Recombinant factor VIII Fc fusion protein for immune tolerance induction in patients with severe haemophilia A with inhibitors-A retrospective analysis.

INTRODUCTION: Immune tolerance induction (ITI) is the gold standard for eradication of factor VIII inhibitors in severe haemophilia A; however, it usually requires treatment for extended periods with associated high burden on patients and healthcare resources. AIM: Review outcomes of ITI with recombinant factor VIII Fc fusion protein (rFVIIIFc) in patients with severe haemophilia A and high-titre inhibitors. METHODS: Multicentre retrospective chart review of severe haemophilia A patients treated with rFVIIIFc for ITI. RESULTS: Of 19 patients, 7 were first-time ITI and 12 were rescue ITI. Of 7 first-time patients, 6 had at least 1 high-risk feature for ITI failure. Four of 7 first-time patients were tolerized in a median of 7.8 months. The remaining 3 patients continue on rFVIIIFc ITI. Of 12 rescue patients, 7 initially achieved a negative Bethesda titre (≤0.6) in a median of 3.3 months, 1 had a decrease in Bethesda titre and continues on rFVIIIFc ITI and 4 have not demonstrated a decrease in Bethesda titre. Of these 4, 3 continue on rFVIIIFc ITI and 1 switched to bypass therapy alone. Two initially responsive patients transitioned to other factors due to recurrence. Overall, 16 of 19 patients remain on rFVIIIFc (prophylaxis or ITI). For those still undergoing ITI, longer follow-up is needed to determine final outcomes. No adverse events reported. CONCLUSIONS: Recombinant factor VIII Fc fusion protein demonstrated rapid time to tolerization in high-risk first-time ITI patients. For rescue ITI, rFVIIIFc showed therapeutic benefit in some patients who previously failed ITI with other products. These findings highlight the need to further evaluate the use of rFVIIIFc for ITI.

Real-world experience with use of Antihemophilic Factor (Recombinant), PEGylated for prophylaxis in severe haemophilia A.

INTRODUCTION: Prophylaxis with extended half-life factor VIII (FVIII) is approved for haemophilia A, but data regarding routine clinical use are limited. AIM: To assess real-world experience of ADYNOVATE® (Antihemophilic Factor (Recombinant), PEGylated prophylaxis in children and adults with haemophilia A. METHODS: A retrospective chart review was conducted in three US haemophilia treatment centres. Records of all patients who began Adynovate prophylaxis in routine clinical practice were identified. Demographic, clinical and patient-reported information beginning 6 months before initiation of Adynovate until the record review was analysed. RESULTS: Fifteen patients (aged 9 months to 28 years), with median 9 months' use of Adynovate (range 1-15 months), were identified. All had switched from another prophylactic regimen, 13 (87%) from standard half-life recombinant FVIII. Nine (60%) patients had ≥1 bleed within 6 months preswitch. The most frequent reason for switching was to reduce infusion frequency (14 patients). After switching, infusion frequency reduced for 13 patients, and overall weekly factor consumption decreased by 19%. Eight (53%) patients had no bleeds postswitch, three (20%) had spontaneous joint bleeds (vs four pre-switch), and three (20%) had only mild traumatic bleeds. Patient/parental satisfaction with Adynovate was documented as positive in 13 of 15 (87%) cases; 2 patients were not satisfied and discontinued Adynovate. No adverse events were considered related to Adynovate. CONCLUSION: In patients who switched from a standard half-life FVIII to Adynovate prophylaxis in routine clinical practice, bleeding control was generally improved or maintained, with a lower infusion frequency and factor consumption in most patients.

Successful combination therapy of a proximal haemophilic pseudotumour with surgery, radiation and embolization in a child with mild haemophilia A.

We describe the management of a young boy with mild haemophilia A and a massive iliac pseudotumour with a multi modality approach involving factor replacement, radiation therapy, embolization and surgery. The patient was initially treated with recombinant factor VIII and radiation therapy. Because of inadequate response and worsening of bony erosion, the patient had a preoperative embolization followed by surgical excision. The surgical procedure was associated with minimal blood loss and the patient had a relatively smooth postoperative course with no physical morbidity. This case illustrates successful aggressive management of a large, proximally located pelvic pseudotumour, which resulted in an excellent outcome despite the need for a normally morbid operation.

Characterization and purification of cytosolic and membrane-bound protease(s) in adults of Haemonchus contortus.

The activity of protease(s) was observed separately in cytosolic and membrane-bound protease(s) from male and female Hae monchus contortus (Nematoda: Trichostrongylidae). Different fractions showed optimum protease activity at 37 degrees C, pH 8.5 and 8.0 mg casein concentration. The female fractions had a particularly high activity of protease(s) in comparison with the male fractions, especially of membrane-bound enzymes in the anterior half. Inhibition, activation studies revealed the presence of four kinds of protease(s) in cytosolic and membrane-bound fractions. Protease(s) in different fractions are purified to a greater extent by higher concentrations of saturated ammonium sulphate solution, i.e. ranging from 50 to 65%. The purification study revealed the presence of multiple forms of protease(s) in cytosolic and membrane-bound extracts of H. contortus.

Haemonchus contortus: characterization and purification of excretory/secretory protease(s).

The activity of protease(s) has been examined separately in excretory/secretory (E/S) products from male and female Haemonchus contortus (Nematoda: Trichostrongylidae). The E/S proteolytic activity indicated the presence of preabsorptive digestion of host blood and/or tissues. Protease activity was optimum at 37 degrees C, pH 8.5 and 8.0 mg casein. These protease(s) were purified to 32.16- and 88.80-folds from male and female E/S products, respectively, by sequential purification with saturated ammonium sulphate followed by ion-exchange chromatography. The purification study revealed the presence of isomeric forms of protease(s) in the E/S products of H. contortus.

Characterization of biliary proteins and composition of bile from sheep and goats.

Higher lipid content in gall bladder bile of sheep and goats as compared with monogastric species was due to higher proportion of glycerides. The volume of bile per gall bladder was higher in sheep than in goats. Bile from sheep and goats has a higher content of proteins and lower contents of dry matter, cholesterol and phospholipids compared with monogastric species. Biliary proteins partially purified by the ammonium sulfate precipitation method were characterized and quantified by polyacrylamide gel electrophoresis (PAGE), fast protein liquid chromatography (FPLC) and immunodiffusion. FPLC and PAGE profiles indicate that biliary proteins of sheep and goats are mainly of low mol. wt. of upto 65 KDa. Proportions of immunoglobulin (Ig) like molecules in biliary proteins precipitated by ammonium sulfate of different degrees of saturation, were 10.4 to 26.7 and 21.9 to 30.7 per cent of total proteins in bile of goats and sheep. IgG and IgA like molecules were the predominant Ig and lacked carbohydrates. On immunodiffusion such Ig like molecules did not show cross reactivity with humoral Ig. It was concluded that sheep and goats appear to secrete Ig into bile after deglycosylation and partial degradation. The proteins of smaller molecular weight may be secretory fragments or degradation products of Ig.

Individual variation in the composition of colostrum and absorption of colostral antibodies by the precolostral buffalo calf.

The total protein and Ig content in the first colostrum was higher in buffalo cows than in dairy crossbred cows. Individual variation in Ig content of first colostrum may be responsible for differences in susceptibility of calves to infections. The decline in Ig content during subsequent milkings was faster in dairy crossbred cows. The major class was IgG in both species. Absorption of colostral proteins and Ig was 14.0 g (68%) and 4.6 g (75.4%), respectively, within 1 h of feeding colostrum to a 7-h-old female buffalo calf. Absorption was decreased to 2.1 and .83 g/h after the first feeding, suggesting that the intestinal absorption of macromolecules declined 1 h after the first feeding. Marginal placental transfer of maternal Ig also was demonstrated in buffalo. An albuminlike protein was predominant in blood plasma of the precolostral buffalo calf. This protein decreased, and IgG in blood plasma increased, after colostrum was fed, indicating that albuminlike protein may be involved in the transfer of maternal nutrients to the fetus.

Biochemical changes of the milk in experimental caprine mastitis induced by Mycoplasma serogroup 11 (2-D).

Biochemical changes of the milk were studied in 7 lactating goats. The right udder halves of the animals were inoculated through the teat canal with 1 ml of Mycoplasma ovine/caprine serogroup 11 (2-D) culture containing 6x10(6) colony forming units ml-1. The left udder halves of these goats were kept as control. The experiment was continued for 35 days during which period one goat was killed every 5th day. All the goats developed clinical mastitis within 24 h and the infected udder halves remained enlarged for 4-5 days. The milk from these halves was yellow-tinged and showed slight separation of proteins. Subsequently, there was marked reduction in the size and secretion of the infected udder halves, followed by agalactia from post-inoculation (PI) day 22. The mycoplasmas were reisolated from milk samples of the infected halves up to PI day 20. Histopathological examination of these halves revealed changes indicative of acute and chronic mastitis. Biochemical analysis of mastitic milk/mammary secretions showed a marked increase in the concentration of total proteins, immunoglobulins, free fatty acids and phospholipids, while the concentration of total lipids and glycerides initially underwent a considerable decrease as compared to the basal and control values. However, the level of total cholesterol decreased sharply during the initial days while later, from PI day 5, it markedly increased. This study shows that Mycoplasma ovine/caprine serogroup 11 is pathogenic to the lactating udder of goats and produces marked biochemical alterations in the milk.

Clinicopathological, hematological, and biochemical studies in some outbreaks of nephritis in poultry.

Hematological, biochemical, histoenzymological, and histopathological changes in serum and tissues were studied in chickens during outbreaks of nephritis. Hematological studies revealed normocytic-normochromic anemia characterized by increased total erythrocyte counts, hemoglobin, packed cell volume, and erythrocyte sedimentation rate. Albumin-to-globulin ratio and sodium levels in serum, glucose in blood, and alkaline phosphatase and glucose-6-phosphatase in liver and kidneys were decreased. Glutamate pyruvate transaminase, uric acid, non-protein-nitrogen, and potassium levels in serum were increased. No significant change in the calcium, phosphorus, and total protein levels in serum was observed. These changes were directly related to the severity of the nephritis.

Hematological changes in nephritis in poultry induced by diets high in protein, high in calcium, containing urea, or deficient in vitamin A.

Nephritis was induced in 300, 18-day-old male Arbor Acre broiler chicks by feeding diets high (42.28%) in protein, high (3.27%) in calcium, containing urea (5%), or deficient in vitamin A. Various hematological parameters were studied at weekly intervals. Normocytic-normochromic anemia, characterized by a decrease in total erythrocyte counts, hemoglobin, packed cell volume, and an increase in erythrocyte sedimentation rate, was evident in the birds kept on diets high in protein, high in calcium, or deficient in vitamin A. Increased total erythrocytes, hemoglobin packed cell volume, and erythrocyte sedimentation rate was observed in birds fed urea. Differential leucocyte counts revealed lymphopenia, heterophilia and monocytosis in birds kept on diets high in protein, containing urea, or deficient in vitamin A. However, lymphocytosis, heteropenia , and monocytosis were recorded in birds fed the high calcium diet.

Renal and biochemical changes produced in broilers by high-protein, high-calcium, urea-containing, and vitamin-A-deficient diets.

Three hundred 18-day-old male chicks (Arbor Acre) were divided into five groups of 60 each and given high-protein (42.28%), high-calcium (3.37%), urea-containing (5%), vitamin-A-deficient, or control diets to study the effect of nutritional imbalances on the development of nephritis and related biochemical changes over 15 weeks. The first four diets increased the levels of glutamate oxaloacetate transaminase, glutamate pyruvate transaminase, uric acid, and nonprotein nitrogen in serum. Blood urea was increased by only the urea diet. Hypoglycemia and a decrease in hepatic glucose-6-phosphatase were also observed in chicks fed the first four diets. The vitamin-A-deficient diet resulted in a depletion of vitamin A in the liver and kidneys. These changes were directly correlated with the prolonged feeding of experimental diets and also with the severity of nephritis and degenerative changes in various organs. It was concluded that increasing the intake of nitrogen or calcium in order to increase production may in fact have the opposite effect, leading to degenerative changes in various tissues and to nephritis.

Effects of feeding mustard oil diets on the collagen biosynthesis in various tissues of male rats.

30 rats were divided into 3 groups to study the effects of erucic-acid-rich mustard oil (MO) and MO plus fish oil and carnitine (MOFOC) diet as compared to groundnut oil (GNO) diet on the collagen biosynthesis in various tissues. Changes in collagen content and monoamine oxidase (MAO) of hearts, aorta, skeletal muscles, lungs and skin were determined after 24, 60 and 80 days of feeding, respectively. Incorporation of U-14C-proline into collagen of these tissues was also studied. MO diets increased the incorporation of U-14C-proline into total and acid-insoluble collagen in heart, skeletal muscles and lungs but this increase was of lower magnitude in heart and skeletal muscles as compared with MOFOC diet. Total and acid-insoluble collagen contents in all these tissues, except skin, were increased by MO diet and MOFOC diet retarded this increase. Similar trends were observed in the activities of MAO. The results suggest that the MO diet increases both the biosynthesis and maturation of collagen and MOFOC diet retards this trend. The response of various tissues being variable in this respect.

Changes in composition and enzyme activities of mitochondrial and post-mitochondrial fractions of tissues of rats given mustard oil diet with carnitine and/or fish oil.

The effects of short-term feeding of a 20% mustard oil (containing 47% erucic acid) diet with 0.1% carnitine and/or fish oil on the activity of enzymes of lipid metabolism in mitochondrial and post-mitochondrial fractions of heart, liver, and skeletal muscles of rats were determined to study the reversal of erucic acid-induced lipidosis. Histopathological changes were also studied. The inclusion of fish oil or fish oil plus carnitine in a mustard oil diet, especially the latter, eliminated the histopathologically detectable lipidosis in heart and skeletal muscles. The reason for this appears to be the increased activity of lipase and phospholipase in tissue homogenates and of cholesterol ester hydrolase in the post-mitochondrial fraction; and also an increase in mitochondrial succinate dehydrogenase which may increase the utilization of the liberated fatty acids. The results suggest that the erucic acid-induced lipidosis is prevented or eliminated by increased lipolysis and oxidation of fatty acids when such a diet is supplemented with carnitine and/or fish oil.