A retrospective study of unexpected deaths with alcohol use disorder from Japanese forensic autopsy cases.

BACKGROUND: Alcohol use disorder (AUD) is an important target for prevention of alcohol-related problems. In this study, we analyzed forensic autopsy cases to reveal the characteristics of the living conditions and death situations of individuals with AUD. METHODS: We retrospectively investigated 486 cases with a history of alcohol consumption for which a forensic autopsy was performed from 2012 to 2021 in Yamaguchi prefecture. Judgement of AUD was made using DSM-5. Various factors were compared statistically between AUD and non-AUD cases. RESULTS: Of the 486 cases, 225 (46.2%) were judged to be AUD, including 89 (18.3%) with advanced AUD, 33 (6.8%) were judged not to be AUD, and a judgement could not be made in the remaining cases. AUD was associated with alcohol consumption prior to death. Only 14.3% of the advanced-AUD cases was in treatment for alcohol dependence. The rates of interpersonal, health, financial and legal problems, receipt of public assistance and an extremely cluttered or hoarding house status were higher in all AUD and advanced AUD cases. Living alone, smoking and BMI were also associated with AUD. CONCLUSIONS: Many cases of alcohol-related deaths may have AUD, and persons with AUD who undergo a forensic autopsy commonly have multiple socioeconomic factors that may be associated with isolation that is involved in exacerbation of AUD. Further studies of these associations are needed because early diagnosis and treatment of AUD and support for the patient may lead to reduction of alcohol-related deaths.

Relationship Between Amplicon Size and Heat Conditions in Polymerase Chain Reaction Detection of DNA Degraded by Autoclaving.

This study examined the influence of heat exposure on DNA samples during polymerase chain reaction (PCR) detection. In this study, λDNA samples, as model DNA, were exposed to 105°C for 3-90 minutes or to 105°C-115°C for 15 minutes by autoclaving. The exposed samples were subjected to real-time PCR using nine primer sets with amplicon sizes of 45-504 bp. Regarding DNA samples exposed to 105°C by autoclaving, the data showed negative correlations between the logarithm of λDNA concentration (log λDNA) calculated using real-time PCR and exposure duration and a good relationship between the slope of the regression line and amplicon size. Regarding λDNA samples exposed to heat for 15 minutes, the data showed negative correlations between the log λDNA and exposure temperature and a good relationship between the slope of the regression line and amplicon size. These results showed that the equations used in this study could predict the degree of degradation in λDNA samples by autoclaving, and the PCR detection levels of the DNA at each amplicon size.

A bHLH heterodimer regulates germ cell differentiation in land plant gametophytes.

Land plants are a monophyletic group of photosynthetic eukaryotes that diverged from streptophyte algae about 470 million years ago. During both the alternating haploid and diploid stages of the life cycle, land plants form multicellular bodies.1,2,3,4 The haploid multicellular body (gametophyte) produces progenitor cells that give rise to gametes and the reproductive organs.5,6,7,8 In the liverwort Marchantia polymorpha, differentiation of the initial cells of gamete-producing organs (gametangia) from the gametophyte is regulated by MpBONOBO (MpBNB), a member of the basic helix-loop-helix (bHLH) transcription factor subfamily VIIIa. In Arabidopsis thaliana, specification of generative cells in developing male gametophytes (pollen) requires redundant action of BNB1 and BNB2.9 Subfamily XI bHLHs, such as LOTUS JAPONICUS ROOTHAIRLESS LIKE1 (LRL1)/DEFECTIVE REGION OF POLLEN1 (DROP1) and LRL2/DROP2 in A. thaliana and the single LRL/DROP protein MpLRL in M. polymorpha, are the evolutionarily conserved regulators of rooting system development.10 Although the role of LRL1/DROP1 and LRL2/DROP2 in gametogenesis remains unclear, their loss leads to the formation of abnormal pollen devoid of sperm cells.11 Here, we show that BNBs and LRL/DROPs co-localize to gametophytic cell nuclei and form heterodimers. LRL1/DROP1 and LRL2/DROP2 act redundantly to regulate BNB expression for generative cell specification in A. thaliana after asymmetric division of the haploid microspore. MpLRL is required for differentiation of MpBNB-expressing gametangium initial cells in M. polymorpha gametophytes. Our findings suggest that broadly expressed LRL/DROP stabilizes BNB expression, leading to the formation of an evolutionarily conserved bHLH heterodimer, which regulates germ cell differentiation in the haploid gametophyte of land plants.

Optimized enhanced acceleration selective arterial spin labeling (eAccASL) for non-gated and non-enhanced MR angiography of the hands.

PURPOSE: Enhanced acceleration selective arterial spin labeling (eAccASL) was introduced as non-enhanced and non-gated magnetic resonance angiography (MRA). This technique has not been applied to hand MRA. The objective of this study was to optimize the eAccASL for MRA of the hands and to investigate the factors for MRA visibility of the hands. METHODS: Twenty healthy volunteers were examined on a 1.5 T MR system. To evaluate arterial visualization, we compared four different acceleration-encoding (AENC) values (i.e., 0.12, 0.29, 0.58, and 0.87 m/s2). Image quality score regarding the MRA depiction of the proximal artery (range, 0-10), the distal artery (0-5), and venous contamination (0-5) was evaluated by three radiologists. We measured the peak to peak arterial blood flow velocity (Vpp) measured by phase contrast cine MRI and hand temperature as the factors for arterial visualization. Qualitative scores were compared with Friedman's tests. Spearman's correlation of qualitative scores with Vpp and hand temperature was performed to analyze influencing factors. RESULTS: For the distal arterial depiction, scores at AENC 0.12 (median, 9.0) and AENC 0.29 (8.0) were significantly better (both P < 0.0001) than those at AENC 0.87 (5.5). For the proximal arterial depiction, scores at AENC 0.12 (2.25) and AENC 0.29 (2.0) were significantly better (P < 0.001 and P < 0.01, respectively) than those at AENC 0.87 (1.5). Conversely, venous contamination scores at AENC 0.12 (3.0) and AENC 0.29 (3.0) were significantly worse (both P < 0.0001) than those at AENC 0.87 (4.0). There were significantly negative correlations between venous contamination and Vpp at AENC 0.12 (ρ = -0.56, P = 0.01), and 0.29 (ρ = -0.68, P = 0.001), whereas hand temperatures were not significantly correlated with scores (all P > 0.05). CONCLUSION: eAccASL MRA of the hands was optimized by using low AENC values (0.12-0.29 m/s2). Venous contamination may increase with elevation of arterial blood flow.

Generative Cell Specification Requires Transcription Factors Evolutionarily Conserved in Land Plants.

Land plants differentiate germ cells in the haploid gametophyte. In flowering plants, a generative cell is specified as a precursor that subsequently divides into two sperm cells in the developing male gametophyte, pollen. Generative cell specification requires cell-cycle control and microtubule-dependent nuclear relocation (reviewed in [1-3]). However, the generative cell fate determinant and its evolutionary origin are still unknown. In bryophytes, gametophytes produce eggs and sperm in multicellular reproductive organs called archegonia and antheridia, respectively, or collectively called gametangia. Given the monophyletic origin of land plants [4-6], evolutionarily conserved mechanisms may play key roles in these diverse reproductive processes. Here, we showed that a single member of the subfamily VIIIa of basic helix-loop-helix (bHLH) transcription factors in the liverwort Marchantia polymorpha primarily accumulated in the initial cells and controlled their development into gametangia. We then demonstrated that an Arabidopsis thaliana VIIIa bHLH transiently accumulated in the smaller daughter cell after an asymmetric division of the meiosis-derived microspore and was required for generative cell specification redundantly with its paralog. Furthermore, these A. thaliana VIIIa bHLHs were functionally replaceable by the M. polymorpha VIIIa bHLH. These findings suggest the VIIIa bHLH proteins as core regulators for reproductive development, including germ cell differentiation, since an early stage of land plant evolution.

Development of an externally controllable sealed isotope generator.

An externally controllable sealed isotope generator has been proposed for radiation education activities. Column (68Ge-68Ga and 137Cs-137mBa) and solvent extraction (68Ge-68Ga)-based isotope generators were applied as radioactive sources. These generators showed high milking efficiencies and low breakthrough after repeated uses, and are expected to promote the use of isotope generators without radioactive contamination or the emission of radioactive waste. This isotope generator provides a new concept for sealed radioisotope sources.

Evaluation of glucose-linked nitroxide radicals for use as an in vivo spin-label probe.

In vivo incorporation and reduction abilities of 4-carboxy-2,2,6,6-tetramethylpiperidine-1-oxyl (4-carboxy-TEMPO) (1), 3-carboxy-2,2,5,5-tetramethylpyrroline-1-oxyl (3-carboxy-dehydro-PROXYL, 3-carboxy-DPRO) (2), 4-hydroxy-TEMPO and 3-hydroxymethyl-DPRO O-ß-D-glucosides (3 and 5), and newly designed forms of 6-O-(TEMPO-4-carbonyl and DPRO-3-carbonyl)-D-glucose (4 and 6) were evaluated using white radish sprouts. For each of these compounds, electron spin resonance (ESR) spectrometry was used to measure two effects: the rate of in vitro reduction via the addition of ascorbic acid; and, the rate of successful incorporation into radish sprouts for a reduction to the corresponding hydroxyl amine. DPRO-radicals 2, 5, and 6 were detected significantly more than TEMPO-radicals 1, 3, and 4 in vitro and in vivo for both experiments. Four glucose-linked nitroxide radicals were reduced faster than the glucose-non-linked ones in the in vitro experiment, but were nonetheless detected more each time in radish sprouts due to the absorbability. Glucose ester-linked radicals 4 and 6 were detected more than glycosides 3 and 5, which suggests that glucose ester-linked DPRO-radical 6 is the best for use as a spin-label probe that a plant will incorporate.

Overexpression of the gibberellin 2-oxidase gene from Torenia fournieri induces dwarf phenotypes in the liliaceous monocotyledon Tricyrtis sp.

Gibberellins (GAs) are the plant hormones that control many aspects of plant growth and development, including stem elongation. Genes encoding enzymes related to the GA biosynthetic and metabolic pathway have been isolated and characterized in many plant species. Gibberellin 2-oxidase (GA2ox) catalyzes bioactive GAs or their immediate precursors to inactive forms; therefore, playing a direct role in determining the levels of bioactive GAs. In the present study, we produced transgenic plants of the liliaceous monocotyledon Tricyrtis sp. overexpressing the GA2ox gene from the linderniaceous dicotyledon Torenia fournieri (TfGA2ox2). All six transgenic plants exhibited dwarf phenotypes, and they could be classified into two classes according to the degree of dwarfism: three plants were moderately dwarf and three were severely dwarf. All of the transgenic plants had small or no flowers, and smaller, rounder and darker green leaves. Quantitative real-time reverse transcription-polymerase chain reaction (PCR) analysis showed that the TfGA2ox2 expression level generally correlated with the degree of dwarfism. The endogenous levels of bioactive GAs, GA1 and GA4, largely decreased in transgenic plants as shown by liquid chromatography-mass spectrometry (LC-MS) analysis, and the level also correlated with the degree of dwarfism. Exogenous treatment of transgenic plants with gibberellic acid (GA3) resulted in an increased shoot length, indicating that the GA signaling pathway might normally function in transgenic plants. Thus, morphological changes in transgenic plants may result from a decrease in the endogenous levels of bioactive GAs. Finally, a possibility of molecular breeding for plant form alteration in liliaceous ornamental plants by genetically engineering the GA metabolic pathway is discussed.