Disclosure of the Mechanisms of Metabolic Disorders in Highly Productive Cows as a Way to Improve Milk Production Technology

The aim of this study was to study the molecular mechanisms of metabolic disorders that develop in highly productive cows. It was found that in the blood serum of animals with subclinical ketosis (SCK), which are in the eighth month of pregnancy, the concentration of β-hydroxybutyrate is increased, the content of iron, calcium and phosphorus is reduced, its iron-binding capacity against the background of the absence of phenomena of hepatocellular insufficiency, violation of the integrity of hepatocytes, hypoplastic anemia. The aggravation of ketoacidosis in fresh cows with rumen acidosis is facilitated by the development of carbohydrate deficiency in them, due to their increased oxidation in the reactions of anaerobic glycolysis, accompanied by excessive catabolism of purine mononucleotides to uric acid. Excessive lipid peroxidation of membrane structures under conditions of increased catabolism of purine nucleotides, along with the accumulation of acylglycerols in the liver caused by cobalamin deficiency, leads to the development of syndromes of impaired hepatocyte integrity and hepatocellular insufficiency. A decrease in the liver's ability to store iron ions and synthesize proteins that provide an increase in the iron-binding capacity of blood plasma in conditions of cobalamin deficiency contributes to the development of hypoplastic anemia not only in cows, but also in calves born from them.

Comparative analysis of hematopoietic activity of total RNA from bone marrow cells and splenocytes of rats with chronic benzene-induced anemia

Цель - оценка лечебных свойств суммарной РНК на модели хронической гипопластической анемии, выявление особенностей гемопоэтического действия суммарных РНК, выделенных из костного мозга и лимфоидных клеток селезенки. Методика. Работа выполнена на 36 белых беспородных крысах-самках массой 130-250 г. Хроническую токсическую анемию моделировали подкожным введением смеси химически чистого бензола (0,05 мл/100 г) и стерильного растительного масла. Смесь вводили трижды с интервалом в 7 сут. Контрольным животным (группа 1 в каждой серии) в эти же сроки внутривенно вводили равное по объему количество 0,9% раствора NaCl. Выполнено 2 серии экспериментов. В 1-й серии оценивали гемопоэтическую активность суммарных РНК лимфоидных клеток селезенки, во 2-й - суммарных РНК костного мозга. Препараты суммарной РНК получали из костного мозга и лимфоидных клеток селезенки интактных крыс, а также из клеток тех же органов крыс, подвергнутых кровопусканию (2% от массы тела) за 17 ч до выделения из них РНК. Суммарную РНК выделяли методом гуанидин тиоцианат-фенол-хлороформной экстракции. Для выяснения влияния суммарных РНК на состояние эритроидного ростка костного мозга в препаратах, окрашенных по Паппенгейму, оценивали количественный и качественный состав эритроидных островков. Результаты. Сравнение гемопоэтической активности суммарных РНК, выделенных из костного мозга и лимфоидных клеток селезенки здоровых животных, при их введении крысам с экспериментальной хронической бензольной анемией, показало, что различия клеточного состава лимфоидных органов-источников РНК отражается на их гемопоэтических свойствах. Суммарная РНК из лимфоидных клеток селезенки интактных животных в первую очередь способствует восстановлению количества лейкоцитов, в то время как суммарная РНК лимфоидных клеток костного мозга в начальный период регенерации не оказывает заметного влияния на лейкопоэз. Показано также, что суммарная РНК, выделенная из костного мозга, в целом обеспечивает более интенсивный процесс гемопоэза. Кроме того, обнаружена более высокая эритропоэтическая активность суммарных РНК, выделенных из клеток обоих лимфоидных органов крыс-доноров, стимулированных кровопотерей, по сравнению с таковой суммарных РНК, полученных из органов интактных крыс. Заключение. Подтверждена гемопоэтическая активность суммарной РНК костного мозга и лимфоидных клеток селезенки при гипопластической анемии. Выявлена более высокая эритропоэтическая активность суммарных РНК обоих лимфоидных органов животных после кровопускания по сравнению с их лейкопоэтической активностью. Aim. To evaluate therapeutic properties of total RNA on a model of chronic hypoplastic anemia and to elucidate features of the hemopoietic effect of total RNA isolated from bone marrow splenic lymphoid cells. Methods. Experiments were performed on 36 white mongrel rats weighing 230-250 g. Chronic toxic anemia was induced by subcutaneous injections of a mixture of chemically pure benzene (0.05 ml/100 g) and sterile vegetable oil. The mixture was injected three times with an interval of 7 days. Control animals (group 1 in each series) received intravenous injections of an equal volume of saline at the same time intervals. Two experimental series were performed. In series 1 and 2, hemopoietic activity of total RNAs from splenic lymphoid cells and from bone marrow, respectively, was evaluated. Total RNA was obtained from splenic lymphoid cells and from bone marrow of intact rats and rats subjected to hemorrhage (2% of body weight) 17 h prior to the RNA isolation. Total RNA was isolated by guanidine-thiocyanate-phenol-chloroform extraction. The effect of total RNA on the erythroid lineage in the bone marrow was evaluated by the quantitative and qualitative composition of erythroid islets in Pappenheim-stained preparations. Results. Comparing the hematopoietic activity of total RNA isolated from bone marrow and splenic lymphoid cells of healthy animals and administered to rats with experimental chronic benzene anemia showed that the difference in cell composition of the lymphoid organs, sources of the RNAs, impacted on the RNA hemopoietic properties. The total RNA from splenic lymphoid cells of intact animals contributed primarily to the recovery of leukocyte count while the RNA from the bone marrow of intact rats did not significantly influence leukopoiesis during the early regeneration period. It was also shown that the total RNA extracted from the bone marrow generally provided a more intensive hemopoiesis. In addition, the erythropoietic activity of total RNA isolated from the cells of both lymphoid organs of the donor rats stimulated by blood loss was higher than that of the total RNA isolated from intact rats. Conclusion. The total RNA isolated from bone marrow and lymphoid cells spleen exerted a hemopoietic effect in hypoplastic anemia. The erythropoietic activity of total RNA from both lymphoid organs of animals after hemorrhage was higher than the RNA leukopoietic activity.

An update on the pathogenesis and diagnosis of Diamond–Blackfan anemia

Diamond–Blackfan anemia (DBA) is a rare congenital hypoplastic anemia characterized by a block in erythropoiesis at the progenitor stage, although the exact stage at which this occurs remains to be fully defined. DBA presents primarily during infancy with macrocytic anemia and reticulocytopenia with 50% of cases associated with a variety of congenital malformations. DBA is most frequently due to a sporadic mutation (55%) in genes encoding several different ribosomal proteins, although there are many cases where there is a family history of the disease with varying phenotypes. The erythroid tropism of the disease is still a matter of debate for a disease related to a defect in global ribosome biogenesis. Assessment of biological features in conjunction with genetic testing has increased the accuracy of the diagnosis of DBA. However, in certain cases, it continues to be difficult to firmly establish a diagnosis. This review will focus on the diagnosis of DBA along with a description of new advances in our understanding of the pathophysiology and treatment recommendations for DBA.

Different Clinical Characteristics of Paroxysmal Nocturnal Hemoglobinuria in Pediatric and Adult Patients

Introduction: Studies of children with paroxysmal nocturnal hemoglobinuria (PNH) are scarce and include a very limited number of patients. The objective of this analysis was to describe characteristics of PNH at enrollment for the largest available registry of pediatric patients, and to compare demographic and clinical characteristics with those of adult patients. Methods: The International PNH Registry is a prospective, multi-center worldwide, observational study of patients with a PNH clone of 0.01-100%. Data are collected from patient medical records at the time of Registry enrollment and every six months thereafter. Adult patients were ³18 years of age at enrollment and disease start and pediatric patients were <18 years at enrollment. Demographics and clinical parameters in patients untreated with eculizumab at enrollment for the two age cohorts were compared using the Wilcoxon-Mann-Whitney test for medians and PearsonÕs chi-square for frequencies. The rate of thrombotic events (TE) between disease start (defined as the earliest reported PNH symptoms, granulocyte clone, or PNH diagnosis) and enrollment was calculated per 100 person-years. Results: As of March 2, 2015, a total of 2,184 patients were eligible for analysis: 94 (4.3%) pediatric patients and 2,090 (95.7%) adult patients. Median age (range) at enrollment was 14.0 years (3-17) in pediatrics and 45.5 years (18-100) in adults; median disease duration was 0.7 years and 2.1 years, respectively (p<0.001). More pediatric than adult patients had a PNH clone of <10% and severe cytopenia (Table). Pediatric patients had lower percent of reticulocytes compared with adults (2.1% vs. 2.6%, respectively; p=0.015). History of aplastic or hypoplastic anemia was more frequent in pediatric than adult patients (76.5% vs 54.4%, respectively; p<0.001). History of TE and any major adverse vascular event was less frequent in pediatrics (2.1% vs 8.7%; p=0.025, and 4.3% vs. 14.4%; p=0.005). The rate of TE between disease start and enrollment was lower in pediatric patients, but not statistically significant: 1.4 per 100 person-years (95%CI 0.2-5.2) compared to adult patients (2.3 per 100 person-years (95%CI 2.0-2.6). More pediatric patients than adults had abdominal pain at enrollment. Conclusions: The International PNH Registry provides the largest available pediatric cohort of patients with a PNH clone to characterize this understudied population and demonstrate an important disease burden. Pediatric patients were more likely to have smaller PNH clones and a higher component of aplastic/hypoplastic anemia. Pediatric patients had fewer vascular events. These findings may reflect the natural evolution of the disease and can be useful in the clinical management of PNH. Table 1. Clinical Characteristics at Enrollment of Pediatric and Adult Patients with PNH Pediatric(n=94) Adult(n=2,090) P-value Clone size (percent GPI-deficient granulocytes), n (%)<10% 10 to < 50% ³50% 47 (55.3) 16 (18.8) 22 (25.9) 550 (38.3) 322 (22.4) 565 (39.3) 0.006* Cytopenia status, n (%)None (neutrophils ³ 1.5 x 109/L and platelets ³100 x 109/L) Moderate (neutrophils <1.5 x 109/L or platelets <100 x 109/L) Severe (neutrophils <0.5 x 109/L or platelets <20x109/L) 22 (29.3) 28 (37.3) 25 (33.3) 735 (42.2) 784 (45.1) 221 (12.7) <0.001* Percent reticulocytesMedian (Q1, Q3) 2.1 (1.1, 3.5) 2.6 (1.6, 4.6) 0.015 Hemolytic status, n (%)Hemolytic (LDH ³1.5 x ULN and/or reticulocytes ³60 x 109/L) Not hemolytic (LDH <1.5 x ULN and reticulocytes <60 x 109/L) 33 (58.9) 23 (41.1) 1,038 (65.3) 551 (34.7) NS LDH Ratio, n (%)<1.5 x ULN³1.5 x ULN 30 (58.8) 21 (41.2) 684 (47.0) 770 (53.0) NS History of TE, n (%)Yes No 2 (2.1) 92 (97.9) 181 (8.7) 1,902 (91.3) 0.025 Rate of TENumber of TE, n Person-years (disease start to enrollment) Rate/100 person-years (95% CI) 2 139.9 1.4 (0.2-5.2) 255 11,119.8 2.3 (2.0-2.6) NS History of MAVE, n (%) 4 (4.3) 300 (14.4) 0.005 GPI, glycosylphosphatidylinositol; LDH, lactate dehydrogenase; MAVE, major adverse vascular event; TE, thrombotic event; ULN, upper limit of normal *P-values for clone size and cytopenia status represent overall comparison of categories. Disclosures Muus: Alexion Pharmaceuticals: Honoraria. Schrezenmeier:Alexion Pharmaceuticals: Membership on an entity's Board of Directors or advisory committees, Research Funding, Speakers Bureau. Almeida:Celgene: Consultancy; Novartis: Consultancy; Bristol Meyer Squibb: Speakers Bureau; Shire: Speakers Bureau. Wilson:Alexion Pharmaceuticals: Employment. Ware:Bayer Pharmaceuticals: Consultancy; Biomedomics: Research Funding; Eli Lilly: Other: DSMB membership; Bristol Myers Squibb: Research Funding.