Human CD45RA(-) FoxP3(hi) Memory-Type Regulatory T Cells Show Distinct TCR Repertoires With Conventional T Cells and Play an Important Role in Controlling Early Immune Activation.

Adoptive immunotherapy with regulatory T cells (Treg) is a new option to promote immune tolerance following solid organ transplantation (SOT). However, Treg from elderly patients awaiting transplantation are dominated by the CD45RA(-) CD62L(+) central memory type Treg subset (TregCM), and the yield of well-characterized and stable naïve Treg (TregN) is low. It is, therefore, important to determine whether these TregCM are derived from the thymus and express high stability, suppressive capacity and a broad antigen repertoire like TregN. In this study, we showed that TregCM use a different T cell receptor (TCR) repertoire from conventional T cells (Tconv), using next-generation sequencing of all 24 Vß families, with an average depth of 534 677 sequences. This showed almost no contamination with induced Treg. Furthermore, TregCM showed enhanced suppressive activity on Tconv at early checkpoints of immune activation controlling activation markers expression and cytokine secretion, but comparable inhibition of proliferation. Following in vitro expansion under mTOR inhibition, TregCM expanded equally as well as TregN without losing their function. Despite relatively limited TCR repertoire, TregCM also showed specific alloresponse, although slightly reduced compared to TregN. These results support the therapeutic usefulness of manufacturing Treg products from CD45RA(-) CD62L(+) Treg-enriched starting material to be applied for adoptive Treg therapy.

Clindamycin-primaquine for pneumocystis jiroveci pneumonia in renal transplant patients.

BACKGROUND: Trimethoprim/sulfamethoxazole (TMP/SMX) is considered first-line therapy for pneumocystis jiroveci pneumonia (PCP) in renal transplant patients. Alternatives have not been formally studied. Clindamycin-primaquine (C-P) is effective in HIV-associated PCP, but data in renal transplant patients are lacking. PATIENTS AND METHODS: Retrospective cohort study of 57 consecutive renal transplant patients who developed PCP and were treated with C-P (n = 23) or TMP/SMX (n = 34). RESULTS: A non-significantly higher failure rate was observed in patients on C-P due to lack of efficacy (30.4 versus 20.6%, p = 0.545). The difference was more pronounced in severe PCP (60 versus 37.5%, p = 0.611) and a significantly lower efficacy of C-P was seen when used as salvage therapy. The two patients who had received C-P after not responding to TMP/SMX failed this regimen, but all seven patients who had failed initial treatment with C-P and had been switched to TMP/SMX were cured (p = 0.028). No treatment-limiting adverse reactions were reported for patients on C-P while six patients (17.6%) on TMP/SMX developed possibly related treatment-limiting toxicity (p = 0.071). However, in only two patients adverse events were definitely related to TMP/SMX (5.9%). CONCLUSIONS: Clindamycin-primaquine appears to be safe and well tolerated for treating PCP in renal transplant patients but is probably less effective than TMP/SMX, the standard regimen. However, our data indicates that C-P represents an acceptable alternative for patients with contraindications or treatment emergent toxicities during TMP/SMX use. Notably, TMP/SMX was also acceptably tolerated in most patients. TMP/SMX remains an effective salvage regimen in case of C-P failure.

Use of sibutramine to assist obese women with weight loss can be successful in dietitian-led clinics: another tool in the dietitian's toolbox.

BACKGROUND: This study investigated differences in weight loss outcomes in obese women with Type 2 diabetes (T2D) compared to those without T2D attending a 6-month dietetic led 'Lifestyle' intervention. In those who had failed to reach weight loss goals using the 'Lifestyle' approach, the study further examined the effect on weight loss with the addition of sibutramine (Reductil, Abbott Laboratories, USA) over a 6-month period. METHODS: The study comprised a case-control analysis of data from 38 obese female patients (18 with T2D) from the 'Lifestyle' and 'Pharmacotherapy' clinics attending a tertiary hospital in the UK. RESULTS: In the 'Lifestyle' treatment groups, those with T2D lost significantly less body weight than those without T2D [5.26 kg (4.54%) versus 9.89 kg (9.55%), respectively]. For subjects who had failed to lose weight via the 'Lifestyle' intervention, the addition of sibutramine resulted in a similar weight loss compared to their pair-matched 'Lifestyle' only 'successful' counterparts (9.66% versus 9.55%). CONCLUSIONS: Not all obese women, and those with T2D in particular, will derive benefit from 'Lifestyle' advice and those who are resistant to this treatment approach may be assisted by pharmacotherapy. Dietitians can play a role in identifying those individuals who may benefit from the use of pharmacotherapy.

Neurotoxic properties of cerebrospinal fluid from behaviorally impaired autoimmune mice.

The chronic, lupus-like autoimmune disease in MRL-lpr mice is associated with leucocyte infiltration into the choroid plexus, brain cell death, and deficits in motivated behavior. The presence of lymphoid cells in the ventricular lumen and the increased number of TUNEL-positive cells in periventricular areas led to the hypothesis that immune cells enter into the cerebrospinal fluid (CSF) and induce primary neuronal damage in regions bordering the cerebral ventricles. Using an in vitro approach, we presently examine the possibility that CSF from autoimmune mice is neurotoxic and/or gliotoxic. The CSF and serum from diseased MRL-lpr mice, less symptomatic MRL +/+ controls, and healthy Swiss/Webster mice (non-autoimmune controls) were frozen until their effects on the viability of pyramidal neurons and astrocytes were assessed in a two-color fluorescence assay. Significant reduction in neuronal viability (in some cases as low as 67%) was observed in the co-cultures of hippocampal neurons and astrocytes incubated for 24 h with CSF from autoimmune MRL-lpr mice. The viability of astrocytes did not differ among the groups, and the CSF from autoimmune mice appeared more toxic than the serum. The behavior of MRL-lpr mice differed significantly from the control groups, as indicated by impaired exploration, reduced intake of palatable food, and excessive immobility in the forced swim test. The present results suggest that CSF from the behaviorally impaired lupus-prone mice is neurotoxic and are consistent with the hypothesis that neuroactive metabolites are produced intrathecally in neuropsychiatric lupus erythematosus.

Disease and nonbattle injury related to peacekeeping operations in South America: summary patient care statistics for CABANAS 2000.

Peacekeeping operations and training for peacekeeping missions currently require far more time and personnel from our armed forces than previously. Although literature exists describing mortality, disease and nonbattle injury (DNBI), and medical support for peacekeeping operations in Europe, Africa, the Caribbean, and the Far East, none was found concerning operations and training for peacekeeping in South America. The present retrospective study presents an analysis of DNBI for forces participating in CABANAS 2000, an eight-nation peacekeeping training exercise held in Argentina. The mean DNBI rate for the 6-week period was 4.1 cases/100 personnel/week. Frequently cited causes for service member presentations for medical treatment were respiratory disease (43%), orthopedic disorders and injuries (25.9%), other miscellaneous medical conditions (8.5%), dermatologic complaints (6.9%), and diarrhea and intestinal complaints (6.5%). These findings indicate that peacekeeping operations and training in South America are relatively safe.

Increased TUNEL staining in brains of autoimmune Fas-deficient mice.

Profound changes in brain morphology and behavior coincide with the spontaneous development of systemic autoimmune/inflammatory disease in Fas-deficient MRL-lpr mice. The dendrites atrophy, the density of hippocampal and cortical neurons decreases, and an anxious/depressive-like behavior emerges while lymphoid cells infiltrate into the choroid plexus of MRL-lpr mice. We hypothesized that the inherited lack of the Fas-dependent anti-inflammatory mechanism would lead to unsuppressed immune activity, characterized by reduced apoptosis in the MRL-lpr brain. Using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeled (TUNEL) method as an indicator of apoptosis, a surprisingly high incidence of TUNEL-positive cells was observed in the hippocampus, choroid plexus and periventricular regions of MRL-lpr mice, 5-10-fold higher than that found in the MRL +/+ control brain. Immunostaining with anti-CD3, CD4 and CD8 monoclonal antibodies showed limited overlap between CD-positive and TUNEL-positive cells, suggesting that the dying cells are for the most part (approximately 70%) not T-lymphocytes. Although further characterization of the phenotype of the dying cells and the mechanism of cell death are required, the present results suggest the involvement of a Fas-independent apoptotic process in neurodegeneration induced by systemic autoimmune disease.

Women's experiences of obtaining emergency contraception: a phenomenological study.

Emergency contraception (EC) has been available since 1984 but has been labelled the 'best kept secret' (Winfield, 1995). Because EC was originally termed 'the morning after pill', many people interpreted this literally and missed an opportunity to use the method. More recent publicity has dropped this term and emphasized that the method is effective up to 72 h after unprotected intercourse or contraceptive failure (Burton & Salvage, 1990). Uptake of EC has steadily increased since 1985 but there is still evidence that younger women in particular are least aware of its existence.

Body composition measurement in elite heavyweight oarswomen: a comparison of five methods.

The study was designed to evaluate the range of body composition in elite heavyweight oarswomen as well as the level of agreement between various methods used to measure this variable. Percent body fat was determined at the start of the competitive season by densitometry, taken to represent the reference standard, and measurement of total body potassium, skinfold thicknesses, bioelectrical impedance analysis and body mass index. The athletes were studied within a two week period with all measurements in any individual taken during one morning. We demonstrated a surprisingly large range of percent fat between these oarswomen, 13.6 to 29.3% by densitometry, which was a feature common to all methods. Percent body fat by total body potassium was lower (p < 0.05) while from body mass index higher (p < 0.01) than the reference value from densitometry. Similar methodologies generated significantly different estimates of % fat (SFT1 versus SFT2, p < 0.01 and BIAv versus BIAB, p < 0.01) highlighting the potential problems that may arise with the use of different regression equations to convert primary measurements into % fat. The limits of agreement between various methods were wide and reflect the large variability about the estimated mean bias. Practically this negates the correction of "non reference" values by adding or subtracting the mean difference or bias between the techniques in individuals. These methodological problems need to be considered when setting specific body composition targets for an athlete.

Can we define indispensable amino acid requirements and assess protein quality in adults?

Adult indispensable amino acid (IAA) requirements, as exemplified by lysine, are examined in a metabolic framework in which dietary IAA are needed to replace both regulatory losses (extrinsic and adaptive IAA oxidation) and obligatory losses (intrinsic metabolic consumption). Thus requirements need to be defined as minimal, operational and optimal. For minimal needs, animal studies show different patterns for maintenance and growth, with lower lysine and leucine in the former compared with the latter pattern, which mirrors tissue protein. Reliable human studies also confirm a low minimum lysine requirement, 15 mg/kg or less, compared with growth needs, supporting the 1985 FAO adult value and lower than the value for preschool children proposed as the basis of a scoring pattern for adults by FAO/WHO in 1991. This pattern derives from studies in children depositing considerable nitrogen and is unsuitable for an adult scoring pattern. Stable isotope lysine balance studies do not provide unequivocal support for an increased minimum requirement value for lysine. As for operational requirements, these are higher because of increased regulatory losses due to the higher intakes of IAA in habitual diets and because of the diurnal feeding pattern. Definition of optimal needs requires value judgements of the beneficial metabolic influence of intakes in excess of minimal. This influence (the anabolic drive) is a tangible phenomenon in growing animals, but is yet to be identified in humans. Thus optimal requirements are not yet definable, and there is no suitable scoring pattern to replace the 1985 FAO values, which may accurately reflect minimal needs.

Nitrogen balance and protein turnover during the growth failure in newly born low-birth-weight infants.

Weight gain, nitrogen balance, protein turnover, and energy and protein intakes were measured during the first week of life of 14 low-birth-weight (LBW) infants, 5 small-for-gestational-age (SGA) infants and 9 appropriate-for-gestational-age (AGA) infants enterally fed at rates determined by the infants ability to assimilate feed. Mean gross intakes were 334 KJ and 1.75 g/kg protein; 4 infants were increasing and 10 were losing weight at rates proportional to gross energy and protein intakes and to nitrogen balance (0.031 g protein balance/g wt gain). Rates of protein synthesis and degradation measured by an intragastric infusion of [1-13C]leucine, averaged 14.5 and 15.9 g protein.kg-1.d-1, some 50% higher than previously reported in older preterm infants and not correlated with nitrogen balance. The growth failure of these infants was not associated with inadequate overall rates of protein turnover, but appeared to reflect an influence of the insufficient energy and protein intakes on the high rates of protein turnover, inducing changes in protein balance in either direction through relatively small changes in protein synthesis and/or degradation.

Psychiatric disorders in the families of borderline patients.

The prevalence of psychiatric disorders was studied among the families of hospitalized borderline patients, defined by Gunderson and Singer's criteria, and compared with the families of schizophrenic and depressed control patients. Among borderline probands, 38.3% have a first-degree relative with depression, 25.5% had one with pathological mood swings, and 23.4% had one with "eccentric or peculiar behavior." There was no significant increase in the prevalence of schizophrenia among the relatives of borderline patients. Depression was more prevalent in the families of schizotypal borderlines compared with unstable or mixed-pattern patients. There were no schizophrenic diagnoses among the impaired relatives of schizotypal borderlines. A relationship is suggested between affective disorder and criteria-defined borderline disorders.