Probiotic Limosilactobacillus Reuteri (Lactobacillus Reuteri) Extends the Lifespan of Drosophila Melanogaster through Insulin/IGF-1 Signaling.

The term probiotic refers to bacteria that provide a beneficial effect to the host. Limosilactobacillus reuteri (Lactobacillus reuteri) is a probiotic isolated from human breast milk. Although L. reuteri has antimicrobial and anti-inflammatory activities occasionally linked to anti-aging effects, there are no reports of the effects of L. reuteri on longevity. This study evaluated the anti-aging effects of L. reuteri on the lifespan and physiology of Drosophila melanogaster. L. reuteri increased the mean lifespan of fruit flies significantly without reducing the reproductive output, food intake, or locomotor activity. Furthermore, the data suggested that the longevity effect of L. reuteri is mediated by the reduction of the insulin/IGF-1 signaling pathway and the action of reuterin, an antimicrobial compound produced by L. reuteri. These results show that L. reuteri can be used as a probiotic that acts as a dietary restriction mimetic with anti-aging effects.

Performance evaluation of the Sysmex CN-6000-automated coagulation analyzers for routine coagulation tests.

The Sysmex CN-6000 is a novel automated multiparameter coagulometer that performs clotting, chromogenic and immunological assays, and platelet aggregation tests in a single system. Here we evaluated its performance of routine coagulation assays. The precision, linearity, carryover and establishment of reference ranges of the CN-6000, as well as correlations between it and the currently used Diagnostica Stago STA-R Max were determined according to Clinical and Laboratory Standards Institute guidelines. The evaluated parameters included prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FBG), antithrombin (AT), d-dimers (DDi), and fibrin and FBG degradation products (FDP). The intra-run and inter-run precisions of the six tests were determined using normal and pathological control materials; all coefficients of variation were acceptable and within the allowable ranges. The CN-6000 showed excellent linearity for FBG, AT, DDi, and FDP (R = 0.999-1.00). Passing-Bablok regression (R2 > 0.95) demonstrated good agreement between the analyzers. In the carryover study, APTT, PT, FBG, AT, DDi, and FDP values were all acceptable. The establishing reference intervals revealed that each manufacturer's range was acceptable. Significant differences were observed in the APTT reference range because of using different detection systems and reagents. The CN-6000 analyzer showed reliable performance and good correlation with the currently used STA-R Max automated hemostatic analyzer. As CN-6000 uses an optical clot-detection method, its reference ranges for PT and APTT are lower than those of the STA-R Max; thus, the difference should be considered before its use.

Sample collection for pre-transfusion crossmatching: Benefits of using an electronic identification system.

BACKGROUND: Transfusion of ABO blood group-mismatched blood or administration to the wrong recipient may result in fatal adverse events. To prevent these types of errors, various strategies have been employed. Recently, we developed a novel sample collection workflow for the pre-transfusion crossmatching test and patient recognition. This study aimed to analyse the usage of the new workflow and improvements in outcomes. METHODS: We analysed the number of crossmatching and wrong-patient errors among the blood transfusion cases during 3 years of data collection (from August 2018 to July 2021). From May 2021 to July 2021, the new workflow was implemented. Outcomes were calculated according to the department type, patient age and processing time. The sample processing time was defined as the time from placing the order to lab arrival. RESULTS: The new workflow utilisation increased from 50.7% to 80.3% and wrong-patient errors decreased annually. The new workflow was used for more adults (3001/3680 samples, 81.5%) than paediatric cases (345/522 samples, 65.5%; p < 0.001) and in general wards than in the emergency room or intensive care unit. The sample processing time differed according to ward type and timing of the request (day: 28.80, 2.43-3889.43 min, night: 3.36, 2.72-1671.47 min; p < 0.001). CONCLUSION: Wrong-patient errors were reduced without increasing sample-processing time after introducing the new workflow which included using an electronic identification system. The time needed for the blood processing differed according to the ward type, patient age, and timing of the request. Patient safety can be promoted by managing these factors and using an electronic identification system.

Potential misdiagnosis of von Willebrand disease caused by insufficient mixing after thawing frozen plasma.

INTRODUCTION: von Willebrand disease (vWD) is a common inherited bleeding disorder caused by a deficiency in von Willebrand factor (vWF), but many laboratories and clinicians continue to struggle with diagnosing or excluding vWD. Its diagnosis requires laboratory testing, which may be compromised by preanalytical events, including poor specimen quality. This study assessed 17 different preanalytical conditions as potential causes of vWD misdiagnosis. METHODS: Specimens from healthy controls (N = 21) were obtained. vWF antigen and vWF activity were analyzed using a newly developed automatic coagulation analyzer according to various preanalytic conditions such as centrifugation conditions, storage room temperature before centrifugation, cold storage temperature after centrifugation, thawing conditions, and inadequate mixing of thawed citrated plasma following the recommendations of the Clinical and Laboratory Standards Institute (CLSI) H21-A5 guidelines. RESULTS: The only condition that was significantly different from the reference condition was lack of mixing after thawing frozen citrated plasma (vWF activity and antigen were reduced by 58.7% and 49.6%, respectively). Our study showed that mixing after thawing was more important than the chosen method of mixing. CONCLUSION: Thawed plasma should be mixed because of the risk of misdiagnosing vWD. Further education regarding the importance of appropriate mixing is warranted to achieve results comparable to those of freshly centrifuged samples.

Platelet Inventory Management Program: Development and Practical Experience.

BACKGROUND: Patients with ongoing or expected bleeding require platelet (PLT) transfusions; however, owing to the testing required after a blood donation, manufacturing PLT products may take 1.5-2.0 days after a request is made. This supply-demand mismatch leads clinicians to retain spare PLTs for transfusions, leading to increased PLT discard rates. We developed a PLT inventory management program to supply PLTs more efficiently to patients requiring PLT transfusions within the expiration date, while reducing PLT discard rates. METHODS: PLT concentrates (58,863 and 58,357 units) and apheresis products (7,905 and 8,441 units) were analyzed from May 2015 to November 2017 and from December 2017 to January 2020, respectively. We developed a program to manage total PLT inventories and prospective PLT transfusion patients based on blood type, blood product, and remaining period of efficacy; the program facilitates PLT preparation transfer to non-designated patients within the remaining period of efficacy. RESULTS: The overall PLT concentrate discard rate was 3,254 (2.78%): 1,811 (3.07%) units before and 1,443 units (2.41%) after program application (P<0.001). The discard rate owing to expiration was reduced from 69 units (3.81%) before to two units (0.14%) after program application (P<0.001). CONCLUSIONS: This program can guide the allocation of PLT preparations based on the remaining period of efficacy, enabling PLT products to be used before their expiration date and reducing PLT product discard rate.

Comparison of three immunoassays for determination of immunity to rubella virus in healthcare workers.

Rubella virus is a critical infectious pathogen to healthcare workers but is preventable by vaccination. In this study, we used three immunoassays - LIAISON Rubella IgG, ARCHITECT Rubella IgG, and AtheNA Multi-Lyte MMRV IgG - to detect rubella virus IgG and tested 182 serum specimens. The percentage of positives with the three Rubella tests were as follows: LIAISON, 71.9%; ARCHITECT, 83.5%; and AtheNA, 99.5%. The three assays showed an overall agreement rate of 71.9%. The rates of seropositive detection with LIAISON, ARCHITECT, and AtheNA among healthcare workers with and without self-reporting history of past infection or vaccination were 70.7% and 90.9%, 83.6% and 81.8%, and 99.4% and 100%, respectively. The three immunoassays showed a low agreement rate for rubella virus IgG. Therefore, choosing accurate and appropriate IgG assay methods is very important for effective infection control and prevention.

Comparative Analysis of Clinical Samples Showing Weak Serum Reaction on AutoVue System Causing ABO Blood Typing Discrepancies.

BACKGROUND: ABO blood typing in pre-transfusion testing is a major component of the high workload in blood banks that therefore requires automation. We often experienced discrepant results from an automated system, especially weak serum reactions. We evaluated the discrepant results by the reference manual method to confirm ABO blood typing. METHODS: In total, 13,113 blood samples were tested with the AutoVue system; all samples were run in parallel with the reference manual method according to the laboratory protocol. RESULTS: The AutoVue system confirmed ABO blood typing of 12,816 samples (97.7%), and these results were concordant with those of the manual method. The remaining 297 samples (2.3%) showed discrepant results in the AutoVue system and were confirmed by the manual method. The discrepant results involved weak serum reactions (<2+ reaction grade), extra serum reactions, samples from patients who had received stem cell transplants, ABO subgroups, and specific system error messages. Among the 98 samples showing ≤1+ reaction grade in the AutoVue system, 70 samples (71.4%) showed a normal serum reaction (≥2+ reaction grade) with the manual method, and 28 samples (28.6%) showed weak serum reaction in both methods. CONCLUSIONS: ABO blood tying of 97.7% samples could be confirmed by the AutoVue system and a small proportion (2.3%) needed to be re-evaluated by the manual method. Samples with a 2+ reaction grade in serum typing do not need to be evaluated manually, while those with ≤1+ reaction grade do.

Correlation of NPM1 Type A Mutation Burden With Clinical Status and Outcomes in Acute Myeloid Leukemia Patients With Mutated NPM1 Type A.

BACKGROUND: Nucleophosmin gene (NPM1) mutation may be a good molecular marker for assessing the clinical status and predicting the outcomes in AML patients. We evaluated the applicability of NPM1 type A mutation (NPM1-mutA) quantitation for this purpose. METHODS: Twenty-seven AML patients with normal karyotype but bearing the mutated NPM1 were enrolled in the study, and real-time quantitative PCR of NPM1-mutA was performed on 93 bone marrow (BM) samples (27 samples at diagnosis and 56 at follow-up). The NPM1-mutA allele burdens (represented as the NPM1-mutA/Abelson gene (ABL) ratio) at diagnosis and at follow-up were compared. RESULTS: The median NPM1-mutA/ABL ratio was 1.3287 at diagnosis and 0.092 at 28 days after chemotherapy, corresponding to a median log10 reduction of 1.7061. Significant correlations were observed between BM blast counts and NPM1-mutA quantitation results measured at diagnosis (γ=0.5885, P=0.0012) and after chemotherapy (γ=0.5106, P=0.0065). Total 16 patients achieved morphologic complete remission at 28 days after chemotherapy, and 14 (87.5%) patients showed a >3 log10 reduction of the NPM1-mutA/ABL ratio. The NPM1-mutA allele was detected in each of five patients who had relapsed, giving a median increase of 0.91-fold of the NPM1-mutA/ABL ratio at relapse over that at diagnosis. CONCLUSIONS: The NPM1-mutA quantitation results corresponded to BM assessment results with high stability at relapse, and could predict patient outcomes. Quantitation of the NPM1-mutA burden at follow-up would be useful in the management of AML patients harboring this gene mutation.

Effects of various toothpastes on remineralization of white spot lesions.

OBJECTIVE: The purpose of this in vitro study was to examine the effects of fluoridated, casein phosphopeptide.amorphous calcium phosphate complex (CPP-ACP)-containing, and functionalized ß-tricalcium phosphate (fTCP)-containing toothpastes on remineralization of white spot lesions (WSLs) by using Quantitative light-induced fluorescence (QLF-D) Biluminator™ 2. METHODS: Forty-eight premolars, extracted for orthodontic reasons from 12 patients, with artificially induced WSLs were randomly and equally assigned to four treatment groups: fluoride (1,000 ppm), CPP-ACP, fTCP (with sodium fluoride), and control (deionized water) groups. Specimens were treated twice daily for 2 weeks and stored in saliva solution (1:1 mixture of artificial and human stimulated saliva) otherwise. QLF-D Biluminator™ 2 was used to measure changes in fluorescence, indicating alterations in the mineral contents of the WSLs, immediately before and after the 2 weeks of treatment. RESULTS: Fluorescence greatly increased in the fTCP and CPP-ACP groups compared with the fluoride and control groups, which did not show significant differences. CONCLUSIONS: fTCP- and CPP-ACP-containing toothpastes seem to be more effective in reducing WSLs than 1,000-ppm fluoride-containing toothpastes.