Thrombalexins: Cell-Localized Inhibition of Thrombin and Its Effects in a Model of High-Risk Renal Transplantation.

Allograft transplantation into sensitized recipients with antidonor antibodies results in accelerated antibody-mediated rejection (AMR), complement activation, and graft thrombosis. We have developed a membrane-localizing technology of wide applicability that enables therapeutic agents, including anticoagulants, to bind to cell surfaces and protect the donor endothelium. We describe here how this technology has been applied to thrombin inhibitors to generate a novel class of drugs termed thrombalexins (TLNs). Using a rat model of hyperacute rejection, we investigated the potential of one such inhibitor (thrombalexin-1 [TLN-1]) to prevent acute antibody-mediated thrombosis in the donor organ. TLN-1 alone was able to reduce intragraft thrombosis and significantly delay rejection. The results confirm a pivotal role for thrombin in AMR in vivo. This approach targets donor organs rather than the recipient and is intended to be directly translatable to clinical use.

Antibody combination therapy targeting CD25, CD70 and CD8 reduces islet inflammation and improves glycaemia in diabetic mice.

Destruction of pancreatic islets in type 1 diabetes is caused by infiltrating, primed and activated T cells. In a clinical setting this autoimmune process is already in an advanced stage before intervention therapy can be administered. Therefore, an effective intervention needs to reduce islet inflammation and preserve any remaining islet function. In this study we have investigated the role of targeting activated T cells in reversing autoimmune diabetes. A combination therapy consisting of CD25-, CD70- and CD8-specific monoclonal antibodies was administered to non-obese diabetic (NOD) mice with either new-onset diabetes or with advanced diabetes. In NOD mice with new-onset diabetes antibody combination treatment reversed hyperglycaemia and achieved long-term protection from diabetes (blood glucose <13·9 mmol/l) in >50% of mice. In contrast, in the control, untreated group blood glucose levels continued to increase and none of the mice were protected from diabetes (P < 0·0001). Starting therapy early when hyperglycaemia was relatively mild proved critical, as the mice with advanced diabetes showed less efficient control of blood glucose and shorter life span. Histological analysis (insulitis score) showed islet preservation and reduced immune infiltration in all treated groups, compared to their controls. In conclusion, antibody combination therapy that targets CD25, CD70 and CD8 results in decreased islet infiltration and improved blood glucose levels in NOD mice with established diabetes.

Neurodevelopmental effects of the FMR-1 full mutation in humans.

Brain dysfunction is the most important sequelae of the fragile X (FMR-1) mutation, the most common heritable cause of developmental disability. Using magnetic resonance imaging (MRI) and quantitative morphometry, we have compared the neuroanatomy of 51 individuals with an FMR-1 mutation with matched controls and showed that subjects with an FMR-1 mutation have increased volume of the caudate nucleus and, in males, the lateral ventricle. Both caudate and lateral ventricular volumes are correlated with IQ. Caudate volume is also correlated with the methylation status of the FMR-1 gene. Neuroanatomical differences between two monozygotic twins with an FMR-1 mutation who are discordant for mental retardation are localized to the cerebellum, lateral ventricles and subcortical nuclei. These findings suggest that the FMR-1 mutation causing the fragile X syndrome leads to observable changes in neuroanatomy that may be relevant to the neurodevelopmental disability and behavioural problems observed in affected individuals.

An antibody combination that targets activated T cells extends graft survival in sensitized recipients.

Memory T cells are the very essence of adaptive immunity with their rapid and efficient response to antigen rechallenge and long-term persistence. However, it is becoming increasingly evident that when primed with self or transplanted tissue, these cells play a key role in causing and perpetuating tissue damage. Furthermore, current treatments, which efficiently control the naive response, have limited effects on primed T cells. We have used a treatment based on a combination of antibodies specific for molecules expressed by activated T lymphocytes to selectively remove these cells. This approach, which we termed multi-hit therapy, leads to cumulative binding of antibodies to the target T cells and a striking prolongation of skin graft survival in presensitized recipients in a stringent skin transplant model. The findings are consistent with the depletion of graft-specific CD4+ and CD8+ T cells, although other modes of action, such as T-cell regulation and altered migration could play a role. In conclusion, our therapeutic strategy controls primed T cells which are a major driving force in the pathology of many autoimmune diseases and in transplant rejection.

The effects of an intensive lifestyle modification program on sleep and stress disorders.

BACKGROUND: To determine if a lifestyle change program can modify behavior to reduce sleep and stress disorders. METHODS: Analyses are based on 2,624 individuals aged 30 to 80 years from the Rockford, Illinois metropolitan area who completed a lifestyle evaluation at baseline and again after four weeks, following participation in a 40-hour educational course given over a four-week period. Participants receive instruction on the importance of making better lifestyle choices related to making long-term improvements in nutrition and physical activity and they learn ways to improve sleep and reduce stress in their lives. RESULTS: Significant percent decreases were observed in the number experiencing selected sleep or stress disorders from baseline to four weeks later for "sleeps restlessly" (-59%), "suffers from insomnia" (-64%), "feels under pressure" (-37%), "easily emotionally upset" (-52%), and "feels fearful or depressed" (-61%). Experiencing a selected sleep or stress disorder after four weeks among those who had the disorder at baseline was significantly more likely in those not physically active and/or not having lowered their BMI after four weeks. Changes in alcohol consumption and smoking did not significantly contribute to changes in the disorders. Those who failed to lower their coffee/tea use after four weeks were significantly more likely to have a sleep disorder and be easily emotionally upset. CONCLUSIONS: Changes in lifestyle behaviors after attending an educational program significantly reduced sleep and stress disorders in as little as four weeks, primarily explained by decreasing BMI and/or increasing exercise.

PARKSERVICE: Home support and walking aid for people with Parkinson's disease.

PARKSERVICE is a telemedical application currently being validated in the EU. The objectives are to provide a combination of home clinical and social support for people with Parkinson's disease with a revolutionary walking aid that uses "visual cues" to enable improved mobility. Early results are presented and the outlook of home telemedicine and visual cueing for people with PD is discussed.

Ga-67 citrate imaging in malignant lymphoma: final report of cooperative group.

In a large cooperative study of Ga-67 uptake in non-Hodgkin's malignant lymphoma, 76% of untreated patients showed positive uptake in one or more lesions. The percentage of known individual lesions seen on scan was significantly lower; thus, negative findings at any one site may have much less significance than positive findings. After treatment, the number of lesions seen decreases sharply, but the role of Ga-67 in evaluating response to therapy is uncertain, especially in view of the fairly large number of lesions undetectable before therapy. Histologic type plays a role in Ga-67 uptake. Large lesions are much more effectively detected than small ones. In spite of numerous false-negative results, Ga-67 scanning is a useful method in evaluating the extent of untreated disease and the presence of lesions posttherapy.

Studies on the chemopreventive potentials of vegetable oils and unsaturated fatty acids against breast cancer carcinogenesis at initiation.

The effect of dietary fat on breast cancer is a longstanding and an unresolved issue. We found that 17beta-estradiol (E2) could be activated by the epoxide-forming oxidant dimethyldioxirane (DMDO) to bind DNA-forming DNA adducts both in vitro and in vivo, and to inhibit nuclear RNA synthesis. We proposed that E2 epoxidation is the underlying mechanism for the initiation of breast cancer carcinogenesis (Carcinogenesis 17, 1957-61, 1996). This report is on the transcriptional and DNA-binding properties of vegetable oils and fatty acids, and on the potentials of these compounds to prevent the formation of E2 epoxide. The results show that vegetable oils, having no effect on nuclear RNA synthesis either before or after DMDO treatment, were all able to prevent the formation of E2 epoxide independent of their mono- or polyunsaturated fatty acid content. Similarly, unsaturated fatty acids, regardless of chain length and number of double bonds, were all able to prevent the formation of E2 epoxide as reflected by the loss of the ability of [3H]E2 to bind DNA. In contrast to vegetable oils, the results indicated that the unsaturated fatty acids palmitoleic, oleic, linoleic, linolenic and arachidonic acid could be activated by DMDO to inhibit nuclear RNA synthesis, and that the mono-unsaturated fatty acids (i.e. palmitoleic and oleic acid) were stronger inhibitors than fatty acids with more than one double bond (e.g. linoleic, linolenic and arachidonic acid). [32P]Post-labeling analysis revealed that under identical DMDO activation, the DNA adducts formed for oleic acid were 17098 adducts/10(8) nucleotides, which was 20-fold more than palmitoleic acid (815), and 120-fold more than alpha-linolenic acid (142). This result strongly suggests that oleic acid could be a potential initiating carcinogen after epoxidation.

Chemotherapy drug dose alteration due to radiation therapy in an adjuvant situation in breast cancer.

Fifty-six patients with T1-T3 and T2-T3 N0 medial lesions of breast carcinoma were randomized after modified radical mastectomy to receive either cytoxan, methotrexate, 5-fluorouracil (5-Fu) (CMF) chemotherapy for 1 year (group A) or CMF for 1 year and postoperative radiation therapy (group B). Thirty-two patients received chemotherapy alone and 25 patients received both chemotherapy and radiation therapy. Twenty-five of 32 group A patients (median age 52) and 20/24 group B patients (median age 50) were evaluable. Leukopenia was the major cause of drug dose reduction in both groups. In group A, 4/25 patients (16%) had leukopenia at less than the 2500 level, whereas 8/20 (40%) group B patients had leukopenia at the same level. If 70% of all three drug dosages are considered as adequate chemotherapy, 21/25 (84%) patients received adequate chemotherapy in group A, and 10/20 (50%) in group B (p approximately equal to 0.029 from contingency table). It appears that radiation therapy in postmastectomy patients hinders adequate drug dose delivery in an adjuvant setting.

A supervised training program in flexible sigmoidoscopy: evaluating skills from residency training to clinical practice.

To assess the effectiveness of a training program in flexible sigmoidoscopy for family practice residents, we prospectively studied the performance of four residents during their training and after graduation. One hundred and four training exams performed with the assistance of an experienced gastroenterologist were compared with 118 unassisted post-training, post-residency exams. The mean depth of insertion for the post-training period was 51.1 +/- 1.2 cm, which was significantly greater (P less than .05, Student's t test) than the mean training period depth of 47.6 +/- 1.2 cm. There was no significant difference in the identification of polyps or cancer between the training and post-training periods. The mean duration of an exam was 17.3 +/- 0.6 minutes in the post-training period. No significant complications were encountered in either period. The residency trained family physicians obtained results similar to those reported by trained endoscopists in depth of examination and pathology detected, although their examinations required more time. We conclude that this model of training was effective in the development of flexible sigmoidoscopy procedural skill for family practice residents.

The effect of real and virtual visual cues on walking in Parkinson's disease.

Patients with Parkinson's disease (PwPD) have a slow, shuffling gait, marked by sporadic freezing of gait (FoG) during which effective stepping ceases temporarily. As these gait problems are not commonly improved by medical and surgical treatments, alternative approaches to manage these problems have been adopted. The aim of this study was to evaluate the effect of real and virtual visual cues on walking in PD. We assessed 26 mid-stage PwPD, on and off medication, on a laboratory-based walking task which simulated real world challenges by incorporating FoG triggers and using appropriate placebo conditions. Cueing interventions were presented via virtual reality glasses (VRG rhythmic, visual flow and static placebo cues), and as transverse lines (TL) on the walkway. Objective measures of gait (task completion time; velocity, cadence, stride length; FoG frequency) and self-rated fear of falling (FoF) were recorded. Cueing intervention affected task completion time only off medication. Whereas placebo VRG cues provided no improvement in walking, visual flow VRG cues marginally reduced the task completion time. TL on the floor elicited more substantial improvements in gait with reduced cadence, increased stride length and reduced FoG frequency. VRG rhythmic cueing impaired overall walking. Notably, a final no-intervention condition yielded quicker task completion, greater walking velocity, increased stride length and less frequent FoG. Although the VRG produced modest improvements only in the visual flow condition, their flexibility is an advantage. These results endorse the use of TL and justify further testing and customisation of VRG cues for individual PwPD.