Flow cytometric characterization of freshwater crayfish hemocytes for the examination of physiological status in wild and captive animals

Enumeration of invertebrate hemocytes is a potentially powerful tool for the determination of physiological effects of extrinsic stressors, such as hypoxia, disease, and toxicant exposure. A detailed flow cytometric method of broad application was developed for the objective characterization and enumeration of the hemocytes of New Zealand freshwater crayfish Paranephrops planifrons for the purpose of physiological health assessment. Hemocyte populations were isolated by flow cytometric sorting based on differential light scatter properties followed by morphological characterization via light microscopy and software image analysis. Cells were identified as hyaline, semigranular, and granular hemocytes based on established invertebrate hemocyte classification. A characteristic decrease in nuclear size, an increase in granularity between the hyaline and granular cells, and the eccentric location of nuclei in granular cells were also observed. The granulocyte subpopulations were observed to possess varying degrees of granularity. The developed methodology was used to perform total and differential hemocyte counts from three lake populations and between wild and captive crayfish specimens. Differences in total and differential hemocyte counts were not observed among the wild populations. However, specimens held in captivity for 14 d exhibited a significant 63% reduction in total hemocyte count, whereas the relative hemocyte proportions remained the same. These results demonstrate the utility of this method for the investigation of subacute stressor effects in selected decapod crustaceans

Ecology of common bully (Gobiomorphus cotidianus) in the Tarawera and Rangitaiki rivers: isolation by inland distance or anthropogenic discharge?

Previous research has identified distinct genetic, life-history and reproductive differences between populations of common bully (Gobiomorphus cotidianus) upstream and downstream of a pulp and paper mill outfall on the Tarawera River in the Bay of Plenty, New Zealand. This study investigated the distribution of common bully in the Tarawera River by examining fish collected from upstream (37 km inland) and downstream (20 km inland) locations and comparing them to fish from similar inland locations (40 km and 17 km inland, respectively) in the nearby Rangitaiki River. Reproductive divergence was observed between upstream and downstream sites of both rivers by differing annual trends in gonadosomatic index. Stable carbon and nitrogen isotopes confirmed residency at each sampling site and otolith microchemistry demonstrated different life-history strategies between upstream and downstream populations. Diadromous recruits dominated in both downstream river populations, with a general disappearance of diadromy upstream. A mixture of diadromous and non-diadromous fish were found in the upstream Rangitaiki River, whereas diadromous recruits were absent in the upstream Tarawera River. A reduction in oculoscapular canal structures also coincided with loss of diadromy in fish from both rivers. A behavioural study to determine whether pulp and paper mill effluent may deter fish migration within the Tarawera River demonstrated a strong avoidance of effluent, but only at concentrations (>25%) greater than those that naturally occur in the river (<15%). The results of this study suggest that combinations of influences coupled with inland distance are likely to be responsible for the isolation of common bully subpopulations within the Tarawera River

Examination of pulp and paper effluent-hypoxia interactions in fish

Recent concerns have focused on the adverse effects of low dissolved oxygen (DO) combined with the potential chronic effects of pulp and paper mill effluent exposure on fishes in the Tarawera River, Bay of Plenty, New Zealand. A degassing system for the removal and control of dissolved oxygen (DO) capable of producing water with-0.5 mg L⁻¹ DO to saturation and two series of five separate DO concentrations each was first constructed. Because the Tarawera River receives effluent discharges from a chemithermomechanical (CTMP) tissue mill and an integrated thermomechanical (TMP)/bleached kraft pulp and paper mill (BK), these effluents were studied. Using the DO control system, the effects of simultaneous effluent and hypoxia exposure on the 48-h DO LC50 (median lethal concentration) were examined in fry and juvenile rainbow trout (Oncorhynchus mykiss) and common bully (Go_biomorphus cotidianus). TMPIBK mill effluent (ME) was diluted to 15 % v/v to reflect the upper concentration in the receiving environment, while CMTP effluent was extracted directly from the river. The presence of either effluent did not significantly increase the lethality of low DO for either species and life stage differences were not detected. Preexposure of juvenile trout to 15 and 70 % TMP/BKME also showed no significant effect on the DO LC50. No significant difference in the oxygen consumption of juvenile trout tested in 15 % v/v effluent were evident, but significant increases in oxygen consumption were observed for fish pre-exposed to 15 and 70 % effluent when tested in reference water. Four-week exposures were performed with juvenile trout to determine the chronic effects ofTMP/BKME (15 % v/v) and CTMP effluent combined with non-lethal DO concentrations (2.5 mg L⁻¹ to fully saturated), revealing excellent survival of fish in the combined effluent/hypoxia exposures, but marred by poor survival of reference water-exposed fish. Relationships for growth were observed for fish exposed to DO and effluent in the CTMP experiment. Several DO dose-response effects were also observed on general hematology in both effluent experiments, but were generally considered minor and typical of hypoxia exposure in fish. Four-week exposures using juvenile trout in 0, 10, 30 and 70 % TMP/BKME and 0, 35, 110 and 250 μg L⁻¹dehydroabietic acid (DHAA) were performed, where swimming performance, oxygen consumption and hematology were investigated. The chief finding was that of reduced swimming performance, indicated by lower critical swimming (Ucrit) speeds, for fish exposed to 70 % TMP/BKME. Other moderate effects on various hematological parameters were also observed. However, all observed effects are considered relatively small in magnitude and at effluent concentrations well above those found in the receiving environment. Forty eight-hour DO LCSOs were determined for juvenile inanga (Galaxias maculatus), common smelt (Retropinna retropinna), shortfin eel elvers (Anguilla australis), rainbow trout, common bully and the freshwater shrimp (Paratya curvirostris). Juvenile inanga were the most sensitive fish species, while eel elvers were the most tolerant fish species tested

In-Situ Load System for Calibrating and Validating Aerodynamic Properties of Scaled Aircraft in Ground-Based Aerospace Testing Applications

An In-Situ Load System for calibrating and validating aerodynamic properties of scaled aircraft in ground-based aerospace testing applications includes an assembly having upper and lower components that are pivotably interconnected. A test weight can be connected to the lower component to apply a known force to a force balance. The orientation of the force balance can be varied, and the measured forces from the force balance can be compared to applied loads at various orientations to thereby develop calibration factors

Capacity limitations of visual memory in two-interval comparison of Gabor arrays

The capacity of short-term visual memory (VSTM) was assessed in a two-interval spatial frequency (SF) discrimination task. The cued Gabor target in a multi-element array either increased or decreased in SF across a 2s interstimulus interval (ISI). Distracters as well as target were made to change across ISI so that memory of the individual SF of Gabor elements was required to solve the discrimination. The dynamics of the information loss from visual memory were analysed by manipulating the timing of spatial cues and masks. Cueing the target position before the first display gave thresholds comparable with those for a single Gabor patch. Cues placed after the first display gave higher thresholds indicating some loss of information. Within the ISI there was little increase in threshold or set size effect with cue delay. However there was a sharp rise in thresholds for cue positions after the second display. Gabor masks placed before a mid-ISI cue were more effective than noise masks or Gabor masks placed after the cue. With a cue placed late in the ISI, preceded by a Gabor mask, the masking effect decreased with increasing delay of the mask after the first display. This suggests a selective, dynamic but increasingly durable representation of the initial stimulus is built up in memory, and there is a graded form of “overwriting” of this representation by new stimuli

Characteristics of the National Applicant Pool for Clinical Informatics Fellowships (2016-2017)

We conducted a national study to assess the numbers and diversity of applicants for 2016 and 2017 clinical informatics fellowship positions. In each year, we collected data on the number of applications that programs received from candidates who were ultimately successful vs. unsuccessful. In 2017, we also conducted an anonymous applicant survey. Successful candidates applied to an average of 4.2 and 5.5 programs for 2016 and 2017, respectively. In the survey, unsuccessful candidates reported applying to fewer programs. Assuming unsuccessful candidates submitted between 2-5 applications each, the total applicant pool numbered 42-69 for 2016 (competing for 24 positions) and 52-85 for 2017 (competing for 30 positions). Among survey respondents (n=33), 24% were female, 1 was black and none were Hispanic. We conclude that greater efforts are needed to enhance interest in clinical informatics among medical students and residents, particularly among women and members of underrepresented minority groups

Revision of the Melanocytic Pathology Assessment Tool and Hierarchy for Diagnosis Classification Schema for Melanocytic Lesions: A Consensus Statement

IMPORTANCE A standardized pathology classification system for melanocytic lesions is needed to aid both pathologists and clinicians in cataloging currently existing diverse terminologies and in the diagnosis and treatment of patients. The Melanocytic Pathology Assessment Tool and Hierarchy for Diagnosis (MPATH-Dx) has been developed for this purpose. OBJECTIVE To revise the MPATH-Dx version 1.0 classification tool, using feedback from dermatopathologists participating in the National Institutes of Health-funded Reducing Errors in Melanocytic Interpretations (REMI) Study and from members of the International Melanoma Pathology Study Group (IMPSG). EVIDENCE REVIEW Practicing dermatopathologists recruited from 40 US states participated in the 2-year REMI study and provided feedback on the MPATH-Dx version 1.0 tool. Independently, member dermatopathologists participating in an IMPSG workshop dedicated to the MPATH-Dx schema provided additional input for refining the MPATH-Dx tool. A reference panel of 3 dermatopathologists, the original authors of the MPATH-Dx version 1.0 tool, integrated all feedback into an updated and refined MPATH-Dx version 2.0. FINDINGS The new MPATH-Dx version 2.0 schema simplifies the original 5-class hierarchy into 4 classes to improve diagnostic concordance and to provide more explicit guidance in the treatment of patients. This new version also has clearly defined histopathological criteria for classification of classes I and II lesions; has specific provisions for the most frequently encountered low-cumulative sun damage pathway of melanoma progression, as well as other, less common World Health Organization pathways to melanoma; provides guidance for classifying intermediate class II tumors vs melanoma; and recognizes a subset of pT1a melanomas with very low risk and possible eventual reclassification as neoplasms lacking criteria for melanoma. CONCLUSIONS AND RELEVANCE The implementation of the newly revised MPATH-Dx version 2.0 schema into clinical practice is anticipated to provide a robust tool and adjunct for standardized diagnostic reporting of melanocytic lesions and management of patients to the benefit of both health care practitioners and patients

Infants with esophageal atresia and right aortic arch: Characteristics and outcomes from the Midwest Pediatric Surgery Consortium

Purpose Right sided aortic arch (RAA) is a rare anatomic finding in infants with esophageal atresia with or without tracheoesophageal fistula (EA/TEF). In the presence of RAA, significant controversy exists regarding optimal side for thoracotomy in repair of the EA/TEF. The purpose of this study was to characterize the incidence, demographics, surgical approach, and outcomes of patients with RAA and EA/TEF. Methods A multi-institutional, IRB approved, retrospective cohort study of infants with EA/TEF treated at 11 children's hospitals in the United States over a 5-year period (2009 to 2014) was performed. All patients had a minimum of one-year follow-up. Results In a cohort of 396 infants with esophageal atresia, 20 (5%) had RAA, with 18 having EA with a distal TEF and 2 with pure EA. Compared to infants with left sided arch (LAA), RAA infants had a lower median birth weight, (1.96 kg (IQR 1.54–2.65) vs. 2.57 kg (2.00–3.03), p = 0.01), earlier gestational age (34.5 weeks (IQR 32–37) vs. 37 weeks (35–39), p = 0.01), and a higher incidence of congenital heart disease (90% vs. 32%, p  0.29). Conclusion RAA in infants with EA/TEF is rare with an incidence of 5%. Compared to infants with EA/TEF and LAA, infants with EA/TEF and RAA are more severely ill with lower birth weight and higher rates of prematurity and complex congenital heart disease. In neonates with RAA, surgical repair of the EA/TEF is technically feasible via thoracotomy from either chest. A higher incidence of anastomotic strictures may occur with a right-sided approach

Rapid ocean acidification and protracted Earth system recovery followed the end-Cretaceous Chicxulub impact

J.W.B.R. was supported by ERC Starting Grant 805246 OldCO2NewArchives.Mass extinction at the Cretaceous–Paleogene (K-Pg) boundary coincides with the Chicxulub bolide impact and also falls within the broader time frame of Deccan trap emplacement. Critically, though, empirical evidence as to how either of these factors could have driven observed extinction patterns and carbon cycle perturbations is still lacking. Here, using boron isotopes in foraminifera, we document a geologically rapid surface-ocean pH drop following the Chicxulub impact, supporting impact-induced ocean acidification as a mechanism for ecological collapse in the marine realm. Subsequently, surface water pH rebounded sharply with the extinction of marine calcifiers and the associated imbalance in the global carbon cycle. Our reconstructed water-column pH gradients, combined with Earth system modeling, indicate that a partial ∼50% reduction in global marine primary productivity is sufficient to explain observed marine carbon isotope patterns at the K-Pg, due to the underlying action of the solubility pump. While primary productivity recovered within a few tens of thousands of years, inefficiency in carbon export to the deep sea lasted much longer. This phased recovery scenario reconciles competing hypotheses previously put forward to explain the K-Pg carbon isotope records, and explains both spatially variable patterns of change in marine productivity across the event and a lack of extinction at the deep sea floor. In sum, we provide insights into the drivers of the last mass extinction, the recovery of marine carbon cycling in a postextinction world, and the way in which marine life imprints its isotopic signal onto the geological record.Publisher PDFPeer reviewe

Tetrabenzoporphyrin and -mono-, - Cis -di- and tetrabenzotriazaporphyrin derivatives: Electrochemical and spectroscopic implications of meso CH Group replacement with nitrogen

Nonperipherally hexyl-substituted metal-free tetrabenzoporphyrin (2H-TBP, 1a) tetrabenzomonoazaporphyrin (2H-TBMAP, 2a), tetrabenzo-cis-diazaporphyrin (2H-TBDAP, 3a), tetrabenzotriazaporphyrin (2H-TBTAP, 4a), and phthalocyanine (2H-Pc, 5a), as well as their copper complexes (1b-5b), were synthesized. As the number of meso nitrogen atoms increases from zero to four, Îmax of the Q-band absorption peak becomes red-shifted by almost 100 nm, and extinction coefficients increased at least threefold. Simultaneously the blue-shifted Soret (UV) band substantially decreased in intensity. These changes were related to the relative electron-density of each macrocycle expressed as the group electronegativity sum of all meso N and CH atom groups, âχR. X-ray photoelectron spectroscopy differentiated between the three different types of macrocyclic nitrogen atoms (the Ninner, (NH)inner, and Nmeso) in the metal-free complexes. Binding energies of the Nmeso and Ninner,Cu atoms in copper chelates could not be resolved. Copper insertion lowered especially the cathodic redox potentials, while all four observed redox processes occurred at larger potentials as the number of meso nitrogens increased. Computational chemical methods using density functional theory confirmed 1b to exhibit a Cu(II) reduction prior to ring-based reductions, while for 2b, Cu(II) reduction is the first reductive step only if the nonperipheral substituents are hydrogen. When they are methyl groups, it is the second reduction process; when they are ethyl, propyl, or hexyl, it becomes the third reductive process. Spectro-electrochemical measurements showed redox processes were associated with a substantial change in intensity of at least two main absorbances (the Q and Soret bands) in the UV spectra of these compounds