Alpha- and gamma-synuclein proteins are present in cerebrospinal fluid and are increased in aged subjects with neurodegenerative and vascular changes.

BACKGROUND: Disease-specific biomarkers should reflect a fundamental feature of neuropathology and be validated in neuropathologically confirmed cases. Several synaptic proteins have been described in cerebrospinal fluid (CSF) of patients with dementia. In Lewy body disease alpha-synuclein is incorporated within Lewy bodies and alpha-, beta- and gamma-synucleins in dystrophic neuritis. These pathological changes are expected to be seen in CSF. METHODS: A total of 25 CSF post-mortem samples (8 control and 17 subjects with dementia) were used to quantify alpha- and gamma-synucleins and IgG. RESULTS: We describe for the first time the presence of gamma-synuclein in CSF. There is an elevation of both alpha- and gamma-synucleins in CSF from elderly individuals with Alzheimer's disease, Lewy body disease (LBD) and vascular dementia (CVD), compared to normal controls. gamma-Synuclein showed a greater elevation in LBD, IgG in CVD. The elevation of alpha- and gamma-synucleins was seen from Braak stage III onwards and remained stable until Braak stage VI. These results were not influenced by age at death or post-mortem delay. CONCLUSIONS: The reported increases in alpha- and gamma-synucleins and IgG in the ventricular CSF of individuals with dementia are novel findings. They now need to be explored further using a greater number of cases in each subgroup, using lumbar CSF samples to determine their applicability and relevance to a clinical diagnostic setting. It needs to be established whether using these markers may help to discriminate LBD from other types of neurodegenerative and vascular dementias.

Increasing selenium concentration in milk: effects of amount of selenium from yeast and cereal grain supplements.

Two experiments were conducted to establish responses in milk Se concentrations in grazing dairy cows to different amounts of dietary Se yeast, and to determine the effects of the Se concentration of the basal diet. The hypothesis tested was that the response in milk, blood, and tissue Se concentrations to supplemental Se would not be affected by whether the Se was from the basal diet or from Se yeast. In addition, by conducting a similar experiment in either early (spring; experiment 1) or late (autumn; experiment 2) lactation, we hypothesized that different Se input-output relationships would result. Both 6-wk experiments involved 60 multiparous Holstein-Friesian cows, all of which had calved in spring. They were allocated to 1 of 10 dietary Se treatments that included 2 types of crushed triticale grain (low Se, approximately 165 microg of Se/kg of DM; or high Se, approximately 580 microg/kg of DM) fed at 4 kg of DM/d, and 1 kg of DM/d of pellets formulated to carry 5 quantities of Se yeast (0, 4, 8, 12, or 16 mg of Se). Daily total Se intakes ranged from <2 to >18 mg/cow in both experiments. Milk Se concentrations plateaued after 15 and 7 d of supplementation in experiments 1 and 2, respectively, and then remained at plateau concentrations. Average milk Se concentrations for the plateau period increased as the amount of Se yeast increased, and low- and high-Se grain treatments were different at all quantities of Se yeast, although there was a tendency for this difference to diminish at the greatest concentrations of yeast. There were significant positive, linear relationships between Se intake and the concentrations of Se in milk, which were not affected by the source of Se, and the relationships were similar for both experiments. Therefore, the output of Se in milk in experiment 1 was greater than that in experiment 2 because the milk yield of the cows in early lactation was greater. The estimated proportions of Se partitioned to destinations other than milk and feces increased with the amount of Se in the diet and were greater in experiment 2 than in experiment 1, a result that was supported by Se concentrations in whole blood and plasma and in semitendinosus muscle tissue. If high-Se products are to be produced for human nutrition, it is important to be able to develop feeding systems that produce milk with consistent and predictable Se concentrations so that products can consistently meet specifications. The results indicate that this objective is achievable.

Rhythmic auditory stimulation improves gait more than NDT/Bobath training in near-ambulatory patients early poststroke: a single-blind, randomized trial.

OBJECTIVES: The effectiveness of 2 different types of gait training in stroke rehabilitation, rhythmic auditory stimulation (RAS) versus neurodevelopmental therapy (NDT)/Bobath- based training, was compared in 2 groups of hemiparetic stroke patients over a 3-week period of daily training (RAS group, n = 43; NDT/Bobath group =35). METHODS: Mean entry date into the study was 21.3 days poststroke for the RAS group and 22.3 days for the control group. Patients entered the study as soon as they were able to complete 5 stride cycles with handheld assistance. Patients were closely equated by age, gender, and lesion site. Motor function in both groups was pre-assessed by the Barthel Index and the Fugl-Meyer Scales. RESULTS: Pre- to posttest measures showed a significant improvement in the RAS group for velocity (P = .006), stride length (P = .0001), cadence (P = .0001) and symmetry (P = .0049) over the NDT/Bobath group. Effect sizes for RAS over NDT/Bobath training were 13.1 m/min for velocity, 0.18 m for stride length, and 19 steps/min for cadence. CONCLUSIONS: The data show that after 3 weeks of gait training, RAS is an effective therapeutic method to enhance gait training in hemiparetic stroke rehabilitation. Gains were significantly higher for RAS compared to NDT/Bobath training.

Whey proteins protect more than red meat against azoxymethane induced ACF in Wistar rats.

Protein type and density have been shown to influence colon cancer risk using a carcinogen-induced rat model. It is suggested that red meat may promote colon cancer risk more than whey proteins. The aim of this study was to evaluate the influence of red meat, whey protein and their density in the diet on the number of aberrant crypt foci (ACF), preneoplastic markers in Wistar rats. The sources of protein, red meat as barbecued kangaroo muscle meat, and whey protein concentrate were fed to rats to provide 8, 16 and 32% protein by weight in a modified AIN-93 diet with low fiber, low calcium and high polyunsaturated fat. Adult Wistar rats (13 weeks of age) were fed these diets for 4 weeks and then two s.c. injections of azoxymethane, 15 mg/kg BW, were administered 1 week apart. Diets were fed for a further 8 weeks, rats were then killed, their colons fixed in formalin saline and stained with methylene blue to quantify ACF number. Fecal samples were collected and the fecal water was isolated for quantification of heme and thiobarbituric acid reactive substances. Increasing red meat density correlated positively, while increasing dairy protein density correlated negatively with rate of weight gain (p<0.05). Dietary intake was not significantly affected by protein type or density. The 32% whey protein group had significantly less ACF in the proximal colon in comparison to the 16 and 32% red meat groups (p<0.05). This reduction in ACF number in the whey protein group may be caused by hormones associated with the reduction in weight gain, and/or by components of whey protein concentrate such as cysteine, lactose and conjugated linoleic acid which have been shown to have anti-cancer effects. Using ACF number as an index, whey protein appeared to be more protective than red meat.

Kinematic optimization of spatiotemporal patterns in paretic arm training with stroke patients.

The effect of rhythmic cueing on spatiotemporal control of sequential reaching movements of the paretic arm was studied in 21 hemispheric stroke patients. Reaching movements were studied with and without rhythmic metronome cuing in a counterbalanced design. Metronome frequencies were entrained to the naturally selected frequency of the patient. Results indicate statistically significant (P<0.05) improvements of spatiotemporal arm control during rhythmic entrainment. Variability of timing and reaching trajectories were reduced significantly. Time series analysis of sequential movement repetitions showed an immediate reduction in variability of arm kinematics during rhythmic entrainment within the first two to three repetitions of each trial. Rhythm also produced significant increases in angle ranges of elbow motion (P<0.05). Analysis of acceleration and velocity profiles of the wrist joint showed significant kinematic smoothing during rhythmic cuing. The link between rhythmic sensory timing and spatiotemporal motor control was investigated using a mathematical optimization model with minimization of peak acceleration as criterion. Rhythmically cued acceleration profiles fit the predicted model data significantly closer (P<0.01) than the self-paced profiles. Since velocity and acceleration are mathematical derivatives of position-time trajectories, the model data suggest that enhanced timing precision via temporal phase and period coupling of the motor pattern to the rhythmic time timekeeper enhances the brain's computational ability to optimally scale movement parameters across time.

Conscious and subconscious sensorimotor synchronization--prefrontal cortex and the influence of awareness.

One of the most compelling challenges for modern neuroscience is the influence of awareness on behavior. We studied prefrontal correlates of conscious and subconscious motor adjustments to changing auditory rhythms using regional cerebral blood flow measurements. At a subconscious level, movement adjustments were performed employing bilateral ventral mediofrontal cortex. Awareness of change without explicit knowledge of the nature of change led to additional ventral prefrontal and premotor but not dorsolateral prefrontal activations. Only fully conscious motor adaptations to a changing rhythmic pattern showed prominent involvement of anterior cingulate and dorsolateral prefrontal cortex. These results demonstrate that while ventral prefrontal areas may be engaged in motor adaptations performed subconsciously, only fully conscious motor control which includes motor planning will involve dorsolateral prefrontal cortex.

The influence of dietary proteins on colon cancer risk.

Experimental evidence is accumulating from animal models and in vitro data which shows that dietary proteins can influence cancer expression, some having a promotional influence, others a preventative effect relative to an arbitrarily established standard diet. This result will to a degree be determined by the nature of the cancer model under study. Dairy proteins have been shown to be relatively protective when compared with defatted soybean meal and cooked red meat in the rat dimethylhydrazine-induced (DMH) colon cancer model. Some epidemiological evidence supports these experimental observations. Both protein and fat appear to be influencing outcome, with potential for interactive effects. A number of possible mechanisms have been postulated as to how these proteins and closely associated factors could be influencing colon cancer risk, an area that deserves more investigation. Combinations of foods such as dairy foods with cereals and/or probiotic bacteria provide potentially interesting alliances in reducing colon cancer risk. The combining of relatively protective agents deserves more investigation as to its potential, in devising functional foods and diets worthy of further evaluation, in animal models of cancer, and human intervention studies using relevant endpoint markers.

Child restraint systems: an update for physicians.

During the past decade, many advances have been made in motor vehicle safety restraint systems, and these advancements have brought changes in the recommendations for child passenger safety. In spite of these advances, a high morbidity and mortality rate continues to be attributed to improper or absent child restraint use. Child transportation safety is a complex issue. Lack of public awareness and incomplete laws contribute to the confusion. The solution involves professional and public education, further technological advances, and advocacy for improved legislation and regulation. Physicians have a unique opportunity to incorporate advice about proper restraint use into their daily practice; however, that requires a clear understanding of the current recommendations. Both the age and weight of the child determine the appropriate child restraint system. Understanding, practicing, and promoting these recommendations will save lives. This article reviews the current recommendations for proper child restraint while traveling in a motor vehicle.

Topoisomerase II alpha and II beta expression in childhood acute lymphoblastic leukaemia: relation to prognostic factors and clinical outcome.

BACKGROUND/AIMS: Many regimens used in the treatment of childhood acute lymphoblastic leukaemia (ALL) include Daunorubicin or Etoposide, which act as topoisomerase poisons. It has been suggested that there may be a relation between topoisomerase expression and response to topoisomerase poisons, based mainly on results from in vitro studies. Therefore, the aim of this study was to investigate this relation in a clinical setting and determine whether topoisomerase II alpha and II beta might be of predictive value in ALL. METHODS: Cellular expression of topoisomerases II alpha and II beta was assessed in 177 cases of ALL by immunohistochemistry using monoclonal antibodies to the two enzymes. The percentages of cell nuclei showing positive staining for topoisomerase II alpha and II beta expression were assessed. RESULTS: Taking the series as a whole, a clear separation of survival curves was seen with the established prognostic markers white blood cell (WBC) count, CD10 status, and sex. However, topoisomerase II alpha and II beta expression showed no relation to survival. No association was found between the topoisomerases and the prognostic markers CD10 and WBC count; however, topoisomerase II alpha expression was found to be related to sex, with expression being lower in girls (p = 0.002). CONCLUSIONS: These results suggest that the response to topoisomerase poisons cannot be predicted by the assessment of topoisomerase II alpha and II beta expression as defined by immunohistochemistry.

Wheat aleurone flour increases cecal beta-glucuronidase activity and butyrate concentration and reduces colon adenoma burden in azoxymethane-treated rats.

Processed wheat aleurone flour (WAF) is a source of insoluble fermentable dietary fiber that comes from the outer layers of the wheat kernel. A study was designed to evaluate WAF, wheat bran (WB) and alpha-cellulose as the source of dietary fiber (5 g/100 g of diet) in a semipurified high fat (20 g/100 g of as 1:1 lard/sunflower seed oil) diet fed to male Sprague-Dawley rats in which intestinal tumors were induced using azoxymethane (AOM). WAF at 33 g/100 g of diet (WAF33) and WB at 16 g/100 g of diet (WB16) increased the weight of feces and produced significantly higher concentrations in the cecum of the short-chain fatty acid butyrate (P < 0.001) than did no fiber (NF) and WAF added at only 10 g/100 g (1.5 g of dietary fiber) (WAF10). Cecal and fecal pH were both significantly lower in the WAF33 and WB16 treatments relative to control and no fiber treatments (P < 0.001). The intestinal tumors in the rats were assessed at 6 mo after the study began, and the WAF33- or WB16-fed rats showed a trend (P = 0.06) with 43% fewer colon adenomas relative to control. There was a significant inverse relationship between ss-glucuronidase activity and colon adenomas in the rat colon (r2 = 0.37, P = 0.001). WAF fiber influenced some metabolic markers of fermentation in the colon in a manner similar to that of WB, which, independent of the bulking effect, was associated with a trend to reduced colon adenomas. Significantly increased cecal ss-glucuronidase activity and/or butyrate concentrations may have protective influences in this context by mechanisms not yet fully elucidated.

Folate deficiency diminishes the occurrence of aberrant crypt foci in the rat colon but does not alter global DNA methylation status.

BACKGROUND AND AIMS: It has been suggested that a diminished folate status may enhance colorectal carcinogenesis by causing DNA hypomethylation. The aims of the present study were to assess the impact of different levels of folate depletion on azoxymethane (AOM)-induced aberrant crypt foci (ACF) formation and DNA hypomethylation in the colon of male Sprague-Dawley rats. METHODS: Rats, aged 4 weeks, were divided into four groups and were fed semipurified diets either containing adequate folate (control), devoid of folate (FD) or FD + 1% succinylsulfathiazole before AOM treatment (FD1) or during the last 4 weeks of the study (FD2). At 8 weeks of age, all animals received subcutaneous injections of AOM once weekly for 3 weeks at a dose rate of 15 mg/kg bodyweight. Animals were necropsied 6 weeks after the last AOM injection and the ACF were visualized under light microscopy in formalin-fixed, methylene blue-stained colons. RESULTS: Blood folate concentrations were significantly depleted (P < 0.001) in the treatment groups consuming folate deplete diets, with the FD2 treatment group having significantly lower folate levels compared with all other groups. Higher plasma homocysteine concentrations (P < 0.001) were observed in the groups that exhibited diminished blood folate levels. There were no significant differences in global DNA methylation in the liver or colonic mucosa between the four groups, despite some groups exhibiting marked folate depletion. Animals with the most severe folate deficiency (FD2) had a lower final bodyweight and had significantly fewer ACF in their colon (P < 0.05) compared with control animals. Total (mean +/- SEM) ACF counts were as follows: control 286+/-24; FD 290+/-25; FD1 218+/-32; and FD2 205+/-27. CONCLUSIONS: In this model, folate deficiency diminished the occurrence of ACF but did not alter global DNA methylation status in the colon.

Dietary butyrate inhibits NMU-induced mammary cancer in rats.

Butyrate has been proposed as an antineoplastic agent, leading to the inhibition of tumorigenesis. The purpose of this study was to examine butyrate, supplied as tributyrin (Tbn) or as a natural component of anhydrous milk fat (AMF), on the development of nitrosomethylurea-induced mammary tumors in female Sprague-Dawley rats. Diets were 1) semipurified rodent diet (AIN-93) with high fat [20% sunflower seed oil (SSO), control], 2) SSO diet with Tbn added at 1%, 3) SSO diet with Tbn added at 3%, and 4) 19% AMF with 1% SSO diet, which contained butyrate equivalent to the 1% Tbn diet. These diets were fed ad libitum from weaning at 21 days of age, and at 24 days of age each rat was injected with nitrosomethylurea (50 mg/kg body wt i.p.). At any one period, there was a relative risk increase of 88% (p < 0.05) that rats in the SSO diet group would develop a mammary tumor compared with those in the AMF diet group. The addition of 1% and 3% Tbn to SSO diets reduced the tumor incidence by 20% and 52%, respectively, in comparison to SSO alone (p < 0.05). There was a linear inverse relationship between Tbn concentration and rats developing a tumor. From 89 days to the end of the experiment, rats fed the diet containing 3% Tbn showed a significantly lower multiplicity of palpable tumors (50% less at Day 118, p < 0.05) than SSO-fed rats. These results indicate that although the AMF diet was effective, particularly early in reducing mammary tumorigenesis, the 3% Tbn diet produced a sustained reduction of tumor multiplicity relative to the control (SSO) group. An inhibitory influence of butyrate on mammary tumorigenesis against a background of high polyunsaturated fat diet has been demonstrated in this animal model of breast cancer.

The molecular epidemiology of four outbreaks of porcine pasteurellosis.

Biochemical profiles, restriction endonuclease analysis (REA) and ribotyping were used to investigate a total of 38 Pasteurella multocida isolates from four separate outbreaks of pasteurellosis in Australian piggeries. Six isolates were obtained from Outbreak 1, 16 from Outbreak 2 and eight each from outbreaks 3 and 4. Outbreaks 1 and 2 were cases of pneumonic pasteurellosis while outbreaks 3 and 4 involved systemic pasteurellosis. Biochemical characterisation established that a number of different types of P. multocida were present in outbreaks 1 and 3 while outbreaks 2 and 4 were associated with a single type of P. multocida. Outbreaks 1 and 3 yielded isolates of P. multocida that belonged to the subspecies multocida and gallicida, with the subspecies multocida isolates being identified as biovar 3 (6 in total) or 12 (1 in total) and the subspecies gallicida isolates (7 in total) being identified as biovar 8. All 24 isolates from outbreaks 2 and 4 belonged to the subspecies multocida and were all biovar 3. REA and ribotyping showed that, in outbreaks 1 and 3, there were three different types of P. multocida in each outbreak with no common strains between the outbreaks. The molecular methods showed that only a single strain of P. multocida was associated with outbreaks 2 and 4, although the outbreaks were associated with strains that differed in REA profiles but shared a ribotype profile. This study has shown that both, systemic and pneumonic pasteurellosis can be associated with either a single strain or multiple strains of P. multocida. The results also indicate that the molecular typing methods of REA and ribotyping are superior to biochemical characterisation for epidemiological investigation of porcine pasteurellosis.

Prognostic factors for time receiving workers' compensation benefits in a cohort of patients with low back pain.

STUDY DESIGN: Prospective inception cohort study. OBJECTIVE: To develop a prognostic model that predicts time receiving workers' compensation benefits for low back pain claimants. SUMMARY OF BACKGROUND DATA: As the cost and difficulty of managing low back pain escalate, any predictor of outcome is advantageous. METHODS: To obtain the outcome and predictor variables, patient data from two separate databases were linked: a clinical database and an administrative (Ontario workers' compensation) database. Claimants injured between January 1 and December 31, 1994, were included and observed for 1 year from the date of accident. The outcome variable was cumulative number of calendar days receiving benefits. RESULTS: Multivariable Cox proportional hazards regression (forward stepwise) showed eight significant predictors; five were associated with increased time receiving benefits compared with their reference groups: 1) working in the construction industry, 2) older age, 3) lag time from injury to treatment, 4) pain referred into the leg, and 5) three or more positive Waddell nonorganic signs. Three predictors were associated with reduced time receiving benefits: 1) higher values of questionnaire score, 2) intermittent pain, and 3) a previous episode of back pain. A predictive score was calculated to categorize claimants as at high or low risk for chronicity. When an arbitrary cutoff point was set at the 75th percentile of predictive score, negative predictive value was 94%. CONCLUSION: This research identified eight factors for time receiving workers' compensation benefits among claimants with low back pain. This model discriminates between high- and low-risk claimants. Few low-risk claimants continued to receive benefits for more than 3 months.

Monoclonal antibodies recognizing CD5, CD10 and CD23 in formalin-fixed, paraffin-embedded tissue: production and assessment of their value in the diagnosis of small B-cell lymphoma.

AIMS: Assessment of the expression of antigens CD5, CD10 and CD23 can be of value in the differential diagnosis of small B-cell lymphoma. Correct subclassification is important since optimal treatment regimes differ between the subtypes. The aim of this study was to generate monoclonal antibodies recognizing these antigens in paraffin-embedded tissue and to assess their efficacy using a panel of cases of small B-cell lymphoma of various subtypes. METHODS AND RESULTS: For each antibody synthetic recombinant protein and conventional murine hybridoma technology was employed. Monoclonal antibodies effective in formalin-fixed, paraffin-embedded tissue were successfully generated, designated NCL-CD5-4C7, NCL-CD10-270 and NCL-CD23-1B12, respectively. A series of 58 cases of small B-cell lymphoma including examples of each subtype (lymphocytic, follicle centre cell, mantle cell, marginal zone and lymphoplasmacytoid) was assembled and immunostaining for the respective antigens carried out using the monoclonal antibodies produced. Our results indicate that the antibodies are specific for their respective antigens and give the predicted phenotypic profile in the small B-cell lymphoma subtypes. CONCLUSIONS: These novel monoclonal antibodies may be of value in routine diagnostic practice.

Folate deficiency reduces the development of colorectal cancer in rats.

Alterations in folate status may play an important role in carcinogenesis. The aim of this study was to examine the effect of a diminished folate status on azoxymethane (AOM)-induced intestinal tumours in Sprague-Dawley rats. A total of 125 weanling male rats were divided into five equal groups and fed semi-purified diets containing either 8 mg/kg folate or no folate. After 4 weeks on experimental diets, all animals received three weekly subcutaneous injections of AOM at a dose rate of 15 mg/kg bodyweight. The animals were necropsied after 26 weeks. Rats with a diminished folate status, evident by significantly reduced blood and colonic folate concentrations and elevated plasma homocysteine levels, had significantly (P < 0.01) lower incidence and number of small intestinal and colonic tumours compared with rats displaying an adequate folate status. There was a significant decrease in the incidence of colonic adenocarcinomas (P < 0.01) and size of colonic tumours observed in the rats displaying a diminished folate status. This study shows that a diminished folate status was associated with a decrease in the development of AOM-induced colorectal cancers. The decrease in risk may be attributed to the known role of folate in cell multiplication.

Graduated driver licensing in Wisconsin: a new law for teens and parents.

The final components of Wisconsin's graduated driver licensing (GDL) statute [Biennial state budget act 1999 Wisconsin Act 9] went into effect on September 1, 2000. The GDL system seeks to reduce the number of teen motor vehicle collisions and fatalities by gradually introducing young drivers to all driving situations, beginning with the easiest and progressing to the more difficult. The stages of licensing are defined by the degree of risk involved. When teen drivers demonstrate the ability to drive safely in a restricted setting with high levels of supervision, they "graduate" to the next licensing level with fewer restrictions. Numerous factors are associated with the high rate of teen motor vehicle crashes and fatalities. Such factors include driver inexperience and easy distractibility, alcohol use and lack of seat belt use. The GDL addresses all of these risk factors. This article reviews the injury prevention strategies incorporated in the GDL and outlines the basic components of Wisconsin's law.

Fatal injuries in adolescents.

Injuries are a major cause of morbidity and mortality among adolescents. Adolescents routinely experiment with high-risk behaviors, increasing their risk of injury. Major modes of injury in adolescents include motor vehicle collisions, drowning, suicide and homicide. This article reviews the risk factors for fatal injuries in adolescents and discusses current prevention strategies. Previous research identifies many risk factors associated with adolescent injuries. Practitioners can help prevent teen injuries by screening all adolescent patients and identifying those at high risk of injury. Patients at high risk need preventive counseling and evaluation for possible referral to other services. Current strategies are useful in preventing many injuries; however, further research is needed to evaluate these strategies and to design new prevention programs.