Intertester reliability of assessing postural sway using the chattecx balance system

In this study, we examined intertester reliability of dynamic and static balance using the Chattecx Dynamic Balance System. Ten female and two male subjects were randomly assigned to two testers and completed ten, 10-second preprogrammed double- and single-leg test conditions. Balance was measured as postural sway in centimeters by the Chattecx Dynamic Balance System. Static balance consisted of the platform remaining stable, whereas dynamic balance consisted of the platform tilting in an anterior and posterior direction during testing. The protocols and average postural sway values were:1. Double-Leg Static Eyes Open ([unk] = .28 cm)2. Double-Leg Static Eyes Closed ([unk] = .37 cm)3. Double-Leg Dynamic Eyes Open ([unk] = .86 cm)4. Double-Leg Dynamic Eyes Closed ([unk] = 1.72 cm)5. Single-Leg Static Dominant Eyes Open ([unk] = .99 cm)6. Single-Leg Static Dominant Eyes Closed ([unk] = 1.70 cm)7. Single-Leg Static Nondominant Eyes Open ([unk] = .65 cm)8. Single-Leg Static Nondominant Eyes Closed ([unk] = 1.48 cm)9. Single-Leg Dynamic Dominant Eyes Open ([unk] = .99cm)10. Single-Leg Dynamic Nondominant Eyes Open ([unk] = .91 cm).The mean values of postural sway ranged from .28 cm to 1.72 cm for double-leg stance measures and from .65 cm to 1.70 cm for the single-leg stance measures. The intraclass correlations (ICCs)(2,1) ranged from poor to excellent (ICCs = .06 to .90) and the standard error of measurement (SEM) ranged from .06 to .34 for the 10 trials. We feel that variability exists between subjects and/or testers between trials. The wide range of reliability values suggests that further research should determine both intratester and intertester reliability using a variety of protocols for assessment of static and dynamic balance.     

Platelet and growth factor concentrations in activated platelet-rich plasma: a comparison of seven commercial separation systems

Platelet-rich plasma (PRP) is blood plasma that has been enriched with platelets. It holds promise for clinical use in areas such as wound healing and regenerative medicine, including bone regeneration. This study characterized the composition of PRP produced by seven commercially available separation systems (JP200, GLO PRP, Magellan Autologous Platelet Separator System, KYOCERA Medical PRP Kit, SELPHYL, MyCells, and Dr. Shin’s System THROMBO KIT) to evaluate the platelet, white blood cell, red blood cell, and growth factor concentrations, as well as platelet-derived growth factor-AB (PDGF-AB), transforming growth factor beta-1 (TGF-β1), and vascular endothelial growth factor (VEGF) concentrations. PRP prepared using the Magellan Autologous Platelet Separator System and the KYOCERA Medical PRP Kit contained the highest platelet concentrations. The mean PDGF-AB concentration of activated PRP was the highest from JP200, followed by the KYOCERA Medical PRP Kit, Magellan Autologous Platelet Separator System, MyCells, and GLO PRP. TGF-β1 and VEGF concentrations varied greatly among individual samples, and there was almost no significant difference among the different systems, unlike for PDGF. The SELPHYL system produced PRP with low concentrations of both platelets and growth factors. Commercial PRP separation systems vary widely, and familiarity with their individual advantages is important to extend their clinical application to a wide variety of conditions.     

两种血细胞分离机干/祖细胞采集和移植效果的比较

背景：外周血造血干/祖细胞通过血细胞分离机进行体外收集,血细胞分离机的性能、工作状况将直接关系到采集物的特性和数量,从而直接影响受者的造血重建。 目的：比较Fenwal CS-3000 Plus与Amicus两种血细胞分离机采集外周血造血干/祖细胞及受者移植效果。 方法：共入选接受异基因外周血干细胞移植患者51例,Fenwal CS-3000 Plus组27例,平均年龄(34.2±10.6)岁;Amicus组24例,平均年龄(35.4±12.1)岁。比较两组采集物有核细胞数、CD34+细胞数、采集效率、红细胞与血小板采集量及造血重建时间。 结果与结论：两组采集物的有核细胞数、CD34+细胞数、采集效率、红细胞采集量及造血重建时间差异均无显著性意义;Fenwal CS-3000Plus组血小板采集量明显高于Amicus组(P < 0.01)。结果显示Fenwal CS-3000 Plus与Amicus血细胞分离机在分离外周血干/祖细胞方面和移植效果没有差异;血小板较低被采集者使用Amicus血细胞分离机采集可能更为安全。     

National French observatory of the quality of blood components for transfusion.

PURPOSE: Since 1998, prestorage leucoreduction of cellular blood components (BC) is mandatory in France. The French Blood Service needs to follow the data on the quality of the BC prepared by blood centers. This article gives an overview of the quality control (QC) data from 2001 to 2006. MATERIAL AND METHODS: QC data are submitted to a central data bank by each centre. The data are stratified according to preparation process for analysis of key performance criteria - residual leukocytes and haemoglobin or platelet content. BC preparation processes, methods for measuring haemoglobin and platelet content, and for counting residual leukocytes are those routinely employed by centers. RESULTS: The preparation process of red cell concentrates (RCC) influences the haemoglobin content: 57.6+/-6.8 g per unit versus 50.9+/-5.4 g per unit for whole blood or RCC filtration, respectively. Apheresis RCC exhibits a reduced variability (51.2+/-3.4 g per unit). For apheresis platelet concentrates, the median residual leukocyte count remains low for all separators (0.019-0.044 x 10(6)leukocytes per unit, in 2006). However, the percentage of units exceeding 1 x 10(6)leukocytes per unit is significantly higher with one separator (1.8% versus 0.8%, in 2006). For pooled buffy-coat derived platelets, we observed a significant increase in platelet recovery throughout the years (0.66-0.77 x 10(11)platelets per buffy-coat in 2001 and 2006, respectively). CONCLUSION: Our QC data show an overall compliance with the requirements for cellular BC. Our data bank is useful to inform on the performance of leucoreduced BC preparation processes carried out with market available devices.     

Seven-day storage of single donor platelets in polyolefin bags: clinical, biochemical, morphological and microbiological evaluation

We compared the in vitro and in vivo function of fresh and stored platelet concentrates (PCs) collected by an automated continuous-flow blood cell separator (CS 3000 Fenwal) in a closed-system apheresis kit in order to evaluate the possibility of extending the storage time to seven days with the polyolefin container (PL-732). The initial 220 ml platelet volume (5.14 +/- 1.23 x 10(11) was divided into two parts. Half was transfused and the other half was stored for 7 days. All cultured units were negative for bacterial contamination. Mean counts for fresh and stored platelets were respectively 2.34 +/- 0.59 and 2.17 +/- 0.50 X 10(11)/100 ml of PCs (mean recovery 88.7 +/- 11.9%). The pO2 levels were maintained during storage (179.9 +/- 30.5 mmHg) but pCO2, pH, LDH, osmolality, glucose consumption, bicarbonates, ATP, and osmotic stress values changed significantly after 7 days storage. From a clinical point of view, in 14 patients receiving a total of 38 PC transfusions no statistically significant change in corrected post-transfusional levels was observed between fresh and stored PC. Biochemical and morphological data and clinical results suggest that PCs collected with CS-3000 blood cell separator in a closed system and stored for 7 days in polyolefin bags (PL-732) can be satisfactorily employed in clinical practice.     

Stem cell collection using the dideco excel continuous flow blood cell separator: parameters for optimal stem cell collection timing

This study evaluates stem cell collection procedures performed with the Dideco Excel blood cell separator, with particular attention given to yields and separator collection efficiencies. Patients' blood precounts and yield parameters related to the harvest capacity of the collection system were investigated. Fifty-five collection procedures were analyzed in 32 patients suffering from hematological malignancies and solid tumors and mobilized with chemotherapy plus G-CSF. The median blood volume processed in each procedure was 15.8 liters (12-19.750), with a blood flow rate of 70 ml/min. Patients had the following median blood precount value: NC 7.81x10(9)/L, CD34+ cells 49.08x10(3)/ml. Leukapheresis procedures gave the following yields: NC 14.95x10(9), MNC 10.83x10(9), CD34+ cells 4.37x10(6); yields/kg, NC 0.21x10(9)kg, MNC 0. 15x10(9)/kg CD34+ cells 4.26x10(6)/kg. Procedures show the following collection efficiencies: NC 10.79%, MNC 29.06%, CD34+ 42.33%, PLT 26.5%. The RBC (red blood cell) contamination of the product was (median value) 20.9 ml for each procedure, and for platelets 1.76x10(11) per procedure. The CD34+ cell precounts strongly correlated with the CD34+ yields/kg (r=0.82. p=0.000). Furthermore the NC and MNC precounts correlated with the CD34+ yields/kg but only the MNC precount correlation is notable (r=0.57, p=0.000). The logistic regression analysis shows that CD34+ (p=0.008) but not NC (po=0.14), MNC (p=0.09), or PLT (p=0.53) precounts significantly influenced the collection of a sufficient dose of CD34+ cells for transplantation (> or =2.5x10(6)/kg). Eleven of the thirty-two patients have been transplanted till now, and all had a prompt and lasting trilineage engraftment NC >1x10(9)/L on day 12 (10-17). Our data show that the collection system analyzed in this report is able to collect large amounts of progenitor cells, harvesting >2.5x10(6)/kg CD34+ cells with a single procedure in 68.8% of patients and assuring complete recovery after stem cell transplantation.     

Modifications and substitutions of the RNA extraction module in the ViroSeq HIV-1 genotyping system version 2: effects on sensitivity and complexity of the assay

Genotypic testing for HIV-1 resistance to anti-retroviral drugs has become accepted widely as a routine method to guide anti-retroviral therapy. However, implementation into routine high-throughput laboratory diagnosis is difficult due to the complexity of the assay. A commercially available assay is the ViroSeq HIV-1 Genotyping System (Applied Biosystems, Weiterstadt, Germany). We modified and substituted the RNA extraction module to optimize the proportion of samples amplified successfully as follows: 1 ml plasma was concentrated by ultracentrifugation and extracted according to the manufacturer's instructions (Kit), by substituting the lysis buffer (Roche, Roche Diagnostics GmbH, Mannheim, Germany), and by using the QIAamp Viral RNA Kit (Qiagen GmbH, Hilden, Germany) with elution volumes of 60 (Q60) or 50 micro l (Q50). Overall Q50 showed a higher success rate (97%) than the other extraction modules used (range 88-91%). In samples with a viral load range of 1,000-4,999 copies/ml, Q50 was superior (95 vs. 65% to 83%), while in samples with a viral load range of 5,000-9,999 copies/ml or those with 10,000 or more copies/ml, the success rate of the extraction procedures showed no significant differences. In 18 samples, which were negative using the Kit or Roche extraction, Q60 resulted in 7/18 positive results; in addition the Q50 was successful in amplifying 7/10 of the Q60 negative samples. When investigating samples with a measurable viral load of less than 1,000 copies/ml or lower, Q50 had the highest success rate with 80% compared to the other procedures (33-63%). A statistically significant new cut-off could be defined for Q50 at a value of 250 copies/ml. The results showed clearly that the ViroSeq System is suitable for analyzing the HIV-1 genotype over a wide range of viral loads but could be improved significantly when substituting the RNA extraction module with Q50 without using a nested PCR protocol. This is of great importance as it avoids further time- and cost-intensive steps.     

The possible cost effectiveness of peripheral blood stem cell mobilization with cyclophosphamide and the late addition of G-CSF

The purpose of this study was to develop a cost-effective protocol for the mobilization of peripheral blood stem cells (PBSC) in patients with malignancy. Thirty consecutive patients were randomized to mobilize PBSC with the late addition of a standard 250 microg dose of G-CSF (Neutrogen) from day 8 or early addition of the same dose of G-CSF from day 2, following cyclophosphamide (CY) 4 g/m2. The median yield of CD34+ cells from evaluated patients was 7.87 x 10(6)/kg (range, 2.06-27.25), collected in a median of four apheresis (range, 2-9). Target CD34 + cell doses > or = 2.0 x 10(6)/kg were achieved in all patients able to be evaluated. There were no statistically significant differences in CD34+ cell yields or toxicities. Overall engraftment occurred with median days to neutrophils > or = 0.5 x 10(9)/L or platelets > 20 x 10(9)/L of 11 and 17 days, respectively. However, the duration of G-CSF administration was markedly shorter in the late use of G-CSF group than in the early use of G-CSF group, with a median of 9 days compared with 15 days (p<0.001). PBSC harvesting after priming with CY plus delayed use of G-CSF made it a safe and cost-effective procedure.     

A new blood donation strategy: automated blood collection (ABC)

BACKGROUND: The aim of this study was to find out if Cobe Trima, a blood cell separator that automatically collects RBC, PLT and plasma, is adequate for routine multiple blood donation by apheresis. MATERIALS and METHODS: Eighty donors underwent multiple blood component donations by Cobe Trima. Blood counts were determined on the apheresis products to analyze their quality. RESULTS: Eighty procedures were performed collecting 193 products. The average platelet yield was 3.5x10(11) (+/- 0.46) in the 54 single product (SP) procedures and 7x10(11) (+/- 0.88) in the 26 double product (DP) procedures. WBC contamination of the PLT products was 1.7 x 10(5) (1.2-4.2). The mean platelet efficiency was 60 +/- 8.35% for SP and 66 +/- 9.59% for DP. The hemoglobin (Hb) content per unit was 46.21 g (+/- 7.84) in 8 DP and 40.82 g (+/- 6.41) in 34 SP procedures. CONCLUSION: The production of standardized blood components with good PLT yield and low WBC contamination plus high efficiency makes Trima one of the best blood cell separators of the new generation.     

Changes in hematologic parameters induced by thermal treatment of human blood

Abstract. To investigate the effects of high temperature on hematologic parameters of human blood, we assessed complete blood cell counts, red cell indices, and platelet aggregability in 72 adult blood samples after exposure to varying degrees of temperature. There were no significant differences in erythrocyte counts, hemoglobin concentration, or mean corpuscular hemoglobin (MCH) between pre- and post-treated groups when blood samples were treated at 50 degrees C for 5 min. Under the same conditions, however, mean corpuscular volume (MCV) and hematocrit became significantly higher in the post-treated group (p <0.01). A dramatic increase was observed in the numbers of platelets and eosinophils measured by electronic counter: the mean numbers of platelets and eosinophils in the post-treated group (2,978.2 x 109/L and 5.58 x 10(9)/L) were significantly higher than those for the pre-treated group (246.6 x 10(9)/L and 0.15 x 10(9)/L, p < 0.01). Platelet clumps of varying size were seen in EDTA-anticoagulated blood after exposure to 46 degrees C for 90 sec; however, these clumps were not detected in citrate-, heparinized-, or washed EDTA-anticoagulated blood. Platelet aggregability to various agonists was profoundly decreased after treating the blood at 43 degrees C for 5 min. In summary, this study shows that critical changes of hematologic parameters occur when blood is exposed to 50 degrees C, and that the anticoagulating property of EDTA is altered upon exposure to high temperature.     

Platelets in blood stored in untreated and siliconed glass bottles and plastic bags: II Survival studies

Platelet survival was determined using untreated and siliconed glass bottles and plastic bags (Fenwal) for collecting and storing blood. The platelets were tagged in vivo with P³² in six polycythaemic patients undergoing treatment with P³². The results showed that fresh ACD blood collected in untreated glass, siliconed glass, and plastic gave the same recovery of platelets in the recipients. The use of EDTA (Fenwal formula) as anticoagulant gave results inferior to those obtained with blood using ACD as anticoagulant. Even after storage up to 24 hours in untreated glass bottles (ordinary bank blood) a satisfactory recovery of platelets was observed. After storage for 72 hours the recovery was less but not negligible.     

Preparation of small volume, leuko and erythrocyte very poor platelet concentrates

Recently developed automated discontinuous flow centrifuge (DFC) separators can produce leuko- and erythrocyte-poor platelet concentrates (PC). According to general experience with these machines it is difficult to obtain more than 4 X 10(11) platelets, though a second program set up by Coffe et al. appears to produce PC containing approximately 5 X 10(11) platelets suspended in a plasma volume of 390 ml. At our center we employed a new Dideco cell separator equipped with the surge pump and a technique developed for the production of small volume, RBC and WBC-very poor PC. In 60 routine procedures we obtained the following results: mean processing time 87 +/- 11 minutes; final volume of PC 136 +/- 19 ml, with a mean platelet yield of 5.21 X 10(11) platelets. WBC contamination was 1.8 X 10(8) (93% lymphocytes) and RBC were 3.1 X 10(8). Plasma volume as well as WBC and RBC contamination were reduced by recirculating PC after the 6th pass. The demand for single donor platelet concentrates (PC) is increasing progressively. Recently developed automated cell separators can produce leukocyte (WBC) and erythrocyte (RBC) poor PC. With these machines it may be difficult to obtain PC containing at least 4 X 10(11) platelets and less than 1 X 10(9) leukocytes (1, 2, 3) since donor variables such as hematocrit, precounts, buffy coat formation and initial plasma light transmission are of paramount importance for the efficiency of the program. At our center a prototype discontinuous flow centrifuge (DFC) cell separator equipped with the surge pump was studied.(ABSTRACT TRUNCATED AT 250 WORDS)     

Les concentrés de plaquettes d'aphérèse:méthodes de préparation

Apheresis platelet concentrates are platelet suspension obtainedby apheresis from a blood cell separator. These techniques are now 20 years old. They are best way to collect great quantities of platelets from only one donor. These blood cell separators (discontinuous or continuous flow), realise extracorporeal circuit in which the blood in mixed with an anticoagulant (mainly citrate). All the procedures used arte automatic and adapted with the donor parameters. The platelet donor is between 18 and 60 years old and can give 5 times per year. The platelet concentration in the product is divided into 3 classes (1=2−4×10¹¹−2=4−6×10¹¹−3>6×10¹¹). Each category has a special price. Actually we can store these platelets only 5 days. In the future, we need new studies and new directions (additive solution, new plastic) to propose a better viability and a longer storage.     

Platelet loss across the hemofilter during continuous hemofiltration

BACKGROUND: Thrombocytopenia is a common finding in patients in the intensive care unit receiving continuous renal replacement therapy (CRRT). It is unknown if the hemofilter itself contributes to the platelet loss. OBJECTIVE: To measure the direct effect of the hemofilter on platelet counts during CRRT. DESIGN: Prospective, observational study. SETTING: Intensive care unit of a University hospital. PATIENTS: Critically ill patients with acute renal failure receiving CRRT. METHODS: Two samples of blood were drawn simultaneously, pre-filter and post-filter, and analyzed for platelet count. A correction factor was applied to the post-filter platelet count to adjust for the hemoconcentrating effect of net ultrafiltration. RESULTS: Forty-eight sets of paired data from 22 patients were studied. There was a small but significant decrease in mean platelet count across the hemofilter. The mean platelet count drop was 2.32 x 10(9)/L (s.e. 1.06, p = 0.0487, 95% CI (0.01, 4.62)). Blood flow was strongly related to degree of platelet loss, with a decreased loss of 0.07 x 10(9)/L for every ml/min increase in blood flow (p = 0.015). There was no overall decrease in concurrently measured red cell counts across the hemofilter. However, there was a machine-specific affect on red cell loss (p < 0.0001). The total calculated daily platelet loss across the filter was 625 x 10(9) cells. CONCLUSION: The hemofilter may contribute to the thrombocytopenia seen during CRRT, by means of either destruction or retention of platelets during passage. This affect appears attenuated by higher blood flows. This information is useful in the assessment of a low platelet count in patients receiving CRRT.     

Matched-pair analysis of hematopoietic progenitor cell mobilization using G-CSF vs. cyclophosphamide, etoposide, and G-CSF: enhanced CD34+ cell collections are not necessarily cost-effective.

Using matched-pair analysis, we compared two popular methods of stem cell mobilization in 24 advanced-stage breast cancer patients who underwent two consecutive mobilizing procedures as part of a tandem transplant protocol. For the first cycle, 10 microg/kg/day granulocyte colony-stimulating factor (G-CSF) was given and apheresis commenced on day 4 and continued for < or =5 days (median 3 days). One week after the first cycle of apheresis, 4000 mg/m2 cyclophosphamide, 400 mg/m2 etoposide, and 10 microg/kg G-CSF were administered for < or =16 days (cycle 2). Apheresis was initiated when the white blood cell (WBC) count exceeded 5000 cells/microL and continued for < or =5 days (median 3 days). Mean values of peripheral blood WBC (31,700+/-3200 vs. 30,700+/-3300/microL) were not significantly different between cycles 1 and 2. Mean number of mononuclear cells (MNC) collected per day was slightly greater with G-CSF mobilization than with the combination of chemotherapy and G-CSF (2.5+/-0.21x10(8) vs. 1.8+/-0.19x10(8) cells/kg). Mean daily CD34+ cell yield, however, was nearly six times higher (12.9+/-4.4 vs. 2.2+/-0.5x10(6)/kg; p = 0.01) with chemotherapy plus G-CSF. With G-CSF alone, 13% of aphereses reached the target dose of 5x10(6) CD34+ cells/kg in one collection vs. 57% with chemotherapy plus G-CSF. Transfusions of red blood cells or platelets were necessary in 18 of 24 patients in cycle 2. Three patients were hospitalized with fever for a median of 3 days after cycle 2. No patients received transfusions or required hospitalization during mobilization with G-CSF alone. Resource utilization (cost of drugs, aphereses, cryopreservation, transfusions, hospitalization) was calculated comparing the median number of collections to obtain a target CD34+ cell dose of 5x10(6) cells/kg: four using G-CSF vs. one using the combination in this data set. Resources for G-CSF mobilization cost $7326 vs. $8693 for the combination, even though more apheresis procedures were performed using G-CSF mobilization. The cost of chemotherapy administration, more doses of G-CSF, transfusions, and hospitalizations caused cyclophosphamide, etoposide, and G-CSF to be more expensive than G-CSF alone. A less toxic and less expensive treatment than cyclophosphamide, etoposide, and G-CSF is needed to be more cost-effective than G-CSF alone for peripheral blood progenitor cell mobilization.     

Early hematopoietic reconstitution after autologous transplantation with blood-derived stem cells in a patient with advanced lymphoma

A 30-year-old man with advanced non-Hodgkin lymphoma underwent repeated leukaphereses for harvesting blood-derived hemopoietic stem cells. Collection was started 8-10 days after the end of L-VAMP therapy (3 cycles). Nine procedures were performed and a total of 65.4 x 10(9) mononuclear cells (0.87 x 10(9)/Kg) were collected, processed, cryopreserved and stored in liquid nitrogen. The yields of CFU-GM, BFU-E and CFU-GEMM were respectively 964 x 10(4) (12.4 x 10(4)/Kg), 249 x 10(4) (3.2 x 10(4)/Kg) and 798 x 10(4) (10.4 x 10(4]. The patient received a myeloblative regimen consisting of fractionated total body irradiation (1200 cGy) and cyclophosphamide (120 mg/kg) followed by infusion of his own thawed cells. Early trilineage hematopoietic recovery was first observed on day +8; 1 x 10(9)/l WBC were reached on day +11, 0.5 x 10(9)/l PMN on day +13 and 50 x 10(9)/l platelets on day +11. Course was uneventful and the patient was discharged from hospital on day +21. Eight months after transplant the patient is in continuous unmaintained complete remission with normal blood cell counts. This reports suggests that complete and sustained engraftment can be achieved with peripheral stem cells recruited after "soft" chemotherapy.     

Hereditary sideroblastic anemia: a rare diagnosis

Hereditary sideroblastic anemia is a very rare disease recessive and X-linked that affect heme biosynthesis by deficit or decreased of delta aminolevulinic acid synthase (ALAS) activity. We report a case of a six-month-old boy, admitted in the hospital for anemic syndrome. The hemogram showed anemia (hemoglobin: 4.5 g/dL), frankly hypochronic microcytic and a regenerated (mean corpuscular hemoglobin concentration: 26 g/dL, mean cell volume: 53 fl, reticulocytes: 10 x 10(9)/L) with red cells morphologic disorders in smears (anisopoikylocytosis) without attack of the other lineages; white blood cells: 11 x 10(9)/L (neutrophils: 64% and lymphocytes: 35%); platelets: 350 x 10(9)/L. Examination of bone marrow showed an important erythroid hyperplasia (about 69%) with dyserythropoiesis. Perls stain revealed intense siderosis with 90% of ringed sideroblasts and a large number of siderocytes. Exploration of ALAS2 and ABC7 genes on the DNA of the infant was not found abnormalities. Treatment with pyridoxine corrects moderately the anemia. By the way, we proposed to remind that iron deficiency, inflammatory syndrome and thalassemia are the common microcytic anemia. However, it's mandatory to explore other causes if diagnosis is not solved.     

Isokinetic Hamstrings:Quadriceps Ratios in Intercollegiate Athletes

OBJECTIVE: To compare the differences in the concentric hamstrings:quadriceps (H:Q) ratio among athletes in different sports at 3 velocities. DESIGN AND SETTING: We measured the H:Q ratio of both knees using the Biodex Pro Isokinetic Device. SUBJECTS: Eighty-one male and female collegiate athletes. MEASUREMENTS: We performed analyses for sport, velocity, and side of body for each sex. To compare the means of the concentric H:Q ratios for mean peak torque and mean total work, a 2 x 3 x 4 mixed-factorial analysis of variance was computed for women and a 2 x 2 x 3 mixed-factorial analysis of variance was computed for men. RESULTS: We observed no significant interactions for men and women for the concentric H:Q ratio for mean peak torque. There was a significant mean difference among velocity conditions and a significant difference for men with respect to velocity. No significant differences were found for side of body or sport. CONCLUSIONS: The H:Q ratio increased as velocity increased. No differences existed for the H:Q ratio for sport or side of body.     

Punch and spindle-shaped biopsies for collecting oral mucosal tissue for the fabrication of transplantable autologous epithelial cell sheets

The oral mucosa is an easily accessible source of cells. Oral mucosal collection will be an essential surgical procedure for regenerative medicine and cell biological research. However, there is no current report that describes the details of the surgical procedure used for oral mucosal collection. Moreover, the number of cells that can be obtained has not been determined. Two different procedures, the punch biopsy and the spindle-shaped biopsy, were performed for the fabrication of transplantable autologous epithelial cell sheets. The mean values of the cells collected per square centimeter of tissue using the punch biopsy and the spindle-shaped biopsy were 76.8 ± 45 × 10(4) cells/cm(2) and 195.7 ± 120 × 10(4) cells/cm(2) , respectively. There was no significant difference between the punch biopsy and the spindle-shaped biopsy. The coefficient of variation of the punch biopsy and the spindle-shaped biopsy was 58.9% and 69.8%, respectively. This result indicated that both procedures showed variations in the number of collected cells. Although the punch biopsy may be easier and simpler than the spindle-shaped biopsy, multiple punch biopsies may result in a more complicated procedure, and the spindle-shaped biopsy may be preferable when a large number of cells is necessary. ? 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A 100A:2849-2854, 2012.     

A simplified automated procedure for generation of human lymphokine-activated killer cells for use in clinical trials

The administration of lymphokine-activated killer (LAK) cells along with interleukin-2 (IL-2) can mediate regression of tumors in selected patients. A closed automated system utilizing commercial blood cell separators has been developed for washing and Ficoll-Hypaque (FH) separation of lymphocytes, for lymphocyte culture in polyolefin bags, and for concentration of LAK cells out of culture prior to infusion. We now demonstrate that preparation of LAK cells can be simplified by elimination of the FH sedimentation step. Patient leukapheresis was performed using Fenwal CS-3000 blood cell separators, with a mean cellular yield per procedure of 54 X 10(9) WBC (95% lymphocytes), 184 X 10(9) RBC, and 306 X 10(9) platelets (n = 22). These cells were then washed in the same apheresis kit with a counter-current flow of saline, thereby eliminating 85% of platelets while retaining 88% of WBC. Aliquots of the washed cells were separated on FH gradients in 50 ml centrifuge tubes, and both FH-separated and washed-only cells were cultured at 3 X 10(6)/ml with 1500 U/ml IL-2 in polyolefin bags. Cytotoxicities of 22 preparations of LAK cells from 14 patients were evaluated in 4 h 51Cr release assays. Cells that were washed-only averaged 47, 35, and 9 lytic units/10(6) cells against K562, Daudi, and fresh tumor, while FH separated cells averaged 46, 33, and 6 lytic units/10(6) cells respectively. Cellular recoveries using the wash-only technique were 25% greater than when using FH sedimentation. Omission of FH separation saves time and expense in preparation and provides greater numbers of LAK cells for use in adoptive immunotherapy.