Erythropoietin attenuates loss of potassium chloride co-transporters following prenatal brain injury.

Therapeutic agents that restore the inhibitory actions of γ-amino butyric acid (GABA) by modulating intracellular chloride concentrations will provide novel avenues to treat stroke, chronic pain, epilepsy, autism, and neurodegenerative and cognitive disorders. During development, upregulation of the potassium-chloride co-transporter KCC2, and the resultant switch from excitatory to inhibitory responses to GABA guide the formation of essential inhibitory circuits. Importantly, maturation of inhibitory mechanisms is also central to the development of excitatory circuits and proper balance between excitatory and inhibitory networks in the developing brain. Loss of KCC2 expression occurs in postmortem samples from human preterm infant brains with white matter lesions. Here we show that late gestation brain injury in a rat model of extreme prematurity impairs the developmental upregulation of potassium chloride co-transporters during a critical postnatal period of circuit maturation in CA3 hippocampus by inducing a sustained loss of oligomeric KCC2 via a calpain-dependent mechanism. Further, administration of erythropoietin (EPO) in a clinically relevant postnatal dosing regimen following the prenatal injury protects the developing brain by reducing calpain activity, restoring oligomeric KCC2 expression and attenuating KCC2 fragmentation, thus providing the first report of a safe therapy to address deficits in KCC2 expression. Together, these data indicate it is possible to reverse abnormalities in KCC2 expression during the postnatal period, and potentially reverse deficits in inhibitory circuit formation central to cognitive impairment and epileptogenesis.

Factors associated with frequency of abortions recorded through Dairy Herd Improvement test plans.

Frequency of abortions recorded through Dairy Herd Improvement (DHI) testing was summarized for cows with lactations completed from 2001 through 2009. For 8.5 million DHI lactations of cows that had recorded breeding dates and were >151 d pregnant at lactation termination, the frequency of recorded abortions was 1.31%. Effects of year, herd-year, month, and pregnancy stage at lactation termination; parity; breed; milk yield; herd size; geographic region; and state within region associated with DHI-recorded abortion were examined. Abortions recorded through DHI (minimum gestation of 152 d required) were more frequent during early gestation; least squares means (LSM) were 4.38, 3.27, 1.19, and 0.59% for 152 to 175, 176 to 200, 201 to 225, and 226 to 250 d pregnant, respectively. Frequency of DHI-recorded abortions was 1.40% for parity 1 and 1.01% for parity ≥ 8. Abortion frequency was highest from May through August (1.42 to 1.53%) and lowest from October through February (1.09 to 1.21%). Frequency of DHI-recorded abortions was higher for Holsteins (1.32%) than for Jerseys (1.10%) and other breeds (1.27%). Little relationship was found between DHI-recorded abortions and herd size. Abortion frequencies for effects should be considered to be underestimated because many abortions, especially those caused by genetic recessives, go undetected. Therefore, various nonreturn rates (NRR; 60, 80, …, 200 d) were calculated to document pregnancy loss confirmed by the absence of homozygotes in the population. Breeding records for April 2011 US Department of Agriculture sire conception rate evaluations were analyzed with the model used for official evaluations with the addition of an interaction between carrier status of the service sire (embryo's sire) and cow sire (embryo's maternal grandsire). Over 13 million matings were examined using various NRR for Holstein lethal recessive traits (brachyspina and complex vertebral malformation) and undesirable recessive haplotypes (HH1, HH2, and HH3) as well as >61,000 matings for a Brown Swiss haplotype (BH1), and 670,000 matings for a Jersey haplotype (JH1). Over 80% of fertility loss occurred by 60 d after breeding for BH1, HH3, and JH1, by 80 d for HH2, by 100 d for BY, and by 180 d for HH1. For complex vertebral malformation, fertility loss increased from 40 to 74% across gestation. Association of undesirable recessives with DHI-recorded abortions ranged from 0.0% for Jerseys to 2.4% for Holsteins.

Genome sequences of Mycoplasma alligatoris A21JP2T and Mycoplasma crocodyli MP145T.

Mycoplasma alligatoris and Mycoplasma crocodyli are closely related siblings, one being highly virulent and the other relatively attenuated. We compared their genomes to better understand the mechanisms and origins of M. alligatoris' remarkable virulence amid a clade of harmless or much less virulent species. Although its chromosome was refractory to closure, M. alligatoris differed most notably by its complement of sialidases and other genes of the N-acetylneuraminate scavenging and catabolism pathway.

Potential consequences of selection to change gestation length on performance of Holstein cows.

Genetic evaluations for gestation length (GL) for Holstein service sires were studied to determine their effectiveness in predicting GL in an independent data set. Consequences of selection on GL were also assessed by examining correlated changes in milk and fitness traits. Holstein bulls with ≥ 300 calvings between 1998 and 2005 were stratified into the following 7 groups using predicted transmitting ability (PTA) for service sire GL: <-3.00, -3.00 to -2.01, …, 1.00 to 1.99, and ≥ 2.00 d. An independent set of 261,598 first-parity cows mated later to the same bulls and calving between 2006 and 2009 were segregated by the service sire PTA GL groups (group had 8,317 to 73,324 gestations), and these mates' GL were examined to determine effectiveness of service sire PTA GL. The model included fixed effects for herd-year and service sire group, plus covariates for conception dates to account for time opportunity among mates. Mean GL for mates by service sire group (from lowest to highest PTA GL) were 275.3, 276.5, 277.8, 278.6, 279.5, 280.6, and 281.7 d. Thus, service sire PTA GL was effective in identifying bulls that modified GL. Subsequent yield and fitness traits were also examined for the (independent) mates with the same service sire groups. Intermediate service sire PTA GL was optimal for yield traits and days open; performance for productive life and culling generally became less favorable as service sire PTA GL increased. A second examination was made by replacing service sire PTA GL groups in the model with phenotypic cow GL groups. Relationships between GL and subsequent performance for milk yield and fitness traits were examined using 9 phenotypic cow GL groups: ≤ 271, 272-273, …, 284-285, and ≥ 286 d. Performance generally improved for subsequent lactation yield as cow GL increased; however, intermediate GL was optimal for productive life, calving ease, stillbirth, culling, and days open. Results indicated that neither shortening nor increasing the mean for GL in the Holstein breed provided much overall benefit when all traits were considered. The same traits examined in the cows for the correlated effect from various GL were also examined in their offspring to determine whether the GL producing the calf had any influence on these same traits when the offspring reached their own productive period. Little carryover occurred from GL on the dam to the other traits observed on the offspring when examined a generation later.

Consequence of alternative standards for bulk tank somatic cell count of dairy herds in the United States.

Noncompliance with current US and European Union (EU) standards for bulk-tank somatic cell count (BTSCC) as well as BTSCC standards recently proposed by 3 US organizations was evaluated using US Dairy Herd Improvement Association (DHI) herds and herds supplying milk to 4 Federal Milk Marketing Orders (FMO). Herds with 15 to 26 tests (frequently monthly) from January 2009 through October 2010 were included. Somatic cell scores (SCS) from 14,854 herds and 164,794 herd-tests were analyzed for DHI herds with ≥10 cows for all tests. Herd test-day SCC was derived as a proxy for BTSCC and was the basis for determining noncompliance and percentage of the milk it represented. For FMO herds, actual milk marketed and BTSCC were available from 27,759 herds and 325,690 herd-tests. A herd was noncompliant for the current EU BTSCC standard after 4 consecutive rolling 3-test geometric means (geometric method) were >400,000 cells/mL. A herd was noncompliant for the current US BTSCC standard after 3 of 5 consecutive monthly BTSCC shipments (frequency method) were >750,000 cells/mL. Alternative proposed standards (600,000, 500,000, or 400,000 cells/mL) also were examined. A third method designated noncompliance when a single 3-mo geometric mean of >550,000 or >400,000 cells/mL and a subsequent test exceeded the same level. Results were examined based on herd size or milk shipped by month. Noncompliance for the current US standard for the 12 mo ending October 2010 in DHI and FMO herds was 0.9 and 1.0%, respectively, compared with 7.8 and 16.1% for the current EU standard. Noncompliance was always greater for the frequency method than for the geometric method and was inversely related to herd size or milk shipped. Using the frequency method at 400,000 cells/mL, noncompliance was 19.1% for DHI herd-tests in herds with <50 cows compared with 1.1% for herds with ≥ 1,000 cows. For FMO herds shipping <900 t, noncompliance was 44.5% using the frequency method at 400,000 cells/mL compared with 8.0% for herds marketing >9,000 t. All methods proposed increased the percentages of herds and shipped milk that exceeded the regulatory limit. Producers will need to place more emphasis on reducing the incidence and prevalence of subclinical mastitis through known management practices such as proper milking techniques, well-functioning milking machines, postmilking teat disinfectant, dry cow treatment, and culling of problem cows to meet any of the proposed new standards.

Use of sexed semen and its effect on conception rate, calf sex, dystocia, and stillbirth of Holsteins in the United States.

Use of sexed semen for artificial insemination of US Holstein heifers (1.3 million breedings) and cows (10.8 million breedings) in Dairy Herd Improvement herds was characterized by breeding year, parity, service number, region, herd size, and herd milk yield. Sexed semen was used for 1.4, 9.5, and 17.8% of all reported breedings for 2006, 2007, and 2008, respectively, for heifers, and for 0.1, 0.2, and 0.4%, respectively, for cows. For 2008 sexed semen breedings, 80.5 and 68.6% of use was for first services of heifers and cows, respectively. For cows, 63.1% of 2008 sexed semen use was for first parity. Mean sexed semen use within herd was the greatest for heifers in the Southwest (36.2%) and for cows in the Mideast (1.3%). Mean sexed semen use increased for heifers but changed little for cows as either herd size or herd mean milk yield increased. Availability of sexed semen was examined for Holstein bulls in active AI service; of 700 bulls born after 1993, 37% had sexed semen marketed by mid August 2009. Active AI bulls with marketed sexed semen were superior to average active AI bulls for evaluations of yield traits, productive life, somatic cell score, daughter pregnancy rate, service-sire calving ease, service-sire stillbirth, final score, sire conception rate, and lifetime net merit. The effect of sexed semen use on conception rate, calf sex, dystocia, and stillbirth also was examined for heifers and cows. Mean conception rate for heifers was 56% for conventional and 39% for sexed semen; corresponding conception rates for cows were 30 and 25%. For single births from sexed semen breedings, around 90% were female. Dystocia and stillbirth were more frequent for heifers (6.0 and 10.4%, respectively, for conventional semen; 4.3 and 11.3%, respectively, for sexed semen) than for cows (2.5 and 3.6%, respectively, for conventional semen; 0.9 and 2.7%, respectively, for sexed semen). Difficult births declined by 28% for heifers and 64% for cows with sexed semen use. Stillbirths were more prevalent for twin births except for sexed semen heifer breedings. Stillbirths of single male calves of heifers were more frequent for breedings with sexed semen (15.6%) than conventional semen (10.8%); a comparable difference was not observed for cows, for which stillbirth frequency of single male calves even decreased (2.6 vs. 3.6%). Overall stillbirth frequency was reduced by sexed semen use for cows but not for heifers.

Response to alternative genetic-economic indices for Holsteins across 2 generations.

Four US genetic-economic indices for dairy cattle were retrofitted to illustrate differences in phenotypic response observed for retrospective selection over 2 generations for currently evaluated traits, even though producers did not have evaluations available at the time for direct selection for those traits. Differences among cows were compared based on ranking of their sires and maternal grandsires (MGS) for the 4 retrofitted indices. Holstein artificial insemination bulls (106,471) were categorized by quintile for each index, and 25 cow groups were formed based on quintiles for sire and MGS (2 generations). Data included records from 1,756,805 cows in 26,106 herds for yield traits, productive life, pregnancy rate, and somatic cell score; 692,656 cows in 9,967 herds for calving difficulty; and 270,564 cows in 4,534 herds for stillbirths. For each index, least squares differences between the 25 cow groups were examined for 8 first-parity traits (milk, fat, and protein yields; productive life; somatic cell score; pregnancy rate; calving difficulty; and stillbirth) that had been standardized for age. Analysis removed effects of herd and cow birth year. Seven of 25 cow groups were consolidated into 3 groups based on index ranking for their male ancestors (low, medium, and high). The cow group with high sire and MGS rankings for the 2006 net merit index produced more milk (219 kg), fat (21 kg), and protein (11 kg) and had longer productive life (6.3 mo), lower somatic cell score (0.21), higher pregnancy rate (1.2 percentage units), fewer difficult births in heifers (3.8 percentage units), and lower stillbirth rate (4.6 percentage units) than did the group with low sire and MGS rankings. For cow groups based on sire and MGS rankings for 1971 (milk and fat) and 1977 (milk, fat, and protein) indices, advantages for the group with high sire and MGS rankings were much larger for yield traits but smaller (and sometimes even unfavorable) for other traits. Cow groups based on sire and MGS rankings for the 1994 net merit index generally had differences that were intermediate to groups based on sire and MGS rankings for the 1977 and 2006 indices. Phenotypic differences revealed retrospectively between genetic-economic indices indicate that genetic improvement should be made for all traits included in recent net merit indices.

Cortisol regulates insulin-like growth-factor binding protein (igfbp) gene expression in Atlantic salmon parr.

The growth hormone (Gh)/insulin-like growth-factor (Igf)/Igf binding protein (Igfbp) system regulates growth and osmoregulation in salmonid fishes, but how this system interacts with other endocrine systems is largely unknown. Given the well-documented consequences of mounting a glucocorticoid stress response on growth, we hypothesized that cortisol inhibits anabolic processes by modulating the expression of hepatic igfbp mRNAs. Atlantic salmon (Salmo salar) parr were implanted intraperitoneally with cortisol implants (0, 10, and 40 µg g-1 body weight) and sampled after 3 or 14 days. Cortisol elicited a dose-dependent reduction in specific growth rate (SGR) after 14 days. While plasma Gh and Igf1 levels were unchanged, hepatic igf1 mRNA was diminished and hepatic igfbp1b1 and -1b2 were stimulated by the high cortisol dose. Plasma Igf1 was positively correlated with SGR at 14 days. Hepatic gh receptor (ghr), igfbp1a, -2a, -2b1, and -2b2 levels were not impacted by cortisol. Muscle igf2, but not igf1 or ghr, levels were stimulated at 3 days by the high cortisol dose. As both cortisol and the Gh/Igf axis promote seawater (SW) tolerance, and particular igfbps respond to SW exposure, we also assessed whether cortisol coordinates the expression of branchial igfbps and genes associated with ion transport. Cortisol stimulated branchial igfbp5b2 levels in parallel with Na+/K+-ATPase (NKA) activity and nka-α1b, Na+/K+/2Cl--cotransporter 1 (nkcc1), and cystic fibrosis transmembrane regulator 1 (cftr1) mRNA levels. The collective results indicate that cortisol modulates the growth of juvenile salmon via the regulation of hepatic igfbp1s whereas no clear links between cortisol and branchial igfbps previously shown to be salinity-responsive could be established.

Foamy virus as a gene transfer vector to the central nervous system.

Engineered foamy virus (FV) vectors have been lauded for their superior safety profiles and stable integration patterns compared to their gammaretroviral counterparts. The drawback has been the belief that FV incorporation is cell cycle-dependent, thereby limiting its utility in post-mitotic tissues such as the central nervous system. In this brief communication, we challenged this theory by examining FV in vivo. We injected equal titers of FV and lentivirus (LV) into the adult rat brain and found that at 1 week, FV transduced a significantly greater volume of bromodeoxyuridine (BrdU)-negative brain parenchyma than did LV. By 8 weeks, however, the volume of transduced tissue was greatly reduced--comparable to LV-and restricted to BrdU+. Taken together, these data implicate a role for FV in short-term gene delivery strategies to the CNS.

Cross-bridges mediate anterograde and retrograde vesicle transport along microtubules in squid axoplasm.

To assay the detailed structural relationship between axonally transported vesicles and their substrate microtubules, vesicle transport was focally cold blocked in axoplasm that was extruded from the squid giant axon. A brief localized cold block concentrated anterogradely and retrogradely transported vesicles selectively on either the proximal or or distal side of the block. Normal movement of the concentrated vesicles was reactivated by rewarming the cold-blocked axoplasm. At the periphery of the axoplasm, moving vesicles were located on individual microtubules that had become separated from the other cytomatrix components. The presence of moving vesicles on isolated microtubules permitted the identification of the structural components required for vesicle transport along microtubules. The results show that 16-18-nm cross-bridges connect both anterogradely and retrogradely moving vesicles to their substrate microtubules. These observations demonstrate that cross-bridges are fundamental are fundamental components of vesicle transport along axonal microtubules. Thus, vesicle transport can now be included among those cell motile systems such as muscle and axonemes that are based on a cross-bridge-mediated mechanism.

Morphology, behavior, and interaction of cultured epithelial cells after the antibody-induced disruption of keratin filament organization.

The organization of intermediate filaments in cultured epithelial cells was rapidly and radically affected by intracellularly injected monoclonal antikeratin filament antibodies. Different antibodies had different effects, ranging from an apparent splaying apart of keratin filament bundles to the complete disruption of the keratin filament network. Antibodies were detectable within cells for more than four days after injection. The antibody-induced disruption of keratin filament organization had no light-microscopically discernible effect on microfilament or microtubule organization, cellular morphology, mitosis, the integrity of epithelial sheets, mitotic rate, or cellular reintegration after mitosis. Cell-to-cell adhesion junctions survived keratin filament disruption. However, antibody injected into a keratinocyte-derived cell line, rich in desmosomes, brought on a superfasciculation of keratin filament bundles, which appeared to pull desmosomal junctions together, suggesting that desmosomes can move in the plane of the plasma membrane and may only be 'fixed' by their anchoring to the cytoplasmic filament network. Our observations suggest that keratin filaments are not involved in the establishment or maintenance of cell shape in cultured cells.

Rat optic nerve oligodendrocytes develop in the absence of viable retinal ganglion cell axons.

Retinal ganglion cell axons and axonal electrical activity have been considered essential for migration, proliferation, and survival of oligodendrocyte lineage cells in the optic nerve. To define axonal requirements during oligodendrogenesis, the developmental appearance of oligodendrocyte progenitors and oligodendrocytes were compared between normal and transected optic nerves. In the absence of viable axons, oligodendrocyte precursors migrated along the length of the nerve and subsequently multiplied and differentiated into myelin basic protein-positive oligodendrocytes at similar densities and with similar temporal and spatial patterns as in control nerves. Since transected optic nerves failed to grow radially, the number of oligodendrocyte lineage cells was reduced compared with control nerves. However, the mitotic indices of progenitors and the percentage of oligodendrocytes undergoing programmed cell death were similar in control and transected optic nerves. Oligodendrocytes lacked their normal longitudinal orientation, developed fewer, shorter processes, and failed to form myelin in the transected nerves. These data indicate that normal densities of oligodendrocytes can develop in the absence of viable retinal ganglion axons, and support the possibility that axons assure their own myelination by regulating the number of myelin internodes formed by individual oligodendrocytes.

Molecular specialization of astrocyte processes at nodes of Ranvier in rat optic nerve.

The HNK-1 and L2 monoclonal antibodies are thought to recognize identical or closely associated carbohydrate epitopes on a family of neural plasma membrane glycoproteins, including myelin-associated glycoprotein, the neural cell adhesion molecule, and the L1 and J1 glycoproteins, all of which have been postulated to play a part in mediating cell-cell interactions in the nervous system. We have used these two antibodies in immunofluorescence and immunogold-electron microscopic studies of semithin and ultrathin frozen sections of adult rat optic nerve, respectively, and we show that they bind mainly to astrocyte processes around nodes of Ranvier. Most other elements of the nerve, including astrocyte cell bodies and large astrocytic processes, are not labeled by the antibodies. To our knowledge, this is the first demonstration that perinodal astrocyte processes are biochemically specialized. We provide evidence that one of the HNK-1+/L2+ molecules concentrated around perinodal astrocyte processes is the J1 glycoprotein; our findings, taken together with previously reported observations, suggest that the other known HNK-1+/L2+ molecules are not concentrated on these processes. Since anti-J1 antibodies previously have been shown to inhibit neuron to astrocyte adhesion in vitro, we hypothesize that J1 may play an important part in the axon-glial interactions that presumably are involved in the assembly and/or maintenance of nodes of Ranvier.

Proteolytic self-cleavage of hepatitis B virus core protein may generate serum e antigen.

A model is proposed to explain the presence of the e antigen (HBeAg) of hepatitis B virus (HBV) in the serum of individuals infected with this virus. The e antigen, which has only recently been characterized, is a fragment of the virus core, or nucleocapsid, protein. Serum HBeAg is a valuable clinical marker for active HBV infection because its appearance correlates both with virus replication in the liver and with the presence of circulating virions. In this study a protease-like amino acid sequence was identified at the amino terminus of the core protein sequence. Experimental evidence indicates that HBeAg may be produced by proteolytic self-cleavage of the core protein.

Human immunodeficiency virus may encode a novel protein on the genomic DNA plus strand.

The genome of the human immunodeficiency virus (HIV) is known to contain eight open reading frames (ORFs) on the minus strand of the double-stranded DNA replicative intermediate. Data presented here indicate that the DNA plus strand of HIV contains a previously unidentified ORF in a region complementary to the envelope gene sequence. This ORF could encode a protein of approximately 190 amino acid residues with a relative molecular mass of 20 kilodaltons if translation began from the first initiation codon. The predicted protein is highly hydrophobic and thus could be membrane associated. It is possible, therefore, that the HIV genome encodes a protein on antisense messenger RNA.

Synodic month: variations in the geologic past.

The value of 31.56 +/- 0.74 days for the synodic month in the Upper Cambrian is used together with a maximum in the expected number of days in a synodic month to argue that solar tidal dissipation is important in the dynamics of the earth-moon system and that the moment of inertia of the earth cannot have been much larger then than it is now. The tendency of the measurements to remain near 30 days may be a resonance effect.

Preferred microtubules for vesicle transport in lobster axons.

The hypothesis that transported vesicles are preferentially associated with a subclass of microtubules has been tested in lobster axons. A cold block was used to collect moving vesicles in these axons; this treatment caused the vesicles to accumulate in files along some of the microtubules. Quantitative analysis of the number of vesicles associated with microtubule segments indicated that lobster axons have two distinct populations of microtubules--transport microtubules that are the preferred substrates for vesicle transport and architectural microtubules that contribute to axonal structure.

Silurian of central Texas: a first record for the region.

Silurian outcrops, not previously recorded from central Texas, have been identified from the Llano uplift, where they occur in collapse structures within the Lower Ordovician Honeycut Formation of the Ellenburger Group. The formation is a pinkish-gray granular limestone, contains fossils of probable Wenlock age, and is named the Starcke Limestone.

Lack of protective immunity against reinfection with hepatitis C virus.

Some individuals infected with hepatitis C virus (HCV) experience multiple episodes of acute hepatitis. It is unclear whether these episodes are due to reinfection with HCV or to reactivation of the original virus infection. Markers of viral replication and host immunity were studied in five chimpanzees sequentially inoculated over a period of 3 years with different HCV strains of proven infectivity. Each rechallenge of a convalescent chimpanzee with the same or a different HCV strain resulted in the reappearance of viremia, which was due to infection with the subsequent challenge virus. The evidence indicates that HCV infection does not elicit protective immunity against reinfection with homologous or heterologous strains, which raises concerns for the development of effective vaccines against HCV.

Impact of genetic merit for milk somatic cell score of sires and maternal grandsires on herd life of their daughters.

A retrospective study of the impact of the estimated breeding values of sires and maternal grandsires for somatic cell score (SCS) on productive life (PL) of Holsteins and Jerseys was conducted. Data included records from 2,626,425 Holstein and 142,725 Jersey cows. The sires and maternal grandsires of cows were required to have been available through artificial insemination and to have predicted transmitting ability (PTA) SCS evaluations based on 35 or more daughters. A weighted function (WPTA) of sire and maternal grandsire PTA for SCS was used: (sire PTA + 0.5 maternal grandsire PTA)/1.5. The 3 dependent variables were PL, frequency of cows culled for mastitis, and first-lactation SCS. The model included effects of herd, birth year, and WPTA (WPTA was categorized into groups: <2.70, 2.70 to 2.79, ..., 3.20 to 3.29, > or =3.30). For analysis of first-lactation SCS, calving year and calving month were substituted for birth year. Differences among WPTA groups were highly significant: as WPTA increased, PL decreased, whereas percentage culled for mastitis and first-lactation SCS increased. The range in PL from lowest to highest WPTA was 5.07 mo for Holsteins and 4.73 mo for Jerseys. Corresponding differences for percentage culled for mastitis were 7.0 and 5.6% and for SCS were 0.95 and 1.04 (for Holsteins and Jerseys, respectively). Although phenotypic studies suggest that cows with extremely low SCS were less resistant to mastitis, our results showed consistent improvements in PL, percentage culled for mastitis, and SCS of daughters when bulls were chosen for low PTA SCS.