Community Pharmacist Telephonic Medication Reviews with Uncontrolled Asthma Patients: A Pilot Study.

This study reports the process of telephonic medication reviews conducted by community pharmacists for patients with asthma. The study occurred at an independent community chain in association with a Missouri Medicaid consulting group. Participants were identified utilizing claims data and met the National Quality Forum criteria for uncontrolled moderate-to-severe persistent asthma. A pharmacist performed the initial encounter via telephone which included a knowledge questionnaire, symptom control assessment, and medication review. Pharmacists identified drug-related problems (DRPs) and faxed recommendations to patients' primary care providers (PCPs). Thirty days later, pharmacists called to follow up with the patients and faxed PCPs to resolve any outstanding DRPs, new DRPs, or recommendations. Questionnaire scores and symptom control assessments were compared and analyzed utilizing a paired t-test, Chi-squared test, or Fisher's exact test. The number and categories of DRPs, recommendations made by pharmacists, and intervention time were reported. Fourteen participants completed initial encounters with twelve completing follow-up. The majority answered 'yes' to at least one symptom control assessment question indicating partially controlled to uncontrolled asthma. The average knowledge assessment score was 5.17 out of 7 initially and 5.42 for the follow-up. Pharmacists identified 43 DRPs and made 41 recommendations with a mean intervention time of 65 min.

Impact of community pharmacist-performed post-discharge medication reviews in transitions of care.

OBJECTIVES: To describe a community pharmacist-led transitions of care process for patients discharged from the inpatient to the outpatient setting. SETTING: Independent community pharmacy chain in northwestern and central Missouri and a rural nonprofit hospital in Marshall, Missouri. PRACTICE DESCRIPTION AND INNOVATION: This innovative transitions of care service model relies on the inpatient pharmacy team for recruitment and referral of patients who use Red Cross Pharmacy. On discharge, patient information was transmitted to Red Cross Pharmacy via direct messaging in the form of a continuity of care document. Pharmacists used the patients' continuity of care documents to perform telephone-based medication reconciliations and comprehensive medication reviews. Drug-related problems and pharmacists' recommendations were documented in a subjective, objective, assessment, and plan (SOAP) note that was transmitted to the hospital and primary care provider. EVALUATION: The number and type of drug-related problems and pharmacists' recommendations were assessed with the use of descriptive statistics. Pharmacists' time spent was tracked, and its relationship to alternate variables was assessed with the use of bivariate correlations. RESULTS: Pharmacists identified 69 drug-related problems and made 145 recommendations for the 35 patients meeting study criteria. The mean time for total service completion was 65 minutes and positively correlated with a number of variables, including identification of the drug-related problem and unnecessary drug therapy and recommendation to decrease the dosage of a medication. CONCLUSION: Community pharmacists have the ability to identify drug-related problems and make recommendations for patients moving from the inpatient to an outpatient setting. In addition, the data suggest that when given adequate time, pharmacists performing service responsibilities may identify more drug-related problems, resulting in additional recommendations.

Community pharmacy transition of care services and rural hospital readmissions: A case study.

OBJECTIVES: To explore community pharmacist involvement in the transition of care (TOC) process for patients discharged with acute myocardial infarction (AMI), heart failure (HF), pneumonia, chronic obstructive pulmonary disease (COPD), or elective total hip or knee arthroplasty (THA/TKA). SETTING: Patients discharged from a 60-bed acute care hospital located in rural Missouri were seen by a community pharmacist in 2 independent community pharmacy locations. PRACTICE INNOVATION: Patients admitted with 1 of the 5 qualifying conditions and identifying the participating pharmacy as their primary pharmacy spoke with a community pharmacist within 72 hours of discharge to complete a comprehensive medication review. A follow-up telephone call occurred 7 days after the encounter to evaluate for drug-related problems, adherence, and key information recalled from the previous visit. A final telephone call occurred on the 30th day after discharge to assess for hospital readmissions and emergency department (ED) visits. EVALUATION: Number of patients readmitted or visiting the ED within 30 days after discharge. RESULTS: Of the 9 patients completing the study, none were readmitted or visited the ED within 30 days after discharge. All of the participants were satisfied with the care and education provided by pharmacists. The majority of patients recalled points related to specific medication education topics. Based on the adherence tool, 8 of the 9 study participants had at least 1 barrier to medication access or adherence that could lead to post-discharge medication-related problems. CONCLUSION: Community pharmacist involvement in the TOC process may help to prevent readmissions for patients with AMI, HF, pneumonia, COPD, and elective THA/TKA. Patients are overall satisfied with community pharmacist involvement as they move from inpatient care to home. In addition, there are multiple barriers affecting access and adherence to medication therapy while at home, providing opportunities for pharmacist intervention and assistance.

Physicians' Preferences for Communication of Pharmacist-Provided Medication Therapy Management in Community Pharmacy.

OBJECTIVES: (1) To identify physicians' preferences in regard to pharmacist-provided medication therapy management (MTM) communication in the community pharmacy setting; (2) to identify physicians' perceived barriers to communicating with a pharmacist regarding MTM; and (3) to determine whether Missouri physicians feel MTM is beneficial for their patients. METHODS: A cross-sectional prospective survey study of 2021 family and general practice physicians registered with MO HealthNet, Missouri's Medicaid program. RESULTS: The majority (52.8%) of physicians preferred MTM data to be communicated via fax. Most physicians who provided care to patients in long-term care (LTC) facilities (81.0%) preferred to be contacted at their practice location as opposed to the LTC facility. The greatest barriers to communication were lack of time and inefficient communication practices. Improved/enhanced communication was the most common suggestion for improvement in the MTM process. Approximately 67% of respondents reported MTM as beneficial or somewhat beneficial for their patients. CONCLUSIONS: Survey respondents saw value in the MTM services offered by pharmacists. However, pharmacists should use the identified preferences and barriers to improve their currently utilized communication practices in hopes of increasing acceptance of recommendations. Ultimately, this may assist MTM providers in working collaboratively with patients' physicians.

Rural health systems' perceptions of referral to community pharmacists during transitions of care.

OBJECTIVES: To identify rural health systems' perceptions of value, benefits, barriers, and opportunities associated with community pharmacist involvement in patient transitions of care. SETTING: Rural health systems in northwest and central Missouri. PRACTICE DESCRIPTION AND INNOVATION: Qualitative descriptive study of key informant interviews with self-identified decision makers of rural health systems within a 50-mile radius of 15 independent community pharmacy chain locations. EVALUATION: Interviews were recorded, transcribed, and coded to evaluate themes in participant responses. RESULTS: Fifteen interviews were conducted at 8 rural health systems. Participants expressed significant value in community pharmacist involvement in transitions of care and highlighted several benefits, barriers, and opportunities related to potential collaboration. Benefits that were identified included medication monitoring, resource for patient information, and desire among health care providers to work with community pharmacists. Barriers included legal and regulatory issues with referral, communication, and prescriber utilization. Opportunities described included: patient education, monitoring, and follow-up; targeted interventions; medication access assistance; bedside medication delivery; and collaboration between community pharmacies and health care entities. CONCLUSION: Rural health system informants perceived community pharmacy involvement to be valuable and were receptive to collaboration during transitional care to improve patient outcomes. They highlighted barriers to overcome to truly incorporate community pharmacists into the transitional care arena. Understanding these rural health systems' perceptions can guide community pharmacies in developing collaborative relationships and patient care services to assist with care transitions.

[Autologous serum and alternative blood products for the treatment of ocular surface disorders]. / Eigenserum und alternative Blutprodukte zur Behandlung von Augenoberflächenerkrankungen.

Eye drops made from autologous serum have been increasingly used in the past decade to treat ocular surface disorders such as persistent epithelial defects and dry eye. Due to biologically active ingredients such as growth factors, vitamins, and nutrients, they can be used to lubricate the ocular surface and support epithelial wound healing. According to current legal requirements, they can be dispensed only for outpatient treatment if the producer has obtained a license from the appropriate local authorities. Therefore, the production and dispensing of autologous serum eye drops in Germany is currently limited to a very few institutions and their patients. We review the current evidence on the use of serum eye drops, recommend a standard protocol for their production, and describe a number of recently emerging alternative blood products for the treatment of ocular surface diseases along with their potential advantages and limitations.

[On the use of autologous serum eyedrops in Germany : results of a survey among members of the Cornea Section of the German Ophthalmological Society (DOG)]. / Zum Stand der Anwendung von Eigenserum-Augentropfen in Deutschland : Ergebnisse einer Umfrage unter den Mitgliedern der Sektion Kornea in der DOG.

PURPOSE: The use of serum eye drops has become more common for treating ocular surface diseases such as persistent epithelial defect and dry eye. For the production and use of blood products, regulatory restrictions apply. We surveyed the practice of production and application of serum eyedrops among the members of the Cornea Section of the German Ophthalmological Society (DOG) in Germany. METHODS: A questionnaire concerning the application of topical blood-derived therapeutic agents for use in the eye was sent to 103 institutions in 2005 and 2007. The questionnaire included 14 groups of questions. RESULTS: The rate of response was 49% in 2005 and 55% in 2007. Sixty-five percent (2005: 48%) of the institutions used blood-derived products as therapeutic agents in the eye (in 96%, serum eye drops). In 2007, 17% of the centers stated that they had a license to produce serum eye drops according to the official regulations (2005: 10%). Three hospitals stopped using serum eyedrops for regulatory reasons. In 2007 1,237 patients were treated (2005: 1,389); 36% of the ophthalmic departments produced the serum eye drops themselves (2005: 55%) and 43% produced them in cooperation together with or exclusively in another institution. Quality controls (e.g., virus serology and sterility checks) were performed in 47% (2005: 24%). In 2005, one corneal fibrin deposit was described. In 2007, no complication was reported. CONCLUSION: Although fewer patients were treated in total in 2007, the number of centers using serum eye drops increased between 2005 and 2007. Serious complications were not described. Some hospitals stopped the production and use of serum eye drops because of regulatory reasons or had the drops produced by nonophthalmic institutions with a license to produce blood-derived products. In view of prospective randomized clinical trials showing that autologous serum support ocular surface wound healing, this therapy should be accepted as a standard of care.

[Legal regulations to produce serum eye drops : when is it necessary, and how can it be obtained?]. / Arzneimittelrechtliche Erlaubnis bei der Herstellung von Serum-Augentropfen : Wann ist sie erforderlich und wie kann sie umgesetzt werden?*

According to European Union and German legislation, the production of medicines such as eye drops from autologous serum requires a license from an appropriate authority. However, an exemption is granted to medical doctors who produce and apply medications under their immediate responsibility. If doctors do not actually produce such medications themselves, they must select appropriate personnel to do so. These individuals have to be sufficiently qualified and reliable and must be carefully instructed on the standard quality and manufacturing procedures, although the doctors are responsible for controlling the production and quality of the final product. The treating physicians are also responsible for the appropriate application of the produced medications. To dispense medication to patients without an appropriate license is considered a criminal offense. We describe how two German hospitals have established the production of serum eye drops in full compliance with legal regulations.

[Experiences with a centralised national service for autologous serum eyedrops in England]. / Erfahrungen mit einem national zentralisierten Herstellungsverfahren für Eigenserum-Augentropfen in England.

The majority of patients with dry eye syndromes respond to conventional treatment aimed at optimising the ocular surface environment. There are some, however, who do not respond adequately to conventional lubricants. The first description of the use of autologous serum as a nutrient tears substitute was published more than 20 years ago. In 1997, NHS Blood and Transplant (NHSBT) developed a reliable and reproducible method for the production of eyedrops derived from autologous serum according to GMP Guidelines. The current cost of a batch of eyedrops (i. e. the product from one donation episode) is approximately 1300 GBP - this covers costs of collection, processing, testing and distribution. One "batch" of eyedrops will last for approximately 5 months if a bottle a day is used. A 6 month shelf life is put on the product and patients keep them in their domestic freezer.

[Epitheliotrophic capacity of serum eye drops from healthy donors versus serum from immunosuppressed patients with rheumatoid arthritis]. / Epitheliotrophe Kapazität von Serum-Augentropfen gesunder versus immunsupprimierter Patienten mit rheumatoider Arthritis.

BACKGROUND: Autologous serum has been advocated for the treatment of persistent corneal epithelial defects and other ocular surface disorders which may be a local manifestation of a systemic disease. Many of these underlying disorders are cytokine-mediated and require immunosuppressive therapy. The systemic disease and medication could potentially influence the epitheliotrophic capacity of serum eye drops. We compared the effect of serum from healthy and immunosuppressed donors in a human corneal epithelial cell culture model. METHODS: Serum was prepared under standardised conditions from full blood samples of 10 healthy donors and 10 patients suffering from rheumatoid arthritis. All patients were treated with prednisolone and methotrexate, one also with azathioprine. In these serum samples EGF, FGF, HGF, PDGF-AB, TGF-beta1, fibronectin, vitamin A and E as well as IL-6 were quantified by means of routine ELISA or HPLC technology. SV-40 immortalised human corneal keratinocytes were cultured in 96-well plates at 37 degrees C, 5 % CO (2) with a fully defined culture medium. At 30 % confluency the culture medium was substituted by one of the test preparations. Proliferation of cell cultures was quantified by means of a luminescence-based ATP assay in dose-response experiments. A colony dispersion assay was used to examine the effect on cell migration and differentiation was assessed by means of scanning electron microscopy. RESULTS: Serum from healthy donors differed from serum of immunosuppressed individuals suffering from rheumatoid arthritis only in that it contained significantly higher amounts of fibronectin and TGF-beta1. Support of proliferation, migration and differentiation of corneal epithelial cells was dose-dependent, but no significant difference was observed between the serum of the two different groups of donors. At a dilution of 25 % serum of healthy donors showed a significantly higher stimulation of migration than serum of immunosuppressed patients. CONCLUSION: The effect of serum on migration but not proliferation is affected by systemic diseases requiring immunosuppression. If an epithelial defect of a patient with rheumatoid arthritis does not respond to treatment with diluted autologous serum, undiluted serum should be tried since the positive effect of serum on cell migration is positively correlated with dose.

The large form of ADAR 1 is responsible for enhanced hepatitis delta virus RNA editing in interferon-alpha-stimulated host cells.

Hepatitis delta virus (HDV) RNA editing controls the formation of hepatitis-delta-antigen-S and -L and therefore indirectly regulates HDV replication. Editing is thought to be catalysed by the adenosine deaminase acting on RNA1 (ADAR1) of which two different forms exist, interferon (IFN)-alpha-inducible ADAR1-L and constitutively expressed ADAR1-S. ADAR1-L is hypothesized to be a part of the innate cellular immune system, responsible for deaminating adenosines in viral dsRNAs. We examined the influence of both forms on HDV RNA editing in IFN-alpha-stimulated and unstimulated hepatoma cells. For gene silencing, an antisense oligodeoxyribonucleotide against a common sequence of both forms of ADAR1 and another one specific for ADAR1-L alone were used. IFN-alpha treatment of host cells led to approximately twofold increase of RNA editing compared with unstimulated controls. If ADAR1-L expression was inhibited, this substantial increase in editing could no longer be observed. In unstimulated cells, ADAR1-L suppression had only minor effects on editing. Inhibition of both forms of ADAR1 simultaneously led to a substantial decrease of edited RNA independently of IFN-alpha-stimulation. In conclusion, the two forms of ADAR1 are responsible almost alone for HDV editing. In unstimulated cells, ADAR1-S is the main editing activity. The increase of edited RNA under IFN-alpha-stimulation is because of induction of ADAR1-L, showing for the first time that this IFN-inducible protein is involved in the base modification of replicating HDV RNA. Thus, induction of ADAR1-L may at least partially cause the antiviral effect of IFN-alpha in natural immune response to HDV as well as in case of therapeutic administration of IFN.

Topical treatment of ocular surface defects: comparison of the epitheliotrophic capacity of fresh frozen plasma and serum on corneal epithelial cells in an in vitro cell culture model.

Accelerated healing of ocular surface disorders was reported using serum for topical application. It is supposed that growth factors, fibronectin and vitamins in serum support the proliferation of corneal epithelial cells. The use of fresh frozen plasma (FFP) instead of serum is theoretically attractive, as it is more easily available from blood banks. In this study, serum and FFP were investigated for composition of epitheliotrophic factors and effect on corneal epithelial cells. Whole blood was taken from five donors. Serum and FFP were prepared, and the concentrations of epithelial growth factor (EGF), Platelet-derived growth factor (PDGF), transforming growth factor-beta1, fibronectin and vitamin A were determined. Immortalized human corneal epithelial cells were used to investigate growth, migration and differentiation in response to both blood products. Significant differences were found regarding the mediator composition of serum and FFP. Serum rather than FFP was significantly superior in stimulating cell growth, migration and differentiation. The epitheliotrophic capacity of blood products depends upon the composition of growth factors and vitamins. Blood clotting strongly influences the growth factor pattern. The superior epitheliotrophic capacity of serum might be due to the higher concentration of proliferation mediators such as EGF and PDGF and its higher content of vitamin A.

An optimised protocol for the production of autologous serum eyedrops.

BACKGROUND: Serum eyedrops have been successfully used in the treatment of severe dry eye, persistent epithelial defects and other severe ocular surface disorders. A number of clinical studies showed a variable efficacy of this approach, but the parameters for the production of this blood product varied significantly. In order to establish an optimised protocol for the production of serum eyedrops, we examined the effect of various clotting times, centrifugation forces, types of diluent and dilutions on the concentration of growth factors, fibronectin, and vitamins in serum and tested the epitheliotrophic capacity of these serum modifications in a cell culture model of human SV-40-immortalised corneal epithelial cells (HCE-T). METHODS: Serum samples were prepared with a clotting time of 20, 60 or 120 min, a centrifugation force of 500 xg or 3,000 xg, and diluted with BSS or isotonic saline. The concentrations of EGF, TGF-beta1, PDGF-AB, FGF, HGF, fibronectin, vitamin A and vitamin E in these samples were evaluated with ELISA and HPLC. HCE-T cells were incubated for 24, 48, 72, 96 and 144 h with 100, 50, 25, 12.5, 6.25 and 3.125% serum in diluent, and cell proliferation, migration and differentiation were evaluated by means of a luminescence-based ATP assay, a colony-dispersion assay and scanning electron microscopy. RESULTS: Using a longer clotting time resulted in an increased concentration of all the epitheliotrophic factors examined in serum; the difference was statistically significant for EGF, TGF-beta1 and HGF. Increasing the g force of centrifugation from 500 xg to 3,000 xg resulted in significantly less TGF-beta1, but more EGF and vitamin A. Cell proliferation was better supported by serum prepared with 3,000 xg and diluted with BSS. Serum prepared with a longer clotting time yielded better cell migration and differentiation. CONCLUSION: Clotting time, centrifugation and diluents have a significant impact on the composition and epitheliotrophic effects of serum. A long clotting time (>or=120 min), a sharp centrifugation (3,000 xg for 15 min) and dilution with BSS improve the ability of serum eyedrops to support proliferation, migration and differentiation of corneal epithelial cells.

Autologous serum eye drops for ocular surface disorders.

Tears have antimicrobial, nourishing, mechanical, and optical properties. They contain components such as growth factors, fibronectin, and vitamins to support proliferation, migration, and differentiation of the corneal and conjunctival epithelium. A lack of these epitheliotrophic factors--for example, in dry eye, can result in severe ocular surface disorders such as persistent epithelial defects. Recently, the use of autologous serum in the form of eye drops has been reported as a new treatment for severe ocular surface disorders. Serum eye drops may be produced as an unpreserved blood preparation. They are by nature non-allergenic and their biomechanical and biochemical properties are similar to normal tears. In vitro cell culture experiments showed that corneal epithelial cell morphology and function are better maintained by serum than by pharmaceutical tear substitutes. Clinical cohort studies have reported its successful use for severe dry eyes and persistent epithelial defects. However, the protocols to prepare and use autologous serum eye drops varied considerably between the studies. As this can result in different biochemical properties protocol variations may also influence the epitheliotrophic effect of the product. Before the definitive role of serum eye drops in the management of severe ocular surface disease can be established in a large randomised controlled trial this has to be evaluated in more detail. In view of legislative restrictions and based upon the literature reviewed here a preliminary standard operating procedure for the manufacture of serum eye drops is proposed.

[Epitheliotrophic capacity of serum and plasma eyedrops. Influence of centrifugation]. / Epitheliotrophe Kapazität von Serum- und Plasmaaugentropfen. Einfluss der Zentrifugation.

BACKGROUND: Serum eyedrops are a new modality for the treatment of ocular surface disorders. We examined the influence of the preparation of blood products in a cell culture model and compared it with plasma. MATERIAL AND METHODS: Serum and plasma were obtained from full blood of ten healthy volunteers and centrifuged at 500 and 3000 G. EGF, PDGF, TGF-beta1, fibronectin, and vitamin A were quantified by means of ELISA and HPLC. Cultures of human corneal epithelial cells were incubated with the four blood products in dose-response experiments and the intracellular ATP quantified. RESULTS: EGF, PDGF, and vitamin A were present in serum in significantly higher concentrations than in plasma. The concentration of fibronectin was not influenced by the preparation. Support of proliferation was best by 25% platelet-poor serum. Serum supported the differentiation and migration of epithelial cells better than plasma. CONCLUSION: The biochemical character of serum eyedrops is determined by the parameters chosen to produce the blood product. Plasma does not seem to offer an epitheliotrophic capacity equivalent to serum eyedrops. Their production should be optimized before any meaningful randomized controlled clinical trial can be attempted.

[Autologous serum-eye-drops for ocular surface disorders. A literature review and recommendations for their application]. / Autologe Serum-Augentropfen zur Therapie der Augenoberfläche Eine Ubersicht zur Wirksamkeit und Empfehlungen zur Anwendung.

The natural tear film has mechanical, optical, antimicrobial and nutritional properties. Tear film components, such as EGF, fibronectin and vitamin A, play a vital role in the proliferation, migration and differentiation of the corneal and conjunctival epithelium. In ocular surface disease, such as severe dry eye, the epithelia may be depleted of these nutritional factors. Replacing the aqueous component of tears alone,by using pharmaceutical tear substitutes,often has little effect on the ocular surface.Eye-drops prepared from autologous serum are a new treatment option for severe ocular surface disease. They can be produced according to the regulations on drug use as an unpreserved blood preparation. Autologous serum eye-drops are non-allergenic and their biomechanical and biochemical properties are similar to normal tears. In cell culture experiments, serum was found to be superior to preserved or unpreserved pharmaceutical products in the maintenance of human keratinocyte morphology and function. It supports the migration of corneal epithelial cells and the differentiation of conjunctival epithelial cells. The first clinical cohort studies report its successful use for severe dry eyes and persistent epithelial defects. In these studies, however, varying methods for the preparation and different concentrations of autologous serum eye-drops were used. These methodological variations determine the biochemical properties and thus the epitheliotrophic effect of serum eye-drops. In this review we summarise the currently available clinical evidence, discuss relevant legislatory restrictions and describe a standard operating protocol for the use of serum eye drops. This has to be evaluated and optimised in more detail before any meaningful, randomised, controlled trial can attempt to establish the role of serum eye-drops in the management of severe ocular surface disease.

Evidence for de novo synthesis of cytokines and chemokines in platelet concentrates.

BACKGROUND AND OBJECTIVES: Inflammatory cytokines in platelet concentrates (PC) may cause side-effects such as febrile non-haemolytic transfusion reactions. The maximum white blood cell (WBC) content tolerable to avoid the accumulation of cytokines, and whether these cytokines originate from degranulating leucocytes or de novo synthesis during storage, had not been investigated prior to this study. MATERIAL AND METHODS: We investigated the secretion of interleukin (IL)-1beta, IL-2, IL-6, IL-8, tumour necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) and quantified the appropriate expression of corresponding mRNA in PC with regard to different levels of WBC contamination and storage times. In addition we tested the viability of WBCs during PC storage (by staining with 7-aminoactinomycin D) and their ability to perform de novo cytokine synthesis (by using superantigen stimulation). RESULTS: We detected a statistically significant increase of IL-1beta, IL-6, IL-8 and TNF-alpha in PC with > or = 108 WBCs. Quantitative reverse transcription-polymerase chain reaction (RT-PCR) showed increasing mRNA expression of the respective cytokines depending on the number of WBC present. On day 5 of storage, WBC viability was > 80% and the leucocytes were still able to produce cytokines de novo. CONCLUSIONS: These data show clear evidence for de novo synthesis of cytokines in PC. The cytokine pattern supports the hypothesis that activated monocytes are responsible for this cytokine synthesis. PC with a WBC contamination of > or = 108 contain inflammatory mediators in clinically relevant concentrations.

Successful long-term erythrocytapheresis therapy in a patient with symptomatic sickle-cell disease using an arterio-venous fistula.

The use of long-term automated erythrocytapheresis via an arterio-venous fistula for the prevention of recurrent ischaemic stroke in a child with sickle-cell disease (SCD) has not been described previously. We report the successful use of this technique in a 13-year-old boy. A procedure was performed every 36 +/- 6 days, transfusing six units of donor packed red blood cells (RBCs) and discarding 1318 +/- 174 mL of exchanged erythrocytes (Hct 60%). After transfusion of 85 units over 17 months, there is no evidence for iron-overload, red cell alloimmunization, transfusion-transmitted infections, or other complications. Until now, no cerebrovascular ischaemia has been observed.

A novel RT-PCR for reliable and rapid HCV RNA screening of blood donations.

BACKGROUND: The objective of this work was to develop a novel and highly sensitive RT-PCR method that is suitable for HCV RNA screening of blood donations according to the criteria released by the Paul Ehrlich Institute, the federal licensing agency of Germany, for routine HCV NAT. STUDY DESIGN AND METHODS: RNA was prepared from plasma pools of up to 20 single blood donations using an automated nucleic acid isolation system (NucliSens Extractor, Organon Teknika). For reverse transcription, amplification, and simultaneous detection of PCR products, a novel approach based on the TaqMan technology was developed. Glyceraldehyde-3-phosphate dehydrogenase messenger RNA, which is detectable in human plasma, was coamplified in each reaction as an internal positive control. RESULTS: The HCV genotypes and subtypes 1a, 1b, 2a, 2b, 2c, 2i, 3a, 4, and 5a were detected in parallel with comparable amplification efficiency. The 95-percent detection limit related to the WHO HCV RNA standard preparation was calculated to be 389 IU per mL of plasma of the single blood donation. Total CVs (%) were <4. The screening of up to 180 blood donations took 5 hours; as a rule, the blood components could be released on the day of donation. CONCLUSION: The TaqMan HCV RT-PCR is an almost completely automated, highly sensitive, specific, and rapid method that is reliable for HCV RNA screening of blood donations. It allows a closed-tube HCV RNA detection without risk of contamination by PCR products.

Improvement of the precision in CFU-GM and BFU-E counting by flow cytometry-based standardization of short-term culture assays.

The counting of colony-forming units granulocyte-macrophage (CFU-GM) and burst-forming units erythrocyte (BFU-E) provides substantial in vitro information about the graft quality after peripheral stem cell transplantation (PBSCT). By using different techniques for culturing and scoring, high inter- and intralaboratory coefficients of variation (CV) are frequently reported. We minimized the imprecision by using flow cytometry-based incorporation of constant numbers of CD34(+) cells per culture dish instead of the formerly used mononuclear cells. Our results show acceptable CVs for CFU-GM (12.3%) and for BFU-E (13.3%) based on this seeding technique, which contributes to fulfilling the demands of a quality assurance system in stem cell laboratories.