CBC Results Research Articles

Does Chronic Ketone Salt Supplementation Alter BP, CBC, or CMP Results in Adults Diagnosed with PTSD?

This item presents the abstract for a presentation at the 2021 Annual Phi Kappa Phi Student Research and Fine Arts Conference at Augusta University.

Comparison of plasma total solids concentration as measured by refractometry and plasma total protein concentration as measured by biuret assay in pet rabbits and ferrets

To determine the agreement between plasma total solids (TS) concentration as measured by refractometry and plasma total protein (TP) concentration as measured by biuret assay in pet rabbits and ferrets. 253 and 146 blood samples from 146 and 121 ferrets and rabbits, respectively, with results of CBC and plasma biochemical analyses. Data were collected from medical records regarding plasma TS and TP concentrations, PCV, plasma biochemical values, plasma appearance, and patient signalment. Agreement was determined between refractometer and biuret assay (reference method) values for plasma TS and TP concentration. Other variables were examined for an impact on this agreement. Mean ± SD plasma TP and TS concentrations were 6.4 ± 0.8 mg/dL and 6.6 ± 0.8 mg/dL, respectively, for rabbits and 6.3 ± 1.2 mg/dL and 6.4 ± 1.1 mg/dL for ferrets. On average, refractometer values overestimated plasma TP concentrations as measured by biuret assay. Plasma cholesterol, glucose, and BUN concentrations and hemolysis and lipemia had significant effects on this bias for ferrets; only BUN concentration had an effect on bias for rabbits given the available data. Other variables had no influence on bias. The limits of agreement were wider than the total allowable analytic error, and > 5% of the data points were outside acceptance limits, indicating that the 2 methods were not in clinical agreement. Refractometer measurements of plasma TS concentration failed to provide a good estimation of biuret assay measurements of plasma TP concentration in rabbits and ferrets, suggesting that these 2 analytic methods and the results they yield cannot be used interchangeably in these species.

Long-Term Outcomes of Bronchiectasis Surgery: 108 Cases

Purpose: We aimed to analyze the cases that we monitored for long term after applying surgical resection. Materials and Methods: Admitted to our clinic between 2005 and 2019, the files of 108 patients (38 male, 70 female, mean age of 23.03 years (18-63)), who visited our clinic, were diagnosed with bronchiectasis and operated on, were retrospectively analyzed. For the monitoring of the patients after surgery, chest radiography, PFT, CBC and CRP follow-up results, and their preoperative complaints (coughing, expectoration, recurrent pulmonary infections) were examined and recorded. While assessing the patients’ status after the surgery, they were categorized as disappearance of the preoperative complaints, decrease in the complaints, and no change in the complaints. Results: 115 surgical operations were carried out on 108 patients. The most frequently encountered complications after surgery were atelectasis and prolonged air leak. Mostly lobectomy was applied as surgery. Postoperative morbidity was seen in 44 patients (41%). Mortality was seen in 5 patients (5%). Complete recovery was observed in 87 patients (81%), while the complaints of 15 patients (14%) were on an acceptable level, and they needed to use antibiotics. The disease relapsed in another lobe in 6 patients (5%). The mean time of monitoring among our patients was 3.71 years (8 months - 12 years). Conclusion: Infections fall into a vicious circle in bronchiectasis patients who are not appropriately treated. The risk factors that require surgery determine morbidity. This cycle may be ended by the surgical resection of the affected tissues.

CLINICAL AND HEMATOLOGICAL FEATURES OF LEISHMANIASIS IN A TERTIARY CARE HOSPITAL OF PESHAWAR

Objective: To evaluate the hematological and clinical features of Visceral and Cutaneous Leishmaniasis.
 Materials and Methods: This was hospital-based retrospective study which included all cases of Cutaneous leishmaniasis and Visceral leishmaniasis that were diagnosed from Jan 2018 till December 2019 at Pathology Department, Rehman Medical Institute, Peshawar. The diagnosed cases were analyzed for clinical and laboratory profile in details, results of CBC, demographic information and physical signs at presentation were noted as well as bone marrow aspirations were performed for LD bodies.
 Results: Out of 104 cases, 36 (34.61%) visceral leishmaniasis and 68 (65.38%) cutaneous leishmaniasis were observed during the study period, in which 82 (78.85%) were male and 22 (21.15%) were females. Age range was from 03-40 years with mean of 23 + 4.21 years. Intermittent fever was observed in almost all the cases of visceral leishmaniasis and 52 (76.47%) of cutaneous leishmaniasis. All patients with visceral leishmaniasis had hepatomegaly and splenomegaly. Patients with CL presented as lesions on face and foot region. In VL all patients were male, while in CL 46 were males and only 22 were females. Majority (83.33%) of patients in VL were children (age group 1-10 years), in cutaneous leishmaniasis 34 (50%) were in age group 31-40 years.
 Conclusion: Our study concludes that leishmaniasis mainly affecting age group 10-20 years. Patients with visceral Leishmaniasis presented with pallor, weight loss fever and splenomegaly while those with cutaneous leishmaniasis presented with lesions on the foot and face especially nose. Anemia and thrombocytopenia were most common hematological parameters. It is essential that the Public Health authorities be more aware of the condition in order to improve environmental sanitation and personal protective measures and to establish diagnostic laboratories for early and correct diagnosis and treatment.

Outcomes of Hydroxyurea Dose Adjustment Via Telehealth

The UAB Pediatric Sickle Cell Clinic cares for patients at both an academic center and satellite clinics. One difference in clinics is that CBC results are not available at the time of patient care during satellite clinics, but are available at the academic center. Therefore, hydroxyurea (HU) dosing adjustments are made within seven days through telehealth; after reviewing faxed CBC results, a sickle cell nurse calls families and educates them on any revised HU dosing plans and reinforces adherence to HU. We hypothesized that this type of telehealth HU dosing adjustments would not impact HU laboratory response. Methods: We performed a one-year retrospective review of 172 HbSS and HbSB0 thalassemia patients prescribed HU during 2019 that were cared for in both the academic center (Birmingham) and satellite clinics (Montgomery, Tuscaloosa, Opelika). We excluded patients on HU and a second sickle cell disease modifying therapy. We abstracted from the EMR: age, sex, clinic location, diagnosis, HU dose (mg/kg), WBC, Hb, MCV, Platelets, Absolute Reticulocyte, HbF, and ANC. We recorded the total number of well-clinic visits and categorized clinic visit/year as ≤ 2or >2; satellite clinic visits are fixed either monthly to every three months which limited our ability to reschedule patients with missed visits. We also recorded the number of ED visits and admissions to the hospital for a) pain and b) all sickle cell complications except for scheduled procedures. We collected acute visit data from the EMR at the academic center for onsite participants and from patient report from satellite participants. We evaluated dosing adjustment as whether dosing was increased for ANC > 4000 per institutional standard of care. We categorized correct dosing adjustments if > 65% of visits had a dose adjustment per institutional protocol; we categorized patients as a deviation from protocol if the physician did not increase the dose based on ANC. We performed descriptive statistics, t-test for continuous variables, chi-square for categorical variables, and regression modeling using JMP Pro 14 (Cary, NC). P-values <0.05 were considered statistically significant. Results: The mean age of the 172 participants was 11±5 years, mean Hb was 8.5±1.1g/dL, mean MCV was 91.3±10.1 fL, mean HbF% was 12.7±6.7, and mean ANC was 4.9±2.3. Fifty two percent of participants were female. We identified 107 participants prescribed HU at the academic center and 65 participants from satellite clinics. We identified no significant differences in HU monitoring laboratory values for Hb (p=0.6), MCV (p=0.7), HbF (p=0.13), ANC (p=0.9) by clinic location. We did not identify a difference in sex of participants by clinic location (p=0.9); also, HbF% (p=0.6) and HU dose (p=0.3) were not associated with sex. We did not identify a difference in all admissions to the hospital (p=0.5) or pain admissions to the hospital (p=0.6) between academic center vs. satellite clinic patients. However, academic center patients did have a higher number of acute pain visits (p=0.03) to either the ED or UAB pediatric pain clinic. As expected, among all participants, non-adherence to HU (HbF <5%) was associated with the highest number of admissions (p=0.02). Among 149 patients with more than one clinic visit, 97 (65%) patients had more than 65% of clinic visits with a correct dose adjustment; we identified no statistical difference in correct dosing adjustment deviations among Birmingham patients (67%) and satellite clinic patients (62%, p=0.5). Patients cared for at the academic center had a statistically higher number of clinic visits (p=0.01) and more patients that attended >2 clinic visits. HbF response was significantly associated with clinic visits (p=0.03). Finally, HbF response was significantly inversely correlated with age (p=0.003). Adjusting for clinic location, number of clinic visits, and age, HbF % was associated with age (p=0.006) and clinic visits (p=0.006), but not associated with clinic location. Conclusion: Our data demonstrate that telehealth for HU dosing provided a similar level of HU response to in-person HU dosing adjustments. This finding importantly suggests that telehealth for monitoring and adjusting HU in patients with poor access to a Pediatric Hematologist or during a pandemic should not impact HU response. Strategies to improve overall adherence need prospective evaluation at our institution, but are not related to telehealth dosing of HU. Disclosures Lebensburger: Novartis: Consultancy.

Targeted Sequencing of 7 Genes Can Help Reduce Pathologic Misclassification of MDS

Targeted Sequencing of 7 Genes Can Help Reduce Pathologic Misclassification of MDS

Clinical signs in dogs attributed to Yersinia enterocolitica antigen 0:9

Canine yersiniosis is currently a scantily researched disease. Two agents predominately cause yersiniosis: Y. enterocolitica (gut yersiniosis), Y. preudotuberculosis (yersiniosis). There are three clinical forms of the disease: intestinal, generalized and secondary-focal. Current available research states the prevalence of Y. enterocolitica against other biovariants in canine infections. The majority of infected dogs demonstrate both asymptomatic clinical course and unspecific symptoms or serve as a carrier. Meanwhile yersiniosis pose a threat to human health causing a severe complex of symptoms. In some cases the disease can be lethal, thus the disease has both epizootological and epidemiological value. The goal of this paper was to generalize clinical signs in dogs that demonstrated positive reaction to Y. enterocolitica antigen 0:9, which is a dominant causative agent of yersiniosis in the northeastern region of Ukraine. The study was conducted based on clinical data, biochemical and hematological laboratory studies. Contamination of canine subjects with Y. enterocolitica 0:9 was conducted using agglutination reaction using respective antigen. The research showed, that the dominant symptoms in canines, affected by Yersinia serovariant 0:9 were gastrointestinal lesions in 100 % of the cases. The clinical signs included melena or bloody stool, general depression, anorexia, cachexia, more rarely – vomiting, tachypnea and breathing irregularities. The results of biochemical blood assays and CBC were heterogeneous and cannot be used as a specific marker of Yersinia infection. The main method of confirmation for Yersinia infection would be a serological agglutination reaction, which can identify positive diagnostic titers in animal blood samples. Further research is planned to study mono- and concurrent course of Yersiniosis with different infectious diseases.

Radiation Induced Lymphopenia is Associated with the Effective Dose to the Circulating Immune Cells (EDIC) for Breast Cancer

Radiation induced lymphopenia (RIL) has been reported to associate with treatment outcome in many types of solid cancers, including breast cancer of small series. In this study, we hypothesized that effective dose to circulating immune cells (EDIC) impacts the severity of radiation-induced lymphopenia in breast cancer. Breast cancer patients who received adjuvant radiation were enrolled in this study. Radiation dosimetric factors such as RT fields, RT fractionation schema, use of electron, mean cardiac dose, mean ipsilateral/bilateral lung dose, integral dose of the total body was collected retrospectively. A previously published EDIC model, which considered that the exposure of circulating immune cells as the proportion of blood flow into blood containing organs including lung, heart, and the volume of the exposed area of the total body, were used for this study. The model is expressed as EDIC = 0.12*MLD+0.08*MHD+0.45+0.35*0.85*n451/2*ITD61.8*103, where MLD, MHD, and ITD is the mean lung dose, mean heart dose and integral dose, n is the number of radiation fractions. The post- and pre-radiation lymphocyte ratio was calculated as the end point to predict lymphopenia. Linear regression between the lymphocyte ratio and EDIC was used for the analysis. The lymphocyte ratios were also compared by dividing patients into 3 EDIC groups. T-test was used for the comparison. Between March 2015 to February 2020, a total of 1288 breast cancers patients received radiation in clinical oncology of Hong Kong University Shenzhen Hospital and 735 patients with complete CBC results were included in this study. The lymphocyte ratio decreased linearly with increasing EDIC (R2 = 0.23, p<0.0001). The median lymphocyte ratio was 73.1%, 62.2% and 36.4% respectively, when EDIC was ≤ 0.75Gy, >0.75Gy but ≤1.25Gy, and >1.25Gy. The average EDIC was 75%±21%, 64%±19%, and 41%±23% correspondingly for the 3 EDIC groups (p<0.0001). EDIC is significantly associated with lymphocyte count changes and NCI defined lymphopenia after radiation. EDIC may be used to predict radiation induced lymphopenia and guide radiation plan optimization for breast cancer patients treated with radiotherapy.

Effect of Nursing Educational Brochure on Incidence and Risk Factors of Septicemia among Burned Patients

Background: The major challenge for a burn care team is nosocomial infection. Infection of the burn injury leading to septicemia is the main cause of death. Aims: Assess the relation between incidence and risk factors of septicemia among burned patients and design a suggested nursing educational brochure. Methods: A descriptive correlational research design was utilized, data were collected using; Tool (I) Structured interview questionnaire, consisted of patients' demographic and medical data, which included: general, local assessment, laboratory investigations, blood culture and patient prognosis. Tool (II) risk factors of septicemia. In addition to a suggested nursing educational brochure. Data collected for a full year (November 2017– October 2018) on (101) convenient burned patients admitted in the burn unit at Assiut University hospital. Results: The prevalence of septicemia, around one third of the studied burned patients had a positive blood culture (staphylococcus). The mean TBSA were 34.73±24.27, all patients had invasive devices, major of them had an abnormal CBC results and discharged but near one quarter dead. Conclusion: There were a statistically significant relation between cause of burn, TBSA, wound changes and dressing type with patient's positive blood culture results (septicemia). Recommendations: Application of the developing strategies for infection control, early detection and treatment of infection (septicemia) among the burned patients

Stability of complete blood count parameters depends on the storage temperature, storage time, transport position and selected stability criterion

Purpose: To evaluate stability of CBC parameters in samples containing K3-EDTA in Hema&Tube brand evacuated tubes stored at 4 °C and room temperature in upright, horizontal and upside-down transport positions for up to 240 min by comparing with different stability criteria. Methods: A total of 450 outpatient samples, 75 for each condition, were stored for 240 min. Blood samples were analyzed at baseline, 60th, 120th and 240th min of storage time using Sysmex® XT-4000i hematology analyzer. CBC results were compared according to the paired samples t-test, one CV% change, CVI, BV DES 2014, RCPA, IQMH, EFLM Biological Variation 2019, ESfEQAand CLIA stability criteria. Results: In all transport conditions, almost all CBC parameters were unstable according to the paired samples t-test. Based on one CV % change RBC, hemoglobin, hematocrit, RDW-CV, neutrophils, platelets, MPV, PDW and plateletcrit were unstable in some transport conditions. However, most parameters were stable at all conditions, except especially for MCH, MPV, PDW and plateletcrit, while just MPV and PDW were unstable at RT in all transport conditions according to the lower and higher CVI, respectively. Conclusions: Paired samples t-test and one CV% change stability criteria are not suitable for decision. Clinical laboratories should establish the impact of storage time, storage temperature and transport positions for each CBC parameter while paying attention to the brand of evacuated tube according to the lower CVI of biological variation data stability criteria since it is more related to the impact of analysis deviation on clinical decisions to prevent misdiagnosis.

Clinical Appropriateness of Serum Folate ordering pattern in a tertiary care hospital in Saudi Arabia

Folate, also known as vitamin B9, is a co-factor necessary for DNA synthesis. Folate deficiency is associated mainly with hematological findings including megaloblastic anemia and pancytopenia. Many countries have mandated grain fortification with micronutrients including folic acid resulting in a reduced prevalence of folate deficiency. Saudi Arabia imports most of the grain products and folate is usually added after milling. There are no local studies to address the folate deficiency prevalence. In this study we aimed to analyse the clinical appropriateness of ordering practice of serum Folate level. MethodWe reviewed all serum folate requests received at our laboratory in Aseer Central Hospital over one-year period (July 2018 June 2019). We collected patients’ demographics from the electronic requests along with biochemical results of serum B12, ferritin and CBC results. We assessed appropriateness of orders against pre-specified criteria and applied statistical tests to explore for any association or significance. ResultsSerum folate requests from 614 patients were received during the study period. Serum B12 (543, 88%), and serum ferritin (511, 83%) were concurrently requested. The most common reason for request, when available, was anemia. Anemia was present in (313, 51%) of the subjects for which microcytic anemia was predominant (199, 63.5%), followed by normocytic anemia (101, 33%) and only 10 subjects had macrocytic anemia (3.2%). The most common hematinics’ deficiency was ferritin (30%) followed by B12 (17.2%). Serum folate deficiency was low, observed in only 2.8%. Low folate levels were not significantly different between the group with anemia and the normal hemoglobin group. ConclusionThis study identifies a commonly inappropriate serum folate ordering practice that includes ordering all hematinics at the same visit without considering the possible anemia etiologies. The excessive requests might be related to doctors attempt to avoid multiple blood extractions and to try to reduce the time for diagnosis. These policies are generating unnecessary costs and time loss. Education, phasing out or restricting some tests and introducing laboratory policies like sample storing could help reduce unnecessary requests.

4172 Introduction to R Programming and GitHub: Developing Automated Analysis of Complete Blood Count Data as a Translational Science Undergraduate Project

OBJECTIVES/GOALS: Introduce students to programming and software development practices in the life sciences by analyzing standard clinical diagnostic bloodwork for differential immune responses. Including lectures and a semester project with the goal of enhancing undergraduate students’ education to prepare them for careers in translational science. METHODS/STUDY POPULATION: The educational content was taught for the first time as a component of the newly developed course BTEC 330 “Software Applications in the Life Sciences” in UMBC’s Translational Life Science Technology (TLST) Bachelor’s degree program at the Universities at Shady Grove campus. Eleven students took the course. All were beginners with no programming background. Lectures provided background on the diagnostic components of the CBC, criteria for differential diagnosis in the clinical setting, and introduction to hematology and flow cytometry, forming underpinnings for interpretation of the CBC results. Weekly computer lab practical sessions provided training fundamentals of R programming language, the R-studio integrated development environment (IDE), and the GitHub.com open-source software development platform. RESULTS/ANTICIPATED RESULTS: The graded assignment consisted of a coding project in which students were each assigned an individual parameter from the CBC results. These include, for example, relative lymphocyte count or hemoglobin readouts. Students each created their own R-language script using R-studio, with functional code which: 1) Read in data from a file provided, 2) Performed statistical testing, 3) Read out statistical results as text, and charts as image files, 4) “Diagnosed” individuals in the dataset as being inside or outside the clinical normal range for that parameter. Each student also registered their own GitHub account and published their open-source code. Grading was performed on code functionality by downloading each student repository and running the code with the instructor as an outside developer using the resource. DISCUSSION/SIGNIFICANCE OF IMPACT: In this curriculum, students with no background in programming learned to code a basic R-language script and use GitHub to automate interpretation of CBC results. With advanced automation now becoming commonplace in translational science, such course content can provide introductory level of literacy in development of clinical informatics software.

Mortality rate and prognostic factors for dogs with severe anaphylaxis: 67 cases (2016–2018)

To determine mortality rates for dogs with severe anaphylaxis and identify potential prognostic factors. 67 dogs with suspected anaphylaxis graded as severe. Dogs were classified on the basis of outcome as survivors and nonsurvivors. Medical records were reviewed, and data were extracted including signalment, examination findings, time to hospital admission from onset of clinical signs, CBC results, serum biochemical analysis results, coagulation testing results, and findings on abdominal ultrasonography. Initial treatment within the first 6 hours after hospital admission was recorded for analysis, specifically including the use of epinephrine, diphenhydramine, corticosteroids, antimicrobials, fresh-frozen plasma, and supplemental dextrose. The overall mortality rate was 14.9% (10/67) for dogs with anaphylaxis graded as severe. Serum phosphorus concentration and prothrombin time (PT) were significantly higher in nonsurvivors, compared with survivors. Nonsurvivors had lower presenting body temperatures than survivors. Serum phosphorus concentration ≥ 12.0 mmol/L, hypoglycemia within 6 hours after hospital admission, high PT value, concurrently high PT and partial thromboplastin time (PTT) values > 50% above the reference range limit, and the need for supplemental dextrose were associated with death. The incidences of coagulopathy and peritoneal effusion were unexpectedly high (85.2% and 65.5% of dogs, respectively) but were not indicative of survival. Despite the poor presenting clinical condition seen in dogs with severe anaphylaxis, the rate of survival with treatment was fairly high. Coagulopathy and the presence of peritoneal effusion were common findings in dogs with severe anaphylaxis. Serum phosphorus concentration ≥ 12.0 mmol/L, high PT value, concurrent increases of PT and PTT values > 50% above reference range limits, hypoglycemia within 6 hours after hospital admission, and the need for supplemental dextrose were associated with death.

Clinical features and radiographic findings in cats with eosinophilic, neutrophilic, and mixed airway inflammation (2011-2018).

BackgroundIdiopathic inflammatory airway disease (IAD) in cats often is described as asthmatic (eosinophilic) or bronchitic (neutrophilic), but this designation requires collection of airway fluid and it fails to consider cats with mixed airway inflammation.ObjectiveTo identify clinical features that would differentiate inflammatory disease types.AnimalsForty‐nine cats with nonspecific airway inflammation identified by bronchoscopic bronchoalveolar lavage (BAL) between 2011 and 2018 were evaluated.MethodsThis is a retrospective study. Cats were categorized by BAL differential cytology as having eosinophilic (eosinophils >20% with neutrophils <14%, or eosinophils >50%), mixed (eosinophils 20%‐50% and neutrophils >14% or discordant inflammation from 2 BAL sites), or neutrophilic (neutrophils >14% and eosinophils <20%) inflammation. Type and duration of presenting complaints, signalment, body condition score, respiratory rate, CBC results, bronchoscopy, BAL results (% recovery, total nucleated cell count, differential cell count), and radiographic findings were compared among groups.ResultsIdiopathic IAD was diagnosed in 49 cats, with BAL eosinophilic inflammation in 23, mixed inflammation in 14, and neutrophilic inflammation in 12. Cough was the predominant presenting complaint with no difference in duration of signs among groups (median, 5.5 months). Respiratory rate and effort also did not differ. Cats with eosinophilic inflammation were significantly younger (4.4 ± 3.3 years) than those with neutrophilic (8.0 ±5.6 years) or mixed inflammation (7.5 ± 4.0 years; P = .03). Results of CBC and interpretation of radiographic findings did not differ among groups.Conclusions and Clinical ImportanceSubstantial overlap exists in clinical and radiographic findings in cats with various forms of idiopathic airway inflammation.

Clinicopathological characteristics of histiocytic sarcoma affecting the central nervous system in dogs.

BackgroundHistiocytic sarcoma affecting the central nervous system (CNS HS) in dogs may present as primary or disseminated disease, often characterized by inflammation. Prognosis is poor, and imaging differentiation from other CNS tumors can be problematic.ObjectiveTo characterize the clinicopathological inflammatory features, breed predisposition, and survival in dogs with CNS HS.AnimalsOne hundred two dogs with HS, 62 dogs with meningioma.MethodsRetrospective case series. Records were reviewed for results of cerebrospinal fluid (CSF) analysis, CBC, treatment, and outcome data.ResultsPredisposition for CNS HS was seen in Bernese Mountain Dogs, Golden Retrievers, Rottweilers, Corgis, and Shetland Sheepdogs (P ≤ .001). Corgis and Shetland Sheepdogs had predominantly primary tumors; Rottweilers had exclusively disseminated tumors. Marked CSF inflammation was characteristic of primary rather than disseminated HS, and neoplastic cells were detected in CSF of 52% of affected dogs. Increased neutrophil to lymphocyte ratios were seen in all groups relative to controls (P <.008) but not among tumor subtypes. Definitive versus palliative treatment resulted in improved survival times (P < .001), but overall prognosis was poor.Conclusions and Clinical ImportanceClinicopathological differences between primary and disseminated HS suggest that tumor biological behavior and origin may be different. Corgis and Shetland Sheepdogs are predisposed to primary CNS HS, characterized by inflammatory CSF. High total nucleated cell count and the presence of neoplastic cells support the use of CSF analysis as a valuable diagnostic test. Prognosis for CNS HS is poor, but further evaluation of inflammatory mechanisms may provide novel therapeutic opportunities.

Does Trimethoprim-Sulfamethoxazole prophylaxis induce myelosuppression in primary immune deficiency disease patients; A retrospective, 3 groups comparative study.

The possible myelosuppression side effect of Trimethoprim-Sulfamethoxazole (TMP-SMX) on primary immune deficiency (PID) patients has not been established yet. Identify if the PID patients are at higher risk of developing myelosuppression secondary to the use of TMPSMX. Retrospective, three groups study, of PID patients (on and off TMP-SMX prophylaxis) and urinary tract infection (UTI) patients received prophylaxis TMP-SMX. Data about CBC results (WBC, ANC, Lymphocytes, RBC, Hemoglobin, and Platelet counts) at baseline, first, and maximum myelosuppression observed during the period of TMP-SMX administration were collected. A total of 122 patients were included in this study (41 PID patients on TMP-SMX prophylaxis, 45 PID patients not on TMP-SMX prophylaxis, and 36 UTI patients on prophylaxis TMP-SMX). There are significant differences noticed in the percentage of patients who developed clinical myelosuppression (i.e. less than normal value for age) in ANC (39.0% vs. 8.9% vs. 16.7%, p = 0.002), RBC (36.6% vs. 13.3% vs. 13.9%, p = 0.014), WBC (41.5% vs. 13.3% vs. 13.9%, p = 0.003), and platelet (24.4% vs. 15.6% vs. 2.8%, p = 0.028) in group 1, 2, and 3, respectively. Significant difference in myelosuppression between the groups was most likely due to the combination of TMP-SMX effect on PID patients rather than the disease or the drug itself. Primary immune deficiency (PID) patients are at higher risk of developing myelosuppression secondary to TMP-SMX prophylaxis (especially ANC) comparing to immune-competent patients or other PID patients who did not receive prophylactic TMP-SMX. Future larger prospective study is required to confirm this association.

Diagnostic value of integral indexes of intoxication to assess the risk of complications development in gastric and intestinal diseases of calves

The assessment of the intoxication degree and the state of general organism resistance was carried by identifying “hematological indices” showing the connections between various cell classes of leukocyte formula. Comparison of integral indices of intoxication has recorded “slight degree of intoxication of the organism” in sick calves in 47.6% of cases. In the case of “moderate severity of organism intoxication”, the tension in the hemopoiesis system was recorded in 85.7% of cases, and the imbalance in the reactions of the cellular and humoral link of immunity – in 48.1% cases. For animals with a “severe degree of organism intoxication” imbalance in the links of immunity was determined in 90.9% of cases. According to the results of the determination of LII, LSI and LI, 31.6% calves were identified as having a risk of gastrointestinal complications development, whereas according to the results of CBC only 9.5% animals were classified as a “risk group”.

Comparison of αLEPRA and αLELY Co-Inheritance with a SPTA1 Whole Gene Deletion, a Family Study

Background: Genetic variants resulting in quantitative defects in the production of α-spectrin are associated with autosomal recessive hereditary spherocytosis (HS). Because erythroid cells produce α-spectrin in a 3-4:1 ratio in relation to β-spectrin, one defective allele is typically well-tolerated and a single null allele combined with a second partially defective allele may display a sub-clinical phenotype. Protein expression can be reduced by nonsense, frameshift, missense or splicing variants but reported large deletion events are extraordinarily rare (n = 1). Splicing variants include the low expression alleles αLELY - Low Expression LYon (SPTA1:c.6531-12C&gt;T, p.?) and αLEPRA-Low Expression PRAgue (SPTA1:c.4339-99C&gt;T, p.?) which reduce protein expression by 50% and 16%, respectively. Here, we present a family study with a whole gene SPTA1 deletion that characterizes the patient phenotype when a full gene mutation is co-inherited with the αLEPRA and αLELY low expression alleles. Methods: The 6 month-old male proband had a history of congenital Coombs negative hemolytic anemia requiring transfusion from 3 months of age and in whom a red cell membrane disorder was clinically suspected. Pretransfusion CBC and peripheral blood smear results were unknown. Transfused CBC indices were as follows: RBC 4 x 1012/L, HGB 11 g/dL, MCV 76 fL, MCH 27 pg, MCHC 36 g/dL and a reticulocyte count of 1.8%. The proband's mother, of Cuban descent, and father, of European descent, had no history of anemia or HS. Targeted high throughput sequencing of genes related to hereditary hemolytic anemia was performed for the detection of single nucleotide variants (SNVs) and small insertion/deletion variants (indels). Relative quantitative real time PCR (RT-PCR) was utilized to confirm large structural variants. Results: Sequencing identified a large heterozygous deletion that spanned the entire SPTA1 gene (NC_000001.11:g.(?_158656337)_(158581024_?) in the proband and his mother. In addition, the proband had inherited hemizygous αLEPRA from his father. The mother had hemizgous αLELY. The father showed heterozygous αLEPRA and no other SPTA1 variants (Table 1). Conclusion: To our knowledge, only Iolascon et al in 2011 has reported a SPTA1 gene large deletion (SpectrinExeter, exon 11-exon 12 deletion, loss of 83 amino acids) which resulted in a structurally abnormal protein and ellipto-poikilocytosis phenotype when in trans to αLELY. Our current case is the first reported SPTA1 full gene deletion and our family study allows comparison of the effects of coinheritance of the low expression alleles αLEPRA and αLELY. When in combination with complete absence of the SPTA1 gene in trans configuration, αLELY manifests in subclinical effects and αLEPRA causes a recessive HS phenotype, which resulted in severe anemia. This contrast further demonstrates that, unlike αLEPRA, αLELY does not decrease the α-spectrin concentration sufficiently to contribute to the baseline phenotype regardless of the severity of the second allele mutation for quantitative disorders such as HS. Disclosures No relevant conflicts of interest to declare.

Exploring Anemia's Impact on Brain Microstructure, Volume, Functional Connectivity, Iron and Cognitive Performance

Introduction: We used MR imaging techniques to investigate the neurological consequences of chronic anemia in human patients with sickle cell disease, non-sickle anemic syndromes (called anemic-controls) and non-anemic controls. We previously demonstrated that hemoglobin level is an independent predictor of white matter volume (WMV) in brain as well as cognitive performance. Importantly, WMV was independent of genotype (sickle vs non-sickle), treatment type, HgB S%, fetal HgB level, Lactate dehydrogenase (LDH) and presence of silent strokes. We also demonstrated that low hemoglobin is associated with abnormal brain functional connectivity and iron levels in select brain regions. In the present study, we examine the effects of anemia on brain integrity using apparent diffusion coefficient (ADC) calculated from diffusion weighted imaging (DWI). ADC measures the magnitude of the motion of water in tissue and can be used to imply tissue damage, making it highly sensitive to neuropathology as altered tissue integrity and loss of cellular structures can change the diffusivity of water. We then explore the associations between ADC, local brain volume, functional connectivity, iron and cognitive performance. Methods: MRI data, CBC and neuropsychological testing results were obtained from 26 sickle cell disease (age = 20.9 ± 11.3, F:M = 13:13, HgB = 9.7 ± 2.1), 20 anemic-control (age = 25.9 ± 11.3, F:M = 10:10, HgB = 10.9 ± 0.5) and 25 control subjects (age = 23.1 ± 8.4, F:M = 16:9, HgB = 13.2 ± 1.2). (Recruited with informed consent or assent; IRB: CHLA CCI#11-00083). DWI (TE = 86ms; TR = 6700ms; resolution=2.5mm3; 30 directions; bvalue=1000m/s2; reverse-gradient b=0) were acquired on a 3T Philips Achieva (v.3.2.1) using an 8-channel head coil. Images were corrected for motion and distortion then voxel-wise calculations of ADC were computed then transferred to a common atlas space. (brainsuite.org, v18). 3D T1 weighted, quantitative susceptibility mapping, and functional MRI (fMRI) images were collected using our previously published protocols. After regressing out age and sex, we determined the effect of hemoglobin level at each voxel of the brain on ADC then corrected for multiple comparison (BH FDR α=0.1). Significant regions were retained, (p&amp;lt;0.05), and a mean ADC was computed for each subject from those regions. Pearson's correlations were used to determine the effects of hemoglobin level on the following measures: (1) mean ADC, (2) WMV, (3) Connectivity Dissimilarity Index (CDI), a measure quantifying the dissimilarity of functional connectivity pattern between each subject and a reference fMRI atlas (4) Iron measured in the substantia nigra of the brain's basal ganglia through R2* and susceptibility images and (5) Matrix Reasoning, a nonverbal measure of novel problems (fluid reasoning). Results: Hemoglobin level significantly correlated with ADC throughout the brain, but most strongly in deep white matter and subcortex, followed by the occipital lobe and cerebellum (Figure 1). The scatterplot between mean ADC and hemoglobin showed no discernable differentiation between anemia subtypes (Figure 2). Mean ADC and our previously developed markers of disease correlated well with each other showing that the severity of anemia correlates with higher ADC, lower white matter volume (WMV), abnormal functional connectivity (CDI), higher brain iron and lower Matrix Reasoning scores (males only; Table 1). Conclusion: Increased ADC, correlating with anemia severity, was observed in subcortical structures of an anemic population at risk for white matter shrinkage and cognitive dysfunction. ADC (but not WMV) correlated with brain iron which is known to accumulate in the presence of cerebral hypoxia. ADC and WMV changes were comparable in males and females but only males showed lower fluid reasoning. This data shows that anemia and brain iron are associated with brain tissue disruption and function. Disclosures Coates: vifor: Consultancy, Honoraria; celgene: Consultancy, Honoraria, Other: steering committee of clinical study; agios pharma: Consultancy, Honoraria; apo pharma: Consultancy, Honoraria, Speakers Bureau. Wood:BiomedInformatics: Consultancy; Imago Biosciences: Consultancy; National Institutes of Health: Research Funding; Apopharma: Consultancy; WorldcareClinical: Consultancy; Philips Healthcare: Research Funding; BluebirdBio: Consultancy; Celgene: Consultancy.

Morphologic and quantitative evaluation of bone marrow aspirates from Hispaniolan Amazon parrots (Amazona ventralis).

Bone marrow aspirate assessments provide valuable information about hematopoietic status and hematologic disease. Hematopoietic cell differentials and morphologies have been anecdotally described in psittacines, but quantitative studies are lacking. We aimed to determine differential cell counts and calculate granulocyte:erythroid (G:E) ratios in bone marrow aspirates from Hispaniolan Amazon parrots and report representative morphologies of the hematopoietic cells. Bone marrow aspirates were collected from 32 clinically healthy, captive, parrots. Peripheral blood was obtained for CBCs. Bone marrow differential cell counts (%) were determined by counting 500 cells on modified Wright's-stained smears. G:E ratios were calculated. Representative images of hematopoietic cells at all stages of development were taken. Of the 32 parrots sampled, 17 bone marrow samples were of sufficient cellularity and quality for evaluation. Erythroid cells comprised 68.9±8.6% (total±SD) of the hematopoietic cells and consisted primarily of early- and late-stage polychromatophilic rubricytes (43.6±2.1% of total erythroid cells). Granulocytic cells comprised 28.1±3.8% of the hematopoietic cells and consisted primarily of mature and band heterophils (11.9±5.2% and 6.5±3.4%, respectively, of total granulocytic cells). A unique morphologic finding in avian progranulocytes was the presence of multiple different granules. The G:E ratio was 0.4±0.2 (median 0.4, range 0.1-0.9). Thrombocyte lineage cells could not be reliably identified and were not counted. CBC results were largely within expected limits. The low G:E ratios observed could be normal in this species; however, these ratios could be affected by factors related to sampling and cell identification. These findings will be a valuable resource for the diagnostic evaluation of clinical bone marrow samples from Hispaniolan Amazon parrots and could serve as a general reference for psittacine bone marrow evaluation.