Flow FISH Research Articles

Impacts of cascade reservoirs on fishes in the mainstream of Pearl River and mitigation measures

2013年6-7月对珠江龙滩库尾至长洲坝下江段干支流鱼类进行了调查，共采集到鱼类6418尾，隶属于10目23科82属122种.分析了渔获物的种类组成与分布、生态类型、珍稀濒危特有鱼类采样情况、各江段群落相似度和鱼类多样性等，并与该区域鱼类历史记录进行了对比分析.总体来看，珠江干流梯级开发后，由于大坝阻隔、水文情势改变等，鱼类资源发生了较大变化，河海洄游性鱼类和河口鱼类受大坝阻隔影响分布范围变窄；珍稀濒危特有鱼类由于生境破坏，种群规模变小，濒危程度加剧；库中江段流水性鱼类种类数和资源量显著下降，静缓流鱼类成为优势种；外来鱼类种类多、分布广.对导致珠江干流鱼类资源衰退的其他原因进行了分析，主要包括过度捕捞、生境破坏、水污染和外来鱼类入侵等.针对珠江干流梯级开发及鱼类资源现状，提出了栖息地保护、河流连通性恢复、鱼类增殖放流站建设、生态调度、渔政管理和生态补偿等措施建议.;The fish resources in the mainstream and its tributaries of the Pearl River from Longtan Dam to Changzhou Dam were investigated in June-July 2013. A total of 6418 fish specimens were collected, belonging to 10 orders, 23 families, 82 genera and 122 species. This study analysed the species composition and distribution, ecological types of fishes, sampling result of rare and endemic fish, the community similarity and diversity of fishes of each sample reach, and compared with the historical record. Overall, after cascade reservoirs in the Pearl River, massive changes in fish resources had taken place, such as river migratory fish and estuarine fish distribution range becoming narrow due to the dam barrier effect; rare and endemic fish being more endangered due to habitat loss; flow water fish species decreasing significantly in the reservoirs, while the static flow fish becoming the dominant species; and exotic fish species multiplying and distributing widely. Some other reasons for the decline of fish resources in the Pearl River mainstream were analysed, including overfishing, habitat destruction, water pollution, invasion of exotic fish and so on. In view of the Pearl River cascade reservoirs and the status of fishery resources, this study proposed mitigation measures such as habitat protection, restoration of river connectivity, fish breeding and releasing station construction, ecological scheduling, fishery management and ecological compensation.

A Rectal Tonsil Like No Other

Rectal tonsil is a rare condition. Its etiology is unknown, but it is comprised of a reactive proliferation of lymphoid tissue. Here we present a case of large, bleeding rectal tonsil. A 33-year-old healthy female presented with a 3-year history of intermittent rectal bleeding. Initial colonoscopy revealed irregular granular-appearing rectal mucosa, which was partially circumferential and extended 10 cm from the anal verge. Biopsy showed atypical lymphoid infiltrate positive for CD-20 and bcl-2, concerning for low-grade B-cell non-Hodgkin lymphoma. Endoscopic ultrasound showed mild, diffuse wall thickening of the mucosal/submucosal layer. Biopsies confirmed reactive lymphoid hyperplasia with flow cytometry negative for clonal population. She continued to have rectal bleeding at 6-month follow-up. Repeat colonoscopy showed a circumferential area of severely granular mucosa from the dentate line to 5 cm from the anal verge. Biopsies again revealed reactive lymphoid hyperplasia with negative flow cytometry and FISH. Currently, the patient continues to be monitored with close follow-up. The rectal tonsil is a rare finding. The most common presentation is rectal bleeding. Our case is the largest lesion presented in the literature as majority of the lesions reported consisted of discrete small (< 6mm) polypoid lesions or nodules. Previous studies reported complete resection of these small discrete lesions and show no recurrence or lymphoma transformation. However, the follow up for these studies is brief. Rectal tonsil should be distinguished from MALT lymphoma and other lymphoid diseases with malignant potential. Our patient is unique as the lesion was large and circumferential; therefore the patient elected to remain under close surveillance. It is important to continue to surveillance of these patients to understand long-term outcomes.Figure: Colonoscopy: Rectal tonsil.Figure: Colonoscopy: Rectal tonsil (retroflexion).

A comparison of genetic diversity and population structure of the endangered scaleshell mussel (Leptodea leptodon), the fragile papershell (Leptodea fragilis) and their host-fish the freshwater drum (Aplodinotus grunniens)

The larvae of freshwater mussels in the order Unionoida are obligate parasites on fishes. Because adult mussels are infaunal and largely sessile, it is generally assumed that the majority of gene flow among mussel populations relies on the dispersal of larvae by their hosts. The objective of this study was to compare the genetic diversity and the degree of congruence between the population structures of two related freshwater mussels Leptodea leptodon and Leptodea fragilis and their fish host, Aplodinotus grunniens. Host specificity in parasites has been shown to result in greater congruence between the population structures of the two interacting species, and assessing the congruence of genetic structure of the endangered L. leptodon with its sister species L. fragilis and their sole host is an important step in understanding the impact of host dispersal on population structure. Analysis of microsatellite data indicated that despite its imperiled status, L. leptodon displayed greater genetic diversity than the more common L. fragilis. However, the population structures of all three species were incongruent even in the presence of substantial gene flow. Other factors such as habitat specificity may play a role in generating the differences in population structure observed. This study indicates that barriers to gene flow or lack of available host fish are not the cause of decline of the federally endangered L. leptodon, and suggests that alternative explanations should be considered.

Genomic signatures of local adaptation reveal source-sink dynamics in a high gene flow fish species

Understanding source-sink dynamics is important for conservation management, particularly when climatic events alter species’ distributions. Following a 2011 ‘marine heatwave’ in Western Australia, we observed high recruitment of the endemic fisheries target species Choerodon rubescens, towards the cooler (southern) end of its distribution. Here, we use a genome wide set of 14 559 single-nucleotide polymorphisms (SNPs) to identify the likely source population for this recruitment event. Most loci (76%) showed low genetic divergence across the species’ range, indicating high levels of gene flow and confirming previous findings using neutral microsatellite markers. However, a small proportion of loci showed strong patterns of differentiation and exhibited patterns of population structure consistent with local adaptation. Clustering analyses based on these outlier loci indicated that recruits at the southern end of C. rubescens’ range originated 400 km to the north, at the centre of the species’ range, where average temperatures are up to 3 °C warmer. Survival of these recruits may be low because they carry alleles adapted to an environment different to the one they now reside in, but their survival is key to establishing locally adapted populations at and beyond the range edge as water temperatures increase with climate change.

Correlation of Leukocyte Telomere Length Measurement Methods in Patients with Dyskeratosis Congenita and in Their Unaffected Relatives

Several methods have been employed to measure telomere length (TL) in human studies. It has been difficult to directly compare the results from these studies because of differences in the laboratory techniques and output parameters. We compared TL measurements (TLMs) by the three most commonly used methods, quantitative polymerase chain reaction (qPCR), flow cytometry with fluorescence in situ hybridization (flow FISH) and Southern blot, in a cohort of patients with the telomere biology disorder dyskeratosis congenita (DC) and in their unaffected relatives (controls). We observed a strong correlation between the Southern blot average TL and the flow FISH total lymphocyte TL in both the DC patients and their unaffected relatives (R2 of 0.68 and 0.73, respectively). The correlation between the qPCR average TL and that of the Southern blot method was modest (R2 of 0.54 in DC patients and of 0.43 in unaffected relatives). Similar results were noted when comparing the qPCR average TL and the flow FISH total lymphocyte TL (R2 of 0.49 in DC patients and of 0.42 in unaffected relatives). In conclusion, the strengths of the correlations between the three widely used TL assays (qPCR, flow FISH, and Southern blot) were significantly different. Careful consideration is warranted when selecting the method of TL measurement for research and for clinical studies.

Flow magnitude and variability influence growth of two freshwater fish species in a large regulated floodplain river

Fish are often targets for environmental watering outcomes under the premise that aspects of the flow regime are linked to key components of their life-history. This study examined the conceptual link between variability in river discharge and fish productivity by measuring annual growth patterns (generated using sclerochronology over a 22-year period) of two native freshwater cod Maccullochella spp. species over a range of flow conditions in a regulated Australian floodplain River. We found a positive relationship between fish growth, flow variability and river discharge. Flow variability during spring and summer-autumn, as well as their antecedent values, was particularly important in explaining annual growth of the nationally endangered Maccullochella macquariensis. Growth of Maccullochella peelii displayed similar patterns, though were more closely aligned with spring discharge. These results are consistent with the general view that increased river regulation, due to its suppression of flow magnitude and variability, has been a major contributing factor in the decline of native fish populations throughout the world. Our results provide support and guidance for the use of environmental water delivery, and have broad application to rivers worldwide for which any quantification of ecological impacts of regulation, and responses to water management remain scarce.

Spatiotemporal response of pelagic fish aggregations in their spawning grounds of middle Yangtze to the flood process optimized by the Three Gorges Reservoir operation

Flood patterns altered by regulation and operation of the Three Gorges Reservoir (TGR) on the Yangtze has negatively affected fish spawning downstream, particularly results in the decline of spawning sizes. The TGR release was optimized by reoperation for enhancing fish reproduction downstream since 2011. Response of spawning sizes to the flood alteration was documented, yet litter is known concerning the adult fish. Accurate estimate of spatiotemporal changes and abundance of adults in the spawning grounds is critical for quantifying the contribution of flood changes to the spawning. This study used hydroacoustic surveys to document the response of fish aggregations to the changes of flood process (i.e., deployed before-, during- and after flood) on the Jingjiang- and Yichang spawning ground. Acoustic signals from fish tracking were allocated into two groups according to frequency distributions of Target strength (TS), defined as small fish (−60 dB < TS < −35 dB) and large fish (TS > −35 dB). Sizes of small fish acquired before- and during flood were significant larger than those acquired after flood whereas no significant different observed for large fish. Both small and large fish aggregated in both spawning grounds during flood and disappeared after flood. Besides, fish numbers presence in both spawning grounds during flood was greater than those before and after flood. This study suggested spatial distributions of fish aggregations with different sizes in both spawning grounds shifted with the flood changes. Fish aggregated into the spawning grounds for spawning during the flood and departed after the flood, resulting in large fish numbers presence during the flood period. Further research is required to reveal relationships between the hydrological and hydraulic changes results from flow alteration and fish responses to realize their precise requirements and behaviors for spawning.

Risk for pulmonary late effects in childhood Hodgkin lymphoma survivors.

112 Background: Survivors of Hodgkin lymphoma (HL) are at risk for pulmonary late effects, including pulmonary fibrosis (PF). Established risk factors such as younger age at diagnosis and specific therapeutic exposures explain a fraction of the risk for these outcomes. Notably, systemic inflammation and impaired telomere maintenance are associated with adverse pulmonary outcomes in adult populations, but have not been explored in cancer survivors. Our aim was to identify pulmonary diffusion defects in HL survivors, and evaluate the impact of short lymphocyte telomere length (LTL) and systemic inflammation on these outcomes. Methods: Blood samples, demographic and treatment data, and lung carbon monoxide diffusion capacity corrected for hemoglobin and alveolar volume (DLCO) were obtained from HL survivors ≥6 months off therapy and without history of relapse or transplant. LTL was measured by telomere flow FISH (Repeat Diagnostics), and age-based percentiles determined from population controls. Plasma elevations in pro-inflammatory cytokines (HCYTMAG panel, EMD Millipore) were detected on a Luminex platform. Associations between clinical features and most recent DLCO were determined by linear regression, adjusted for age, sex, radiation, and race/ethnicity. In a subset of subjects, cytokine levels and LTL were compared in survivors with DLCO above or below the mean using a Student t-test or Fisher’s Exact test, respectively. Results: Seventy-two HL survivors met inclusion criteria (mean age at diagnosis=14 years, range: 3-18 years). Mean off-therapy DLCO was 75% (range: 52-98%), below the lower limit of normal (76%) in healthy populations. DLCO was inversely associated with female sex (p = 0.002). Five of 24 survivors (21%) had LTL ≤10th percentile for age, but there was no difference in LTL relative to DLCO. Survivors with DLCO ≤75% had higher IL1α and IL1β levels (n = 30, p = 0.07). Conclusions: HL survivors had lower than expected DLCO, with females at highest risk for impaired pulmonary diffusion. Twice as many survivors as expected had LTL ≤10th percentile, and PF-related cytokine profiles were observed in survivors with lower DLCO, suggesting inflammation and telomere maintenance defects may contribute to late effects in HL survivors.

Life history theory predicts long-term fish assemblage response to stream impoundment

Life history theory predictions for hydrologic filtering of fish assemblages are rarely tested with historical time series data. We retrospectively analyzed flow regime and fish assemblage data from the Sabine River, USA, to test relationships between life history strategies and hydrologic variability altered by impoundment construction. Downstream flow variability, but not magnitude, was altered by completion of Toledo Bend Reservoir (TBR) in 1966. Consistent with life history theory, occurrence of opportunistic strategists declined while equilibrium strategists increased as the fish assemblage was transformed between periods immediately after (1967–1973) and approximately one decade after (1979–1982) completion of TBR. Assemblage transformation was related to decline of opportunistic strategists throughout 250 km of river downstream of TBR. Temporal trajectories for opportunistic and intermediate strategist richness modelled as a function of flow variability converged 12 years postimpoundment. The spatiotemporal scaling of our study is novel among tests of life history theory, and results suggest impoundment-induced alteration to natural hydrologic filtering of fish assemblages can operate on the scale of hundreds of stream kilometres and manifest within approximately one decade.

Progress of marginal zone lymphoma

Marginal zone lymphoma (MZL) accounts for approximately 10% of non-Hodgkin lymphoma (NHL). It can be divided into three specific entities: extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT lymphoma), nodal marginal zone lymphoma(NMZL) and splenic marginal zone lymphoma (SMZL). MALT lymphoma is the most frequent overall, representing 7.5% of all NHLs. Reports on research progress of MZL in the 58th American Society of Hematology Annual Meeting covered multiple respects which ranged from basic research to clinical prognosis and treatment. Based on the technology such as flow cytometry, cytogenetics and FISH, further study on pathogenesis of MZL is developing, and new prognostic index system can help to stratify patients more exactly and give a guidance to treatment. What's more, the change of therapy and new drugs will benefit to the clinical efficacy and safety of MZL patients. Key words: Lymphoma, marginal zone; American Society of Hematology Annual Meeting

Optimal undulatory swimming for a single fish-like body and for a pair of interacting swimmers

We establish through numerical simulation conditions for optimal undulatory propulsion for a single fish, and for a pair of hydrodynamically interacting fish, accounting for linear and angular recoil. We first employ systematic two-dimensional (2-D) simulations to identify conditions for minimal propulsive power of a self-propelled fish, and continue with targeted 3-D simulations for a danio-like fish; all at Reynolds number 5000. We find that the Strouhal number, phase angle between heave and pitch at the trailing edge, and angle of attack are principal parameters. For 2-D simulations, imposing a deformation based on measured displacement for carangiform swimming provides, at best, efficiency of 35 %, which increases to 50 % for an optimized motion; for a 3-D fish, the efficiency increases from 22 % to 34 %. Indeed, angular recoil has significant impact on efficiency, and optimized body bending requires maximum bending amplitude upstream of the trailing edge. Next, we turn to 2-D simulation of two hydrodynamically interacting fish. We find that the upstream fish benefits energetically only for small distances. In contrast, the downstream fish can benefit at any position that allows interaction with the upstream wake, provided its body motion is timed appropriately with respect to the oncoming vortices. For an in-line configuration, one body length apart, the efficiency of the downstream fish can increase from 50 % to 60 %; for an offset arrangement it can reach 80 %. This proves that in groups of fish, energy savings can be achieved for downstream fish through interaction with oncoming vortices, even when the downstream fish lies directly inside the jet-like flow of an upstream fish.

Prognostic Significance of Pulmonary Function Test Abnormalities in Patients with Dyskeratosis Congenita

▪Introduction: Pulmonary fibrosis (PF) is a known complication in patients with dyskeratosis congenita (DC), reported in 15-20% of patients. PF is more common in older individuals with DC who may or may not have the classical clinical features of DC. PF in younger DC patients has been reported mainly in the setting of hematopoietic cell transplantation (HCT). There are no prospective studies of pulmonary function tests (PFT) in patients with DC.Objectives: To determine baseline pulmonary function status in patients with DC, association of PFT abnormalities with development or progression of pulmonary disease, and pulmonary outcomes in patients who did or did not receive HCT.Methods: DC patients older than 5 years of age underwent PFTs as part of a prospective IRB-approved cohort study at the NIH from 2002-2015. Data were obtained from spirometry [forced expiratory volume in 1 second (FEV1), forced vital capacity (FVC)], lung volumes [total lung capacity (TLC)] and carbon monoxide diffusion capacity (DLCO). PFT abnormalities were classified as restrictive (normal FEV1/FVC and low TLC), obstructive (low FEV1/FVC and normal TLC) or mixed (low FEV1/FVC and low TLC) according to the American Thoracic Society Task Force guidelines. A low value was < lower limit of normal adjusted for age, sex, race and height. DLCO was also adjusted for hemoglobin. PFT data were analyzed in relation to the DC triad (nail dystrophy, skin pigmentation, leukoplakia), severe bone marrow failure (BMF needing treatment) and telomere length by flow FISH. Patients were followed through 2015. Pulmonary outcomes were compared in those with or without PFT abnormalities at evaluation and in those who did or did not receive HCT. P value <0.05 was significant.Results: Fifty-one patients with DC had PFTs, median age 19 yrs (range 6 - 69). Spirometry was abnormal in 16 patients (31%): 13 had restriction, 2 had obstruction, and 1 had a mixed pattern. DLCO was reduced (<80% of the predicted) in 32/49 patients (65%): 17 had mild (>60%), 13 had moderate (40-60%) and 2 had severely reduced (<40%) DLCO. All patients with abnormal spirometry had reduced DLCO compared with 18/35 (48%) with normal spirometry (p<0.0001). Patients with abnormal spirometry were more likely to have moderate/severe diffusion defects (10/16 vs 5/35; p=0.02), and severe BMF (12/16 vs 10/35; p=0.003) than those with normal spirometry. Telomere length and 2 or more features of the DC triad were not associated with PFT abnormality.Fourteen patients (27%) developed pulmonary disease at a median follow-up of 6 yrs (range 0.1 - 12 yrs) and a median age of 17 yrs (11 - 65): 10 had PF with interstitial honeycomb pattern on chest CT and 4 had pulmonary arteriovenous malformation (AVM) and mild fibrosis. Patients with abnormal spirometry and moderate/severely reduced DLCO were more likely to develop pulmonary disease than those with normal spirometry (12/16 vs 2/35; p<0.0001). Fourteen patients received a HCT (3 prior to and 11 after PFT evaluation) at a median age of 12 yrs (range 3 - 29). Eight of the 14 (57%) developed pulmonary disease 1 - 5 yrs after HCT (median 4 yrs) while 6/37 (16%) non-HCT patients developed PF (p=0.01). Transplanted patients developed pulmonary disease at a younger age than non-transplanted (median age 15 yrs, range 11 - 22 vs 54 yrs, 25 - 65; p=0.001), and had mutations in TINF2 (n=3), RTEL1, DKC1, WRAP53 (1 each); gene unknown (n=2). DC genes mutated in older non-HCT patients with PF were TERC (n=4), TERT (n=1), and unknown (n=1). The independent impact of severe DC phenotype, BMF, and HCT on pulmonary outcome could not be distinguished in young patients as most patients with severe disease had BMF and received HCT. There was no difference in PF among those who did or did not receive radiation for HCT. Four of the 10 deaths were due to PF.Conclusion: More than 30% of patients with DC have restriction and moderate/severe diffusion defect on PFTs, and these are more common in patients with severe BMF. PFT abnormalities are significantly associated with progression to PF, although they may also signal the presence of AVMs, a less common pulmonary manifestation of DC. PF in older patients is a known manifestation of the telomere biology disorder. It is unclear whether pulmonary disease in young patients is a feature of severe DC or the toxicity of HCT preparative regimens. Larger prospective studies are needed to confirm the predictive value of PFTs for future lung disease in patients with DC. DisclosuresNo relevant conflicts of interest to declare.

Comparative Analysis of Japanese and European Typical CLL Patients

Background: Chronic lymphocytic leukemia (CLL) is the most frequent leukemia in the Western countries but very rare in the East Asia including Japan. The previous reports from Japan showed that CLL patients in Japan have different characteristics including atypical morphology and immunophenotypes compared to those of Western countries. We started the cooperative study of CLL between Japan and Austria from 2012.Aim: The aim of this retrospective study is to compare the characteristics of typical CLL patients in Japan and Austria.Material and Methods: Diagnostic procedures were first harmonized between sites and laboratories. Samples were exchanged between Japan and Austria, and morphology, flow cytometry (FCM), FISH and IGVH gene sequence were independently assessed. After technical agreement, we selected 86 Austrian CLL patients from the database of the Medical University of Vienna, and 80 Japanese patients from the CLLRSG-01 prospective observational study. All CLL cases had typical immunophenotypes (Matutes score 4 or 5). The analyses of morphology, FCM and FISH including the deletion in 11q22, 13q14, and 17p13, and trisomy 12 were performed. Immunoglobulin heavy chain variable region (IGHV) gene mutation was also analysed.Results: Median age of CLL patients was 62.5 years old (range: 35-93) in Austria and 66.4 years (range: 34-93) in Japan. Japanese patients were significantly older at diagnosis than Austrian patients (P = 0.027). No differences were found for sex and the Binet stage. Also, there were no differences in laboratory findings (lymphocyte and platelet counts and serum LDH and beta-2-microglobulin levels) except for hemoglobin (Hb) level (median: 13.8g/dL in Austria vs. 13.1g/dL in Japan, P = 0.005) between two countries. Immunophenotypic study by FCM showed higher proportion of CD38 expression in Austrian CLL patients (34% in Austria vs. 10% in Japan, P < 0.001). No differences were found for chromosomal aberrations by FISH analyses between Austria and Japan. del(13q14) was found in 62% and 56% (P = 0.456), trisomy 12 in 7% and 13% (P = 0.315), del(11q22) in 13% and 10% (P = 0.513) and del(17p13) in 7% and 3% (P = 0.179), in Austria and Japan, respectively. The IGHV mutation rate was higher in Japan (P = 0.002). Mutated IGHV was found in 61% of Austrian and 84% of Japanese patients. IGHV family usage profile was significantly different. In Austrian patients, VH1, 2, 3, 4, 5, 6, and 7 were 24%, 1%, 54%, 19%, 1%, 1% and 0%, respectively. However, in Japanese patients, they were 2%, 0%, 75%, 20%, 2%, 0% and 2%, respectively. The proportion of VH1 was surprisingly lower in Japan (P = 0.006). Notably, none of the Japanese patients used VH1-69, in contrast to the use of 10% in Austrian patients.Conclusion: Our analysis indicates that there are clear differences between Japanese and European CLL patients, in age, Hb level, CD38 expression and IGHV usage and mutations. The reason for particular IGHV usage should further be investigated. DisclosuresTakizawa:Teijin: Research Funding; Takeda: Honoraria, Research Funding; Celgene: Honoraria; Chugai: Honoraria, Research Funding; Kyowa Hakko Kirin: Honoraria, Research Funding; Janssen: Honoraria; Sumitomo Dainippon: Honoraria, Research Funding; MSD: Honoraria, Research Funding. Suzuki:Bristol-Myers Squibb: Honoraria; Kyowa Hakko kirin: Honoraria; Chugai: Honoraria. Hoermann:Gilead: Research Funding; Novartis: Honoraria; Amgen: Honoraria; Ariad: Honoraria. Aoki:SymBio Pharmaceuticals: Consultancy. Suzumiya:Kyowa Hakko kirin: Research Funding; Eisai: Honoraria, Research Funding; Astellas: Research Funding; Takeda: Honoraria; Toyama Chemical: Research Funding; Chugai: Honoraria, Research Funding. Jaeger:Janssen: Consultancy, Honoraria, Other: Travel, Accommodations, Expenses.

Taking "Cellfies": Blood Morphology in the Era of Smartphones

Background: Microscopic examination of blood and marrow smears by hematologists is an essential clinical skill at risk of extinction in this high-tech era. This skill is on the decline being replaced by the reliance on flow cytometry, FISH, and molecular diagnostics. Five years ago, we observed that photographs of the oil-immersion field (OIF) in focus could be taken by positioning the smartphone camera lens close to either eyepiece. The captured image could be viewed on the smartphone, swiped to view detail, stored, and shared. We coined the term "Cellfie" for this smartphone image of the OIF. Cellfies can be taken with any smartphone at any microscope, with most learners taking a few minutes for their first Cellfie, and experts able to take Cellfies in a few seconds. Since 2012, we have taught fellows to use Cellfies at the microscope in the hospital and in clinic, and we present the experience of our hematology fellows using Cellfies.Methods: We surveyed past and current hematology fellows (Class of 2015-to Class of 2019) regarding their experience with Cellfies.Results: Nineteen of the twenty fellows invited to participate in the survey responded (95%). Two were PGY IV, 4 were PGY V, and 6 were PGY-VI. Four were fellows who graduated and in practice for one year (3 fellows did not provide level of training). Fifteen of 19 fellows (79%) had taken a Cellfie at some point during their training. Five took <10 Cellfies, 4 took 10-20 Cellfies, and one fellow had taken 50 or more Cellfies. Eight (53%) fellows rated the technique as ’easy’, and 6 as ’difficult’ (40%); one patient found the technique ’very difficult’. Cellfies were commonly used for consultation (80% of responders) and morphology teaching or use in conferences (67%). Only 2 responders had ever used Cellfies to paste in the electronic medical record (EMR). Overall, 80% of the responders thought Cellfies as a very useful tool in their everyday hematology practice.Conclusion: Cellfie use by hematology fellows has resurrected this essential clinical skill of expertise in blood morphology. We have found a way to rekindle a passion for morphology and renew enthusiasm amongst hematology fellows, residents, and students to use information from microscopy as an essential component of their clinical skills. Taking and sharing Cellfies has the potential to put microscope skills back at the bedside, enable rapid diagnosis and treatment, shorten time to intervention in emergencies (acute promyelocytic leukemia, microangiopathy syndromes for example), expedite consultation after-hours and weekends, improve the quality of documentation in EMRs, create a portfolio of stored images for review, and teaching, and enhance patient-care. DisclosuresNo relevant conflicts of interest to declare.

Incorporating thermal requirements into flow regime development for multiple Pacific salmonid species in regulated rivers

It has long been recognized that altered flow regimes and warm water temperature are among major stressors that have contributed to the precipitous decline of salmonid populations in California’s Central Valley, resulting in many of them being listed as endangered or threatened pursuant to the U.S. Endangered Species Act. While it is widely acknowledged that water temperature affects all freshwater life stages of salmonids, it has not been a focus when developing instream flow requirements. In this study, we developed a framework that incorporates water temperature requirements into flow regime development for multiple Pacific salmonid species. We applied this framework to Clear Creek, an upper Central Valley stream that supports three salmonid species and has been regulated by reservoir releases for instream flow since 1963 when Whiskeytown dam was built. The analysis of flow data indicated that from the pre-project (i.e., pre-dam) period to post- project period, flow magnitude was reduced by 45–76% for flood flows, 43% for spring pulse flows, and 27–53% for monthly average flows from January through May. Water temperature from 1996 to 2013 exceeded the U.S. Environmental Protection Agency recommended criteria for salmonids from May to October, with most exceedances occurring in September, followed by May and October. Statistical analyses suggested that the abundances of both adult and juvenile salmonids in Clear Creek were strongly correlated to flow magnitude, water temperature, or both. Higher flows that coincided with adult immigration were correlated to higher numbers of adults spawning in Clear Creek, which in turn produced a higher number of juveniles; however, higher flows that occurred during the early life stages (e.g., fry) appeared to reduce the number of outmigrating juveniles. Higher water temperatures in the immigration, spawning, and juvenile rearing periods were correlated with reduced numbers of both adult and juvenile spring-run Chinook salmon in Clear Creek. High water temperatures in June and July were correlated with reduced number of outmigrating juvenile steelhead. Adult or juvenile fall-run Chinook salmon showed either a curvilinear or linear relationship with water temperature during their immigration, spawning, and rearing periods. Based on the pre-project flow and strong correlation between fish abundance and flow or water temperature, we developed a flow regime that incorporates water temperature requirements and other needs of different life stages of salmonids. The flow regime includes the following elements: winter and spring base flow, summer and fall water temperature sustaining flow, winter floodplain inundation flow, and spring pulse flow. This approach can be applied to other regulated streams for developing flow regimes to conserve and recover listed salmonid species.

Hydrodynamic interaction of swimming organisms in an inertial regime.

We numerically investigate the hydrodynamic interaction of swimming organisms at small to intermediate Reynolds number regimes, i.e., Re∼O(0.1-100), where inertial effects are important. The hydrodynamic interaction of swimming organisms in this regime is significantly different from the Stokes regime for microorganisms, as well as the high Reynolds number flows for fish and birds, which involves strong flow separation and detached vortex structures. Using an archetypal swimmer model, called a "squirmer," we find that the inertial effects change the contact time and dispersion dynamics of a pair of pusher swimmers, and trigger hydrodynamic attraction for two pullers. These results are potentially important in investigating predator-prey interactions, sexual reproduction, and the encounter rate of marine organisms such as copepods, ctenophora, and larvae.

Brilliant violet fluorochromes in simultaneous multicolor flow cytometry–fluorescence in situ hybridization measurement of monocyte subsets and telomere length in heart failure

Conventional analytical methods to determine telomere length (TL) have been replaced by more precise and reproducible procedures, such as fluorescence in situ hybridization coupled with flow cytometry (flow–FISH). However, simultaneous measurement of TL and cell phenotype remains difficult. Relatively expensive and time-consuming cell-sorting purification is needed to counteract the loss, due to stringent FISH conditions, of prehybridization fluorescence by the organic fluorochromes conventionally used in the phenotyping step. Here, we sought to assess whether the newly developed Brilliant Violet (BV) dyes are valuable to specifically and simultaneously assess the distribution and telomere attrition of monocyte subsets circulating in the blood of a cohort of patients with heart failure. We performed flow–FISH on blood samples from 28 patients with heart failure. To differentiate among monocyte subsets, we used BV and conventional fluorochromes conjugated to antibodies against CD86, CD14, CD16, and CD15. We simultaneously assessed the TLs of the monocyte subsets with a telomere-specific peptide nucleic acid probe labeled with fluorescein isothiocyanate. The BV dyes completely tolerated the harsh conditions required for adequate DNA denaturation and simultaneously provided accurate identification of monocyte subpopulations and respective TLs. The presented protocol may be faster and less expensive than those used currently for purposes such as establishing associations among patient categories, disease progression, monocyte heterogeneity, and aging in the context of heart failure.

MEASUREMENT AND ANALYSIS OF THE ECONOMIC EFFECTS OF PRICE POLICIES FOR FISH IN EGYPT

Prices reflect the relative scarcity of resources in terms of planning is based under the free-market mechanisms for price and market signals as an essential mechanism in the allocation of resources and guidance system, Price is also working on consumer directed toward maximizing satisficing the resources available to them, which leads to maximizing the well-being of the community. The research problem is in direct government intervention in price policy, which may give rise to a variation in the local and indirect world prices lead to non-optimal utilization of the resources used in the production process, which affects the welfare of producers and consumers, as well as on government revenue and the outcome of the state foreign trade. research aims to identify the price differences between local and world prices resulting from price, The policies fish and by estimating the partial equilibrium model to examine the effect influence of the policies of government intervention at all stages of fish flow in the market, by measuring the efficiency of the product, the consumer, the community level, and measurement of well-being at the community level, as well as measurement of government revenues and this may help to measure the efficiency of these policies in achieving its objectives and examine the effects on producers, consumers and the overall economy. The results showed the following The average nominal protection for fish rate during the study period (1998-2013) amounted to 0.67 i.e. it is greater than zero, which means that the state supports the producers of fish by about at 67% in favor of the producers of fish on the expense of the consumer represented in the higher domestic price for the world price, The results indicate that the concern of the State on promoting fish producers to reduce the size of the gap and reduce the import of foreign currency into force. The study showed also that the expansion of fish production leads to increased foreign exchange earnings by reducing imports and reduce the food gap, where the maximum fish production reached amounted to 1372.0, 1454.4 during the years 2012.2013, respectively, while the maximum foreign exchange earnings reached a maximum in 2012, 2013 amounted to 27067.03. The results of the welfare check gains in producer excess as the average change in the product excess amounted to 3.1336 billion pounds, which reflects the impact of price policy applied, while under the negative values of consumer excess has been achieved loss in consumer surplus, bringing the average loss during the study period of about 5566.88 million pounds. Decline in consumer surplus was due to the local consumer bear the superiority of market forces and the presence of protectionist policy to support the product and taxing the consumer, leading to increased consumer spending as a result of the purchase of small quantities at high prices, leading to lower prices of consumer welfare.

Evaluation of lemon balm (Melissa officinalis) collections

Lemon balm (Melissa officinalis L.) is a well-known medicinal and aromatic plant and of increasing importance resulting in rising growth area in Germany. Because of its proven sedative, spasmolytic and antiviral effects, it is often used in watery or alcoholic extracts for self-medication or pharmaceutical and medical purposes. This therapeutic effect is due to the content of essential oil and phenolic carbon acids, like rosmarinic acid. Improved knowledge on the genome structure, number of chromosomes in connection with the taxonomical structure of balm is indispensable for improved new varieties. A set of 120 balm accessions was evaluated for the variability of essential oil content and composition as well as the content of rosmarinic acid. These accessions came from the Bavarian State Institute for Agriculture at Freising, Germany (LfL), the federal ex-situ collection of agricultural and horticultural plants of the Leibniz Institute of Plant Genetics and Crop Plant Research at Gatersleben, Germany (IPK) and the N.I. Vavilov Institute of Plant Industry at St. Petersburg, Russia (VIR). Out of these 120 accessions 40 balm accessions (M. officinalis) were characterized by flow cytometry and FISH (18/25S and 5S rDNA) to determine the chromosome number and ploidy level. Three different types were found: diploid genotypes with 2n = 2× = 32 chromosomes; tetraploid 2n = 4× = 64 chromosomes and triploid 2n = 3× = 48 chromosomes. Therefore a haploid base number of × = 16 chromosomes is likely. For the first time triploid accessions are described, which were sterile but cytologically and morphologically stable for many years. Triploids express better winter hardiness and regeneration after harvesting cuts as well as bigger leaves and internodes. We characterized three chemotypes (ct.) of essential oil: ct. citral, ct. germacrene D and ct. β-caryophyllene oxide. In addition autotetraploid material from diploid ct. citral was developed for this characterization and belongs also to ct. citral.

The swimming patterns and energy-saving mechanism revealed from three fish in a school

Fish in nature are believed to favor their swimming performance by swimming in a school. We investigated numerically the thrust and power consumption of a school of fish with an arrangement of one upstream and two downstream. The upstream fish enhances the thrust with an area of high pressure at the heads of the two downstream fish, but the region of low pressure created between the two downstream fish generates a strong suction and is detrimental to the swimming of the upstream fish. The thrust forces increase but the power consumptions of the downstream fish decrease. The downstream fish in this arrangement can avoid the jet flow of the upstream fish and benefit from the counter flow at the verge of the upstream wake. The power-saving mechanism is obvious and sensitive to the longitudinal distance when the fish are in a school within a small lateral distance. The maximum average power consumption in a school is 20% less than not in a school. The mechanisms of thrust enhancement and energy saving in a school of fish interact in manifold ways, and provide insight into the bio-inspired design for arrays of underwater vehicles.