Ultrasound-guided injection through the rotator cuff interval: a clinical perspective of one institution's results and description of technique.

CONTEXT: Fluoroscopic injection through the rotator cuff interval (RCI) is a common technique for diagnostic arthrography and therapeutic intervention. Ultrasound approaches through the RCI have been less commonly studied, but there is a growing body of literature. OBJECTIVES: The purpose of this study was to present a standardized technique of ultrasound-guided injection into the glenohumeral joint utilizing the RCI in magnetic resonance imaging (MRI) arthrography (MRA) and to report one medical group's experience with the technique. METHODS: A retrospective chart review of all ultrasound-guided injections into the glenohumeral joint utilizing the RCI was performed from July 1, 2014 through June 1, 2021. Data were compiled for age, gender, body mass index (BMI), and prior surgery on the shoulder. The primary endpoint was successful administration of intra-articular dilute gadolinium contrast adequate for radiologic interpretation. A total of 487 injections into the glenohumeral joint via the RCI were performed. One hundred and fifty-five patients had previous shoulder surgery, with the remainder naive to intervention. RESULTS: The success rate of injections into the glenohumeral joint was 99.4 %, with only three injections considered unsuccessful. The three unsuccessful injections did not succeed because of a lack of intra-articular contrast media present. This success rate is impressive and promising, particularly when considering that 155 of the patients had previous surgery, which could potentially cause complications, and because these injections were performed over a long period of 7 years. CONCLUSIONS: Accessing the RCI under ultrasound guidance is a very successful technique for injection within the glenohumeral joint.

Laparoscopic Heller Myotomy With EGD and Balloon Dilation: A Durable Solution for Achalasia.

BACKGROUND: Laparoscopic Heller myotomy (LHM) and esophageal balloon dilation (BD) are the two mainstays of achalasia treatment-this study examines the outcomes when they are performed simultaneously without fundoplication. METHODS: All patients undergoing LHM&BD were reviewed for demographic and procedural data, and to see if additional procedures for achalasia had been performed. Patients were surveyed using the Eckardt score and the GERD quality-of-life score (GERD-HRQL) to assess the durability of repair. RESULTS: From 2013-2020, 66 patients underwent LHM&BD. There were no esophageal perforations and a median LOS of 1 day. Seven patients have required additional operations or procedures at median 4-years follow up. 31 patients (47%) responded to the survey. The average Eckardt score was 2.9 (goal<4) with mean GERD-HRQL of 14.4 (goal<25). CONCLUSIONS: LHM&BD allows for a safe, durable repair of achalasia. Reflux symptoms are manageable with PPI without fundoplication and the re-intervention rate similar to published values.

Effects of Freshly Irradiated vs Irradiated and Stored Red Blood Cell Transfusion on Cerebral Oxygenation in Preterm Infants: A Randomized Clinical Trial.

Importance: Gamma irradiation of leukoreduced red blood cells (RBCs) prevents transfusion-associated graft-vs-host disease but also exacerbates storage lesion formation in RBCs. It is unknown whether freshly irradiated RBCs are more efficacious than irradiated and stored RBCs in preterm infants with high transfusion requirements. Objective: To examine whether transfusion of freshly irradiated vs irradiated and stored RBC components improves cerebral oxygen delivery in preterm infants with anemia. Design, Setting, and Participants: This single-center, double-blinded, proof-of-concept randomized clinical trial was conducted at the neonatal intensive care unit of Wellington Regional Hospital in Wellington, New Zealand, between December 1, 2017, and November 30, 2018. Participants were preterm infants (<34 weeks' gestation at birth) who were at least 14 days of age and had anemia. Participants underwent nonurgent transfusions, and these episodes were randomized to the intervention group (in which the infants received a transfusion of RBCs that were freshly irradiated on the day of transfusion) or control group (in which the infants received a transfusion of RBCs that were irradiated and stored for up to 14 days). Data were analyzed using the evaluable population approach. Intervention: Transfusion of freshly irradiated RBCs. Main Outcomes and Measures: The prespecified primary outcome was the change in cerebral regional oxygen saturation (crSO2) from baseline (immediately before) to immediately after the transfusion. The prespecified secondary outcomes were the change in cerebral fractional tissue oxygen extraction (cFTOE) at different time points (immediately after, 24 hours after, and 120 hours or 5 days after transfusion). Outcomes were measured by blinded clinicians using near-infrared spectroscopy. A covariate-adjusted linear mixed model was used to quantify mean treatment effects and account for multiple transfusions in some infants. Results: A total of 42 infants (mean [SD] gestational age, 26 [10] weeks and 3 days; 29 [69%] boys) were enrolled in the trial and underwent 64 transfusion episodes, which were randomized to the intervention (n = 31) or control (n = 33) group. Compared with infants in the control group, those in the intervention group showed a covariate-adjusted mean increase in crSO2 (2.0 percentage points; 95% CI, 1.2-2.8 percentage points) and a mean decrease in cFTOE (0.02; 95% CI, 0.01-0.04) immediately after transfusion. These differences were sustained up to 120 hours or 5 days after transfusion. There were negligible mean changes in crSO2 or cFTOE in infants in the control group at any of the follow-up time points. Conclusions and Relevance: Results of this trial showed that transfusion of freshly irradiated RBCs conferred a small advantage in cerebral oxygenation for at least 5 days after transfusion compared with transfusion of irradiated and stored RBC components. On-demand irradiation of RBC components may be considered to optimize oxygen delivery in the recipient, but this physiological finding requires further research. Trial Registration: ANZCTR Identifier: ACTRN12617001581358.

Storage after gamma irradiation affects in vivo oxygen delivery capacity of transfused red blood cells in preterm infants.

BACKGROUND: Gamma irradiation of red blood cells (RBCs) is well recognized to exacerbate storage lesion formation, but the effect of storage after irradiation on in vivo oxygen delivery capacity of transfused RBCs is currently not known. STUDY DESIGN AND METHODS: In 24 preterm infants with anemia receiving nonurgent transfusion of irradiated RBCs, we examined cerebral regional tissue oxygenation (crSO2 ) and time spent with peripheral arterial saturation (SpO2 ) less than 88%. Physiologic data were obtained immediately before, immediately after, and 5 days after transfusion. RESULTS: We observed linear negative moderate correlations between time since irradiation and the magnitude of change in crSO2 (r = -0.60; 95% CI, -0.81 to -0.27; p = 0.0018) and time spent with SpO2 of less than 88% (r = -0.42; 95% CI, -0.71 to 0.003; p = 0.04) immediately after transfusion. In infants (n = 9) who received fresher RBCs (irradiated <10 days before transfusion), there was a sustained increase in mean crSO2 up to 5 days after transfusion (3.0%; 95% CI, 0.3% to 5.7%; p = 0.04). Conversely, in infants (n = 15) who received older RBCs (irradiated ≥10 days before transfusion), there were negligible changes in crSO2 after transfusion at any time point. CONCLUSION: Our findings indicate that storage after gamma irradiation may have a detrimental effect on the oxygen delivery capacity of RBCs given to anemic preterm infants.

Hepatitis E virus infection in the Irish blood donor population.

BACKGROUND: Hepatitis E virus (HEV) Genotype 3 (G3) infection is a zoonosis that may be transmitted during the acute phase by transfusion. The aim of this study was to determine the incidence of HEV and seroprevalence among Irish blood donors. STUDY DESIGN AND METHODS: Anonymized samples from 1076 donations collected in 2012 were tested for HEV immunoglobulin (Ig)G using the Wantai enzyme-linked immunosorbent assay. A total of 24,985 anonymized donations collected between December 2013 and June 2014 were individually tested for HEV RNA using the Procleix HEV assay; reactive donations were confirmed by an in-house real-time polymerase chain reaction (PCR) test. RESULTS: Seroprevalence for anti-IgG was 5.3% (95% confidence interval [CI], 4.0%-6.8%), ranging from 1.1% in the 18- to 29-years age group to 33.3% in males over 60 years. HEV RNA screening of 24,985 samples yielded five PCR-confirmed donations (1:4997, 0.02%; 95% CI, 0.0065%-0.0467%), only one of which was serologically reactive (HEV IgM reactive only). Viral loads ranged from 10 to 44,550 IU/mL. Genotype analysis on three samples identified HEV G3 virus. Four of the five viremic donations were from donors in the 18- to 29-years age group (p = 0.01). CONCLUSION: Seroprevalence for anti-HEV IgG was low compared to some European countries, but 1 in 5000 donations was viremic. Viremia was predominantly in younger Irish donors. After Department of Health approval the Irish Blood Transfusion Service implemented individual blood donation HEV RNA screening initially for a 3-year period from January 2016.

Serum ferritin in plateletpheresis and whole blood donors.

BACKGROUND AND OBJECTIVES: We performed a prospective analysis of iron status in plateletpheresis donors, using whole blood donors as a control group, to assess the haematinic effects of regular anti-coagulated extracorporeal circulation and platelet collection. MATERIALS AND METHODS: Ferritin levels were measured in samples from 31 regular male plateletpheresis donors and from 14 first time male whole blood donors, immediately before and immediately after donation, and immediately before the next donation. An additional 33 regular male plateletpheresis donors and 17 first time male whole blood donors had serum ferritin levels checked predonation. RESULTS: Male plateletpheresis donors had a statistically significant fall in serum ferritin after donation (P = 0.005)*. In addition, male platelet donors had significantly lower serum ferritin levels than first time male blood donors: ferritin <20 µg/L was found in 6/64 (9%) of regular platelet donors and 1/31 (3%) of first time blood donors (P < 0.001)*. DISCUSSION: Our studies support the value of serum ferritin measurement in apheresis donor management.

Measuring dissolved oxygen to track erythroid differentiation of hematopoietic progenitor cells in culture.

As stem cell technologies move from the developmental to the commercial stage strategies must be developed to monitor culture operations. These will ensure consistency of differentiation programs and maintenance of optimum cell viability during production runs. Due to the sensitivity of stem cells to their environment, and their variability in response to external stimuli, accurate monitoring of in vitro conditions will be crucial for effective large-scale culturing of therapeutic stem cells. Here we describe a simple method to monitor the expansion and maturation of adult human haematopoietic stem/progenitor cells into red blood cells in vitro by measuring the oxygen consumption rate of cultures. Cell cultures followed a characteristic pattern of oxygen consumption that is reflective of in vivo erythroid maturation. This method could be easily developed as an online system to map erythroid differentiation and maturation of cultured cells as effectively as the more time consuming process of flow cytometric analysis of surface marker expression patterns.

The sex difference in haemoglobin levels in adults - mechanisms, causes, and consequences.

Men and women have different mean haemoglobin levels in health in venous blood - women have mean levels approximately 12% lower than men. A similar sex-related difference in haemoglobin levels in adult animals is found in many species of mammals, birds and reptiles, indicating that it is an important physiological phenomenon. It is probably a direct effect of sex hormones, both oestrogen and androgens, on erythropoiesis. However, since there is no difference in erythropoietin levels between the sexes, this effect most likely takes place in the kidney, rather than in the bone marrow. Oestrogens dilate and androgens constrict the renal microvasculature: dilation and vasoconstriction in vessels below 300 µm in diameter respectively increase and decrease the haematocrit in blood in arterioles, capillaries and venules, altering the oxygen delivery per unit red cell mass, and providing a mechanism for varying the red cell mass without compensatory changes in erythropoiesis.

Blood transfusion practice in a rural hospital in Northern Ghana, Damongo, West Gonja District.

BACKGROUND: Blood transfusion in rural sub-Saharan Africa presents special challenges. Transfusions are primarily given for emergencies--life-threatening blood loss or anemia; blood is usually collected from family or replacement donors; and facilities to store an adequate reserve in a hospital bank are constrained. We report the everyday and organizational practices in a medium-sized district hospital in Northern Ghana. STUDY DESIGN AND METHODS: Information and data on blood transfusion practices at West Gonja Hospital, Damongo, were available from the laboratory reports, from day books and workbooks, and from direct observation in the following four areas: blood collection and blood donors; blood donation testing; blood storage and logistics; and clinical transfusion practice, adverse events, and follow-up. RESULTS: The hospital serves a rural community of 86,000. In 2009, a total of 719 units of whole blood were collected, a rate of 8.36 units per 1000 population. All donors were family or replacement donors. Positivity rates for infectious disease markers were 7.5% (64/853) for hepatitis B surface antigen, 6.1% (50/819) for hepatitis C virus, 3.9% (33/846) for human immunodeficiency virus, and 4.7% (22/468) for syphilis. Supply of laboratory materials was sometimes problematic, especially for temperature-critical materials. Difficulties in sample labeling, storage of blood and laboratory supplies, and disposal of waste were also incurred by operational, material, and financial constraints. Follow-up for outcomes of transfusion is not currently feasible. CONCLUSIONS: The operational, demographic, and financial environment pertaining in a rural hospital in Northern Ghana differs substantially from that in which much of current blood transfusion practice and technology evolved. Considerable effort and innovation will be needed to address successfully the challenges posed.

Testing platelet components for bacterial contamination.

Bacteria in transfused platelets can cause serious morbidity and, rarely, death. Most contaminating bacteria enter the blood at the time of venepuncture. While many of these contaminants fail to grow in the platelet unit, storage of platelets at 20-24°C facilitates growth of some organisms, and the cumulative risk of severe sepsis increases with the storage age of platelet components. Several methods have been developed or adapted to attempt to detect contaminating bacteria with high sensitivity and specificity, but the perfect test has yet to be found. Testing early in the platelet component's shelf life, even using exquisitely sensitive culture-based tests, is compromised by major problems of sample error - there may be too few bacteria present at this stage to ensure that any practical sample volume contains even one of them. Culture techniques are too slow to be useful as a release test. On the other hand, available rapid tests are too insensitive to use early in the shelf life, and have yet to show convincingly that they are sensitive enough for testing close to the time of transfusion. Nevertheless testing for bacteria in platelet components represents a significant advance in blood transfusion safety, and prevents the transfusion of many hundreds of bacterially-contaminated platelet units each year.

A novel panel of protein biomarkers for predicting response to thalidomide-based therapy in newly diagnosed multiple myeloma patients.

Multiple myeloma (MM) is a heterogeneous group of disorders both genotypically and phenotypically. Response to thalidomide-based induction therapy in newly diagnosed patients varies significantly in published clinical trials. Proteomic analysis was performed on 39 newly diagnosed MM patients treated with a thalidomide-based regimen (22 responders; 17 non-responders) using immunodepletion, 2-D DIGE analysis and mass spectrometry. Zinc-α-2-glycoprotein (ZAG), vitamin D-binding protein (VDB), serum amyloid-A protein (SAA) and ß-2-microglobulin (B2M) had statistically significant higher concentrations in non-responders compared to responders, while haptoglobin (Hp) had a lower concentration. ELISAs were used to validate the candidate protein biomarkers using unfractionated serum from 51 newly diagnosed MM patients (29 responders; 22 non-responders). Using logistic regression, the best possible area under the curve (AUC) was 0.96 using ZAG, VDB and SAA in combination. Leave-one-out-cross-validation (LOOCV) indicated an overall predictive accuracy of 84% with associated sensitivity and specificity values of 81.8 and 86.2%, respectively. Subsequently, 16 of 22 thalidomide-refractory patients successfully achieved complete response or very good partial response using second-line treatment suggesting that the biomarker profile is specific to thalidomide response rather than identifying patients with MM refractory to all therapies. Using a novel panel of predictive biomarkers, the feasibility of predicting response to thalidomide-based therapy in previously untreated MM has been demonstrated.

The effect of mild agitation on in vitro erythroid development.

The cultivation of erythroid cells at large scale would have to be performed in suitable bioreactors which would most likely employ some mode of agitation to ensure optimal mass and gas transfer and prevent culture inhomogeneity. The effect of low agitation at 15-20 rpm on ex vivo erythropoiesis of PB CD34+ derived cultures was investigated and found to have significant impact on erythroid development. Agitated cultures showed a reduced lag phase and increased cell expansion during the early stages of culture. Additionally, agitation accelerated erythroid differentiation as seen by the loss of early development markers, acquisition of late erythroid markers and premature cell cycle arrest, although not yielding higher fractions of terminally differentiated cells in comparison to stationary culture. However, agitation at 20 rpm led to significantly increased loss of cell viability after day 15 in culture, an effect that could be reduced by decreasing the agitation rate to 15 rpm. On the one hand these results imply that agitation may improve cell yields and reduce expensive cytokine-dependent early culture stages but on the other hand it might introduce the risk of increased cell death in large scale culture.

The potential of human peripheral blood derived CD34+ cells for ex vivo red blood cell production.

The potential of peripheral blood derived CD34+ cells for ex vivo erythropoiesis was investigated in a stroma-free culture system using a novel strategy of daily passaging. By expanding PB-derived CD34+ cells up to 1.5 x 10(6)-fold this method achieved expansion factors previously only reported for CD34+ cells derived from more potent stem cell sources such as cord blood, bone marrow and mobilized peripheral blood. Analysis of cell surface markers showed differentiation of immature CD34+ cells to populations with 80% CD71-/GpA+ cells and up to 45% enucleated cells, indicating a significant amount of terminal maturation. Cell crowdedness was found to have decisive effects on in vitro erythropoiesis. Cell density per surface area rather than cell concentration per media volume determined cell expansion during exponential growth where more crowded cells showed reduced overall expansion. In late stage erythropoiesis, however, when cells no longer proliferating, increased cell density was seen to enhance cell viability. These results indicate that peripheral blood derived haematopoietic stem cells can be an alternative to cells sourced from bone marrow, cord blood or leukapheresis in terms of expansion potential. This provides distinct advantages in terms of availability for studies of conditions for scale-up and maturation, and may have particular clinical applications in the future.

Circulating microparticles: pathophysiology and clinical implications.

Microparticles (MP) derived from vascular endothelium or circulating blood cells circulate in the peripheral blood. They originate from blebbing and shedding from cell membrane surfaces in physiological and pathological conditions and are present in low concentrations in normal plasma. Increased levels are generated by a number of mechanisms including platelet activation, direct vascular endothelial damage, thrombin activity on the cell surface, C5b-9 activation, and PF4-heparin-antibody interaction. Several techniques are currently used to study the generation and nature of circulating microparticles. In particular, the genesis and role of microparticles, derived from platelets, endothelial cells and monocytes, in sepsis (especially meningococcal-induced), heparin-induced thrombocytopenia (HIT), thrombotic thrombocytopenic purpura (TTP), aplastic anaemia, paroxysmal nocturnal haemoglobinuria (PNH) and sickle cell disease (SCD) have been well studied, and provide important insights into the underlying diseases. A defect in the ability to form microparticles leads to the severe bleeding disorder of Scott syndrome, which in turn provides a revealing insight into the physiology of coagulation. In addition the complex role of microparticles in vascular and cardiovascular diseases is an area of immense interest, that promises to yield important advances into diagnosis and therapy.