Non‐Invasive Measurements of Skin Pigmentation In Situ

Objective in situ measurements of skin pigmentation are needed for accurate documentation of pigmentation disorders, in studies of constitutive and induced skin pigmentation, for testing of the efficacy of pro‐pigmentation or de‐pigmentation agents, etc. Non‐invasive instrumental measurements of skin pigmentation have been used for many decades. All are based on the ability of melanin to attenuate light. However, hemoglobin in dermal capillaries also attenuates light and needs to be accounted for when pigmentation is assessed. The methods under consideration include: (a) single point measurements, in which light reflected from a defined skin area is collected and a pigment index is calculated representing the average pigmentation over the examined area, and (b) imaging methods that attempt to generate a concentration distribution map of melanin pigment for the skin area being imaged. In this article, we describe the potentials and the limitations of the different approaches to both single point and imaging methods.     

Mass and Density Measurements of Live and Dead Gram-Negative and Gram-Positive Bacterial Populations

Monitoring cell growth and measuring physical features of food-borne pathogenic bacteria are important for better understanding the conditions under which these organisms survive and proliferate. To address this challenge, buoyant masses of live and dead Escherichia coli O157:H7 and Listeria innocua were measured using Archimedes, a commercially available suspended microchannel resonator (SMR). Cell growth was monitored with Archimedes by observing increased cell concentration and buoyant mass values of live growing bacteria. These growth data were compared to optical density measurements obtained with a Bioscreen system. We observed buoyant mass measurements with Archimedes at cell concentrations between 10⁵ and 10⁸ cells/ml, while growth was not observed with optical density measurements until the concentration was 10⁷ cells/ml. Buoyant mass measurements of live and dead cells with and without exposure to hydrogen peroxide stress were also compared; live cells generally had a larger buoyant mass than dead cells. Additionally, buoyant mass measurements were used to determine cell density and total mass for both live and dead cells. Dead E. coli cells were found to have a larger density and smaller total mass than live E. coli cells. In contrast, density was the same for both live and dead L. innocua cells, while the total mass was greater for live than for dead cells. These results contribute to the ongoing challenge to further develop existing technologies used to observe cell populations at low concentrations and to measure unique physical features of cells that may be useful for developing future diagnostics.     

A First Record of Obligate Halophilic Aspergilli from the Dead Sea

The isolation of obligate halophilic aspergilli from the Dead Sea and the range of salt tolerance of halophilic fungi isolated, are reported here for the first time. The mycobiota of the Dead Sea isolated in this study, was dominated by Aspergillus and Penicillium species; Cladosporium were found in lesser numbers. All three genera were obtained from the water sample; however, Aspergillus was the only genus obtained from the sediment. There was significant difference in growth of each isolate at different salt concentrations and intraspecies analysis revealed dissimilarity in response of strains to different salt concentrations in the growth medium The isolates were euryhaline, with halotolerance up to 20–25% solar salt, Aspergillus and Penicillium species showing a higher level of halotolerance, as compared to that of Cladosporium. Halophilic fungi were found in greater numbers in the sediment sample as compared to that in the water sample. Penicillium and Cladosporium species were exclusively facultative halophiles, while some species of Aspergillus were facultative halophiles. All the obligate halophiles isolated, belonged to the genus Aspergillus and were identified as A. penicillioides and A unguis, the latter being a first record of the species from the Dead Sea.     

Using Environmental DNA to Estimate the Distribution of an Invasive Fish Species in Ponds

Knowledge of the presence of an invasive species is critical to monitoring the sustainability of communities and ecosystems. Environmental DNA (eDNA), DNA fragments that are likely to be bound to organic matters in the water or in shed cells, has been used to monitor the presence of aquatic animals. Using an eDNA-based method, we estimated the presence of the invasive bluegill sunfish, Lepomis macrochirus, in 70 ponds located in seven locales on the Japanese mainland and on surrounding islands. We quantified the concentration of DNA copies in a 1 L water sample using quantitative real-time polymerase chain reaction (qPCR) with a primer/probe set. In addition, we visually observed the bluegill presence in the ponds from the shoreline. We detected bluegill eDNA in all the ponds where bluegills were observed visually and some where bluegills were not observed. Bluegills were also less prevalent on the islands than the mainland, likely owing to limited dispersal and introduction by humans. Our eDNA method simply and rapidly detects the presence of this invasive fish species with less disturbance to the environment during field surveys than traditional methods.     

有创-无创序贯机械通气与传统有创机械通气治疗重症肺炎合并呼吸衰竭的疗效对比研究及院内死亡的危险因素分析

摘要 目的：对比传统有创机械通气与有创-无创序贯机械通气治疗重症肺炎合并呼吸衰竭的疗效,并分析院内死亡的危险因素。方法：病例源于成都市第五人民医院2019年8月~2022年1月期间收治的重症肺炎合并呼吸衰竭患者120例,根据机械通气治疗方案不同将患者分为两组,常规组（n=60,传统有创机械通气）和序贯组（n=60,有创-无创序贯机械通气）。对比两组临床指标、血气分析指标、炎症因子指标、心功能指标。120例患者根据院内死亡情况分为死亡组（n=31）和存活组（n=89）,统计死亡组和存活组的一般资料情况,多因素Logistic分析院内死亡的危险因素。结果：序贯组的重症监护室（ICU）住院时间、总住院时间、有创通气时间、机械通气总时间均短于常规组,院内死亡率低于常规组（P<0.05）。序贯组治疗后氧分压（PaO₂）、pH值高于常规组,二氧化碳分压（PaCO₂）低于常规组（P<0.05）。序贯组治疗后白介素-10（IL-10）高于常规组,肌钙蛋白Ｉ（cTnＩ）、Ｂ型氨基端利钠肽原（NT-proBNP）、可溶性髓系细胞触发受体-１（sTREM-1）、肿瘤坏死因子-α（TNF-α）低于常规组（P<0.05）。重症肺炎合并呼吸衰竭院内死亡与急性生理与慢性健康评分Ⅱ( APACHEⅡ评分) 、多重耐药菌感染、白蛋白（Alb）、合并基础疾病、多器官功能障碍评分（MODS评分）、年龄、肺炎严重程度评分（PSI）、红细胞比容（Hct）、社区获得性肺炎评分（CURB-65）、空腹血糖（FBG）有关（P<0.05）。合并基础疾病、存在多重耐药菌感染、高CURB-65评分、高PSI评分、高MODS评分、低Alb、高APACHEⅡ评分是重症肺炎合并呼吸衰竭院内死亡的危险因素（P<0.05）。结论：有创-无创序贯机械通气可缩短ICU住院时间、总住院时间、有创通气时间、机械通气总时间,降低院内死亡率,调节血气分析指标、心功能指标和炎症因子水平。重症肺炎合并呼吸衰竭患者院内死亡受到高MODS评分、低Alb、高PSI评分、合并基础疾病、高APACHEⅡ评分、存在多重耐药菌感染、高CURB-65评分等多种因素的影响。     

A thermophilic amyloglucosidase fromHalobacterium sodomense,a halophilic bacterium from the Dead Sea

Halobacterium sodomense, a halophilic bacterium from the Dead Sea, degraded starch to glucose by means of an extracellular amyloglucosidase with a temperature optimum of around 65°C in the presence of 1.4 M NaCl, and around 75°C in the presence of 3.9 M NaCl. The enzyme required salt concentrations higher than 1 M for optimal activity, NaCl, KCl, and MgCl₂ being equally suitable as activators. The optimum pH was 7.5.H. sodomense culture supernatants showed only a very low maltose degrading activity. H. sodomense excreted amyloglucosidase constitutively, and relatively high activities were found in cultures grown in the absence of starch; when glucose was added to the growth medium, the amount of enzyme excreted into the medium decreased.     

Evaluation of Extracellular,High-affinity β-N-acetylglucosaminidase Measurements from Freshwater Lakes: An Enzyme Assay to Estimate Protistan Grazing on Bacteria and Picocyanobacteria

Protistan community grazing rates upon both bacterioplankton and autotrophic picoplankton were estimated using fluorescently-labeled prey and by measurement of extracellular hydrolysis of 4-methylumbelliferyl (MUF) beta-N-acetylglucosaminide in a eutrophic reservoir and an oligo-mesotrophic lake during phytoplankton blooms. In addition, enzyme methods were optimized in bacterivorous flagellate cultures by two enzyme assays, based on fluorometric detection of protistan digestive activity, which were compared and calibrated independently against flagellate bacterivory. Enzymatic hydrolyses of MUF beta-N,N',N'-triacetylchitotriose and MUF beta-N-acetylglucosaminide were measured in cell-free (sonicated) and whole-cell (unsonicated) samples. The hydrolysis of both substrates, using the whole-cell enzyme assay at in situ pH, was correlated significantly with total grazing rate of Bodo saltans. Thus the whole-cell enzyme assay with MUF beta-N-acetylglucosaminide was used for freshwater samples. High-affinity (Km < 1 μmol l-1) and low-affinity (Km > 100μmol l-1) enzymes were distinguished kinetically in most samples from both systems studied. Activities (Vmax) of the high-affinity enzyme varied from 0.24 to 1.43 nmol l-1 h-1. Protistan community grazing on bacterioplankton was in the range of 0.15-1.36 μg C l-1 h-1 both for lake and reservoir, the differences being observed in grazing on picocyanobacteria (lake, 0.03-0.22 μg C l-1 h-1; reservoir, 0.35-1.56 μg C l-1 h-1). The enzyme activities were correlated significantly with the protistan grazing both on bacterioplakton (rs=0.62, P<0.001) and total procaryotic picoplankton (the sum of organic carbon grazed from bacteria and picocyanobacteria, rs=0.73, P<0.001) in the eutrophic reservoir. Weaker relationships (rs=0.42) with a lower slope were found for the oligo-mesotrophic lake. Ingestion rate studies are time-consuming and the digestive enzyme assay with MUF beta-N-acetylglucosaminide presents a rapid alternative for estimating total protistan prokaryotic picoplanktivory in freshwaters.     

Comparison of Invasive Blood Pressure Measurements from the Caudal Ventral Artery and the Femoral Artery in Male Adult SD and Wistar Rats

Background and Purpose

Studies have suggested that the caudal ventral artery is a potential site for continuous arterial blood pressure monitoring in rats. However, the agreement of mean arterial pressure values between the femoral artery and the caudal ventral artery has not been investigated. This study was performed to identify whether the caudal ventral artery could be safely used for continuous blood pressure monitoring as an alternative site to the femoral artery.

Methods

Rats were randomized into four groups: Sprague Dawley rats under normothermia; Wistar rats under normothermia; Sprague Dawley rats under hypothermia; Wistar rats under hypothermia. Each rat underwent simultaneous monitoring of blood pressure using femoral artery and caudal ventral artery catheterization during a stable hemodynamic state and three periods of acute severe hemodynamic changes. The effects of rat strain, rectal temperature, experimental time course and hemodynamic factors on pressure gradients, the concordance of mean arterial pressure values between the femoral artery and the caudal ventral artery, and the rates of distal ischemia after surgery were determined.

Results

There was a significant difference in the rate of distal ischemia between femoral and caudal ventral arteries after catheterization (25% vs 5%, P<0.05). The overall mean gradient and the mean gradient under a steady hemodynamic state were 4.9±3.7 mm Hg and 5.5±2.5 mm Hg, respectively. The limits of agreement (bias±1.96 SD) were (−2.5 mm Hg, 12.3 mm Hg) and (-0.5 mm Hg, 10.5 mm Hg), respectively. Although the concordance decreased during the first 30 sec of each period of severe hemodynamic changes, the degree of agreement was acceptable regardless of the effects of rat strain and rectal temperature.

Conclusions

Based on the degree of agreement and the safety of catheterization, the caudal ventral artery may be a preferred site for continuous arterial blood pressure monitoring without acute severe hemodynamic changes.     

Role of Ultrasonography in the Differential Diagnosis of Thyrotoxicosis: A Non Invasive,Cost-Effective,and Widely Available But Underutilized Diagnostic Tool

ObjectiveTo explore the use of ultrasonography as a diagnostic alternative to the traditional “gold standard” imaging study of nuclear scintigraphy in the evaluation of thyrotoxicosis.MethodsWe review the relevant literature and share our own experience to highlight the promising role of ultrasonography in thyrotoxicosis. In addition, we present a diagnostic algorithm suggesting liberal use of ultrasonography in the evaluation of thyrotoxicosis.ResultsUltrasonography has proved effective not only in the differentiation of Graves disease from other types and causes of thyrotoxicosis but also in the detection of subtle thyroid nodules. The latter role is emphasized in light of the recent observation of an increased risk of occurrence of papillary thyroid carcinoma in patients with Graves disease.ConclusionUltrasonography is a cost-effective, noninvasive, portable, and safe imaging modality in the evaluation of thyrotoxicosis, both for physiologic assessment and for detection of nonpalpable thyroid cancers that may elude identification on physical examination and nuclear imaging studies. Although thyroid scintigraphy remains a standard radiologic study, thyroid ultrasonography can be a practical alternative in many cases and the primary imaging modality in some situations such as during pregnancy and lactation and for evaluation and management of amiodarone-induced thyrotoxicosis. (Endocr Pract. 2012;18: 567-578)     

Use of Nitrifier Activity Measurements To Estimate the Efficiency of Viable Nitrifier Counts in Soils and Sediments

A procedure for estimating the efficiency of the most-probable number (MPN) technique for counting ammonium-oxidizing bacteria was tested on sediments and soils collected from Delaware Inlet, Nelson, New Zealand. The procedure involved estimating the nitrifier populations required to produce observed activities and comparing these estimates with the MPN-countable populations. MPN counts ranged between 0.15 × 10³ to 3.0 × 10³ cells g⁻¹ in sediments and between 4.4 × 10³ to 19 × 10³ cells g⁻¹ in soils. These counts were only 0.1 to 5.0% of the estimated populations that would be required to produce the observed activity. Similar efficiency calculations were made for data already in the literature, and these calculations gave much higher percentages. Thus, we concluded that for the soils and sediments we studied, the MPN counting technique greatly underestimated the populations present and that the efficiency calculation could be used as a counting efficiency index.     

吗啡联合有创呼吸机辅助通气治疗重症急性左心衰竭的疗效

目的:探讨吗啡联合有创呼吸机辅助通气治疗重症急性左心衰竭的临床疗效。方法:将2012年5月-2014年5月我院收治的82例重症急性左心衰竭患者随机分为观察组和对照组,各41例。两组患者均因常规药物治疗无效再行经口气管插管机械通气及对症治疗,观察组在此基础上加用吗啡静脉注射。对比两组患者的体征、症状、动血气指标及临床疗效。结果:两组治疗后血压、心率(HR)、呼吸频率(BR)、血气指标明显优于治疗前,比较差异有统计学意义(P0.05),且观察组各指标改善明显优于对照组,比较差异有统计学意义(P0.05);治疗后观察组总有效率为92.7%(38/41),高于对照组的82.9%(34/41),比较差异有统计学意义(P0.05)。结论:吗啡联合有创呼吸机辅助通气治疗重症急性左心衰竭可有效缓解患者症状,改善临床体征,临床疗效显著,值得推广。     

Early Detection of Myocardial Bioenergetic Deficits: A 9.4 Tesla Complete Non Invasive 31P MR Spectroscopy Study in Mice with Muscular Dystrophy

Background

Duchenne muscular dystrophy (DMD) is the most common fatal form of muscular dystrophy characterized by striated muscle wasting and dysfunction. Patients with DMD have a very high incidence of heart failure, which is increasingly the cause of death in DMD patients. We hypothesize that in the in vivo system, the dystrophic cardiac muscle displays bioenergetic deficits prior to any functional or structural deficits. To address this we developed a complete non invasive 31P magnetic resonance spectroscopy (31P MRS) approach to measure myocardial bioenergetics in the heart in vivo.

Methods and Results

Six control and nine mdx mice at 5 months of age were used for the study. A standard 3D -Image Selected In vivo Spectroscopy (3D-ISIS) sequence was used to provide complete gradient controlled three-dimensional localization for heart 31P MRS. These studies demonstrated dystrophic hearts have a significant reduction in PCr/ATP ratio compare to normal (1.59±0.13 vs 2.37±0.25, p<0.05).

Conclusion

Our present study provides the direct evidence of significant cardiac bioenergetic deficits in the in vivo dystrophic mouse. These data suggest that energetic defects precede the development of significant hemodynamic or structural changes. The methods provide a clinically relevant approach to use myocardial energetics as an early marker of disease in the dystrophic heart. The new method in detecting the in vivo bioenergetics abnormality as an early non-invasive marker of emerging dystrophic cardiomyopathy is critical in management of patients with DMD, and optimized therapies aimed at slowing or reversing the cardiomyopathy.