Differentiation of ruminant transmissible spongiform encephalopathy isolate types, including bovine spongiform encephalopathy and CH1641 scrapie

With increased awareness of the diversity of transmissible spongiform encephalopathy (TSE) strains in the ruminant population, comes an appreciation of the need for improved methods of differential diagnosis. Exposure to bovine spongiform encephalopathy (BSE) has been associated with the human TSE, variant Creutzfeldt–Jakob disease, emphasizing the necessity in distinguishing low-risk TSE types from BSE. TSE type discrimination in ruminants such as cattle, sheep, goats and deer, requires the application of several prion protein (PrP)-specific antibodies in parallel immunochemical tests on brain homogenates or tissue sections from infected animals. This study uses in a single incubation step, three PrP-specific antibodies and fluorescent Alexa dye-labelled anti-mouse Fabs on a Western blot. The usual amount of brain tissue needed is 0.5 mg. This multiplex application of antibodies directed towards three different PrP epitopes enabled differential diagnosis of all established main features of classical scrapie, BSE and Nor98-like scrapie in sheep and goats, as well as the currently known BSE types C, H and L in cattle. Moreover, due to an antibody-dependent dual PrP-banding pattern, for the first time CH1641 scrapie of sheep can be reliably discriminated from the other TSE isolate types in sheep

A chemical ionization study of deuteron transfer initiated propene loss from propoxypyridines

AbstractThe mechanism of propene loss from the metastable [M + D]+ ions of isomeric 2-, 3-, and 4-n-propoxypyridines and the related isopropoxypyridines has been examined by chemical ionization (CI) and tandem mass spectrometry in combination with deuterium labeling. The [M + D]+ ions were generated with CD3OD, CD3CN, (CD3)2CO, or pyrrole-D5 (listed in order of increasing proton affinity) as the CI reagent. The results reveal that the deuteron added in the CI process is not interchanged with the hydrogen atoms of the propyl group prior to propene loss from the metastable [M + D]+ ions of the propoxypyridines. The site selective labeling of the α-, β-, or γ-position of the propyl group indicates that the [M + D]+ ions of 2-n-propoxypyridine expel propene with formation of an ion-neutral complex composed of a propyl carbenium ion and 2-pyridone. By contrast, the [M + D]+ ions of 3-n-propoxypyridine expel propene by: (1) Formation of ion-neutral complexes, and (2) a conventional 1,5-hydride shift from the β-position of the n-propyl group to the ring and/or a 1,2-elimination type process. For the 4-isomer, the results suggest the occurrence of propene loss by a 1,2-elimination in addition to the intermediate formation of ion-neutral complexes. Loss of propene with one deuterium atom is the only reaction of the [M + D]+ ions of the isopropoxypyridines labeled at the α-position of the isopropyl group. The results for the isopropoxypyridines labeled with three deuterium atoms at the β-position are consistent with: (1) The loss of propene by ion-neutral complex formation and the occurrence of a substantial isotope effect in the subsequent proton/deuteron transfer within the complex, and/or (2) the loss of propene by a 1,2-elimination type reaction

Role of focused assessment with sonography for trauma as a screening tool for blunt abdominal trauma in young children after high energy trauma

Background: The objective of the study was to review the utility of focused assessement with sonography for trauma (FAST) as a screening tool for blunt abdominal trauma (BAT) in children involved in high energy trauma (HET), and to determine whether a FAST could replace computed tomography (CT) in clinical decision-making regarding paediatric BAT. Method: Children presented at the Trauma Unit of the Red Cross War Memorial Children's Hospital, Cape Town, after HET, and underwent both a physical examination and a FAST. The presence of free fluid in the abdomen and pelvis was assessed using a FAST. Sensitivity, specificity, and positive and negative predictive values (PPV and NPV) for identifying intraabdominal injury were calculated for the physical examination and the FAST, both individually and when combined. Results: Seventy-five patients were included as per the criteria for HET as follows: pedestrian motor vehicle crashes (MVCs) (n = 46), assault (n = 14), fall from a height (n = 9), MVC passenger (n = 4) and other (n = 2). The ages of the patients ranged from 3 months to 13 years. The sensitivity of the physical examination was 0.80, specificity 0.83, PPV 0.42 and NPV 0.96. The sensitivity of the FAST was 0.50, specificity 1.00, PPV 1.00 and NPV 0.93. Sensitivity increased to 0.90 when the physical examination was combined with the FAST. Nonoperative management was used in 73 patients. Two underwent an operation. Conclusion: A FAST should be performed in combination with a physical examination on every paediatric patient involved in HET to detect BAT. When both are negative, nonoperative management can be implemented without fear of missing a clinically significant injury. FAST is a safe, effective and easily accessible alternative to CT, which avoids ionising radiation and aids in clinical decision-making

Skin measurement devices to assess skin quality:A systematic review on reliability and validity

Background Many treatments aim to slow down or reverse the visible signs of skin aging and thereby improve skin quality. Measurement devices are frequently employed to measure the effects of these treatments to improve skin quality, for example, skin elasticity, color, and texture. However, it remains unknown which of these devices is most reliable and valid. Materials and methods MEDLINE, Embase, Cochrane Central, Web of Science, and Google Scholar databases were searched. Instruments were scored on reporting construct validity by means of convergent validity, interobserver, intraobserver, and interinstrument reliability. Results For the evaluation of skin color, 11 studies were included describing 16 measurement devices, analyzing 3172 subjects. The most reliable device for skin color assessment is the Minolta Chromameter CR-300 due to good interobserver, intraobserver, and interinstrument reliability. For skin elasticity, seven studies assessed nine types of devices analyzing 290 subjects in total. No intra and interobserver reliability was reported. Skin texture was assessed in two studies evaluating 72 subjects using three different types of measurement devices. The PRIMOS device reported excellent intra and interobserver reliability. None of the included reviewed devices could be determined to be valid based on construct validity. Conclusion The most reliable devices to evaluate skin color and texture in ordinary skin were, respectively, the Minolta Chromameter and PRIMOS. No reliable device is available to measure skin elasticity in ordinary skin and none of the included devices could be determined to be designated as valid

Variability in disease phenotypes within a single PRNP genotype suggests the existence of multiple natural sheep scarpie strains within Europe

Variability of pathological phenotypes within classical sheep scrapie cases has been reported for some time, but in many instances it has been attributed to differences in the PRNP genotype of the host. To address this issue we have examined by immunohistochemistry (IHC) and Western blotting (WB) for the disease-associated form of the prion protein (PrPd), the brains of 23 sheep from five European countries, all of which were of the same ARQ/ARQ genotype. As a result of IHC examinations, sheep were distributed into five groups with different phenotypes and the groups were the same regardless of the scoring method used, ‘long’ or ‘short’ PrPd profiling. The groups made did not respond to the geographical origin of the cases and did not correlate with the vacuolar lesion profiles, which showed a high individual variability. Discriminatory IHC and WB methods coincided to detect a ‘CH1641-like’ case but otherwise correlated poorly in the classification of disease phenotypes. No other polymorphisms of the PRNP gene were found that could account for the pathological differences, except perhaps for a sheep from Spain with a mutation at codon 103 and a unique pathological phenotype. Preliminary evidence indicates that those different IHC phenotypes correlate with distinct biological properties on bioassay, suggesting that they are indicative of strain diversity. We therefore conclude that natural scrapie strains exist and that they can be revealed by detailed pathological examinations, which can be harmonized between laboratories to produce comparable results

Reply:The Effects of Facial Lipografting on Skin Quality: A Systematic Review

BACKGROUND: Autologous lipografting for improvement of facial skin quality was first described by Coleman in 2006. The current dogma dictates that adipose tissue-derived stromal cells that reside in the stromal vascular fraction of lipograft contribute to skin rejuvenation (e.g., increased skin elasticity), a more homogenous skin color, and softening of skin texture. Nowadays, many studies have been reported on this "skin rejuvenation" effect of autologous fat grafting. This systematic review was undertaken to assess the efficacy of autologous lipografting on skin quality.METHODS: The MEDLINE, Embase, Cochrane Central, Web of Science, and Google Scholar databases were searched for studies evaluating the effect of autologous lipografting on facial skin quality (May 11, 2018). Outcomes of interest were skin texture, color, and elasticity in addition to histologic outcomes and number of complications.RESULTS: Nine studies were included, with 301 patients treated in total. No meta-analysis could be performed because of heterogeneity of the metrics and outcomes. Eight studies reported increased skin elasticity; improvement in skin texture; and a more homogeneous skin color after treatment with lipografting, cellular stromal vascular fraction, or nanofat. One study reported no increased skin elasticity after lipografting. Histologic improvement was seen after lipografting and adipose tissue-derived stromal cell injections. However, in general, the level of evidence of the included studies was low. No serious complications were reported.CONCLUSION: Autologous facial lipografting and cellular stromal vascular fraction and adipose tissue-derived stromal cell injections hardly seem to improve facial skin quality but can be considered a safe procedure.</p

Early start of enzyme replacement therapy in pediatric male patients with classical Fabry disease is associated with attenuated disease progression

Background Enzyme replacement therapy (ERT) slows disease progression of Fabry disease (FD), especially when initiated before the onset of irreversible organ damage. However, with the clinically asymptomatic progression of renal, cardiac and cerebral disease manifestations spanning decades, optimal timing of ERT initiation remains unclear. Methods In this cross-sectional retrospective study, seven male FD patients with a classical disease phenotype (cFD) who started treatment with agalsidase-beta in childhood were evaluated after 10 years of treatment (median age at evaluation 24 years, range 14–26). Cardiac imaging (echocardiography and MRI), electrophysiological and biochemical data of these patients were compared to those of untreated male cFD patients (n = 23, median age 22 years, range 13–27). Results Albuminuria was less common and less severe in treated patients (albumin to creatinine ratio, ACR 0–8.8 mg/mmol, median 0.4) compared to untreated patients (ACR 0–248 mg/mmol, median 3.7, p = 0.02). The treated group had a lower left ventricular mass, measured using echocardiography (median 80 g/m2 versus 94 g/m2, p = 0.02) and MRI (median 53 g/m2 versus 68 g/m2, p = 0.02). Myocardial fibrosis was absent in all included patients. eGFR was normal in all treated patients whereas 7/23 (30%) of untreated patients had abnormal eGFR. Cerebral manifestations did not differ. Conclusions Start of treatment with ERT before age 16, in male cFD patients is associated with reduced occurrence of renal and cardiac manifestations of FD, as assessed by intermediate endpoints. Confirmation that this approach delays or even prevents renal failure and cardiac events requires another decade of follow-up.publishedVersio

Multiproxy analysis of permafrost preserved faeces provides an unprecedented insight into the diets and habitats of extinct and extant megafauna

The study of faecal samples to reconstruct the diets and habitats of extinct megafauna has traditionally relied on pollen and macrofossil analysis. DNA metabarcoding has emerged as a valuable tool to complement and refine these proxies. While published studies have compared the results of these three proxies for sediments, this comparison is currently lacking for permafrost preserved mammal faeces. Moreover, most metabarcoding studies have focused on a single plant-specific DNA marker region. In this study, we target both the commonly used chloroplast trnL P6 loop as well as nuclear ribosomal ITS (nrITS). The latter can increase taxonomic resolution of plant identifications but requires DNA to be relatively well preserved because of the target length (∼300–500 bp). We compare DNA results to pollen and macrofossil analyses from permafrost and ice-preserved faeces of Pleistocene and Holocene megafauna. Samples include woolly mammoth, horse, steppe bison as well as Holocene and extant caribou. Most plant identifications were found using DNA, likely because the studied faeces contained many vegetative remains that could not be identified using macrofossils or pollen. Several taxa were, however, identified to lower taxonomic levels uniquely with macrofossil and pollen analysis. The nrITS marker provides species level taxonomic resolution for commonly encountered plant families that are hard to distinguish using the other proxies (e.g. Asteraceae, Cyperaceae and Poaceae). Integrating the results from all proxies, we are able to accurately reconstruct known diets and habitats of the extant caribou. Applying this approach to the extinct mammals, we find that the Holocene horse and steppe bison were not strict grazers but mixed feeders living in a marshy wetland environment. The mammoths showed highly varying diets from different non-analogous habitats. This confirms the presence of a mosaic of habitats in the Pleistocene ‘mammoth steppe’ that mammoths could fully exploit due to their flexibility in food choice