In vitro stability of lyophilized and reconstituted recombinant activated factor VII formulated for storage at room temperature

BACKGROUND: Recombinant activated factor VII (rFVIIa) is indicated to treat bleeding episodes or prevent bleeding related to surgery in patients with hemophilia A or B who have antibodies to coagulation factors VIII or IX. The first-generation rFVIIa formulation is stable when stored under refrigeration. A new formulation has been developed for storage at room temperature, which may improve patients' access to treatment during bleeding episodes. OBJECTIVE: These in vitro experiments were conducted to evaluate the stability of the new formulation of rFVIIa, both lyophilized and reconstituted, under the expected storage conditions, as well as at higher temperatures. METHODS: The stability of the new rFVIIa formulation when stored under various conditions before and after reconstitution was evaluated in terms of retained activity (clotting assay), rFVIIa content (high-performance liquid chromatography [HPLC]), and rFVIIa degradation products (HPLC), including aggregates (dimer/oligomer). Activity was analyzed within specific limits representing the allowable minimum/maximum for each test parameter at the end of the product's shelf-life, as adopted by the European Medicines Agency and the US Food and Drug Administration. Before reconstitution, vials from 9 lots of the new rFVIIa formulation, 3 of each size (1, 2, and 5 mg/vial), were stored at refrigerated temperature (5 degrees C) and at room temperature (25 degrees C) for 24 months, and at 30 degrees C for 12 months. To simulate short-term exposure to temperatures higher than recommended, samples were stored at 40 degrees C for 6 months, followed by storage at 25 degrees C for 12 months. To simulate the home setting, in which the product may be alternately stored in and out of the refrigerator, samples were stored at 30 degrees C for 8 hours and then at 5 degrees C for 16 hours, repeated daily for 5 days. To analyze the effect of storage at extremely elevated temperatures, samples were exposed to temperatures of 50 degrees C, 60 degrees C, and 70 degrees C for 12 hours. After reconstitution, samples were maintained at 25 degrees C for up to 6 hours or at 5 degrees C for up to 24 hours. RESULTS: The specific activity and rFVIIa content of the new lyophilized formulation remained stable after storage for 24 months at 5 degrees C and 25 degrees C, and for 12 months at 30 degrees C; after 5 days of daily alternation between storage at 5 degrees C and 30 degrees C; and after storage for 6 months at 40 degrees C followed by 12 months at 25 degrees C. When stored at 50 degrees C and 60 degrees C for 12 hours, activity remained constant, whereas rFVIIa aggregates increased within the specified limits; after storage at 70 degrees C for 12 hours, rFVIIa activity decreased in parallel with the formation of aggregates, which exceeded the specified limit for the 5-mg product. After reconstitution, samples of all vial sizes of the new rFVIIa formulation retained their activity when stored at 25 degrees C for 6 hours and at 5 degrees C for 24 hours. CONCLUSIONS: In these in vitro experiments, the new lyophilized formulation of rFVIIa was stable when stored for 24 months at 25 degrees C, 12 months at 30 degrees C, 6 months at 40 degrees C, and 12 hours at 50 degrees C and 60 degrees C without compromise to its activity or rFVIIa content. The reconstituted product retained activity when stored at 25 degrees C for 6 hours and at 5 degrees C for 24 hours.     

Associations between physical and mental health problems and sexual dysfunctions in sexually active Danes

IntroductionStudies have shown a high prevalence of sexual dysfunctions among individuals with a variety of health problems.AimTo obtain a population‐based assessment of these matters, we studied associations between indicators of physical and mental health problems and sexual dysfunctions in Denmark.MethodsWe used questionnaire data from 4,415 sexually active men (mean age 48 years) and women (mean age 45 years) who participated in a nationally representative survey in 2005. Cross‐sectional associations of overall health and physical and mental health problems with sexual dysfunctions in the last year were estimated by logistic regression‐derived, confounder‐adjusted odds ratios (OR_adj).Main Outcome MeasuresWe calculated OR_adj with 95% confidence intervals (CI) for erectile dysfunction, anorgasmia, premature ejaculation, and dyspareunia in men, for lubrication insufficiency, anorgasmia, dyspareunia, and vaginismus in women, and for sexual dysfunction and sexual difficulties overall in both sexes.ResultsSexual dysfunction was significantly more common among participants with poor self‐rated health (OR_adj 1.86, 95% CI 1.05‐3.33 in men, OR_adj 1.91, 1.08–3.37 in women). Physical health problems were significantly associated with male sexual dysfunctions (OR_adj 1.75, 1.18–2.61), whereas mental health problems were significantly associated with female sexual dysfunctions (OR_adj 2.59, 1.60–4.22).ConclusionAmong sexually active Danes, poor self‐reported overall health is associated with increased rates of sexual dysfunction in both sexes, with physical health problems mainly affecting men's sex lives, and mental health problems being strongly associated with female sexual dysfunction. Christensen BS, Grønbæk M, Osler M, Pedersen BV, Graugaard C, and Frisch M. Associations between physical and mental health problems and sexual dysfunctions in sexually active Danes. J Sex Med 2011;8:1890–1902.     

Discordance between changes in bone mineral density measured at different skeletal sites in perimenopausal women--implications for assessment of bone loss and response to therapy: The Danish Osteoporosis Prevention Study.

Assessing bone loss and gain is important in clinical decision-making, both in evaluating treatment and in following untreated patients. The aim of this study was to correlate changes in bone mineral density (BMD) at different skeletal sites during the first 5 years after menopause and determine if forearm measurements can substitute for dual-energy X-ray absorptiometry (DXA) of the spine and hip. BMD was measured at 0, 1, 2, 3, and 5 years using Hologic 1000/W and 2000 densitometers in 2,016 perimenopausal women participating in a national cohort study. This analysis comprises 1,422 women remaining in the study after 5 years without changes to their initial treatment (hormone-replacement therapy [HRT], n = 497, or none, n = 925). Despite correlated rates of change between forearm and spine (r2 = 0.11; p < 0.01), one-half of those who experienced a significant decrease in spine BMD at 5 years showed no significant fall in forearm BMD (sensitivity, 50%; specificity, 85%; kappa = 0.25). The total hip had significant better agreement with spine (sensitivity, 63%; specificity, 85%; kappa = 0.37; p < 0.01). Analysis of quartiles of change also showed significant better agreement with spine and whole body for the total hip than for the femoral neck or ultradistal (UD) forearm. In a logistic regression analysis for identification of group (HRT or control), the prediction was best for whole body (82.6%) and spine (80.9%), followed by total hip (78.5%) and forearm (74.7%). In conclusion, changes at the commonly measured sites are discordant, and DXA of the forearm is less useful than DXA of the hip or spine in determining the overall skeletal response to therapy or assessing bone loss in untreated women.     

Renal function of patients in long-term treatment with lithium citrate alone or in combination with neuroleptics and antidepressant drugs

Renal function was studied in patients given lithium citrate alone or in combination with neuroleptics or tricyclic antidepressants or both. No other drugs were given. None of the groups given lithium citrate with other drugs had lithium ion clearances that differed significantly from the groups given lithium citrate alone, nor was there any difference in the clearances of sodium, potassium, or creatinine between these groups. The 24-hour urine volume of patients receiving antidepressant drugs was similar to that of the patients receiving lithium citrate monotherapy but was significantly higher in patients given neuroleptics. The increase in urine volume could not be ascribed to alterations in glomerular filtration rate or proximal tubular resorption but could be accounted for entirely by lowered resorption of water in the distal tubules. We concluded that no change of lithium citrate dose is required when patients so treated are given additional neuroleptic or tricyclic antidepressant drugs.     

Activities in the Danish Year of the Brain 1997

In this paper we report about the Danish Year of the brain 1997 because it may serve as a model for other countries. It was possible in Denmark to organize a very large scale effort which has been well received by the general public and which has significantly improved the image of the neurological disciplines. The resources spent on nationwide initiatives in the Danish Year of the Brain are more than 2 million ECU in country with 5 million inhabitants. In addition however considerably amounts were spent in each country of Denmark. Since all professionals have worked for free innumerable hours of work have not been budgeted. In fact the Danish Year of the Brain has been the largest privately organized health campaign ever held in Denmark and it has been so successful that the Danish Heart Association. Which is old well organized and extremely strong financially has been worried about the shift of attention from the heart to the brain.     

Granulocyte colony-stimulating factor crosses the placenta and stimulates fetal rat granulopoiesis

We studied the effect of recombinant human granulocyte colony- stimulating factor (rhG-CSF) administration to pregnant rats upon fetal and neonatal myelopoiesis. Pregnant rats were treated with rhG-CSF twice daily for 2, 4, and 6 days before parturition. rhG-CSF crossed the placenta and reached peak fetal serum concentrations 4 hours after administration. Peak fetal serum levels were 1,000-fold lower than levels detected in the dam. Hematopoietic effects of rhG-CSF were assessed by cytologic analysis of the newborn blood, spleen, bone marrow, thymus, and liver. White blood cell counts were increased twofold to fourfold in newborns. This increase was due to circulating numbers of polymorphonuclear cells (PMN). rhG-CSF induced a myeloid hyperplasia in the newborn marrow consisting of immature and mature myeloid cells in the day-2 and day-4 treated pups. Bone marrow of pups treated for 6 days contained mostly hyper-segmented PMN with little or no increase in myeloid precursors. An increase in the number of postmitotic (PMN, bands, and metamyelocytes) and mitotic (promyeloblasts, myeloblasts, and metamyeloblasts) myeloid cells in the spleen of neonates was observed. No change was detected in splenic lymphocytes or monocytes. No effect of rhG-CSF was noted in the newborn liver or thymus. These results demonstrate that maternally administered rhG-CSF crosses the placenta and specifically induces bone marrow and spleen myelopoiesis in the fetus and neonate. The significant myelopoietic effects of rhG-CSF at low concentrations in the fetus suggest an exquisite degree of developmental sensitivity to this cytokine and may provide enhanced defense mechanisms to the neonate.