Dam parity affects fetal growth, placental size, and neonatal metabolism in spring-born beef calves.

To determine effects of dam parity on perinatal nutrient availability in beef cattle, data and samples were collected from 18 primiparous and 35 multiparous spring-calving Sim-Angus dams and their calves. Time to stand was recorded and neonatal vigor assessed. Jugular blood was collected from a subset of calves at 0 (post-standing and pre-suckling) 6, 12, 24, 48, and 72 h of age, and blood chemistry panels were completed. Expelled placentas were dissected, dried, and weighed. Prepartum maternal circulating glucose, non-esterified fatty acids (NEFA), triglycerides, and urea N were analyzed. All statistical models included the fixed effect of dam parity, and calf sex (when P ≤ 0.25) was included for calf and placental variables. Effects of sampling hour, and parity × hour were included for calf metabolites over time using repeated measures. Multiparous dams had greater body weight prepartum (P < 0.001) but similar (P = 0.25) body condition score. Maternal circulating urea N and triglycerides were greater (P ≤ 0.05) in multiparous dams pre-calving. Calves born to primiparous dams weighed 10% less (P ≤ 0.04) at birth with smaller (P ≤ 0.01) heart and abdominal girths. Cotyledonary, intercotyledonary, and total placental masses were less (P ≤ 0.05) for primiparous dams. Dam parity did not affect (P ≥ 0.58) calf time to stand, vigor score at 10 min, or rectal temperature. Serum glucose was greater (P = 0.03) at 0 h but less (P ≤ 0.04) at all other hours in calves from primiparous dams. Calves from primiparous dams had greater (P ≤ 0.02) serum NEFA at 6, 12, and 24 h although plasma triglycerides were greater (P < 0.001) at 6 h. Calves from primiparous dams had greater (P ≤ 0.04) serum urea N at 12 h and creatinine at 12 and 24 h. Plasma insulin was greater (P ≤ 0.04) in calves from multiparous dams at 12, 48, and 72 h, but parity did not affect (P ≥ 0.18) serum total protein or plasma cortisol. Serum aspartate aminotransferase was greater (P ≤ 0.04) at 6 and 24 h, creatine kinase was greater at 24 h, and gamma-glutamyl transpeptidase was less (P ≤ 0.04) at 6, 12, and 24 h, for calves from primiparous dams. Calves born to primiparous dams had greater (P ≤ 0.02) total bilirubin and direct bilirubin at 12 and 24 h. Data indicate that calves born to first-parity heifers had decreased perinatal nutrient availability, resulting in reduced fetal and placental growth, as well as greater energy reserve mobilization and metabolic indicators of stress as neonates.

Approximately two-thirds of beef calf deaths prior to weaning occur within the first 3 wk after birth. The goal to have heifers produce their first calf by 2 yr of age likely contributes to factors that limit nutrients available for fetuses and calves immediately after birth. However, little is known about differences in heifers (first parity) and cows (later parities) regarding factors affecting calf resilience, such as fetal growth and calf metabolism shortly after birth. Our data show that calves born to first-parity heifers had altered nutrient availability, demonstrated through smaller placentas, lower birth weights, and altered metabolites in early life. Although calves had similar vigor and ability to maintain body temperature, calves born to first-parity heifers had to mobilize more energy and had lower insulin during the first 3 d post-birth. Calves born to first-parity heifers had greater indicators of stress during the first 72 h of life not associated with calving difficulties. Overall, these effects may have increased morbidity and mortality of calves born to first-parity heifers if they were in a less intensively-managed system. Better understanding of challenges faced by calves born to first-parity dams provides opportunities for their improved management.

Effects of spring- versus fall-calving on perinatal nutrient availability and neonatal vigor in beef cattle.

To determine the effect of calving season on perinatal nutrient availability and neonatal beef calf vigor, data were collected from 4 spring- (average calving date: February 14; n = 203 total) and 4 fall- (average calving date: September 20; n = 179 total) calving experiments. Time to stand was determined as minutes from birth to standing for 5 s. After birth, calf weight and size (length, heart and abdominal girth, and cannon circumference) were recorded. Jugular blood samples and rectal temperatures were obtained at 0, 6, 12, and 24 h postnatally in 6 experiments and at 48 h postnatally in Exp. 2 to 8. Data were analyzed with fixed effects of season (single point) or season, hour, and their interaction (over time, using repeated measures). Experiment was a random effect; calf sex was included when P ≤ 0.25. Within calving season, correlations were determined between calf size, vigor, and 48-h serum total protein. Fall-born calves tended to have lighter (P = 0.09) birth weight and faster (P = 0.05) time to stand than spring-born calves. Season did not affect (P ≥ 0.18) gestation length, other calf size measures, or 48-h serum total protein. Fall-born calves had greater (P ≤ 0.003) rectal temperature at 0, 24, and 48 h postnatal. Spring-born calves had greater (P ≤ 0.009) circulating glucose at 0 h, serum non-esterified fatty acids at 0 and 6 h, and plasma triglycerides at 0, 6, 12, and 48 h. Fall-born calves had greater (P ≤ 0.03) sodium from 6 to 48 h and magnesium from 0 to 24 h of age. Phosphorus was greater (P ≤ 0.02) at 6 and 12 h of age in spring-born calves. Spring-born calves had greater (P ≤ 0.04) aspartate aminotransferase at 12 and 24 h and creatine kinase at 0 and 12 h of age. Fall-born calves had greater (P ≤ 0.03) albumin, calcium, and chloride, had lower (P ≤ 0.03) bicarbonate and direct bilirubin, and tended to have greater (P = 0.10) anion gap (all main effects of calving season). Calf birth weight had a weak positive relationship (P ≤ 0.03) with 48-h serum total protein and time to stand in fall-born, but not spring-born, calves. Overall, fetal growth was restricted and neonatal dehydration was increased by warm conditions for fall-born calves, but vigor and metabolism were negatively affected by cold conditions in spring-born calves. These data suggest that calving season influences perinatal nutrient availability, which may impact the transition of beef calves to postnatal life.

Blood chemistry and rectal temperature changes in a population of healthy, fall-born, suckling beef calves from birth to 72 h of age.

To determine changes in suckling neonatal beef calf blood chemistry and body temperature during the first 72 h of life, jugular blood samples and rectal temperatures were obtained from 24 healthy, fall-born Angus-cross and Hereford calves (average calving date = September 11) at 0 (after standing, but pre-suckling), 6, 12, 24, 48, and 72 h postnatally. Serum chemistry panels were conducted, and plasma triglycerides and serum non-esterified fatty acids (NEFA) were also determined. Data were analyzed using sampling hour as a repeated effect, and individual data points were compared to adult bovine reference intervals. All serum chemistry measures were affected by hour of age except bicarbonate (P = 0.48). Serum glucose, total protein, and globulin concentrations increased (P < 0.001) from 0 to 24 h. Plasma triglycerides increased (P < 0.001) from 0 to 6 h and 24 to 72 h. Serum urea nitrogen increased (P < 0.001) from 0 to 6 h and decreased (P = 0.01) from 48 to 72 h. Serum NEFA and creatine kinase (CK) increased (P < 0.001) from 0 to 6 h, but decreased (P ≤ 0.02) from 12 to 72 h. Serum magnesium, aspartate aminotransferase (AST), and total and direct bilirubin increased (P < 0.001) during the first 12 h, then decreased (P < 0.001) from 24 to 72 h. Gamma-glutamyl transpeptidase (GGT) increased (P < 0.001) from 0 to 12, and then decreased (P < 0.003) from 12 to 48 h. Serum creatinine decreased (P < 0.001) from 0 to 72 h, and albumin decreased (P < 0.01) from 0 to 12 h but increased (P < 0.001) from 24 to 48 h. Anion gap decreased (P ≤ 0.05) from 6 to 24 h. No serum components measured were within adult bovine reference intervals for all calves at all sampling times. All serum albumin concentrations were outside of reference intervals, and the majority of serum glucose, calcium, phosphorus, potassium, CK, GGT, and total bilirubin were outside of reference intervals. More serum chemistry measures diverged from reference intervals in the first 24 h of neonatal life. Rectal temperature decreased (P = 0.003) from 0 to 6 h of age, then increased (P ≤ 0.02) from 12 to 48 h. In summary, blood chemistry profiles of healthy, suckling neonatal beef calves change over the first 72 h of life, indicating rapid changes in metabolism and physiology. Bovine reference intervals based on adults do not appear to represent neonatal calves; thus, neonatal status and animal age should be taken into consideration when interpreting serum chemistry in a clinical or research setting.

A Feasibility Pilot Study of Online Modules of Hydroxyurea and Sickle Cell Disease Care for Adolescents and Young Adults for Family Medicine Residents.

Family medicine (FP) residency programs are located throughout Indiana, and most adults with sickle cell disease (SCD) in Indiana have access to a primary care clinic administered by a FP program. Allen County ranks third in SCD incidence in Indiana, but has few providers for adolescents, young adults (AYAs) and adults with SCD. Initiation of a novel partnership between Indianapolis-based adult hematologists (130 miles distant), and the FP program in Allen County aimed to educate FP residents about SCD, hydroxyurea, transition, and SCD complications. To determine the feasibility of utilizing online learning modules to educate FP residents about SCD care in AYA and adults, 3 online learning modules (comprehensive care of AYAs with SCD, hydroxyurea, and best practices in AYA transition) were developed and continuing medical education-accredited. Electronic pretest and posttest were distributed to 32 FP residents to test the retention of content through an Institutional Review Board approved protocol. This pilot study demonstrates that it is feasible to utilize online educational modules to educate providers about SCD care.

Factors affecting placental size in beef cattle: Maternal and fetal influences.

Our objectives were to determine the effects of dam body condition score (BCS), age of dam, and calf sex on placental size and the relationships between dam body weight (BW) and calf size with placental size. Expelled placentas and calf size at birth were collected from crossbred beef heifers and cows during four experiments (n = 22 to 39/experiment). Placentas deemed complete by visual inspection were dissected; dry weights were determined for cotyledonary and intercotyledonary tissues. Mixed linear models were used to individually determine main effects of peripartum BCS category [Thin (<5), Moderate (=5), or Fleshy (≥6)], age of dam category [Primiparous (2 yr), Young (3-4 yr), or Mature (≥5 yr)], and calf sex on placental measures. Correlations were determined for placental characteristics with prepartum dam BW, gestation length, and calf size. Thin BCS dams had lower (P ≤ 0.05) cotyledonary, total placental, and average cotyledon weights and greater placental efficiency (calf birth BW/placental weight) than moderate and fleshy dams. Intercotyledonary weight was lower (P < 0.01) in thin BCS dams compared with fleshy dams. Thin and moderate BCS dams had smaller (P ≤ 0.04) calf birth BW than fleshy dams. Primiparous dams had lower (P ≤ 0.05) total placental and average cotyledon weights than young and mature dams, yet calf birth BW was unaffected (P = 0.17). Male calves were heavier (P = 0.01) than females, yet there were no differences (P ≥ 0.59) in placental weights. Calf birth BW and heart girth had moderate positive correlations (P < 0.01) and shoulder to rump length and abdominal girth had weak positive correlations (P < 0.01) with all placental weights. Dam prepartum BW and calf flank girth had moderate positive correlations (P < 0.01) with total placental weights and weak positive correlations (P < 0.01) with cotyledonary and average cotyledon weights. Intercotyledonary weight had moderate positive correlations (P < 0.01) with gestation length and calf flank girth and a weak positive correlation (P < 0.01) with dam prepartum BW. Gestation length had a weak positive correlation (P = 0.02) with total placental weight. Number of cotyledons was not correlated (P ≥ 0.28) with any dam or offspring characteristics. In conclusion, these data suggest that both maternal age and BCS affected placental size. Calf size at birth and placental weight were positively correlated, but it is still unknown which controls and signals for the growth of the other.

Adherence to Quality of Care Indicators and Location of Sickle Cell Care Within Indiana.

Newborn screening (NBS) follow-up programs for infants with sickle cell disease (SCD) are highly variable among states. Initiated in 2009, Sickle SAFE, the NBS follow-up program for infants with SCD in Indiana, follows infants through home visits and phone contact. The current study assessed the attainment rates for recently published quality indicators of pediatric SCD care for Sickle SAFE participants. Using retrospective data, we determined the proportion of children who received transcranial Doppler (TCD) screening, influenza, and pneumococcal vaccination and were prescribed hydroxyurea. We calculated the mean age at confirmatory testing, time to receipt of penicillin prophylaxis, and mean age when genetic counseling was offered. One hundred ninety-eight children born with SCD in Indiana between July 1, 2009 and June 30, 2017 were followed for at least 1 year. While 97.5% received at least one dose of conjugated pneumococcal vaccine, vaccination with the 23 valent pneumococcal vaccine varied by location (county) of care (Allen: 14.3%, Lake: 26.7%, St. Joseph: 40.0%, Marion: 73.3%). Overall TCD screening rate for eligible children was 53%; TCD screening rate varied widely by location of care (Lake: 25% vs. Marion: 63.8%). Similarly, hydroxyurea prescribing practices varied significantly by location of care (p < 0.001). Identified gaps in adherence to quality indicators in SCD care will serve as the basis for future quality improvement initiatives.

Serum Chemistry and Hematology Changes in Neonatal Stock-Type Foals During the First 72 Hours of Life.

Limited research has characterized blood chemistry in healthy stock-type foals. Our objectives were to determine foal serum chemistry and hematology changes during the first 72 hours postnatal and compare these with adult horse reference ranges. Over 3 foaling seasons, serum chemistry and hematology were determined for 16 healthy stock-type foals born to similarly managed mares. Blood was collected at 0 hours (before nursing but after standing, within 85 minutes of birth) and 6, 12, 24, 48, and 72 hours after initial sampling. Data were analyzed with sampling hour, year, and their interaction as fixed effects. Serum glucose and triglyceride concentrations increased (P < .001) from 0 to 24 hours. Triglycerides increased (P < .001) and glucose decreased (P = .01) between 24 to 48 hours. Serum nonesterified fatty acids increased (P = .04) from 0 to 6 hours and decreased (P ≤ .01) from 6 to 24 hours. Serum urea nitrogen increased (P < .001) between 0 and 6 hours and decreased (P < .001) from 12 to 72 hours. Serum total protein and globulin concentrations increased (P < .001) from 0 to 12 hours, albumin decreased (P ≤ .01) from 0 to 24, and creatinine decreased (P ≤ .02) from 0 to 72 hours. Hour also affected (P ≤ .03) electrolytes, metabolic enzymes, and all hematological components except for mean corpuscular hemoglobin (P = .77). These data demonstrate dynamic blood biochemical and hematological changes throughout the first 72 hours in foals, suggesting that sampling time should be considered when evaluating individual neonates.

Locomotion behavior changes in peripartum beef cows and heifers.

Changes in locomotor activity of beef females during the 72 h prepartum were determined in 3 experiments: (i) a 2-yr study with spring-calving multiparous cows (Exp. 1; n = 34 and 27 for years 1 and 2, respectively); (ii) spring-calving primiparous (first pregnancy; n = 13) and multiparous (n = 21) dams (Exp. 2); and (iii) fall-calving multiparous cows (Exp. 3; n = 33). For all experiments, IceQube activity monitors (iceRobotics, Edinburgh, UK) were placed above the left hind fetlock of pregnant females ≥3 d prepartum. During the calving season, females were housed in 18 × 61 m drylots with ad libitum access to hay or haylage. Parturition was closely monitored, and time of birth was noted. Motion index, standing and lying time, step count, and the number of lying bouts for each dam (summed per hour) were determined for the 72 h preceding calving. Within experiment, data were analyzed by day (days -3 to -1 prepartum), by 6-h period during the final 24 h prepartum, and by hour during the final 6 h prepartum using a mixed model with time as a repeated effect. Year was also included as a fixed effect in Exp. 1. Fixed effects of parity and time prepartum × parity were included for Exp. 2. In all 3 experiments, motion index, standing time, step count, and the number of lying bouts were greater (P < 0.001) on day -1 compared with days -2 and -3 prepartum. In the 24 h prepartum, dams had greater (P < 0.01) motion index, standing time, step count, and the number of lying bouts during 6 h preceding parturition compared with -11 to -6 h in all experiments. Motion index, step count, and lying bouts changed (P ≤ 0.02) during the last 6 h in all experiments. Primiparous dams had more (P ≤ 0.01) lying bouts than multiparous dams during the last day and -11 to -6 prepartum. In all experiments, the number of lying bouts more than doubled (P < 0.001) from -2 to -1 prepartum, with no effect of year (P = 0.57) in Exp. 1 or parity (P ≥ 0.29) in Exp. 2. This suggests that lying bout changes may be the most reliable of parameters measured in detection of calving. Moreover, fall-calving cow behavioral patterns were similar to changes observed in spring-calving females, suggesting that calving season may have minimal effects on pre-calving behavior. Overall, electronic locomotor activity monitors can detect behavioral changes peripartum in beef heifers and cows. More research is necessary to determine if these can be used to remotely sense early signs of parturition in beef cattle.

Validation of the Brazilian version of the VERITAS-Pro scale to assess adherence to prophylactic regimens in hemophilia.

INTRODUCTION: Prophylaxis is the treatment of choice for patients with severe hemophilia. Low adherence may limit the effectiveness of the prophylactic regimen, thereby compromising outcomes. OBJECTIVE: The objective of this study was to validate the Brazilian version of the VERITAS-Pro prophylaxis adherence scale, originally an American questionnaire that can be answered by the individual responsible for prophylaxis as well as by an observer. METHODS: The scale has 24 questions divided into six subscales: Routine, Dosage, Plan, Remember, Skip and Communicate. Participants were recruited at a blood center in southeastern Brazil for validation and reliability analyses. Validation measures included the results obtained using analog visual scales of adherence, interval between medication dispensed by the treatment center pharmacy and the percentage of recommended doses administered and infusions registered in the patients' logs. RESULTS: The study included 32 individuals responsible for prophylaxis and five observers. The internal consistency was very good for the VERITAS-Pro total score, excellent for the Remember, Skip and Communicate subscales, good for the Dosage subscale, and acceptable for the Routine and Plan subscales. Twelve participants answered the questionnaire on more than one occasion to evaluate reproducibility. The intraclass correlation coefficient was excellent. Regarding convergent validity, the VERITAS-Pro scores were moderately correlated with the global adherence scale and with infusion log records, but showed a weak correlation with pharmacy dispensation records. CONCLUSION: The Brazilian version of VERITAS-Pro is a valid and reliable instrument, enabling the understanding of specific factors related to non-adherence and allowing targeted interventions for proper treatment.

Validation of the Brazilian version of the VERITAS-Pro scale to assess adherence to prophylactic regimens in hemophilia

Abstract Introduction: Prophylaxis is the treatment of choice for patients with severe hemophilia. Low adherence may limit the effectiveness of the prophylactic regimen, thereby compromising outcomes. Objective: The objective of this study was to validate the Brazilian version of the VERITAS-Pro prophylaxis adherence scale, originally an American questionnaire that can be answered by the individual responsible for prophylaxis as well as by an observer. Methods: The scale has 24 questions divided into six subscales: Routine, Dosage, Plan, Remember, Skip and Communicate. Participants were recruited at a blood center in southeastern Brazil for validation and reliability analyses. Validation measures included the results obtained using analog visual scales of adherence, interval between medication dispensed by the treatment center pharmacy and the percentage of recommended doses administered and infusions registered in the patients' logs. Results: The study included 32 individuals responsible for prophylaxis and five observers. The internal consistency was very good for the VERITAS-Pro total score, excellent for the Remember, Skip and Communicate subscales, good for the Dosage subscale, and acceptable for the Routine and Plan subscales. Twelve participants answered the questionnaire on more than one occasion to evaluate reproducibility. The intraclass correlation coefficient was excellent. Regarding convergent validity, the VERITAS-Pro scores were moderately correlated with the global adherence scale and with infusion log records, but showed a weak correlation with pharmacy dispensation records. Conclusion: The Brazilian version of VERITAS-Pro is a valid and reliable instrument, enabling the understanding of specific factors related to non-adherence and allowing targeted interventions for proper treatment.

Treatment adherence in hemophilia.

Prophylactic clotting-factor regimens reduce the occurrence of bleeding episodes and maintain joint health in individuals with moderate and severe hemophilia. However, these outcomes are only achieved with adherence to prescribed prophylaxis regimens. There are several types of barriers to adherence related to key patient, condition, treatment, health-care system, and/or socioeconomic variables. Notably, health-care professionals may not prescribe prophylaxis if they perceive that a patient will be nonadherent. Prophylactic treatment strategies should be developed with the patient and family, focused on individualized treatment goals. Personalized strategies are needed to reinforce the importance of and encourage confidence in administering the regular infusions required for prophylactic therapy. These strategies may include verbal and written information delivered by health-care professionals, peers, and inter-active media. The advent of extended half-life clotting factors requiring less frequent infusion may improve adherence.

Validation of the VERITAS-Pro treatment adherence scale in a Spanish sample population with hemophilia.

PURPOSE: We aimed to conduct a validation in Spanish of the Validated Hemophilia Regimen Treatment Adherence Scale - Prophylaxis (VERITAS-Pro) questionnaire for use in patients with hemophilia under prophylactic treatment. PATIENTS AND METHODS: The VERITAS-Pro scale was adapted through a process of back translation from English to Spanish. A bilingual native Spanish translator translated the scale from English to Spanish. Subsequently, a bilingual native English translator translated the scale from Spanish to English. The disagreements were resolved by agreement between the research team and translators. Seventy-three patients with hemophilia, aged 13-62 years, were enrolled in the study. The scale was applied twice (2 months apart) to evaluate the test-retest reliability. RESULTS: Internal consistency reliability was lower on the Spanish VERITAS-Pro than on the English version. Test-retest reliability was high, ranging from 0.83 to 0.92. No significant differences (P>0.05) were found between test and retest scores in subscales of VERITAS-Pro. In general, Spanish patients showed higher rates of nonadherence than American patients in all subscales. CONCLUSION: The Spanish version of the VERITAS-Pro has high levels of consistency and empirical validity. This scale can be administered to assess the degree of adherence of prophylactic treatment in patients with hemophilia.

Variable bleeding phenotype in an Amish pedigree with von Willebrand disease.

Through a cross-sectional study design, the bleeding phenotype in the Amish in Indiana (IN) and Wisconsin (WI) was described using two different bleeding scores. von Willebrand factor (VWF) testing was performed and bleeding questionnaires from Centers for Disease Control and Prevention (CDC) and European MCMDM-1 (Tosetto bleeding score (BS)) were administered to the IN and WI cohort respectively. Seven hundred and seventy nine subjects were recruited, 17% were diagnosed with VWD based on Ristocetin cofactor, VWF:RCo < 30 IU/dl. Majority of the affected (AF), 67%, were tested and had a common mutation c.4120 C > T. The WI AF were much younger at a mean age 15 years vs 26 years in IN AF cohort. The AF subjects had a median VWF:RCo of 13IU/dl with a statistically significant higher median BS 1 versus 0 in the WI AF vs WI Unaffected (UA), 2 vs 1 in the IN AF vs IN UA, P < 0.01. Adults had a higher median BS compared to children in the WI and IN cohort, 2 vs 1 and 3 vs 1 respectively (P < 0.05) but there was no statistically significant difference in the BS between males and females in either cohort. The common symptoms reported were epistaxis and gingival oozing. BS ≥ 3 and BS ≥ 4 were observed in 46% of AF IN and 16.6% of AF WI, respectively. There was significant variability in the bleeding phenotype, with an overall low BS in the affected Amish with VWD, despite a unifying mutation. Am. J. Hematol. 91:E431-E435, 2016. © 2016 Wiley Periodicals, Inc.

A validated measure of adherence to antibiotic prophylaxis in children with sickle cell disease.

BACKGROUND: Antibiotic prophylaxis is a mainstay in sickle cell disease management. However, adherence is estimated at only 66%. This study aimed to develop and validate a Sickle Cell Antibiotic Adherence Level Evaluation (SCAALE) to promote systematic and detailed adherence evaluation. METHODS: A 28-item questionnaire was created, covering seven adherence areas. General Adherence Ratings from the parent and one health care provider and medication possession ratios were obtained as validation measures. RESULTS: Internal consistency was very good to excellent for the total SCAALE (α=0.89) and four of the seven subscales. Correlations between SCAALE scores and validation measures were strong for the total SCAALE and five of the seven subscales. CONCLUSION: The SCAALE provides a detailed, quantitative, multidimensional, and global measurement of adherence and can promote clinical care and research.

Examination of Reticulocytosis among Chronically Transfused Children with Sickle Cell Anemia.

Sickle cell anemia (SCA) is an inherited hemolytic anemia with compensatory reticulocytosis. Recent studies have shown that increased levels of reticulocytosis during infancy are associated with increased hospitalizations for SCA sequelae as well as cerebrovascular pathologies. In this study, absolute reticulocyte counts (ARC) measured prior to transfusion were analysed among a cohort of 29 pediatric SCA patients receiving chronic transfusion therapy (CTT) for primary and secondary stroke prevention. A cross-sectional flow cytometric analysis of the reticulocyte phenotype was also performed. Mean duration of CTT was 3.1 ± 2.6 years. Fifteen subjects with magnetic resonance angiography (MRA) -vasculopathy had significantly higher mean ARC prior to initiating CTT compared to 14 subjects without MRA-vasculopathy (427.6 ± 109.0 K/µl vs. 324.8 ± 109.2 K/µl, p<0.05). No significant differences in hemoglobin or percentage sickle hemoglobin (HbS) were noted between the two groups at baseline. Reticulocyte phenotyping further demonstrated that the percentages of circulating immature [CD36(+), CD71(+)] reticulocytes positively correlated with ARC in both groups. During the first year of CTT, neither group had significant reductions in ARC. Among this group of children with SCA, cerebrovasculopathy on MRA at initiation of CTT was associated with increased reticulocytosis, which was not reduced after 12 months of transfusions.

Noninvasive buccal swab antigen sample and molecular testing provides extended antigen typing for patients with hemoglobinopathies.

OBJECTIVE: To demonstrate the feasibility of performing a noninvasive, molecular-based red blood cell (RBC) antigen test on infants and very young children with sickle cell disease as part of a statewide newborn screening follow-up program. STUDY DESIGN: A prospective pilot project was conducted using a noninvasive buccal swab and test kit to perform DNA-based, extended RBC phenotyping in 92 children participating in a newborn hemoglobinopathy screening follow-up program. Reported data include the extended panel of antigens detected by molecular analysis compared with unaffected population estimates. RESULTS: Molecular-based RBC antigen testing was successful, with extended RBC typing generated for all subjects. Molecular testing detected several rare negative or rare positive phenotypes, demonstrating the utility of obtaining an extended antigen panel. CONCLUSION: This study demonstrates the feasibility of performing antigen testing on buccal swab specimens from children with sickle cell disease as part of a newborn screening follow-up program with the aim of allowing specific unit matching to prevent alloimmunization with RBC transfusions. The general applicability of testing may be limited by a lack of uniform insurance coverage for buccal swab testing, however.

The Protein Maker: an automated system for high-throughput parallel purification.

The Protein Maker is an automated purification system developed by Emerald BioSystems for high-throughput parallel purification of proteins and antibodies. This instrument allows multiple load, wash and elution buffers to be used in parallel along independent lines for up to 24 individual samples. To demonstrate its utility, its use in the purification of five recombinant PB2 C-terminal domains from various subtypes of the influenza A virus is described. Three of these constructs crystallized and one diffracted X-rays to sufficient resolution for structure determination and deposition in the Protein Data Bank. Methods for screening lysis buffers for a cytochrome P450 from a pathogenic fungus prior to upscaling expression and purification are also described. The Protein Maker has become a valuable asset within the Seattle Structural Genomics Center for Infectious Disease (SSGCID) and hence is a potentially valuable tool for a variety of high-throughput protein-purification applications.