Exploring the Role of Cereal Dietary Fiber in Digestion.

Increasing the dietary fiber of staple foods such as bread is an attractive way to promote healthy eating in a large part of the population, where dietary fiber consumption is reportedly below the recommended values. However, many consumers prefer white breads, which are typically low in dietary fiber. In this work, white bread was made from two wheat cultivars with differing fiber contents. The resulting breads showed similar quality parameters (volume, specific volume, firmness, inner structure characteristics) with any differences maintained below 7%. Bread digestibility was evaluated using a novel dynamic in vitro digestion model. Reduced digestion rates of 30% were estimated for the high-fiber white bread compared to that in the control. Overall, this work demonstrates the potential to produce healthy, high-fiber white breads that are acceptable to consumers, with a reduced rate of starch digestion, by exploiting a genetic variation in the dietary fiber content of wheat cultivars.

Evaluation of dry heat treatment of soft wheat flour for the production of high ratio cakes.

An accurate method to heat treat flour samples has been used to quantify the effects of heat treatment on flour functionality. A variety of analytical methods has been used such as oscillatory rheology, rheomixer, solvent retention capacity tests, and Rapid Visco Analysis (RVA) in water and in aqueous solutions of sucrose, lactic acid, and sodium carbonate. This work supports the hypothesis that heat treatment facilitates the swelling of starch granules at elevated temperature. Results furthermore indicated improved swelling ability and increased interactions of flour polymers (in particular arabinoxylans) of heat treated flour at ambient conditions. The significant denaturation of the proteins was indicated by a lack of gluten network formation after severe heat treatments as shown by rheomixer traces. Results of these analyses were used to develop a possible cake flour specification. A method was developed using response surfaces of heat treated flour samples in the RVA using i) water and ii) 50% sucrose solution. This can uniquely characterise the heat treatment a flour sample has received and to establish a cake flour specification. This approach might be useful for the characterisation of processed samples, rather than by baking cakes. Hence, it may no longer be needed to bake a cake after flour heat treatment to assess the suitability of the flour for high ratio cake production, but 2 types of RVA tests suffice.

Using discrete multi-physics for detailed exploration of hydrodynamics in an in vitro colon system.

We developed a mathematical model that describes the motion of viscous fluids in the partially-filled colon caused by the periodic contractions of flexible walls (peristalsis). In-vitro data are used to validate the model. The model is then used to identify two fundamental mechanisms of mass transport: the surfing mode and the pouring mode. The first mechanism is faster, but only involves the surface of the liquid. The second mechanism causes deeper mixing, and appears to be the main transport mechanism. Based on the gained understanding, we propose a series of measures that can improve the reliability of in-vitro models. The tracer in PET-like experiments, in particular, should not be injected in the first pocket, and its viscosity should be as close as possible to that of the fluid. If these conditions are not met, the dynamics of the tracer and the fluid diverge, compromising the accuracy of the in-vitro data.

In silico modelling of mass transfer & absorption in the human gut.

An in silico model has been developed to investigate the digestion and absorption of starch and glucose in the small intestine. The main question we are aiming to address is the relative effect of gastric empting time and luminal viscosity on the rate of glucose absorption. The results indicate that all factors have a significant effect on the amount of glucose absorbed. For low luminal viscosities (e.g. lower than 0.1 Pas) the rate of absorption is controlled by the gastric emptying time. For viscosities higher than 0.1 Pas a 10 fold increase in viscosity can result in a 4 fold decrease of glucose absorbed. Our model, with the simplifications used to develop it, indicate that for high viscosity luminal phases, gastric emptying rate is not the controlling mechanism for nutrient availability. Developing a mechanistic model could help elucidate the rate limiting steps that control the digestion process.

Digestion of starch in a dynamic small intestinal model.

PURPOSE: The rate and extent of starch digestion have been linked with important health aspects, such as control of obesity and type-2 diabetes. In vitro techniques are often used to study digestion and simulated nutrient absorption; however, the effect of gut motility is often disregarded. The present work aims at studying fundamentals of starch digestion, e.g. the effect of viscosity on digestibility, taking into account both biochemical and engineering (gut motility) parameters. METHODS: New small intestinal model (SIM) that realistically mimics gut motility (segmentation) was used to study digestibility and simulated oligosaccharide bio accessibility of (a) model starch solutions; (b) bread formulations. First, the model was compared with the rigorously mixed stirred tank reactor (STR). Then the effects of enzyme concentration/flow rate, starch concentration, and digesta viscosity (addition of guar gum) were evaluated. RESULTS: Compared to the STR, the SIM showed presence of lag phase when no digestive processes could be detected. The effects of enzyme concentration and flow rate appeared to be marginal in the region of mass transfer limited reactions. Addition of guar gum reduced simulated glucose absorption by up to 45 % in model starch solutions and by 35 % in bread formulations, indicating the importance of chyme rheology on nutrient bioaccessibility. CONCLUSIONS: Overall, the work highlights the significance of gut motility in digestive processes and offers a powerful tool in nutritional studies that, additionally to biochemical, considers engineering aspects of digestion. The potential to modulate food digestibility and nutrient bioaccessibility by altering food formulation is indicated.

Prediction of small-for-gestational-age neonates: screening by biophysical and biochemical markers at 30-34 weeks.

OBJECTIVE: To investigate the potential value of combined screening by maternal characteristics and medical history (maternal factors), estimated fetal weight (EFW), uterine artery pulsatility index (UtA-PI), mean arterial pressure (MAP) and serum levels of placental growth factor (PlGF) and soluble fms-like tyrosine kinase-1 (sFlt-1) at 30-34 weeks' gestation in the prediction of delivery of small-for-gestational-age (SGA) neonates, in the absence of pre-eclampsia (PE). METHODS: This was a screening study in 9472 singleton pregnancies at 30-34 weeks' gestation, comprising 469 that delivered SGA neonates and 9003 cases unaffected by SGA, PE or gestational hypertension. Multivariable logistic regression analysis was used to determine if UtA-PI, MAP and serum PlGF or sFlt-1, individually or in combination, improved the prediction of SGA neonates provided from screening by maternal factors and EFW. RESULTS: Compared to the normal group, mean log10 multiples of the median (MoM) values of UtA-PI, MAP and serum sFlt-1 were significantly higher and log10 MoM PlGF was lower in the SGA group. Multivariable logistic regression analysis demonstrated that in the prediction of SGA neonates with a birth weight < 5(th) percentile, delivering < 5 weeks and ≥ 5 weeks after assessment, there were significant independent contributions from maternal factors, EFW, UtA-PI, MAP, and serum PlGF and sFlt-1, but the best performance was provided by a combination of maternal factors, EFW, UtA-PI, MAP and serum PlGF, excluding sFlt-1. Combined screening predicted, at a 10% false-positive rate, 89%, 94%, 96% of SGA neonates delivering at 32-36 weeks' gestation with birth weight < 10(th) , < 5(th) and < 3(rd) percentiles, respectively; the respective detection rates of combined screening for SGA neonates delivering ≥ 37 weeks were 57%, 65% and 72%. CONCLUSION: Combined screening by maternal factors and biophysical and biochemical markers at 30-34 weeks' gestation could identify a high proportion of pregnancies that will deliver SGA neonates.

Prediction of small-for-gestational-age neonates: screening by maternal biochemical markers at 30-34 weeks.

OBJECTIVE: To investigate the potential value of serum placental growth factor (PlGF), soluble fms-like tyrosine kinase-1 (sFlt-1), pregnancy-associated plasma protein-A (PAPP-A), free ß-human chorionic gonadotropin (ß-hCG) and α-fetoprotein (AFP) at 30-34 weeks' gestation in the prediction of delivery of small-for-gestational-age (SGA) neonates, in the absence of pre-eclampsia (PE). METHODS: This was a screening study in singleton pregnancies at 30-34 weeks' gestation, including 490 that delivered SGA neonates and 9360 cases that were unaffected by SGA, PE or gestational hypertension (normal outcome). Multivariable logistic regression analysis was used to determine if screening by serum PlGF, sFlt-1, PAPP-A, free ß-hCG and AFP, individually or in combination, improved the prediction of SGA neonates provided by screening with maternal characteristics and medical history (maternal factors), and estimated fetal weight (EFW) from fetal head circumference, abdominal circumference and femur length. RESULTS: Compared to the normal group, the mean log10 multiples of the median (MoM) values of PlGF and AFP were significantly lower and the mean log10 MoM values of sFlt-1 and free ß-hCG were significantly higher in the SGA group with a birth weight < 5(th) percentile (SGA < 5(th)) delivering < 5 weeks following assessment. The best model for prediction of SGA was provided by a combination of maternal factors, EFW and serum PlGF. Such combined screening, predicted, at a 10% false-positive rate, 85%, 93% and 92% of SGA neonates delivering < 5 weeks following assessment with birth weight < 10(th), < 5(th) and < 3(rd) percentiles, respectively; the respective detection rates of combined screening for SGA neonates delivering ≥ 5 weeks following assessment were 57%, 64% and 71%. CONCLUSION: Combined screening by maternal factors, EFW and serum PlGF at 30-34 weeks' gestation can identify a high proportion of pregnancies that subsequently deliver SGA neonates.

Umbilical and fetal middle cerebral artery Doppler at 30-34 weeks' gestation in the prediction of adverse perinatal outcome.

OBJECTIVE: To investigate the potential value of cerebroplacental ratio (CPR) at 30-34 weeks' gestation in the prediction of adverse perinatal outcome. METHODS: This was a screening study in 30 780 singleton pregnancies at 30-34 weeks' gestation. Umbilical artery (UA) and fetal middle cerebral artery (MCA) pulsatility index (PI) were measured and the values were converted to multiples of the median (MoM) after adjustment from variables in maternal characteristics and medical history that affect the measurements. CPR was calculated by dividing MCA-PI MoM by UA-PI MoM. Multivariable logistic regression analysis was used to determine if measuring CPR improved the prediction of adverse perinatal outcome provided by screening with maternal characteristics, medical history and obstetric factors. The detection rate (DR) and false-positive rate (FPR) of screening by CPR were estimated for stillbirth, Cesarean section for fetal distress, umbilical arterial cord blood pH ≤ 7.0, umbilical venous cord blood pH ≤ 7.1, 5-min Apgar score < 7 and admission to the neonatal unit (NNU) and neonatal intensive care unit (NICU). RESULTS: There was a significant association between CPR and birth-weight Z-score. In addition to maternal characteristics, medical history and obstetric factors, measuring CPR provided a significant contribution to the prediction of arterial cord blood pH ≤ 7.0, venous cord blood pH ≤ 7.1 and admission to NNU. The performance of CPR in screening for each adverse outcome was poor, with DR of 5-11% and a FPR of about 5%. In the small subgroup of the population delivering within 2 weeks following assessment, the DR improved to 20-50%, but with a simultaneous increase in FPR to 10-23%. CONCLUSION: The performance of CPR in routine screening for adverse perinatal outcome at 30-34 weeks' gestation is poor.

Prediction of small-for-gestational-age neonates: screening by uterine artery Doppler and mean arterial pressure at 30-34 weeks.

OBJECTIVE: To investigate the potential value of uterine artery (UtA) pulsatility index (PI) and mean arterial pressure (MAP) at 30-34 weeks' gestation in the prediction of small-for-gestational-age (SGA) neonates, in the absence of pre-eclampsia (PE). METHODS: This was a screening study in singleton pregnancies at 30-34 weeks' gestation, including 1727 that delivered SGA neonates with a birth weight < 5(th) percentile and 29 122 that were unaffected by SGA, PE or gestational hypertension (normal group). Multivariable logistic regression analysis was used to determine if measuring the UtA-PI and MAP improved the prediction of SGA neonates provided by screening with maternal characteristics and medical history (maternal factors), and estimated fetal weight (EFW) calculated from fetal head circumference, abdominal circumference and femur length. RESULTS: Combined screening by maternal factors and EFW Z-scores predicted 79%, 87% and 92% of SGA neonates delivering < 5 weeks following assessment, with a birth weight < 10(th) , < 5(th) and < 3(rd) percentiles, respectively, at a false-positive rate of 10%. The addition of UtA-PI and MAP improved the respective detection rates to 83%, 91% and 93%. Screening by maternal factors and EFW Z-scores predicted 53%, 58% and 61% of SGA delivering ≥ 5 weeks following assessment and these rates increased to 53%, 60% and 63% with the addition of UtA-PI and MAP. CONCLUSION: Combined testing by maternal factors, fetal biometry, UtA-PI and MAP at 30-34 weeks' gestation could identify a high proportion of pregnancies that deliver SGA neonates.

Prediction of small-for-gestational-age neonates: screening by fetal biometry at 30-34 weeks.

OBJECTIVE: To investigate the value of fetal biometry at 30-34 weeks' gestation in the prediction of delivery of small-for-gestational-age (SGA) neonates, in the absence of pre-eclampsia (PE). METHODS: This was a screening study in 30 849 singleton pregnancies at 30-34 weeks' gestation, comprising 1727 that delivered SGA neonates with a birth weight < 5(th) percentile and 29 122 cases unaffected by SGA, PE or gestational hypertension. Multivariable logistic regression analysis was used to determine if screening by a combination of maternal factors and Z-scores of fetal head circumference (HC), abdominal circumference (AC) and femur length (FL) or estimated fetal weight (EFW) had a significant contribution to the prediction of SGA neonates. RESULTS: Combined screening by maternal characteristics and obstetric history, with Z-scores of EFW at 30-34 weeks, predicted 79%, 87% and 92% of the SGA neonates that delivered < 5 weeks following assessment, with a birth weight < 10(th) , < 5(th) and < 3(rd) percentiles, respectively, at a 10% false-positive rate. The respective detection rates for the prediction of SGA neonates delivering ≥ 5 weeks from the time of assessment were 53%, 58% and 61%. The performance of screening by a combination of Z-scores of fetal HC, AC and FL was similar to that achieved by the EFW Z-score alone. CONCLUSION: Combined testing by maternal characteristics and fetal biometry at 30-34 weeks could identify a high proportion of pregnancies that will deliver SGA neonates.

Mass transfer and nutrient absorption in a simulated model of small intestine.

There is an increasing need to understand how food formulations behave in vivo from both food and pharma industries. A number of models have been proposed for the stomach, but few are available for the other parts of the gastrointestinal tract. An experimental rig that simulates the segmentation motion occurring in the small intestine has been developed. The objective of developing such an experimental apparatus was to study mass transport phenomena occurring in the lumen and their potential effect on the concentration of species available for absorption. When segmentation motion was applied the mass transfer coefficient in the lumen side was increased up to a factor of 7. The viscosity of the lumen, as influenced by guar gum concentration, had a profound effect on the mass transfer coefficient. The experimental model was also used to demonstrate that glucose available for absorption, resulting from starch hydrolysis, can be significantly reduced by altering the lumen viscosity. Results suggest that absorption of nutrients could be controlled by mass transfer. Practical Application: To address health-related diseases such as obesity, novel foods that provide advanced functions are required. To achieve the full potential offered by the latest developments in the field of food material science, a fundamental understanding of the behavior of food structures in vivo is required. Using the developed gut model we have demonstrated that absorption of nutrients can be controlled by mass transfer limitations.

Outcome of prenatally diagnosed agenesis of the corpus callosum.

OBJECTIVE: To investigate the natural history, associated abnormalities and outcome in fetuses diagnosed prenatally with agenesis of the corpus callosum (ACC). METHODS: A retrospective study of all cases of prenatally detected ACC was performed in patients referred to two tertiary units between January 1993 and October 2003. Associated abnormalities, pregnancy outcome and infant follow-up were recorded. RESULTS: ACC was diagnosed in 117 cases. In 82 (70%) cases this was associated with other fetal structural (n = 49) or chromosomal abnormalities (n = 33). ACC was classified as an isolated prenatal finding in 35 (30%) cases. Assuming normal development in all cases lost to follow-up, significant developmental delay was present in 36% (95% CI, 15-65%) of isolated ACC. Furthermore, developmental delay was present in all cases with ventriculomegaly of at least 15 mm and in one of four cases with ventricular measurements less than 15 mm. CONCLUSIONS: The outcome of prenatally detected ACC is mainly dependent on the presence or absence of associated anomalies. The full assessment of fetal ACC mandates karyotyping, MRI and a search for more subtle ultrasound features of certain genetic syndromes. In this series, at least 36% (95% CI, 15-65%) of cases with isolated ACC exhibited significant developmental delay when assessed postnatally.

Outcome of antenatally diagnosed talipes equinovarus in an unselected obstetric population.

OBJECTIVE: To investigate the natural history and outcome of fetal talipes diagnosed by routine ultrasound scanning at 18-23 weeks' gestation. PATIENTS AND METHODS: This was a retrospective study of 103 228 pregnancies undergoing routine ultrasound scanning at 18-23 weeks' gestation. A computer search was made to identify all cases of fetal talipes and the records of these patients were examined to determine the incidence of other defects and pregnancy outcome. RESULTS: The incidence of fetal talipes following routine ultrasound examination was 0.10% (107/103 228) and was bilateral in 64 (59.8%) and unilateral in 43 (40.2%) cases. In 52 (48.6%) cases, talipes was of complex etiology, as it was associated with other defects, while, in 55 (51.4%) cases, it was classified as idiopathic. In 19% of cases, an initial diagnosis of idiopathic talipes was changed to complex, because of the subsequent development of associated features. Perinatal death and long-term neurodevelopmental or musculoskeletal problems were significantly more common when the talipes was complex rather than idiopathic (odds ratio, 150; 95% confidence interval, 34-665). Adverse outcomes were also seen more frequently with bilateral compared to unilateral talipes (odds ratio, 3.44; 95% confidence interval, 1.50-7.90). CONCLUSION: The outcome of antenatally detected talipes is mainly dependent on the presence or absence of other defects. A significant proportion of cases, thought to be idiopathic at presentation, will develop associated complex features when reassessed on subsequent scans or postnatally.

Outcome of pregnancy in chromosomally normal fetuses with increased nuchal translucency in the first trimester.

OBJECTIVES: To study the outcome of chromosomally normal pregnancies with increased nuchal translucency at the 10-14-week scan. DESIGN: Retrospective study of 1320 chromosomally normal singleton pregnancies with nuchal translucency of > or = 3.5 mm. In addition to fetal karyotyping these patients were managed with follow-up scans at 14-16 and 20-22 weeks, specialist fetal echocardiography and in selected cases by infection screening and further genetic testing. RESULTS: In the 1320 pregnancies there were 68 (5.15%) spontaneous abortions or intrauterine deaths, 18 (1.36%) neonatal and infant deaths and 154 (11.67%) terminations of pregnancy. In the 1080 (81.82%) survivors, 60 (5.56%) had abnormalities requiring medical or surgical treatment or leading to mental handicap. The chance of a livebirth with no defects in the group with nuchal translucency of 3.5-4.4 mm was 86%, for those with translucency of 4.5-5.4 mm it was 77%, for those with translucency of 5.5-6.4 mm it was 67%, and for those with translucency of > or = 6.5 mm it was 31%. CONCLUSIONS: Increased fetal nuchal translucency is associated with chromosomal abnormalities, many fetal defects and genetic syndromes. In the majority of cases a series of antenatal investigations, including fetal karyotyping, detailed scans, fetal echocardiography, as well as genetic testing and infection screening, that can be completed by 20 weeks of gestation would distinguish between the pregnancies destined to result in adverse outcome and those leading to the delivery of infants without major defects.

Tau confers drug stability but not cold stability to microtubules in living cells.

We previously defined two classes of microtubule polymer in the axons of cultured sympathetic neurons that differ in their sensitivity to nocodazole by roughly 35-fold (Baas and Black (1990) J. Cell Biol. 111, 495-509). Here we demonstrate that virtually all of the microtubule polymer in these axons, including the drug-labile polymer, is stable to cold. What factors account for the unique stability properties of axonal microtubules? In the present study, we have focused on the role of tau, a microtubule-associated protein that is highly enriched in the axon, in determining the stability of microtubules to nocodazole and/or cold in living cells. We used a baculovirus vector to express very high levels of tau in insect ovarian Sf9 cells. The cells respond by extending processes that contain dense bundles of microtubules (Knops et al. (1991) J. Cell Biol. 114, 725-734). Cells induced to express tau were treated with either cold or 2 micrograms/ml nocodazole for times ranging from 5 minutes to 6 hours. The results with each treatment were very different from one another. Virtually all of the polymer was depolymerized within the first 30 minutes in cold, while little or no microtubule depolymerization was detected even after 6 hours in nocodazole. Based on these results, we conclude that tau is almost certainly a factor in conferring drug stability to axonal microtubules, but that factors other than or in addition to tau are required to confer cold stability.

Phosphorylated baculovirus p10 is a heat-stable microtubule-associated protein associated with process formation in Sf9 cells.

Insect ovarian Sf9 cells extend processes with complex morphologies when infected with a recombinant baculovirus encoding the catalytic subunit of protein kinase A. Within the shafts of the processes are abundant microtubules, which, in contrast to those in Sf9 cells expressing the microtubule-associated protein tau, are generally not organized into parallel bundles. During infection the late viral polypeptide p10 becomes phosphorylated by the protein kinase A catalytic subunit at its penultimate residue, Ser92. The expression or phosphorylation of other major host cell or viral polypeptides does not change, compared with polypeptides from a wild-type viral infection. Once phosphorylated, p10 associates with microtubules in the infected cells and may thereby play a role in process formation.

Alz-50 recognizes a phosphorylated epitope of tau protein.

Alz-50 is a monoclonal antibody that detects antigens enriched in the brain tissue of Alzheimer's disease (AD) patients. Although Alz-50 recognizes tau, an identified integral constituent of the AD paired helical filament (PHF), the exact nature of the antigenic site is unknown. An immunoblot analysis demonstrated that the antigenic sites to Alz-50 are diminished by acid phosphatase treatment. Consistent with this finding, Alz-50 antigens were more concentrated in brain homogenates prepared with phosphatase inhibitors. The epitope in tau with which Alz-50 reacts is located in the carboxy terminus within a 14-amino acid region from just beyond the microtubule-binding repeats to the carboxy terminus. An isolated carboxy-terminal chymotryptic peptide from bovine brain tau reactive with Alz-50 was analyzed by fast-atom-bombardment mass spectroscopy (FAB-MS) and was found to be present as both a monophosphopeptide and a nonphosphorylated peptide. The immunohistological analysis has demonstrated that Alz-50 staining of neurofibrillary tangles (NFTs) is sensitive to acid phosphatase but not to alkaline phosphatase. Furthermore, Alz-50 staining of NFTs was effectively adsorbed by a high concentration of phosphoserine but not by serine or phosphothreonine. These results strongly suggest that Alz-50 recognizes a phosphorylated epitope in the carboxy terminus of tau which has not been previously detected by using alkaline phosphatase. The strong Alz-50 staining in AD samples may represent another association between a phosphorylation state and neurofibrillary lesions. As a marker of the inchoate tangle-bearing neuron, the characterization of the Alz-50 epitope in tau offers a partial molecular basis for the modifications that contribute to the assembly of PHFs.

Developmentally regulated expression of specific tau sequences.

Tau protein undergoes a shift in its molecular mass and its electrophoretic complexity during early postnatal development. We have sequenced a tau cDNA from an adult rat brain expression library and have found two inserted sequences. One of these inserts predicts a fourth repeated sequence homologous to the other three in the carboxyl end of tau that have the property of microtubule binding. Oligonucleotide probes directed against the insert hybridized only to tau mRNA at postnatal time points, even though tau is first expressed as early as embryonic day 13. A probe directed against the junction revealed expression of non-insert-containing tau mRNA from embryonic day 14 until postnatal day 8, after which time there was an abrupt decline in the expression of this immature form. Comparison of the developmentally expressed tau sequences with those sequences obtained directly from Alzheimer paired helical filaments revealed the presence of both the mature and the immature tau mRNA sequences.

Partial sequence of MAP2 in the region of a shared epitope with Alzheimer neurofibrillary tangles.

A 3.3-kilobase DNA complementary to human microtubule-associated protein 2 (MAP2) was sequenced by the dideoxy method. The 3' end terminates at an internal EcoRI site before the polyA tail. Due to the arrangement of the cDNA insert in the lambda gt11 vector, the MAP2 fragment is not fused to beta-galactosidase when expressed. The Chou Fasman algorithm for the initial 58 amino acids from the first in-frame methionine predicts an alpha helix. Beyond this point, a series of turns is predicted until amino acid 160. The frequent presence of basic residues in proximity to serines or threonines is consistent with multiple phosphorylation sites. The minimum specificity determinant for Ca2+/calmodulin-dependent kinase is repeated 13 times. The sequence of a region containing a MAP2 epitope that is shared with the Alzheimer neurofibrillary tangle was determined by DNase treatment of the cDNA and antibody selecting the small resultant clones in a lambda gt11 sublibrary. Likewise, a MAP2 epitope that is not shared with the neurofibrillary tangle also has been located. Both epitopes are in the projection portion of the molecule. A bovine MAP2 cyanogen bromide fragment, which contains the epitope shared with the neurofibrillary tangle, is partially insoluble under aqueous conditions, probably due to the aggregation of oppositely charged residues. Thus, rapid cleavage of MAP2 to small peptides is probably necessary in vivo to prevent the aggregation of larger cleavage fragments.