anti-allele-specific HLA determinants and anti-LFA-3 antibodies. We demonstrate a highly specific autologous lytic activity of cervical carcinoma TIL, in which a CD3-associated surface antigen recognition is involved. These results may prove useful in further studies on adoptive immunotherapy of cervical cancer patients. © 1994 Wiley-Liss, Inc.     

Nitric oxide modulates renal vasoconstrictor effect of endothelin-1 in conscious lambs

To test the hypothesis that nitric oxide (NO) buffers the renal vasoconstrictor effects of endothelin-1 (ET-1) early in life, renal haemodynamic responses to ET-1 were measured in the presence and absence of endogenously produced NO in conscious lambs. Renal haemodynamic effects of ET-1 were measured for 5 min before (control) and 20 min after intraarterial injection of ET-1 before and after pretreatment with 20 mg/kg of the l-arginine analogue N^G-nitro-l-arginine methyl ester (l-NAME), (experiment 1) and its inactive isomer D-NAME (experiment 2) in conscious lambs aged ~1 week (N=7) and ~6 weeks (N=6). The two experiments were carried out in random order at intervals of 24–48 h. In lambs aged ~6 weeks, a marked increase in renal vascular resistance (RVR) was elicited by ET-1 administration; this response was enhanced twofold following pretreatment with l-NAME. In 1-week-old lambs, however, an increase in RVR in response to ET-1 occurred only after pretreatment with l-NAME. Therefore, we accept our hypothesis and conclude that NO buffers the renal vasoconstrictor effects of ET-1 early in life.The current address of Dr. Liesbeth van der Velde is UCLA Medical Center, Los Angeles, CA, USA. The current address of Dr. Alp Sener is University of Western Ontario, London, ON, Canada     

Inhibition of the development of collagen-induced arthritis in rhesus monkeys by a small molecular weight antagonist of CCR5

OBJECTIVE: Collagen-induced arthritis (CIA) in the rhesus monkey is a nonhuman primate model of rheumatoid arthritis (RA). The close phylogenetic relationship between humans and the rhesus monkey makes this model useful for the preclinical safety and efficacy testing of new therapies that are inactive in animals more distinctly related to humans. In this study, we tested the therapeutic potential of a novel, small molecular weight antagonist of CCR5, SCH-X, in this model. METHODS: CIA was induced in 10 rhesus monkeys. The animals were allocated to receive SCH-X or saline as the control (n = 5 in each group). Treatment was initiated on the day of CIA induction and continued for 45 days. Monkeys were monitored before and 63 days after CIA induction for macroscopic signs of clinical arthritis, such as soft-tissue swelling and body weight. Furthermore, markers of inflammation and joint degradation were monitored to follow the disease course. RESULTS: Only 2 of 5 animals in the SCH-X-treated group displayed prominent soft-tissue swelling, compared with all 5 saline-treated monkeys. In addition to the suppression of joint inflammation, treatment with SCH-X resulted in a reduction in joint destruction, as demonstrated by lower rates of urinary excretion of collagen crosslinks, with confirmation by histology. Whereas in all saline-treated monkeys, marked erosion of joint cartilage was observed, this was absent in 4 of the 5 SCH-X-treated monkeys. CONCLUSION: The systemic effects of treatment with SCH-X were a suppressed acute-phase reaction (reduction in C-reactive protein level) in the 3 treated monkeys with CIA that remained asymptomatic, and an altered antibody response toward type II collagen. The results suggest that the CCR5 antagonist SCH-X might have a strong clinical potential for treatment during periods of active inflammation, as seen in RA.