Use of the refractive index to measure the adequacy of glycerol removal from previously frozen erythrocytes

A common method for measuring deglycerolization of previously frozen erythrocytes is to determine the osmolality of the supernatant fluid after completion of the wash cycle. We attempted to compare this type of examination with the simpler, faster, and cheaper determination of refractive index (RI) using the Goldberg refractometer. One hundred twenty units of deglycerolized red blood cells, some with glycerol added so as to exceed an acceptable 1% glycerol content, had measurements made of the post-wash supernatant fluid by refractive index and osmometry. The responsiveness of refractive index to residual glycerol was essentially the same as osmolality. Although there is more interference from supernatant hemoglobin, this did not preclude the use of RI to measure glycerol removal. Readings up to 28 had glycerol levels of less than 1%. Readings between 29 and 31 should be checked by osmometry, and readings over 31 should cause the unit of blood to be rejected.     

冰冻红细胞解冻后残留甘油含量测定方法初探

目的　建立快速测定冰冻红细胞中残留甘油含量的方法。方法根据GPO PAP法测定甘油三脂的部分原理 ,将甘油用甘油激酶及三磷酸腺苷 (ATP)磷酸化 ,再以磷酸甘油氧化酶 ,氧化三磷酸甘油 ,生成H2 O2 ,并以4 氨基安替比林 (4 AAP)与 4 氯酚显色 ,显色程度与甘油含量成正比。结果含甘油 2 0 0 /L与 1 2 0 0 /L的样本批内CV值分别为 4 .4 9%、3.5 3% ,日间CV值为 4 .6 % ;甘油含 75 0～ 2 0 0 0 /L样本的回收率为 96 %～ 1 0 3% ;甘油浓度在 2 5 0～ 2 0 0 0 /L有较好的线性关系 ,r =0 .9999,Y =0 .0 0 0 5 6 2X +0 .0 0 0 7。结论甘油磷酸氧化酶法测定冰冻红细胞中残留甘油含量 ,具有方法简单、快速、特异、灵敏之特点 ,结果客观、准确、稳定     

In vitro and in vivo measurements of gamma-radiated, frozen, glycerolized RBCs

BACKGROUND: Transfusion-associated GVHD results from the presence of viable lymphocytes in transfused allogeneic blood components. Viable immunocompetent lymphocytes have been detected in RBCs that were frozen with glycerol and washed before transfusion. STUDY DESIGN AND METHODS: The study reported here assessed the effect of irradiation on human RBCs frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. In vitro and in vivo testing was done on human RBCs that were frozen with 40-percent (wt/vol) glycerol at -80 degrees C, with some units exposed to 2500 cGy of gamma radiation and others not irradiated, and that, after thawing and washing, were stored in a sodium chloride-glucose solution at 4 degrees C for 3 days before autologous transfusion. RESULTS: The glycerol-frozen RBCs treated with 2500 cGy before deglycerolization had a mean freeze-thaw-wash recovery of 87 percent and a mean 24-hour posttransfusion survival of 86 percent after storage for 3 days at 4 degrees C in a 0.9-percent NaCl and 0.2-percent glucose solution. For the nonirradiated units, the mean freeze-thaw-wash recovery was 85 percent and the mean 24-hour posttransfusion survival was 83 percent. CONCLUSION: These data show similar, acceptable results for RBCs frozen with 40-percent (wt/vol) glycerol at -80 degrees C and treated in the frozen state with 2500 cGy of gamma radiation and for RBCs that were not irradiated, all of which were washed and then stored in a sodium chloride-glucose solution for 3 days before autologous transfusion.     

兰州市RhD(-)冰冻红细胞库运行情况调查分析

目的了解兰州市RhD(-)冰冻红细胞库建立以来运行情况,以便提高性改进,更好保障临床RhD(-)血型患者用血。方法调查2011—2016年本中心所有RhD(-)血液采供血数据,统计分析RhD(-)血型检出率、RhD(-)RBC临床应用情况及冰冻红细胞库发展动态。结果兰州市2011—2016年无偿献血检验合格的RhD(-)RBC,81.60%入(4±2)℃动态贮存库供应临床日常所需,18.40%制备成冰冻红细胞入RhD(-)冰冻红细胞库,RhD(-)冰冻红细胞入库量呈逐年下降趋势,解冻去甘油红细胞出库量则大幅增长。结论兰州市RhD(-)冰冻红细胞库2011—2013年运行良好,完全可以保障临床RhD(-)血型患者急救用血,2014年开始进入入不敷出的状态,需采取各种措施来保障RhD(-)冰冻红细胞库的健康发展。     

In vivo survival of apheresis RBCs, frozen with 40-percent (wt/vol) glycerol, deglycerolized in the ACP 215, and stored at 4°C in AS-3 for up to 21 days

BACKGROUND: The FDA has approved the storage of frozen RBCs at -80 degrees C for 10 years and the postwash storage at 4 degrees C for no more than 24 hours. The 4 degrees C postwash storage period is limited to 24 hours, because the current deglycerolization systems are functionally open systems. STUDY DESIGN AND METHODS: Two units of RBCs were collected from each of 13 healthy male volunteers. The RBCs were collected in CP2D by the FDA-approved protocol for an automated apheresis device (MCS, LN8150, Haemonetics) and were stored at 4 degrees C in AS-3 for 6 days. Using a single disposable glycerolization set in an automated, functionally closed system (ACP 215, Haemonetics) each unit was transferred to a 1000-mL PVC plastic bag and glycerolized to a concentration of 40-percent (wt/vol) glycerol and frozen at -80 degrees C. A single disposable deglycerolization set in the ACP 215 was used to deglycerolize the 2 units from the same donor. The deglycerolized RBCs were stored at 4 degrees C in AS-3 for as long as 21 days. RESULTS: The mean +/- SD freeze-thaw-wash recovery value was 89.4 +/- 3 percent. The residual hemolysis in the RBCs stored at 4 degrees C in AS-3 for 21 days after deglycerolization was 0.9 +/- 0.2 percent, and the units were negative for both aerobic and anaerobic bacteria. The mean Nageotte WBC count was 9 x 10(6) per unit. When the deglycerolized RBCs were given as an autologous transfusion after storage at 4 degrees C in AS-3 for the 7- to 18-day period, the mean +/- SD 24-hour posttransfusion survival was 77 +/- 7 percent, and the index of therapeutic effectiveness was 69 +/- 8 percent. CONCLUSION: Two units of human RBCs collected from a single donor by apheresis in the MCS using an LN8150 set can be glycerolized sequentially with a single disposable set and deglycerolized sequentially with another single disposable set in the ACP 215. The previously frozen RBCs stored in AS-3 for 7 to 18 days at 4 degrees C had acceptable hemolysis and an acceptable mean 24-hour posttransfusion survival value and index of therapeutic effectiveness.     

Posttransfusion survival (24-hour) and hemolysis of previously frozen, deglycerolized RBCs after storage at 4 degrees C for up to 14 days in sodium chloride alone or sodium chloride supplemented with additive solutions

BACKGROUND: Previously frozen human RBCs currently are glycerolized and deglycerolized by the use of open systems that limit storage of the deglycerolized RBCs at 4 degrees C to only 24 hours. STUDY DESIGN AND METHODS: Healthy male volunteers who met AABB requirements for blood donors (n = 38) were studied. A volume of 450 mL of blood was collected into CPDA-1. The RBC concentrates were stored at 4 degrees C for 3 to 6 days before being frozen with 40-percent (wt/vol) glycerol and stored at -80 degrees C. The RBCs were deglycerolized, resuspended in 0.9-percent sodium chloride and 0.2-percent glucose (SG) solution or SG solution supplemented with AS-1, AS-3, or AS-5, and stored in the resuspension medium at 4 degrees C for 14 days. RESULTS: The mean +/- SD freeze-thaw-wash process recovery was 90.0 +/- 4.0 percent for all 38 units. The mean 24-hour posttransfusion survival value was 79 percent for deglycerolized RBC stored at 4 degrees C for 7 days in SG alone, SG plus AS-3, or SG plus AS-5. Deglycerolized RBC that were stored at 4 C for 14 days in SG supplemented with AS-1, AS-3, or AS-5 had a mean 24-hour posttransfusion survival of 74 percent. After 7 days of storage of deglycerolized RBCs in SG alone, the mean hemolysis was 3. 7 percent. After 14 days of storage of deglycerolized RBCs in SG supplemented with AS-1, AS-3, or AS-5, the mean hemolysis was 2.5 percent. CONCLUSIONS: The levels of hemolysis did not correlate with the 24-hour posttransfusion survival values.     

Use of Medicare hospital and physician data to assess breast cancer incidence.

OBJECTIVES: Health claims data have the potential of being an inexpensive, timely, and nationally representative source of information about cancer. This study examined the utility of Medicare hospital and physician data as an independent source to identify incident breast cancer cases. METHODS: Data came from Medicare and the National Cancer Institute's SEER cancer registries. From 1992, for women residing in the SEER states (n = 659,260), Medicare hospital and physician claims were reviewed to identify women with a breast cancer diagnosis on a claim (n = 6,784). These women were matched with women in the SEER data who had been diagnosed with breast cancer in 1992 (n = 3,230). The sensitivity, specificity, and positive predictive value (PPV) of the Medicare data were calculated. Logistic regression models were used to identified cancer related procedures reported to Medicare that could distinguish true cases from false positive cases. Predicted values from these models were included to create plots of sensitivity versus false positive rates and sensitivity versus PPV. RESULTS: Medicare hospital data had 62% sensitivity, 99.9% specificity, and 88% PPV. Physician claims increased sensitivity by 14%, with specificity of 99.4%, and a PPV of 10%. Inclusion of additional cancer related diagnoses and procedures improved the ability to distinguish true cases from false positives, although the number of false positive cases remained high. CONCLUSIONS: The Medicare data overall offer limited potential to assess breast cancer incidence, largely because of low sensitivity and poor PPV. The Medicare data may have utility to identify women undergoing selected breast cancer treatments. In addition, the data may be used to help registries focus case-finding efforts, particularly for persons undergoing cancer related treatments.     

Use of videotaping to assess competencies and course outcomes

A constant challenge in nursing education is effectively teaching competencies and allowing students to safely practice essential nursing skills. Faculty members must develop sound methods of instruction to achieve program outcomes, which determine program effectiveness, assure professional readiness, and meet accreditation standards. To this end, the use of videotaping is suggested. Videotaping offers a safe way for students to practice skills and develop confidence prior to actual performance. This article reviews the literature on videotaping and discusses videotaping as used at Central Missouri State University baccalaureate school of nursing. Videotaping is a teaching-learning strategy used to help students develop effective communication, physical assessment, and selected psychomotor skills. Videotaping also provides students with a mechanism for detailed instructor feedback to improve performance.     

Methodology to assess the oxygen concentration of red cells and stroma-free hemoglobin solutions

Stroma-free hemoglobin (SFH) solution is an acellular oxygen carrier under development as a temporary red blood cell (rbc) substitute. SFH will be of value if it contributes to oxygen loading (delivery) and unloading (consumption). If there is no interaction between red cell hemoglobin and SFH, then the composite oxygen dissociation curve for whole blood (WB) can be written as: SWB = rSSFH + (1 - r) Srbc, where S is the fractional saturation, and r is the SFH/WB hemoglobin ratio. This composite whole-blood curve was compared with the calculated curve, and the root mean square difference was 0.9%. Direct and indirect methods of measuring oxygen concentration ([O2]) of whole blood or plasma were compared. The regression line was [O2]indirect = 0.995 [O2]direct + 0.07 (r = 0.995, N = 16, p less than .01). The adequacy of anaerobic separation of plasma was evaluated by measuring the changes in PO2 and PCO2. The PO2 increased by less than 2 torr and the PCO2 decreased by less than 1 torr during in vitro tests. Both the direct and indirect methodology yielded consistent [O2] values for each carrier.     

Osmoles, osmolality and osmotic pressure: clarifying the puzzle of solution concentration

Nurses are routinely involved in the collecting and testing of urine and plasma, dialysis, the administration of intravenous fluids and the treatment of osmolar disorders, all of which require an understanding of solution concentration. This article discusses the various ways in which the concentration of solutions are stated, how they differ and why the different ways of expressing concentration are useful in human physiology. It also explains the similarities and differences between the terms used to describe solution concentration: tonicity, percentage concentration, density, specific gravity, molarity, osmolarity, osmolality and osmotic pressure. The terms osmolarity, osmolality and osmotic pressure appear routinely in textbooks used in undergraduate nursing courses but often are used incorrectly as synonyms. The usefulness and the appropriate context to use the different ways of expressing solution concentration is discussed. Osmolality (or osmolarity) should be used instead of osmotic pressure to describe the movement of water between compartments while the use of osmotic pressure should be reserved for situations where filtration and osmosis are operating together.     

Use of imaging to assess normal and adaptive muscle function

Physical therapists must be able to determine the activity and passive properties of the musculoskeletal system in order to accurately plan and evaluate therapeutic measures. Discussed in this article are imaging methods that not only allow for the measurement of muscle activity but also allow for the measurement of cellular processes and passive mechanical properties noninvasively and in vivo. The techniques reviewed are T1- and T2-weighted magnetic resonance (MR) imaging, MR spectroscopy, cine-phase-contrast MR imaging, MR elastography, and ultrasonography. At present, many of these approaches are expensive and not readily available in physical therapy clinics but can be found at medical centers. However, there are ways of using these techniques to provide important knowledge about muscle function. This article proposes creative ways in which to use these techniques as evaluative tools.     

Use of the chronic pain coping inventory to assess older adults' pain coping strategies.

Little is known about the strategies that older adults use to cope with persistent pain. The purpose of this study was to describe strategies used by older, retirement community-dwelling adults to cope with persistent, noncancer pain, as assessed by the Chronic Pain Coping Inventory (CPCI), to examine the associations of these strategies with disability and depression, and to compare the 65-item and 42-item versions of the CPCI in this population. Two hundred fifty residents of 43 retirement communities in the Pacific Northwest completed baseline measures for a randomized controlled trial of a pain self-management intervention, including the CPCI and measures of demographics, comorbidity, pain-related disability, and depression. The most frequently reported strategies, as assessed by the CPCI, were Task Persistence, Pacing, and Coping Self-Statements. The least frequently used strategies were Asking for Assistance and Relaxation. Regression analyses demonstrated that coping strategies explained 26%, 19%, and 18% additional variance in physical disability, depression, and pain-related interference, respectively, after controlling for age, gender, comorbidity, and pain intensity. Internal consistency for most CPCI-65 and CPCI-42 subscales was adequate. This study clarifies strategies used by older adults to cope with persistent pain and provides preliminary validation of the CPCI in this population. PERSPECTIVE: Findings from this study on pain coping strategies in older adults might suggest potentially useful coping strategies clinicians could explore with individual patients. Investigators can use study findings to design trials of interventions to help older adults cope more effectively with pain.     

Comparison of triglyceride concentration with lipemic index in disorders of triglyceride and glycerol metabolism.

BACKGROUND: In plasma, triglycerides (TG) are transported in lipoprotein particles (mainly chylomicrons, very low-density and low-density lipoprotein). Turbidimetry (bichromatically at 660 and 700 nm) allows measurement of the lipemic (L) index. We explored the use of this index, in combination with a TG assay, to detect errors due to non-fasting, to assess abnormalities in TG metabolism and to detect patients with glycerol kinase deficiency (GKD). METHODS: We collected 2441 patient samples. Normolipidemic (n=2347), type IV hyperlipidemic (n=80), postprandial samples (n=22) and serial dilutions of Intralipid with saline (n=6) were selected. One patient presenting with GKD was included, as well as two patients with type I and type V hyperlipoproteinemia, respectively. RESULTS: We introduced the use of the ratio between the logarithm of serum triglycerides and that of the L-index (TG/L ratio). CONCLUSION: Although the proposed TG/L-index ratio cannot be regarded as an alternative for the accurate diagnosis of lipid disorders, it provides additional information about TG-containing particles.     

Comparison of radioisotope methods and a nonradioisotope method to measure the RBC volume and RBC survival in the baboon

BACKGROUND: RBC volume, 24-hour posttransfusion survival, and life span can be measured with radio-isotopes and nonradioactive procedures. STUDY DESIGN AND METHODS: RBC volume was measured directly with autologous baboon RBCs labeled with biotin-X-N-hydroxysuccinimide (NHS), 51Cr, 99mTc, and 111In-oxine and indirectly from the 125I plasma volume and the total body Hct. Twenty-four-hour posttransfusion survival and life span were measured in autologous fresh baboon RBCs labeled with 51Cr, 111In-oxine, 99mTc, and biotin-X-NHS. RESULTS: Significantly larger RBC volumes were observed when the fresh autologous RBCs were labeled with 51Cr, 111In-oxine, or 99mTc than when they were labeled with the nonradioactive biotin-X-NHS. Twenty-four-hour posttransfusion survival values were significantly lower in the RBCs labeled with 111In-oxine or 99mTc than in the RBCs labeled with 51Cr. CONCLUSIONS: The greater in vivo elution of 51Cr, 111In-oxine, and 99mTc than that of biotin-X-NHS influenced the measurements of RBC volume, 24-hour posttransfusion survival, and life span of the fresh baboon RBCs.     

Use of enzyme treatment to enhance reactivity of HLA and platelet-specific antibodies in solid-phase RBC adherence assays

BACKGROUND: Solid-phase RBC adherence (SPRCA) assays (Immucor) detect HLA and/or platelet-specific antibodies. Pretreatment of reagent platelets with enzymes was investigated to determine whether the sensitivity of the assay could be increased. STUDY DESIGN AND METHODS: SPRCA testing, following the manufacturer's directions, was performed on 51 sera from patients with either a positive SPRCA antibody screen, suboptimal clinical responses to platelet transfusions, and/or suspected immune thrombocytopenic purpura; testing was also performed following pretreatment of the reagent platelets with bromelin, papain, or ficin. Sera from 23 patients having negative routine SPRCA antibody screens and good clinical responses to transfusion were tested as controls. Lymphocytotoxic antibody testing was also performed on selected samples. The effectiveness of enzyme treatment was judged by the increase in the proportion of reagent platelets reacting with the sample and the observed reaction strengths. RESULTS: Pretreatments of reagent platelets with all three enzymes increased the reactivity of known antibodies and detected some HLA and platelet-specific antibodies that had not reacted in routine testing. The clinical significance of the antibody specificities detected only after enzyme pretreatment was verified by a correlation with results from transfusing antigen-negative units. Only occasional false-positive results after enzyme pretreatment were observed. CONCLUSIONS: The use of enzyme pretreatment of SPRCA screening strips can provide information that is useful in selecting appropriate units for transfusion.     

Effects of sodium concentration and osmolality on water and electrolyte absorption from the intact human colon

The influence of sodium concentration and osmolality on net water and monovalent electrolyte absorption from or secretion into the intact human colon was studied in healthy volunteers.