Triangulation supports agricultural spread of the Transeurasian languages.

The origin and early dispersal of speakers of Transeurasian languages-that is, Japanese, Korean, Tungusic, Mongolic and Turkic-is among the most disputed issues of Eurasian population history1-3. A key problem is the relationship between linguistic dispersals, agricultural expansions and population movements4,5. Here we address this question by 'triangulating' genetics, archaeology and linguistics in a unified perspective. We report wide-ranging datasets from these disciplines, including a comprehensive Transeurasian agropastoral and basic vocabulary; an archaeological database of 255 Neolithic-Bronze Age sites from Northeast Asia; and a collection of ancient genomes from Korea, the Ryukyu islands and early cereal farmers in Japan, complementing previously published genomes from East Asia. Challenging the traditional 'pastoralist hypothesis'6-8, we show that the common ancestry and primary dispersals of Transeurasian languages can be traced back to the first farmers moving across Northeast Asia from the Early Neolithic onwards, but that this shared heritage has been masked by extensive cultural interaction since the Bronze Age. As well as marking considerable progress in the three individual disciplines, by combining their converging evidence we show that the early spread of Transeurasian speakers was driven by agriculture.

Multiple lines of evidence of early goose domestication in a 7,000-y-old rice cultivation village in the lower Yangtze River, China.

Poultry are farmed globally, with chicken (Gallus gallus domesticus) being the leading domesticated species. Although domestic chicken bones have been reported from some Early Holocene sites, their origin is controversial and there is no reliable domestic chicken bone older than the Middle Holocene. Here, we studied goose bones from Tianluoshan­a 7,000-y-old rice cultivation village in the lower Yangtze River valley, China­using histological, geochemical, biochemical, and morphological approaches. Histological analysis revealed that one of the bones was derived from a locally bred chick, although no wild goose species breed in southern China. The analysis of oxygen-stable isotope composition supported this observation and further revealed that some of the mature bones were also derived from locally bred individuals. The nitrogen-stable isotope composition showed that locally bred mature birds fed on foods different from those eaten by migrant individuals. Morphological analysis revealed that the locally bred mature birds were homogenous in size, whereas radiocarbon dating clearly demonstrated that the samples from locally bred individuals were ∼7,000 y old. The histological, geochemical, biochemical, morphological, and contextual evidence suggest that geese at Tianluoshan village were at an early stage of domestication. The goose population appears to have been maintained for several generations without the introduction of individuals from other populations and may have been fed cultivated paddy rice. These findings indicate that goose domestication dates back 7,000 y, making geese the oldest domesticated poultry species in history.

Is preoperative glenoid defect size a reliable indicator of postoperative recurrence after arthroscopic Bankart repair in teenage competitive athletes?

BACKGROUND: Being younger than 20 years of age at the time of arthroscopic Bankart repair (ABR) is known to be one of the most important risk factors for postoperative recurrence of instability. When deciding on the appropriate surgical approach, surgeons generally consider only the size of a critical glenoid defect, and most of them do not take into account factors such as the size of bone fragments and possible bone union after arthroscopic bony Bankart repair (ABBR). Therefore, this retrospective study aimed to clarify the risk factors for postoperative recurrence after ABR in teenage competitive athletes by focusing on glenoid rim morphologies and bone union. METHODS: Participants were 115 teenage competitive athletes without a capsular injury who underwent primary ABR for chronic traumatic anterior instability and were followed up for a minimum of 2 years. Possible risk factors for postoperative recurrence were investigated by univariate and multivariate analysis. In shoulders with a glenoid defect and bone fragment, the influence of glenoid defect size and bone fragment size on bone union after ABBR was also investigated. RESULTS: Postoperative recurrence was seen in 16 patients (13.9%). Regarding glenoid defect size, recurrence was seen in 1 (3.2%) of 31 shoulders with a glenoid defect smaller than 5% (including those with a normal glenoid), 15 (22.1%) of 68 shoulders with a glenoid defect of 5%-20%, and 0 (0%) of 16 shoulders with a glenoid defect of 20% or larger (P = .009). Regarding bone union, recurrence was seen in 4 (6.9%) of 58 shoulders with complete or partial bone union after ABBR and 8 (40%) of 20 shoulders with nonunion or disappearance of the bone fragment (P = .001). Regarding bone fragment size, recurrence was seen in 12 (20.7%) of 58 shoulders with a small or no bone fragment (<7.5%) and in 3 (8.6%) of 35 shoulders with a large bone fragment (≥7.5%; P = .154). Multivariate analysis identified non-union or disappearance of the bone fragment after ABBR as a significant risk factor for recurrence. Complete or partial bone union was seen in 25 (58.1%) of 43 shoulders with a small bone fragment (<7.5%) and 33 (94.3%) of 35 shoulders with a large bone fragment (≥7.5%; P < .001). CONCLUSIONS: In teenage competitive athletes, bone union after ABBR affects postoperative recurrence after ABR, regardless of the preoperative glenoid defect size, and bone union rate after ABBR is significantly influenced by bone fragment size.

Utility of ultrasonography in the diagnosis of lumbar spondylolysis in adolescent patients.

BACKGROUND: This study aims to investigate the utility of the Doppler effect on ultrasonography for the diagnosis of very early- and early-stage lumbar spondylolysis in adolescent patients. METHODS: In total, 76 adolescent patients with acute and subacute low back pain were prospectively enrolled, with 46 having lumbar spondylolysis and the remaining 30 having low back pain without spondylolysis. MRI and/or computed tomograms scans revealed very early- and early-stage lumbar spondylolysis. Furthermore, positive Doppler findings in ultrasonography around the area from the facet joint to the laminae were investigated. RESULTS: There were no significant differences in age (p > 0.99) and body mass index (p = 0.11) between cases with and without spondylolysis. Very early- and early-stage spondylolysis were observed in 27.6% and 72.4% of patients, respectively. Positive power Doppler was 91.3% and 33.3% in cases with and without spondylolysis, respectively, which was significantly higher in spondylolysis (p < 0.001). The sensitivity and specificity of this positive power Doppler were 91.4% and 66.7%, respectively. Furthermore, the rate of positive power Doppler was significantly higher in early-stage spondylolysis (p = 0.02), with 75.0% and 97.6% sensitivity in very early- and early-stage spondylolysis, respectively. CONCLUSIONS: A positive Doppler effect on ultrasonography is effective for screening very early- and early-stage spondylolysis in adolescent patients in an outpatient clinic.

An intermediate crocodylian linking two extant gharials from the Bronze Age of China and its human-induced extinction.

A solid phylogenetic framework is the basis of biological studies, yet higher level relationships are still unresolved in some major vertebrate lineages. One such group is Crocodylia, where the branching pattern of three major families (Alligatoridae, Crocodylidae and Gavialidae) has been disputed over decades due to the uncertain relationship of two slender-snouted lineages, gavialines and tomistomines. Here, we report a bizarre crocodylian from the Bronze Age of China, which shows a mosaic of gavialine and tomistomine features across the skeleton, rendering support to their sister taxon relationship as molecular works have consistently postulated. Gavialine characters of the new Chinese crocodylian include a novel configuration of the pterygoid bulla, a vocal structure known in mature male Indian gharials. Extinct gavialines have repeatedly evolved potentially male-only acoustic apparatus of various shapes, illuminating the deep history of sexual selection on acoustic signalling in a slender-snouted group of crocodylians. Lastly, a cutmark analysis combined with accelerator mass spectrometry (AMS) radiocarbon dating of bone remains demonstrated that two individuals from Shang and Zhou dynasties in Guangdong, China, suffered head injuries and decapitation. Archaeological evidence together with historical accounts suggests the human-induced extinction of this unique crocodylian only a few hundred years ago.

Modern human teeth unearthed from below the ∼128,000-year-old level at Punung, Java: A case highlighting the problem of recent intrusion in cave sediments.

The emergence of modern humans in the eastern edge of the Eurasian Continent is debated between two major models: early (∼130-70 ka) and late (∼50 ka) dispersal models. The former view is grounded mainly on the claims that several cave sites in Southeast Asia and southern China yielded modern human fossils of those early ages, but such reports have been disputed for the lack of direct dating of the human remains and insufficient documentation of stratigraphy and taphonomy. By tracing possible burial process and conducting direct dating for an early Late Pleistocene paleontological site of Punung III, East Java, we here report a case that demonstrates how unexpected intrusion of recent human remains into older stratigraphic levels could occur in cave sediments. This further highlights the need of direct dating and taphonomic assessment before accepting either model. We also emphasize that the state of fossilization of bones and teeth is a useful guide for initial screening of recent intrusion and should be reported particularly when direct dating is unavailable. Additionally, we provide a revised stratigraphy and faunal list of Punung III, a key site that defines the tropical rainforest Punung Fauna during the early Late Pleistocene of the region.

Conservative treatment of spondylolysis involving exercise initiated early and sports activities resumed with a lumbar-sacral brace.

BACKGROUND: Conservative treatment including the use of a brace and cessation of sports activities is known to be effective in spondylolysis. However, there is some controversy regarding [1] the type of brace, and [2] the endpoint for bracing, and [3] the timing of resumption of sports activities. The aim of the current study was to investigate the appropriateness of early exercise and resumption of sports activities with a lumbar-sacral brace in very early- and early-stage lumbar spondylolysis. METHODS: This prospective cohort study enrolled 45 patients with very early- and early-stage lumbar spondylolysis. A lumbar-sacral brace, structured to be a hard brace in the back and a mesh brace in the front, was used. Isometric exercises/stretching was started immediately after the initial visit. The number of patients for whom bone fusion was fully achieved on CT and the disappearance of signal intensity change on MRI were investigated. RESULTS: In 12 cases of very early-stage spondylolysis, the average elapsed time until signal intensity disappearance as confirmed on MRI was 4.3 ± 1.6 months. Bony union on CT was confirmed in all cases. In 33 cases of early spondylolysis, signal intensity change disappeared on MRI within 5.2 ± 2.4 months. The rate of bony union was 94.3%, and the average period required to achieve bony union observable on CT was 3.7 ± 1.0 months. CONCLUSIONS: Sufficient bony union can be achieved by conservative treatment with early exercise and a lumbar-sacral brace in cases of very early and early spondylolysis.

Penetration factor and indoor deposition rate of elementary and particulate iodine in a Japanese house for assessing the effectiveness of sheltering for radiation exposures.

Sheltering is one of the countermeasures for protection against radiation exposures in nuclear accidents. The effectiveness of sheltering is often expressed by the reduction factor, that is the ratio of the indoor to the outdoor cumulative radioactivity concentrations or doses. The indoor concentration is mainly controlled by the air exchange rate, penetration factor, and indoor deposition rate. The penetration factor and indoor deposition rate depend on the surface and the materials and structure of windows and doors as it is these openings in the building envelope that control penetration. We investigated experimentally these parameters of I2and particles. The experiment was performed in two apartment houses, three single-family houses, and chambers. The obtained penetration factor ranged from 0.3 to 1 for particles of 0.3-1µm and 0.15-0.7 for I2depending on the air exchange rate. The indoor deposition rate for a house room ranged from 0.007 to 0.2 h-1for particles of 0.3-1µm and 0.2-1.5 h-1for I2.

Benefits of bone graft augmentation to arthroscopic Bankart repair for recurrent anterior shoulder instability with glenoid bone loss.

PURPOSE: Glenoid bone loss contributes to recurrent instability after arthroscopic Bankart repair alone. With significant glenoid bone loss, better results have been reported after arthroscopic Bankart repair with glenoid arc reconstruction. However, no reports compare augmentation using bone graft with non-augmentation for glenoid bone loss. The purpose of this study was to assess clinical results of an arthroscopic Bankart repair with or without arthroscopic bone graft augmentation. It was hypothesized that such bone graft augmentation would restore shoulder stability, and lead to excellent outcomes. METHODS: Of 552 patients treated for anterior glenohumeral instability with arthroscopic Bankart repair, 68 met this study's inclusion criteria of glenoid bone loss over 20% and follow-up of at least 2 years. Patients were divided into 2 groups based on whether with bone graft augmentation for glenoid bone loss [Group A: n = 35, median age; 21 years (range 13-72 years)], or not (Group B: n = 33, median age; 21 years (range 13-50 years)]. For grafting, either autologous iliac bone or artificial bone made of hydroxyapatite was used. Rowe score, recurrence rate, and return to sport were used to assess the results. RESULTS: Mean Rowe score was 95.0 (SD 10.6) in Group A and 69.7 (SD 27.2) in Group B (p < 0.05). The recurrence rate was 2.9% (1/36) in Group A and 48.5% (16/33) in Group B (p < 0.05). Regarding contact/collision athletes, 24 were contained in Group A and 22 in Group B. Of the patients with recurrence in Group B, 13 (59.1%) were contact/collision athletes. Finally, 50% of the contact/collision sports athletes for both groups returned to their sports at the same as pre-injury level. Of the 11 patients who returned to the same level of contact/collision sports in Group B, seven returned with residual instability. Nine athletes in Group A and 3 in Group B quit their sports for personal or social reasons. CONCLUSIONS: Bone graft augmentation was beneficial when used with Arthroscopic Bankart repair for recurrent anterior shoulder instability with glenoid bone loss. Especially, for recurrent anterior shoulder instability with glenoid bone loss in contact/collision sports athletes, bone graft augmentation should be strongly considered as beneficial. LEVEL OF EVIDENCE: Level IV.

On-lay grafting of a calcium hydroxyapatite bone substitute: A preliminary animal experimental study.

BACKGROUND: Bone substitutes are widely accepted for various clinical applications. However, the usage is predominantly intraosseous implantation, whereas extraosseous on-lay grafting is rare and lacks scientific evidence. The purpose of this study is to elucidate whether osteoconduction occurs in on-lay grafted bone substitute. METHODS: Custom-made interconnected porous calcium hydroxyapatite ceramic (IPCHA) was on-lay grafted with screw or anchor fixation (S- and A-groups, respectively) at the anterior aspect of the femur of skeletally mature Japanese white rabbits. At 3, 6 and 12 weeks postoperatively, 4 samples for each time point and each group were evaluated by microfocus computed tomography (micro-CT) and histology. RESULTS: Volume-rendered three-dimensional micro-CT images showed a high-density calcified area infiltrating IPCHA from the femoral cortex as of 6 weeks. When quantified, the calcified volume per unit volume first showed no difference between the two groups at 3 weeks but increased over time, and became significantly greater in the S-group than in the A-group (p = 0.012 and 0.004 at 6 and 12 weeks, respectively). Histologically, IPCHA pores were first occupied by fibrous tissue at 3 weeks; then, the pores adjacent to the femoral cortex were gradually replaced by bony tissue as of 6 weeks for both fixations. CONCLUSIONS: IPCHA allowed new bone formation inside the material even though it was implanted in an on-lay fashion on the cortical bone. Our results suggested that on-lay grafted IPCHA exerted its osteoconductivity well, with more new bone forming in screw-fixated samples than in anchor-fixated samples.

Morphological evaluation of the scapula by three-dimensional computed tomography to elucidate shoulder joint function.

INTRODUCTION: There are many variations of scapula morphology. Evaluation of scapula morphology is necessary to elucidate shoulder joint movement. We aimed to analyze the bone morphology of various parts constituting the scapula to identify certain conserved features among them. MATERIALS AND METHODS: Thirty-one healthy individuals were the subjects. We created a scapular three-dimensional (3D) model using computed tomography (CT). X and Y axes were set on a glenoid surface. We measured the approximate plane of the upper and lower scapular bodies and scapular spine and the central axis of the coracoid and acromion. The anatomical position of the scapular spine, upper and lower bodies, coracoid, and acromion was measured. The positional relationship between the upper and lower bodies and scapular spine and the coracoid and acromion was evaluated. The average angle of the upper and lower scapular bodies and the scapular spine with respect to the XZ plane was calculated. The average angle of the coracoid and acromion with the X-axis on the XY plane and Z-axis on the YZ plane was calculated. RESULTS: On the XY plane, approximate planes of the upper and lower part of the body and the scapular spine were significantly correlated to each inclination. On the XY plane, inclinations of the central axes of the coracoid and acromion were significantly correlated. CONCLUSIONS: The findings revealed for the first time the correlation between the inclinations of the scapular body and scapular spine and the inclination angle between the coracoid and acromion.

Interpretation of bulk nitrogen and carbon isotopes in archaeological foodcrusts on potsherds.

RATIONALE: Foodcrust, the charred deposit adhering to the surface of containers, is a possible source of information on the function of ancient vessels and the subsistence of prehistoric humans. While the carbon isotope ratios in those materials are useful in detecting the usage of C4 plants, the reliability of nitrogen isotopic signatures has not been fully investigated. METHODS: The validity of bulk nitrogen isotope ratios has previously been investigated in coastal or riverine environments, where multiple resources from terrestrial and aquatic ecosystems were available, but not in terrestrial settings which provide a simpler mixing of terrestrial animals and plants. Hence, we conducted an exhaustive study on charred deposits on potsherds at two inland archaeological sites belonging to prehistoric Jomon hunter-gathers in central Japan, focusing on δ15 N values and atomic N/C ratios determined using an isotope ratio mass spectrometer and an elemental analyzer, respectively. RESULTS: For both sites, the δ15 N values showed significant correlations with the N/C ratios among samples from the inner surface, suggesting that these have recorded animal contribution. Furthermore, previous studies of Neolithic pottery from North Europe and Far East Russia bearing strong marine signatures had shown reasonably higher δ15 N values and N/C ratios in comparison with our data from terrestrial settings. On the other hand, some charred materials probably originating from plant starch showed lower values with both parameters. Samples from the outer surface produced less meaningful isotopic and elemental ratios altered by a thermal effect and/or contamination from soot. CONCLUSIONS: When the samples of foodcrusts were selected carefully from the inner surface, bulk nitrogen isotopes and N/C ratios reflect the composition of what was cooked or processed in containers. This will provide useful information for understanding the human adaptation from the Late Pleistocene to the Holocene in conjunction with residual lipid analyses.

Societal perceptions and lived experience: Infant feeding practices in premodern Japan.

OBJECTIVES: A change in how children were treated and valued occurred in premodern Japan, as popularized ideas of an inheritance-based family system led to more careful and affectionate child-rearing practices by lower social-status groups. A number of books were written, advising that breastfeeding should last approximately 3 years. The objective of this study is to reconstruct and compare breastfeeding and weaning practices before and after the transition, to illuminate the impact of documented changes in child-rearing practices on subadults' lived experience. MATERIALS AND METHODS: Carbon and nitrogen stable isotopic data were obtained from 40 subadult skeletons excavated from the Sakai Kango Toshi 871 (SKT871) site (late 17th-19th century, Osaka, Japan). Isotopic results from SKT871 were compared with previously reported results from the Hitotsubashi site (AD 1657-1683, Tokyo, Japan). Hitotsubashi and SKT871 represent urban populations of lower status before and after the transition of societal perception of subadults. RESULTS: The most probable age at the end of weaning reconstructed in SKT871 was 1.9 years (1.4-2.7 years with a 95% credible interval) and was lower than that in Hitotsubashi (2.1-4.1 years with a 95% credible interval). DISCUSSION: The age at the end of weaning became younger after the transition of societal perception toward subadults, and this younger weaning age is inconsistent with written recommendations for the duration of weaning in premodern Japan. It is possible that an increased need for inheritors under the inheritance-based family system led to earlier weaning and shorter inter-birth intervals, but authorities recommended an ideal practice of a longer breastfeeding period.

Amino acid 15 N analysis reveals change in the importance of freshwater resources between the hunter-gatherer and farmer in the Neolithic upper Tigris.

OBJECTIVE: The inhabitants of several sites in the Upper Tigris Valley, such as Hakemi Use, domesticated animals and cereals during the Pottery Neolithic period, while the inhabitants in this valley were hunter-gatherers in the Pre-Pottery Neolithic period, consuming freshwater and terrestrial food resources. However, there is considerable uncertainty surrounding whether or not changes in dietary food composition accompanied the shift in food production away from foraging. In order to reveal the impact of the development of agriculture on the human diet over the Pre-Pottery and Pottery Neolithic periods in this region, we analyzed the isotopic compositions of amino acids from the farmers at the Hakemi Use Pottery Neolithic site, and compared them with those from the Pre-Pottery hunter-gatherers in the close region. MATERIALS AND METHODS: Herein, we report the nitrogen isotopic compositions of amino acids, as well as both carbon and nitrogen isotopic compositions of bulk collagen, from human and faunal remains collected from Hakemi Use. RESULTS: Whereas freshwater resources were consumed by hunter-gatherers in this region during the Pre-Pottery period, the δ15 N values of glutamic acid (δ15 NGlu ) and phenylalanine (δ15 NPhe ) suggest that freshwater food resources were rarely consumed by inhabitants following the development of agriculture. DISCUSSION: Despite living in similar settings by the Tigris as its inhabitants during the Pre-Pottery period, the farmers of the Pottery Neolithic period depended less on freshwater resources for their diets relative to the hunter-gatherers of the Pre-Pottery Neolithic period.

A Probabilistic Approach to Assess External Doses to the Public Considering Spatial Variability of Radioactive Contamination and Interpopulation Differences in Behavior Pattern.

Dose assessment is an important issue from the viewpoints of protecting people from radiation exposure and managing postaccident situations adequately. However, the radiation doses received by people cannot be determined with complete accuracy because of the uncertainties and the variability associated with any process of defining individual characteristics and in the dose assessment process itself. In this study, a dose assessment model was developed based on measurements and surveys of individual doses and relevant contributors (i.e., ambient dose rates and behavior patterns) in Fukushima City for four population groups: Fukushima City Office staff, Senior Citizens' Club, Contractors' Association, and Agricultural Cooperative. In addition, probabilistic assessments were performed for these population groups by considering the spatial variability of contamination and interpopulation differences resulting from behavior patterns. As a result of comparison with the actual measurements, the assessment results for participants from the Fukushima City Office agreed with the measured values, thereby validating the model and the approach. Although the assessment results obtained for the Senior Citizens' Club and the Agricultural Cooperative differ partly from the measured values, by addressing further considerations in terms of dose reduction effects due to decontamination and the impact of additional exposure sources in agricultural fields, these results can be improved. By contrast, the measurements obtained for the participants from the Contractors' Association were not reproduced well in the present study. To assess the doses to this group, further investigations of association members' work activities and the related dose reduction effects are needed.

Loss of renal SNX5 results in impaired IDE activity and insulin resistance in mice.

AIMS/HYPOTHESIS: We hypothesised that renal sorting nexin 5 (SNX5) regulates the insulin-degrading enzyme (IDE) and, thus, circulating insulin levels. We therefore studied the dynamic interaction between SNX5 and IDE in human renal proximal tubule cells (hRPTCs), as well as in rat and mouse kidneys. METHODS: The regulation of IDE by SNX5 expressed in the kidney was studied in vitro and in vivo. Snx5 or mock siRNA was added to immortalised hRPTCs (passage <20) in culture or selectively infused, via osmotic mini-pump, into the remnant kidney of uninephrectomised mice and rats. RESULTS: SNX5 co-localised with IDE at the plasma membrane and perinuclear area of hRPTCs and in the brush border membrane of proximal tubules of human, rat, and mouse kidneys. Insulin increased the co-localisation and co-immunoprecipitation of SNX5 and IDE in hRPTCs. Silencing SNX5 in hRPTCs decreased IDE expression and activity. Renal-selective silencing of Snx5 (SNX5 protein: 100 ± 25 vs 29 ± 10, p < 0.05 [% of control]) in C57Bl/6J mice decreased IDE protein (100 ± 13 vs 57 ± 6, p < 0.05 [% of control]) and urinary insulin excretion, impaired the responses to insulin and glucose, and increased blood insulin and glucose levels. Spontaneously hypertensive rats (SHRs) had increased blood insulin and glucose levels and decreased renal SNX5 (100 ± 27 vs 29 ± 6, p < 0.05 [% of control]) and IDE (100 ± 5 vs 75 ± 4, p < 0.05 [% of control]) proteins, compared with normotensive Wistar-Kyoto (WKY) rats. Kidney Snx5-depleted WKY rats also had increased blood insulin and glucose levels. The expression of SNX5 and IDE was decreased in RPTCs from SHRs and hypertensive humans compared with cells from normotensive volunteers, indicating a common cause for hyperinsulinaemia and hypertension. CONCLUSIONS/INTERPRETATION: Renal SNX5 positively regulates IDE expression and function. This study is the first to demonstrate the novel and crucial role of renal SNX5 in insulin and glucose metabolism.

Characteristics of Carbapenemase-Producing Enterobacteriaceae in Wastewater Revealed by Genomic Analysis.

Wastewater is considered a major source of antibiotic-resistant bacteria released into the environment. Here, we characterized carbapenemase-producing Enterobacteriaceae (CPE) in wastewater by whole-genome analysis. Wastewater samples (n = 40) were collected from municipal wastewater treatment plants and hospital wastewater in Japan and Taiwan. Samples were screened for CPE using selective media, and the obtained isolates were sequenced using an Illumina MiSeq. The isolates (n = 45) included the following microorganisms: Klebsiella quasipneumoniae (n = 12), Escherichia coli (n = 10), Enterobacter cloacae complex (n = 10), Klebsiella pneumoniae (n = 8), Klebsiella variicola (n = 2), Raoultella ornithinolytica (n = 1), Citrobacter freundii (n = 1), and Citrobacter amalonaticus (n = 1). Among the 45 isolates, 38 harbored at least one carbapenemase-encoding gene. Of these, the blaGES (blaGES-5, blaGES-6, and blaGES-24) genes were found in 29 isolates. The genes were situated in novel class 1 integrons, but the integron structures were different between the Japanese (In1439 with blaGES-24 and In1440 with blaGES-5) and Taiwanese (In1441 with blaGES-5 and In1442 with blaGES-6) isolates. Other carbapenemase-encoding genes (blaVIM-1, blaNDM-5, blaIMP-8, blaIMP-19, and blaKPC-2) were found in one to three isolates. Notably, class 1 integrons previously reported among clinical isolates obtained in the same regions as the present study, namely, In477 with blaIMP-19 and In73 with blaIMP-8, were found among the Japanese and Taiwanese isolates, respectively. The results indicate that CPE with various carbapenemase-encoding genes in different genetic contexts were present in biologically treated wastewater, highlighting the need to monitor for antibiotic resistance in wastewater.

Improved Method for Isolation and Purification of Underivatized Amino Acids for Radiocarbon Analysis.

We have improved a method for isolation and purification of individual amino acids for compound-specific radiocarbon analysis (CSRA). To remove high-performance liquid chromatography (HPLC) eluent blanks from isolated amino acid fractions prior to the radiocarbon (Δ14C) measurement, each fraction was filtered through a membrane filter and then washed with diethyl ether twice. Radiocarbon measurements on standard amino acids processed and purified with the above method using elemental analyzer-accelerator mass spectrometry resulted in Δ14C values that were in strong agreement ( R2 = 0.998) with the original Δ14C value of each amino acid standard. From these measurements, we calculate dead and modern carbon contamination contributions as 1.2 ± 0.2 and 0.3 ± 0.1 µgC, respectively, which are consistent with direct assessments of HPLC procedural blanks of 1.0 ± 0.8 µgC per sample. These contamination constraints allow correction of measured Δ14C values for accurate and precise CSRA and are widely applicable to future archeological and biogeochemical studies.

Multiparameter Phenotypic Profiling in MCF-7 Cells for Assessing the Toxicity and Estrogenic Activity of Whole Environmental Water.

Multi-parameter phenotypic profiling of small molecules is a powerful approach to their toxicity assessment and identifying potential mechanisms of actions. The present study demonstrates the application of image-based multi-parameter phenotypic profiling in MCF-7 cells to assess the overall toxicity and estrogenic activity of whole environmental water. Phenotypic profiling of 30 reference compounds and their complex mixtures was evaluated to investigate the cellular morphological outcomes to targeted biological pathways. Overall toxicity and estrogenic activity of environmental water samples were then evaluated by phenotypic analysis comparing with conventional bioassays and chemical analysis by multivariate analysis. The phenotypic analysis for reference compounds demonstrated that size and structure of cells related to biological processes like cell growth, death, and communication. The phenotypic alteration and nuclei intensity were selected as potential biomarkers to evaluate overall toxicity and estrogenic activities, respectively. The phenotypic profiles were associated with the chemical structure profiles in environmental water samples. Since the phenotypic parameters revealed multiple toxicity endpoints, it could provide more information that is relevant to assessing the toxicity of environmental water samples in compare with conventional bioassays. In conclusion, the image-based multi-parameters phenotypic analysis with MCF-7 cells provides a rapid and information-rich tool for toxicity evaluation and identification in whole water samples.

Ecological and cultural shifts of hunter-gatherers of the Jomon period paralleled with environmental changes.

OBJECTIVES: Holocene hunter-gatherers adapted to climatic and environmental changes over time. Carbon and nitrogen stable isotope analysis of human skeletal remains from the Inariyama shell mound of the Final Jomon period have revealed large dietary variations in the population. This study analyzed radiocarbon dates of these individuals to test temporal changes in diet and its relationship with tooth ablation. MATERIALS AND METHODS: Twenty-nine human skeletal remains from Inariyama were included in this study. Extracted bone collagen samples were purified to graphite. Then, radiocarbon dating of these samples was performed using the accelerator mass spectrometer. RESULTS: The radiocarbon ages of Inariyama ranged about, 3,230-2,140 cal BP and showed three peaks of occupation. In the early and late phases, terrestrial resource consumption and incisor extraction were observed, while marine resource consumption and canine extraction were observed in the middle phase. DISCUSSION: These temporal changes of diet and tooth ablation types occurred in parallel with climatic cooling and environmental change and help reveal how Holocene hunter-gatherers adapted to the changing environments.