UV embossed polymeric chip for protein separation and identification based on capillary isoelectric focusing and MALDI-TOF-MS.

This paper demonstrates a ultraviolet (UV)-embossed polymeric chip for protein separation by capillary isoelectric focusing (CIEF) and identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The polymeric chip was replicated by a UV-embossing technique using a soft rubber mold. Five diverse widely investigated families of UV-cured formulations were examined for MALDI ionization efficiency of bovine serum albumin (BSA) samples spotted on the polymer surfaces. The signal and signal-to-noise ratio from the polyester formulation were each 12 times those obtained with PMMA (a commonly used polymer material for MALDI) at picomole sample concentration. A polyester chip was successfully used to carry out CIEF to separate proteins, followed by MALDI-TOF-MS identification. Issues related to the successful chip fabrication and protein separation and identification are discussed.

Capillary isoelectric focusing in pseudo-closed channel coupled to matrix assisted laser desorption/ionization mass spectrometry for protein analysis.

Capillary isoelectric focusing (CIEF) was performed in pseudo-closed channel to separate proteins on a plastic chip. Pseudo-closed channel provided a novel way to couple protein separation by CIEF to MALDI mass spectrometry without eluting the focused proteins.

Chromosome 14 linked familial Alzheimer's disease. A clinico-pathological study of a single pedigree.

The clinical features of three affected members of a British pedigree with familial Alzheimer's disease are presented. This pedigree is one of six included in an earlier study which demonstrated linkage to chromosome 14. The individuals were investigated clinically and neuropsychologically, using both PET and MRI over a 4-year period. Further information from three deceased individuals was obtained, including histopathological confirmation of Alzheimer's disease in one case which came to autopsy. The mean age at onset for this family was 43 years. Neurological examination revealed myoclonic jerks in all cases, and one patient was documented to have seizures. Strikingly similar neuropsychological profiles were observed, characterized by an initial memory deficit with early dyscalculia and an impairment in speech production with relative absence of anomia. All individuals showed mild degrees of cerebral atrophy and two individuals had periventricular white matter lesions. PET scanning using [18F]fluorodeoxyglucose showed parieto-temporal hypometabolism in all cases and the two severely affected patients with speech production changes had additional left-sided frontal hypometabolism involving Broca's area. The least affected case initially had a more asymmetrical reduction in metabolism in the left inferior temporal and supramarginal gyri; a follow-up scan showed that this deficit had become bilateral and more severe. These clinical and neuroimaging features have not been previously reported in chromosome 14 linked pedigrees; the phenotypic variability between families suggests allelic heterogeneity at the chromosome 14 locus.

Deficits in cerebral glucose metabolism demonstrated by positron emission tomography in individuals at risk of familial Alzheimer's disease.

In order to establish whether positron emission tomography (PET) can identify metabolic changes in Alzheimer's disease at a presymptomatic stage, we have examined 24 asymptomatic at risk individuals from families with Alzheimer's disease. A significant reduction in global cerebral metabolic rate for glucose was found when compared with 16 age-matched controls. There was also a focal, parieto-temporal deficit similar to, although less extensive than, that found in 18 symptomatic individuals from familial Alzheimer's disease (FAD) pedigrees. Follow up of this cohort will establish whether these metabolic changes relate to a presymptomatic stage of the disease.

The earliest cognitive change in a person with familial Alzheimer's disease: presymptomatic neuropsychological features in a pedigree with familial Alzheimer's disease confirmed at necropsy.

Comprehensive, longitudinal neuropsychological assessments are reported in a person "at risk" from autosomal dominant, necropsy confirmed familial Alzheimer's disease. The first assessment showed a moderately selective verbal memory deficit in the context of mild general intellectual impairment. Subsequent testing showed the progressive deterioration of visual memory and a mild decline of perceptual and spatial skills. Language and literacy skills, however, remained comparatively intact. The neuropsychological profiles obtained at each assessment are presented in profile maps. These permit direct longitudinal comparison of cognitive function, and may serve in the comparison of different potential cases of familial Alzheimer's disease. This case sought medical attention for memory difficulties 26 months after the first neuropsychological assessment. These results mark the first cognitive manifestations in a pedigree with familial Alzheimer's disease which, in this case, were seen presymptomatically. The findings are discussed in relation to neuropsychological studies of affected cases, and in terms of their reflecting the heterogeneous nature of familial Alzheimer's disease.

Management factors affecting reproductive performance of dairy cows in the northeastern United States.

Dairy herds (476) in seven states in the northeastern United States were surveyed to determine effects of various management factors on reproductive performance. Error of estrus detection (greater than 1 ng/ml of milk progesterone) on the day of service was 5.1% for 4558 cows but was as high as 60% in some herds. Error was not affected by herd size but was greater in freestall (6.8%) than in conventional (5.2%) housing. "Standing" and "riding other cows" were the most accurate signs of estrus. Of cows in or near estrus when serviced, 28.1% were open 3 wk later, 12.9% were probably open, and 59% were probably pregnant based on analysis of milk progesterone. Conception rate, not affected by herd size or housing type, was greater for cows in estrus during the morning and serviced the same afternoon (52.2%) than for cows observed in the afternoon and serviced the next morning (47.1%). Fifty-five percent of cows open to first service were serviced again within 3 days of expected return to estrus. Days to first service and conception rates were correlated positively. Milk progesterone concentration 21 to 24 days after service predicted a cow will calve with 88.6% accuracy and that she will not with 93.9% accuracy. Veterinarians predicted a calf with 92.5% accuracy by rectal palpation.

Uterine contractions and fertility following clitoral massage of dairy cattle in estrus.

Lactating and nonlactating Holstein cows received 30 s of gentle clitoral massage to determine its effect on uterine contractility and oxytocin concentrations in blood. Clitoral stimulation caused an immediate single uterine contraction, and this could be repeated at 2.5-min intervals. Oxytocin concentrations did not change during intensive blood sampling following clitoral massage. Thus, the uterine response is interpreted as neurally mediated. Eighteen full-time inseminators were allotted to two groups, balanced so that each day half were inseminating control and half treated animals (5 s of clitoral massage following insemination). The 56-day nonreturns for 2090 controls were 74.3% versus 74.0% for 2049 animals receiving clitoral massage. Heifers were more fertile than cows (81.9% versus 70.6% nonreturns), but there was no indication that clitoral massage altered the nonreturn rate in either age group. Also, the time of day animals were inseminated did not affect nonreturn rate.

Changes throughout estrous cycles of variables that might indicate estrus in dairy cows.

Two experiments examined physiological changes throughout estrous cycles of Holstein cows. In experiment one, changes were characterized for physical activity, vaginal pH, vaginal temperature, milk yield, heart rate, and concentration in blood plasma of progesterone and 13,14-dihydro-15-keto-prostaglandin F2 alpha. Variables were measured daily in 11 cows through a total of 28 estrous cycles. Cows in estrus at least once postpartum were used. Observations for estrus were at least twice daily. Variables changed significantly with day of estrous cycle. Changes of progesterone, physical activity, and 13,14-dihydro-15-keto-prostaglandin F2 alpha were more marked around estrus than were changes of the other variables. Peaks of physical activity coincided with estrus in 75% of the cases. Experiment two was to determine if increased physical activity, measured with pedometers, coincided with estrus in commercial dairy herds. Cows on 14 farms in New York were fitted with mechanical pedometers, and the pedometers were read at each milking. Readings from 3 days before until 3 days after estrus were used from 55 estrous periods identified by the producers. Physical activity was maximal on day of estrus in 73% of the estrous periods. Increased physical activity of commercial dairy cows agreed well with the producers' diagnoses of estrus. Pedometers could be a valuable component of an estrous detection program.