Multiomics Blood-Based Biomarkers Predict Alzheimer's Predementia with High Specificity in a Multicentric Cohort Study.

BACKGROUND: The primary criteria for diagnosing mild cognitive impairment (MCI) due to Alzheimer's Disease (AD) or probable mild AD dementia rely partly on cognitive assessments and the presence of amyloid plaques. Although these criteria exhibit high sensitivity in predicting AD among cognitively impaired patients, their specificity remains limited. Notably, up to 25% of non-demented patients with amyloid plaques may be misdiagnosed with MCI due to AD, when in fact they suffer from a different brain disorder. The introduction of anti-amyloid antibodies complicates this scenario. Physicians must prioritize which amyloid-positive MCI patients receive these treatments, as not all are suitable candidates. Specifically, those with non-AD amyloid pathologies are not primary targets for amyloid-modifying therapies. Consequently, there is an escalating medical necessity for highly specific blood biomarkers that can accurately detect pre-dementia AD, thus optimizing amyloid antibody prescription. OBJECTIVES: The objective of this study was to evaluate a predictive model based on peripheral biomarkers to identify MCI and mild dementia patients who will develop AD dementia symptoms in cognitively impaired population with high specificity. DESIGN: Peripheral biomarkers were identified in a gene transfer-based animal model of AD and then validated during a retrospective multi-center clinical study. SETTING: Participants from 7 retrospective cohorts (US, EU and Australia). PARTICIPANTS: This study followed 345 cognitively impaired individuals over up to 13 years, including 193 with MCI and 152 with mild dementia, starting from their initial visits. The final diagnoses, established during their last assessments, classified 249 participants as AD patients and 96 as having non-AD brain disorders, based on the specific diagnostic criteria for each disorder subtype. Amyloid status, assessed at baseline, was available for 82.9% of the participants, with 61.9% testing positive for amyloid. Both amyloid-positive and negative individuals were represented in each clinical group. Some of the AD patients had co-morbidities such as metabolic disorders, chronic diseases, or cardiovascular pathologies. MEASUREMENTS: We developed targeted mass spectrometry assays for 81 blood-based biomarkers, encompassing 45 proteins and 36 metabolites previously identified in AAV-AD rats. METHODS: We analyzed blood samples from study participants for the 81 biomarkers. The B-HEALED test, a machine learning-based diagnostic tool, was developed to differentiate AD patients, including 123 with Prodromal AD and 126 with mild AD dementia, from 96 individuals with non-AD brain disorders. The model was trained using 70% of the data, selecting relevant biomarkers, calibrating the algorithm, and establishing cutoff values. The remaining 30% served as an external test dataset for blind validation of the predictive accuracy. RESULTS: Integrating a combination of 19 blood biomarkers and participant age, the B-HEALED model successfully distinguished participants that will develop AD dementia symptoms (82 with Prodromal AD and 83 with AD dementia) from non-AD subjects (71 individuals) with a specificity of 93.0% and sensitivity of 65.4% (AUROC=81.9%, p<0.001) during internal validation. When the amyloid status (derived from CSF or PET scans) and the B-HEALED model were applied in association, with individuals being categorized as AD if they tested positive in both tests, we achieved 100% specificity and 52.8% sensitivity. This performance was consistent in blind external validation, underscoring the model's reliability on independent datasets. CONCLUSIONS: The B-HEALED test, utilizing multiomics blood-based biomarkers, demonstrates high predictive specificity in identifying AD patients within the cognitively impaired population, minimizing false positives. When used alongside amyloid screening, it effectively identifies a nearly pure prodromal AD cohort. These results bear significant implications for refining clinical trial inclusion criteria, facilitating drug development and validation, and accurately identifying patients who will benefit the most from disease-modifying AD treatments.

Novel genetic parameters for genetic residual feed intake in dairy cattle using time series data from multiple parities and countries in North America and Europe.

Residual feed intake is viewed as an important trait in breeding programs that could be used to enhance genetic progress in feed efficiency. In particular, improving feed efficiency could improve both economic and environmental sustainability in the dairy cattle industry. However, data remain sparse, limiting the development of reliable genomic evaluations across lactation and parity for residual feed intake. Here, we estimated novel genetic parameters for genetic residual feed intake (gRFI) across the first, second, and third parity, using a random regression model. Research data on the measured feed intake, milk production, and body weight of 7,379 cows (271,080 records) from 6 countries in 2 continents were shared through the Horizon 2020 project Genomic Management Tools to Optimise Resilience and Efficiency, and the Resilient Dairy Genome Project. The countries included Canada (1,053 cows with 47,130 weekly records), Denmark (1,045 cows with 72,760 weekly records), France (329 cows with 16,888 weekly records), Germany (938 cows with 32,614 weekly records), the Netherlands (2,051 cows with 57,830 weekly records), and United States (1,963 cows with 43,858 weekly records). Each trait had variance components estimated from first to third parity, using a random regression model across countries. Genetic residual feed intake was found to be heritable in all 3 parities, with first parity being predominant (range: 22-34%). Genetic residual feed intake was highly correlated across parities for mid- to late lactation; however, genetic correlation across parities was lower during early lactation, especially when comparing first and third parity. We estimated a genetic correlation of 0.77 ± 0.37 between North America and Europe for dry matter intake at first parity. Published literature on genetic correlations between high input countries/continents for dry matter intake support a high genetic correlation for dry matter intake. In conclusion, our results demonstrate the feasibility of estimating variance components for gRFI across parities, and the value of sharing data on scarce phenotypes across countries. These results can potentially be implemented in genetic evaluations for gRFI in dairy cattle.

Genetic (co-)variation of methane emissions, efficiency, and production traits in Danish Holstein cattle along and across lactations.

Methane emissions in ruminant livestock has become a hot topic, given the pressure to reduce greenhouse gas emissions drastically in the European Union over the next 10 to 30 yr. During the 2021 United Nations Climate Change conference, countries also made collective commitments to curb methane emissions by 2050. Genetic selection for low-methane-emitting animals, particularly dairy cows, is one possible strategy for mitigation. However, it is essential to understand how methane emissions in lactating animals vary along lactation and across lactations. This understanding is useful when making decisions for future phenotyping strategies, such as the frequency and duration of phenotyping within and across lactations. Therefore, the objectives of this study were to estimate (1) genetic parameters for 2 methane traits: methane concentration (MeC) and methane production (MeP) at 2 parity levels in Danish Holstein cows across the entire lactation using random regression models; (2) genetic correlations within and between methane traits across the entire lactation; and (3) genetic correlations between the methane traits and economically important traits throughout first lactation. Methane concentration (n = 19,639) records of 575 Danish Holstein cows from a research farm measured between 2013 and 2020 were available. Subsequently, CH4 production in grams/day (MeP; n = 13,866) was calculated; MeP and MeC for first and second lactation (L1 and L2) were analyzed as separate traits: MeC_L1, MeP_L1, MeC_L2, and MeP_L2. Heritabilities, variance components, and genetic correlations within and between the 4 CH4 traits were estimated using random regression models with Legendre polynomials. The additive genetic and permanent environmental effects were modeled using second-order Legendre polynomial for lactation weeks. Estimated heritabilities for MeP_L1 ranged between 0.11 and 0.49, for MeC_L1 between 0.10 and 0.28, for MeP_L2 between 0.14 and 0.36, and for MeC_L2 between 0.13 and 0.29. In general, heritability estimates of MeC traits were lower and more stable throughout lactation and were similar between lactations compared with MeP. Genetic correlations (within trait) at different lactation weeks were generally highly positive (0.7) for most of the first lactation, except for the correlation of early lactation (<10 wk) with late lactation (>40 wk) where the correlation was the lowest (<0.5). Genetic correlations between methane traits were moderate to highly correlated during early and mid lactation. Finally, MeP_L1 has stronger genetic correlations with energy-corrected milk and dry matter intake compared with MeC_L1. In conclusion, both traits are different along (and across) lactation(s) and they correlated differently with production, maintenance, and intake traits, which is important to consider when including one of them in a future breeding objective.

Genome-wide association study for methane emission traits in Danish Holstein cattle.

Selecting for lower methane emitting cows requires insight into the most biologically relevant phenotypes for methane emission, which are close to the breeding goal. Several methane phenotypes have been suggested over the last decade. However, the (dis)similarity of their underlying genetic architecture and correlation structures are poorly understood. Therefore, the objective of this study was to test association of SNP and genomic regions through GWAS on 8 CH4 emission traits in Danish Holstein cattle. The traits studied were methane concentration (MeC; ppm), methane production (MeP ; g/d), 2 definitions of residual methane (RMETc and RMETp: MeC and MeP regressed on metabolic body weight and energy-corrected milk, respectively), 2 definitions of methane intensity (MeI; MeIc = MeC/ECM and MeIp = MeP/ECM); 2 definitions of methane yield per kilogram of dry matter intake (MeY; MeYc = MeC/dry matter intake and MeYp = MeP/dry matter intake). A total of 1,962 cows with genotypes (Illumina BovineSNP50 Chip or Eurogenomic custom SNP chip) and repeated records of the above-mentioned 8 methane traits were analyzed. Strong associations were found with 3 traits (MeC, MeP, and MeYc) on chromosome 13 and with 5 traits (MeC, MeP, MeIp, MeYp, and MeYc) on chromosome 26. For MeIc, MeIp, RMETc, MeYc, and MeYp, some suggestive association signals were identified on chromosome 1. Genomic segments of 1 Mbp (n = 2,525) were tested for their association with these traits, which identified between 33 to 54 significantly associated regions. In a pairwise comparison, MeC and MeP were the traits that shared the highest number of significant segments (17). The same trend was observed when comparing SNP significantly associated with the traits MeC and MeP shared from 23 to 25 SNP (most of which were located in chromosomes 11, 13, and 26). Based on our results on GWAS and genetic correlations, we conclude that MeC is (genetically) more closely linked to MeP than any of the other methane traits analyzed.

Breeding for reduced methane emission and feed-efficient Holstein cows: An international response.

Selecting for lower methane (CH4) emitting animals is one of the best approaches to reduce CH4 given that genetic progress is permanent and cumulative over generations. As genetic selection requires a large number of animals with records and few countries actively record CH4, combining data from different countries could help to expedite accurate genetic parameters for CH4 traits and build a future genomic reference population. Additionally, if we want to include CH4 in the breeding goal, it is important to know the genetic correlations of CH4 traits with other economically important traits. Therefore, the aim of this study was first to estimate genetic parameters of 7 suggested methane traits, as well as genetic correlations between methane traits and production, maintenance, and efficiency traits using a multicountry database. The second aim was to estimate genetic correlations within parities and stages of lactation for CH4. The third aim was to evaluate the expected response of economically important traits by including CH4 traits in the breeding goal. A total of 15,320 methane production (MeP, g/d) records from 2,990 cows belonging to 4 countries (Canada, Australia, Switzerland, and Denmark) were analyzed. Records on dry matter intake (DMI), body weight (BW), body condition score, and milk yield (MY) were also available. Additional traits such as methane yield (MeY; g/kg DMI), methane intensity (MeI; g/kg energy-corrected milk), a genetic standardized methane production, and 3 definitions of residual methane production (g/d), residual feed intake, metabolic BW (MBW), BW change, and energy-corrected milk were calculated. The estimated heritability of MeP was 0.21, whereas heritability estimates for MeY and MeI were 0.30 and 0.38, and for the residual methane traits heritability ranged from 0.13 to 0.16. Genetic correlations between different methane traits were moderate to high (0.41 to 0.97). Genetic correlations between MeP and economically important traits ranged from 0.29 (MY) to 0.65 (BW and MBW), being 0.41 for DMI. Selection index calculations showed that residual methane had the most potential for inclusion in the breeding goal when compared with MeP, MeY, and MeI, as residual methane allows for selection of low methane emitting animals without compromising other economically important traits. Inclusion of residual feed intake in the breeding goal could further reduce methane, as the correlation with residual methane is moderate and elicits a favorable correlated response. Adding a negative economic value for methane could facilitate a substantial reduction in methane emissions while maintaining an increase in milk production.

Multitrait genomic prediction of methane emissions in Danish Holstein cattle.

In dairy cattle, selecting for lower methane-emitting animals is one of the new challenges of this decade. However, genetic selection requires a large number of animals with records to get accurate estimated breeding values (EBV). Given that CH4 records are scarce, the use of information on routinely recorded and highly correlated traits with CH4 has been suggested to increase the accuracy of genomic EBV (GEBV) through multitrait (genomic) prediction. Therefore, the objective of this study was to evaluate accuracies of prediction of GEBV for CH4 by including or omitting CH4, energy-corrected milk (ECM), and body weight (BW) as well as genotypic information in multitrait analyses across 2 methods: BLUP and single-step genomic BLUP (SSGBLUP). A total of 2,725 cows with CH4 concentration in breath (14,125 records), BW (61,667 records), and ECM (61,610 records) were included in the analyses. Approximately 2,000 of these cows were genotyped or imputed to 50K. Ten cross-validation groups were formed by randomly grouping paternal half-sibs. Five scenarios were performed: (1) base scenario with only CH4 information; (2) without CH4, but with information from BW, ECM, or BW+ECM only in reference population; (3) without CH4, but with information from BW, ECM, or BW+ECM in both validation and reference population; (4) with CH4 information and BW, ECM, or BW+ECM information only in the reference population; and (5) with CH4 information and BW, ECM, or BW+ECM information in both validation and reference population. As a result, for each method (BLUP, SSGBLUP), 13 sub-scenarios were performed, 1 from scenario 1, and 3 for each of the subsequent 4 scenarios. The average accuracy of GEBV for CH4 in the base scenario was 0.32 for BLUP and 0.42 for SSGBLUP, and it ranged from 0.10 in scenario 2 to 0.78 in scenario 5 across methods. In terms of bias, the base scenario 1 was unbiased for SSGBLUP; similar results were achieved with scenario 5. Including information on ECM increased the accuracy of GEBV for CH4 by up to 61%, whereas adding information on both traits (BW and ECM) increased the accuracy by up to 90%. Scenarios that did not include CH4 in the reference population had the lowest correlations (0.17-0.33) with single-trait CH4 GEBV, and scenarios with CH4 in the reference population had the highest correlations (0.41-0.81). Thus, failure to include CH4 in future reference populations results in predicted CH4 GEBV, which cannot be used in practical selection. Therefore, recording CH4 in more animals remains a priority. Finally, multiple-trait genomic prediction using routinely recorded BW and ECM leads to higher prediction accuracies than traditional single-trait genomic prediction for CH4 and is a viable solution for increasing the accuracies of GEBV for scarcely recorded CH4 in practice.

Accuracies of breeding values for dry matter intake using nongenotyped animals and predictor traits in different lactations.

Given the interest of including dry matter intake (DMI) in the breeding goal, accurate estimated breeding values (EBV) for DMI are needed, preferably for separate lactations. Due to the limited amount of records available on DMI, 2 main approaches have been suggested to compute those EBV: (1) the inclusion of predictor traits, such as fat- and protein-corrected milk (FPCM) and live weight (LW), and (2) the addition of genomic information of animals using what is called genomic prediction. Recently, several methodologies to estimate EBV utilizing genomic information (EBV) have become available. In this study, a new method known as single-step ridge-regression BLUP (SSRR-BLUP) is suggested. The SSRR-BLUP method does not have an imposed limit on the number of genotyped animals, as the commonly used methods do. The objective of this study was to estimate genetic parameters using a relatively large data set with DMI records, as well as compare the accuracies of the EBV for DMI. These accuracies were obtained using 4 different methods: BLUP (using pedigree for all animals with phenotypes), genomic BLUP (GBLUP; only for genotyped animals), single-step GBLUP (SS-GBLUP), and SSRR-BLUP (for genotyped and nongenotyped animals). Records from different lactations, with or without predictor traits (FPCM and LW), were used in the model. Accuracies of EBV for DMI (defined as the correlation between the EBV and pre-adjusted DMI phenotypes divided by the average accuracy of those phenotypes) ranged between 0.21 and 0.38 across methods and scenarios. Accuracies of EBV for DMI using BLUP were the lowest accuracies obtained across methods. Meanwhile, accuracies of EBV for DMI were similar in SS-GBLUP and SSRR-BLUP, and lower for the GBLUP method. Hence, SSRR-BLUP could be used when the number of genotyped animals is large, avoiding the construction of the inverse genomic relationship matrix. Adding information on DMI from different lactations in the reference population gave higher accuracies in comparison when only lactation 1 was included. Finally, no benefit was obtained by adding information on predictor traits to the reference population when DMI was already included. However, in the absence of DMI records, having records on FPCM and LW from different lactations is a good way to obtain EBV with a relatively good accuracy.

Genomewide association study of methane emissions in Angus beef cattle with validation in dairy cattle.

Methane (CH) is a product of enteric fermentation in ruminants, and it represents around 17% of global CH emissions. There has been substantial effort from the livestock scientific community toward tools that can help reduce this percentage. One approach is to select for lower emitting animals. To achieve this, accurate genetic parameters and identification of the genomic basis of CH traits are required. Therefore, the objectives of this study were 1) to perform a genomewide association study to identify SNP associated with several CH traits in Angus beef cattle (1,020 animals) and validate them in a lactating Holstein population (population 1 [POP1]; 205 animals); 2) to validate significant SNP for DMI and weight at test (WT) from a second Holstein population, from a previous study (population 2 [POP2]; 903 animals), in an Angus population; and 3) to evaluate 2 different residual CH traits and determine if the genes associated with CH also control residual CH traits. Phenotypes calculated for the genotyped Angus population included CH production (MeP), CH yield (MeY), CH intensity (MI), DMI, and WT. The Holstein population (POP1) was multiparous, with phenotypes on CH traits (MeP, MeY, and MI) plus genotypes. Additionally, 2 CH traits, residual genetic CH (RGM) and residual phenotypic CH (RPM), were calculated by adjusting MeP for DMI and WT. Estimated heritabilities in the Angus population were 0.30, 0.19, and 0.15 for MeP, RGM, and RPM, respectively, and genetic correlations of MeP with DMI and WT were 0.83 and 0.80, respectively. Estimated heritabilities in Holstein POP1 were 0.23, 0.30, and 0.42 for MeP, MeY, and MI, respectively. Strong associations with MeP were found on chromosomes 4, 12, 14, 20, and 30 at < 0.001, and those chromosomes also had significant SNP for DMI in Holstein POP1. In the Angus population, the number of significant SNP for MeP at < 0.005 was 3,304, and approximately 630 of those SNP also were important for DMI and WT. When a set (approximately 3,300) of significant SNP for DMI and WT in the Angus population was used to estimate genetic parameters for MeP and MeY in Holstein POP1, the genetic variance and, consequently, the heritability slightly increased, meaning that most of the genetic variation is largely captured by these SNP. Residual traits could be a good option to include in the breeding goal, as this would facilitate selection for lower emitting animals without compromising DMI and WT.

Genetic parameters between feed-intake-related traits and conformation in 2 separate dairy populations--the Netherlands and United States.

To include feed-intake-related traits in the breeding goal, accurate estimates of genetic parameters of feed intake, and its correlations with other related traits (i.e., production, conformation) are required to compare different options. However, the correlations between feed intake and conformation traits can vary depending on the population. Therefore, the objective was to estimate genetic correlations between 6 feed-intake-related traits and 7 conformation traits within dairy cattle from 2 countries, the Netherlands (NL) and the United States (US). The feed-intake-related traits were dry matter intake (DMI), residual feed intake (RFI), milk energy output (MilkE), milk yield (MY), body weight (BW), and metabolic body weight (MBW). The conformation traits were stature (ST), chest width (CW), body depth (BD), angularity (ANG), rump angle (RA), rump width (RW), and body condition score (BCS). Feed intake data were available for 1,665 cows in NL and for 1,920 cows in US, from 83 nutritional experiments (48 in NL and 35 in US) conducted between 1991 and 2011 in NL and between 2007 and 2013 in US. Additional conformation records from relatives of the animals with DMI records were added to the database, giving a total of 37,241 cows in NL and 28,809 in US with conformation trait information. Genetic parameters were estimated using bivariate animal model analyses. The model included the following fixed effects for feed-intake-related traits: location by experiment-ration, age of cow at calving modeled with a second order polynomial by parity class, location by year-season, and days in milk, and these fixed effects for the conformation traits: herd by classification date, age of cow at classification, and lactation stage at classification. Both models included additive genetic and residual random effects. The highest estimated genetic correlations involving DMI were with CW in both countries (NL=0.45 and US=0.61), followed by ST (NL=0.33 and US=0.57), BD (NL=0.26 and US=0.49), and BCS (NL=0.24 and US=0.46). The MilkE and MY were moderately correlated with ANG in both countries (0.33 and 0.47 in NL, and 0.36 and 0.48 in US). Finally, BW was highly correlated with CW (0.77 in NL and 0.84 in US) and with BCS (0.83 in NL and 0.85 in US). Feed-intake-related traits were moderately to highly genetically correlated with conformation traits (ST, CW, BD, and BCS) in both countries, making them potentially useful as predictors of DMI.

Zinc transporter 8 autoantibodies: what is their clinical relevance in gestational diabetes?

AIMS: To investigate the prevalence, clinical significance and antepartum to postpartum trajectory of zinc transporter 8 autoantibodies, a novel marker of islet autoimmunity, in women with gestational diabetes mellitus. METHODS: A total of 302 consecutive women attending a multi-ethnic Australian gestational diabetes clinic were prospectively studied. Zinc transporter 8 autoantibodies were measured at gestational diabetes diagnosis and 3 months postpartum using an enzyme-linked immunosorbent assay, and were correlated with maternal phenotype, antepartum and postpartum glucose tolerance, treatment and perinatal outcomes. RESULTS: Of the 302 women, 30 (9.9%) were positive for one islet autoantibody antepartum. No participant had multiple islet autoantibodies. Zinc transporter 8 autoantibodies were the most prevalent autoantibody [zinc transporter 8 autoantibodies: 13/271 women (4.8%); glutamic acid decarboxylase 7/302 women (2.3%); insulinoma-associated antigen-2: 6/302 women (2.0%); insulin: 4/302 women (1.3%)]. Zinc transporter 8 autoantibody positivity was associated with a higher fasting glucose level on the antepartum oral glucose tolerance test, but not with BMI, insulin use, perinatal outcomes or postpartum glucose intolerance. Five of the six women who tested positive for zinc transporter 8 autoantibodies antepartum were negative for zinc transporter 8 autoantibodies postpartum, which corresponded to a significant decline in titre antepartum to postpartum (26.5 to 3.8 U/ml; P=0.03). This was in contrast to the antepartum to postpartum trajectory of the other islet autoantibodies, which remained unchanged. CONCLUSIONS: Zinc transporter 8 autoantibodies were the most common islet autoantibody in gestational diabetes. Zinc transporter 8 autoantibody positivity was associated with slightly higher fasting glucose levels and, unlike other islet autoantibodies, titres declined postpartum. Zinc transporter 8 autoantibodies may be a marker for islet autoimmunity in a proportion of women with gestational diabetes, but the clinical relevance of zinc transporter 8 autoantibodies in pregnancy and gestational diabetes requires further investigation.

Genetic parameters across lactation for feed intake, fat- and protein-corrected milk, and liveweight in first-parity Holstein cattle.

Breeding values for dry matter intake (DMI) are important to optimize dairy cattle breeding goals for feed efficiency. However, generally, only small data sets are available for feed intake, due to the cost and difficulty of measuring DMI, which makes understanding the genetic associations between traits across lactation difficult, let alone the possibility for selection of breeding animals. However, estimating national breeding values through cheaper and more easily measured correlated traits, such as milk yield and liveweight (LW), could be a first step to predict DMI. Combining DMI data across historical nutritional experiments might help to expand the data sets. Therefore, the objective was to estimate genetic parameters for DMI, fat- and protein-corrected milk (FPCM) yield, and LW across the entire first lactation using a relatively large data set combining experimental data across the Netherlands. A total of 30,483 weekly records for DMI, 49,977 for FPCM yield, and 31,956 for LW were available from 2,283 Dutch Holstein-Friesian first-parity cows between 1990 and 2011. Heritabilities, covariance components, and genetic correlations were estimated using a multivariate random regression model. The model included an effect for year-season of calving, and polynomials for age of cow at calving and days in milk (DIM). The random effects were experimental treatment, year-month of measurement, and the additive genetic, permanent environmental, and residual term. Additive genetic and permanent environmental effects were modeled using a third-order orthogonal polynomial. Estimated heritabilities ranged from 0.21 to 0.40 for DMI, from 0.20 to 0.43 for FPCM yield, and from 0.25 to 0.48 for LW across DIM. Genetic correlations between DMI at different DIM were relatively low during early and late lactation, compared with mid lactation. The genetic correlations between DMI and FPCM yield varied across DIM. This correlation was negative (up to -0.5) between FPCM yield in early lactation and DMI across the entire lactation, but highly positive (above 0.8) when both traits were in mid lactation. The correlation between DMI and LW was 0.6 during early lactation, but decreased to 0.4 during mid lactation. The highest correlations between FPCM yield and LW (0.3-0.5) were estimated during mid lactation. However, the genetic correlations between DMI and either FPCM yield or LW were not symmetric across DIM, and differed depending on which trait was measured first. The results of our study are useful to understand the genetic relationship of DMI, FPCM yield, and LW on specific days across lactation.

Abiotic stress enhances androgenesis from isolated microspores of some legume species (Fabaceae).

To induce androgenesis in field pea, grass pea and the model legume species Medicago truncatula, isolated microspores of various genotypes of these three species were submitted to a range of abiotic stresses prior to and during their initial culture, in order to stimulate them to divide and form embryos. Some stress agents had a positive effect on androgenesis from the treated microspores. Submission of flower buds to a cold period prior to anther excision or microspore isolation, modifying the osmotic pressure of the medium during initial culture and electroporation of isolated microspores were the three major individual stress agents to have an impact on the efficiency of androgenetic proliferation and subsequent differentiation from the microspores of pea, grass pea and M. truncatula genotypes. A combination of osmotic and electric shocks significantly improved responses from isolated microspores and yielded microcalluses and then calluses, but only few underwent morphogenesis. Further work is under way to improve responses and extend them to other genotypes. The results reported here are, to the best of our knowledge, the first successful results from isolated microspores of these species.

[Velopharyngeal sequels in labial-alveolar-velopalatine clefts. Pharyngoplasty by pharynx Lipostructure]. / Les séquelles vélopharyngées dans les fentes labioalvéolopalatovélaires. Pharyngoplastie par Lipostructure du pharynx.

Advancing the posterior pharyngeal wall is a classic technique to treat velopharyngeal insufficiency. Injection of autologous fat behind the posterior pharyngeal mucosa according to the Coleman Lipostructure technique is a recent development. The authors report their experience in six cases using this modification. The preoperative work-up was performed by a speech therapist with a physical examination and measurement of the nasal air loss was performed using an aerophonometer. Fat was harvested either on the abdominal wall or on a buttock and then centrifuged. Fat injection was performed using a curved blunt cannula under the mucosa of the lateral and posterior pharyngeal walls. Injecting fat is an autologous graft of fat tissue: after a postoperative period of three months, the volume of fat becomes permanently stable. In five out of the six patients who presented moderate velopharyngeal insufficiency, speech improvement was significant. The single failure was a patient with bilateral cleft lip and palate sequels after previous pharyngoplasty using the Orticochea procedure. Treatment of moderate velopharyngeal insufficiency using fat injection is an efficient method. The advantages are its innocuousness and that scaring of the pharynx is avoided.

Factor Xa and thrombin, but not factor VIIa, elicit specific cellular responses in dermal fibroblasts.

UNLABELLED: Coagulation factors (F)VIIa, FXa and thrombin are implicated in cellular responses in vascular, mesenchymal and inflammatory cells. Fibroblasts are the most abundant cells in connective tissue, and damage to blood vessels places coagulation factors in contact with these and other cell types. OBJECTIVES: To investigate cellular responses of primary dermal fibroblasts to FVIIa, FXa and thrombin by following changes in expression of candidate proteins: monocyte chemotactic protein-1 (MCP-1), interleukin-8 (IL-8), interleukin-6 (IL-6) and vascular endothelial growth factor (VEGF), and to determine the expression of receptors implicated in signaling by these coagulation factors. METHODS: Steady-state mRNA levels were quantified by RNase protection assay, and protein secretion by ELISA. PAR gene expression was assessed by ribonuclease protection assay and conventional and quantitative reverse-transcription-polymerase chain reaction. RESULTS: FVIIa did not induce the candidate genes. In contrast, FXa and thrombin induced MCP-1 mRNA and protein secretion strongly, IL-8 moderately, and IL-6 weakly. Neither FXa nor thrombin induced VEGF mRNA or protein secretion, although FXa induced VEGF protein secretion in lung fibroblasts. Comparison of the presence of candidate receptors in the two fibroblast subtypes demonstrated higher levels of PAR-1 and PAR-3 in lung fibroblasts relative to their dermal counterparts and the additional expression of PAR-2. CONCLUSIONS: FXa and thrombin induce expression of MCP-1, IL-8 and IL-6, and distribution and expression of PARs on dermal fibroblasts is reduced relative to their lung counterparts. Tissue origin may influence the cellular response of fibroblasts to coagulation proteases.

[Management of cleft lip and palate by the Doctor Magalon's team in Marseille]. / Prise en charge des fentes labio-maxillo-palatines au centre hospitalier universitaire de Marseille.

The goal of the treatment of patients with cleft lip and/or cleft palate was: obtain a normal appearance, a normal speech and a normal growth without severe pertubations of the quality of life. Millard technique was used to repair the cleft lip in neonatal period. Between 6 and 18 months, the palate cleft was closed by wardill-kilner technique. A gingivoperiostoplasty with osseous graft was performed between 8 and 12 years. The orthodontic treatment began at 7 years. Pharyngoplasty was necessary in several cases. Sequellae and rhinoplasty was performed at the end of the growth. The authors explained their choices.

[Cricotracheal resection in children: indications, technique and results]. / La résection crico-trachéale chez l'enfant.

The most significant advance in the surgical treatment of laryngotracheal stenosis has been the changes in external procedures, notably in laryngotracheoplasty aimed at widening the regional stenosis with prosthetic material. In opposition with this therapeutic method, cricotracheal resection which removes the regional stenosis, and a large portion of the cricoid cartilage, has been proven to be a reliable technique in adults. Between June 1993 and June 1998, 10 children underwent cricotracheal resection. There were 5 boys and 5 girls with 9 acquired and 1 congenital stenosis (grade II =5, grade III =2, grade IV =3). At the time of the procedure, the patients' mean weight was 19 kg and mean age was 7.5 years. A tracheotomy present in 5 children prior to the procedure was left in situ postoperatively. In these children a rolled silastic sheet was used to maintain the caliber for 23 days and the tracheotomy canula was removed a mean 58 days later. In the 5 children operated on without tracheaotomy, the nasotracheal tube was removed a mean 2.5 days after the procedure. Mean follow-up was 43 months, with clinical and endoscopic surveillance. No growth retardation was observed among the 5 children presenting 4.5 years after the procedure. The choice between laryngotracheoplasty enlargement and cricotracheal resection is not based on documented evidence but on case-by-case decision making. We discuss here the points which appear to us to be the most relevant in terms of indication, surgical procedure, potential complications, and outcome.

5' sequences direct developmental expression and hormone responsiveness of tyrosine aminotransferase in primary cultures of fetal rat hepatocytes.

Tyrosine aminotransferase (TyrAT) is one of several gluconeogenic enzymes which appear postnatally in humans and rodents in response to increased glucocorticoid and glucagon levels and decreased insulin. Primary cultured fetal rat hepatocytes older than day 15 of gestation (>E15) transcribe the TyrAT gene in response to the synergistic effect of dexamethasone and N6,2'-O-dibutyryl-adenosine 3',5'-monophosphate (Bt2cAMP), whereas less mature hepatocytes (E15 hepatocytes, and not

[Phonetic results after surgery of laryngotracheal stenoses in children]. / Résultats phoniatriques après chirurgie des sténoses laryngo-trachéales chez l'enfant.

The quality of the results of laryngotracheoplasties is clear evidence of the progress made in the treatment of stenoses involving the larynx and the trachea in children. the surgeon is no longer limited to helping the child breath correctly via the natural airways but can also concentrate on phonatrics since dysphonia is frequent after laryngotracheoplasty. Vocal production of 23 children who had undergone surgery for laryngotracheal stenosis was evaluated subjectively by a panel of listeners and objectively on the basis of the pitch and intensity of the voice, rate of speech and maximal time of phonation. In 11 children over the age of 5 years, assisted assessment of the voice completed the study. In 69.5% of the cases, the voice was subjectively considered to be satisfactory. Voice was considered clear in 52% of the cases. Speech flow was impaired in 39% due to frequent breath taking. Objective parameters revealed that pitch was slightly impaired (256 Hz) with a tendancy to normalize near puberty. The intensity of the voice (77 dB) and mean maximum phonation time (7 s) decreased. The phonotary quotient (270 ml/sec), jitter (1.4%) and leakage of the glottis (2.02 cm3/dB/s) were slightly above the normal range. The spectra of the voices were richly furnished with harmonics up to about 1 000 Hz above which air noises increased. An attempt was made to correlate the analysis of the voice with the characteristics of the stenosis and with the therapeutic methods used. The results would suggest the iatrogenic nature of prolonged canulation and of increasing the size the size of the posterior cricoid.(ABSTRACT TRUNCATED AT 250 WORDS)

[Laryngeal stenosis in children. Evaluation of 10 years of treatment]. / Les sténoses laryngées de l'enfant. Bilan de dix ans de traitement.

Over a 10 year period from 1984 to 1994, 98 children underwent curative treatment for sub-glottal stenosis of the larynx at La Timone Hospital in Marseille, France. Eighty-two patients were operated on via an external approach. An endoscopic procedure with the CO2 laser was used in 16 cases. The majority of the children had acquired stenosis (77%), with greater than 70% obstruction (65%), and were under 5 years of age (60%). The details of the different therapeutic techniques used are presented. The laryngotracheoplasty technique used to widen the larynx (n = 58) with an autologous rib cartilage (n = 53) is the most important current progress. The possibilities for laryngotracheofissure (n = 21) and cricotracheal resection (n = 3) are also discussed. After decannulation, 95% of the children could breath normally via the natural airways. The quality of the outcome was not dependent on the therapeutic method used. Perspectives for new methods or modifications of existing methods are discussed.