Zinc deficiency-induced defensin-like proteins are involved in the inhibition of root growth in Arabidopsis.

The depletion of cellular zinc (Zn) adversely affects plant growth. Plants have adaptation mechanisms for Zn-deficient conditions, inhibiting growth through the action of transcription factors and metal transporters. We previously identified three defensin-like (DEFL) proteins (DEFL203, DEFL206 and DEFL208) that were induced in Arabidopsis thaliana roots under Zn-depleted conditions. DEFLs are small cysteine-rich peptides involved in defense responses, development and excess metal stress in plants. However, the functions of DEFLs in the Zn-deficiency response are largely unknown. Here, phylogenetic tree analysis revealed that seven DEFLs (DEFL202-DEFL208) were categorized into one subgroup. Among the seven DEFLs, the transcripts of five (not DEFL204 and DEFL205) were upregulated by Zn deficiency, consistent with the presence of cis-elements for basic-region leucine-zipper 19 (bZIP19) or bZIP23 in their promoter regions. Microscopic observation of GFP-tagged DEFL203 showed that DEFL203-sGFP was localized to the apoplast and plasma membrane. Whereas a single mutation of the DEFL202 or DEFL203 genes only slightly affected root growth, defl202 defl203 double mutants showed enhanced root growth under all growth conditions. We also showed that the size of the root meristem was increased in the double mutants compared with the wild type. Our results suggest that DEFL202 and DEFL203 are redundantly involved in the inhibition of root growth under Zn-deficient conditions through a reduction in root meristem length and cell number.

Relationship between amikacin blood concentration and ototoxicity in low birth weight infants.

Amikacin (AMK) is used as empiric therapy for severe infections such as sepsis in low birth weight (LBW) infants. AMK administered once daily (OD) in adults is reported to be therapeutically effective and prevent side effects, however, evidence on AMK administration in LBW infants is limited, with no clear indications of effectiveness. We performed therapeutic drug monitoring analysis of 20 infants treated with AMK OD for severe infections such as bacteremia. Treatment effectiveness was admitted by the patients' medical records, and side effects of renal dysfunction and ototoxicity were investigated. The mean gestational age was 30.4 ± 5 weeks and mean body weight (Bw) was 1280.2 ± 809.8 g. The mean AMK dose was 14.1 ± 2.6 mg/kg and mean administration period was 10.1 ± 4.1 days. Blood concentration was measured 6.3 ± 2.3 days after AMK administration; mean peak and trough concentrations were 29.1 ± 7.5 µg/mL and 7.6 ± 6.9 µg/mL, respectively. Additionally, therapeutic effect was observed in all patients, and no significant change in serum creatinine (CRE) concentration (a marker of renal dysfunction) was observed, suggesting no renal dysfunction. Ototoxicity was observed in 4 patients, 3 of whom had trough concentrations ≥10 µg/mL. When we categorized patients into two groups using a trough cut-off value of 10 µg/mL, no difference in AMK dose was observed. However, there were significant differences in peak concentration, Bw, volume of distribution and CRE. Our findings suggest AMK trough concentration ≥10 µg/mL significantly affects ototoxicity in neonates.

Safety of cold polypectomy for small colorectal neoplastic lesions: a prospective cohort study in Japan.

BACKGROUND: Cold polypectomy has been widely accepted for removal of small colorectal polyps. However, no large-scale prospective study exists as for its safety in Japan. We investigated this issue in this single-center, prospective cohort study in a total of 1198 colorectal polyps resected with cold polypectomy. PATIENTS AND METHODS: Four hundred and seventy-four patients who underwent cold polypectomy for colorectal neoplastic lesions less than 10-mm diameter between September 2014 and October 2016 were enrolled. Primary outcome was the incidence of delayed bleeding within 2 weeks after the procedure. Secondary outcomes were the rate of immediate bleeding, perforation, endoscopic en bloc resection, and advanced histology. RESULTS: Cold polypectomy was performed on 1198 polyps in the 474 patients. No delayed bleeding or colonic perforation was observed. Immediate bleeding during the procedure, requiring endoscopic hemostasis, occurred in 97 lesions (8.1%), and all of them were successfully managed endoscopically. The endoscopic en bloc resection rate was 97.2%. Twenty-eight lesions (2.3%) were histologically diagnosed as advanced neoplasia; among them, three lesions were well-differentiated adenocarcinomas, and in two of them, a negative margin was not histologically confirmed. CONCLUSIONS: Cold polypectomy for small colorectal polyps is a safe technique without significant complication, but careful endoscopic diagnosis at cold polypectomy is necessary to identify advanced neoplasia. The reliability of cold polypectomy in excision of polyps with high-grade neoplasia should be established before the procedure becomes standard in the excision of small colorectal polyps. CLINICAL TRIAL REGISTRATION NUMBER: UMIN000014812.

Identification of putative target genes of bZIP19, a transcription factor essential for Arabidopsis adaptation to Zn deficiency in roots.

Zinc (Zn) depletion adversely affects plant growth. To avoid lethal depletion of cellular Zn, plants have evolved mechanisms to adjust the expression of genes associated with Zn homeostasis, the details of which are poorly understood. In the present study, we isolated an Arabidopsis thaliana T-DNA insertion mutant that exhibited hypersensitivity to Zn depletion. By monitoring root development under Zn-deficient conditions, we isolated a single mutant lacking the basic-region leucine-zipper transcription factor gene bZIP19. To identify proteins whose expression is affected by bZIP19, an iTRAQ-based quantitative proteomics analysis was performed using microsomal proteins from wild-type and the bzip19 mutant A. thaliana roots grown on Basal and Zn-deficient media. Of the 797 proteins identified, expression of two members of the Zrt- and Irt-related protein family, ZIP3 and ZIP9, and three defensin-like family proteins was markedly induced in wild-type but not in the bzip19 mutant under Zn-deficient conditions. Furthermore, selected reaction monitoring and quantitative real-time PCR revealed that ZIP9 expression is mediated by bZIP19 and may be partly supported by bZIP23, a homolog of bZIP19. Mutant analysis revealed that ZIP9 is involved in uptake of Zn by the roots, and the mutant lacking ZIP9 was significantly more sensitive to Zn depletion than the wild-type. These results demonstrate that bZIP19 mainly contributes to expression of genes, such as ZIP9, under Zn-deficient conditions.

Constitutively high-level expression of TGFß isoforms in cord blood and its relationship to perinatal findings.

BACKGROUND: The clinical significance of TGFß isoforms in cord blood is not well understood. METHODS: We obtained cord blood samples from 37 term infants and 85 preterm infants who were born in several clinical settings. The serum levels of 3 TGFß isoforms and of the other 17 cytokines in cord blood were investigated using cytometric bead array technology. RESULTS: Very high levels of TGFß1 and TGFß2 isoforms compared to the level of other cytokines were found; mean levels were 44,180 and 1871pg/mL, respectively. The levels of all 3 isoforms of TGFß were significantly correlated with birth weight, and the levels of TGFß1 and TGFß3 were correlated with gestational age. The levels of TGFß1 and ß2 isoforms were strongly correlated with each other, but not with levels of other cytokines. The levels of TGFß1 and TGFß2 were significantly higher in male infants and significantly lower in infants with fetal growth restriction. The prevalence of chronic lung disease was related to a low level of TGFß1, and that of patent ductus arteriosus was related to a high level of TGFß1 in preterm infants. CONCLUSIONS: TGFß1 and TGFß2 appeared to play a significant role in physiological and pathological conditions in the fetus. TGFß isoform levels appear to be regulated independently of those of other cytokines and do not appear to be influenced by inflammation in the fetal period. The role of TGFß3 in cord blood and the postnatal chronological changes of the TGFß isoforms should be investigated in the future.

Correlation analysis of proteins responsive to Zn, Mn, or Fe deficiency in Arabidopsis roots based on iTRAQ analysis.

KEY MESSAGE: For discovering the functional correlation between the identified and quantified proteins by iTRAQ analysis, here we propose a correlation analysis method with cosine correlation coefficients as a powerful tool. iTRAQ analysis is a quantitative proteomics approach that enables identification and quantification of a large number of proteins. In order to obtain proteins responsive to Zn, Mn, or Fe mineral deficiency, we conducted iTRAQ analysis using a microsomal fraction of protein extractions from Arabidopsis root tissues. We identified and quantified 730 common proteins in three biological replicates with less than 1 % false discovery rate. To determine the role of these proteins in tolerating mineral deficiencies and their relation to each other, we calculated cosine correlation coefficients and represented the outcomes on a correlation map for visual understanding of functional relations among the identified proteins. Functionally similar proteins were gathered into the same clusters. Interestingly, a cluster of proteins (FRO2, IRT1, AHA2, PDR9/ABCG37, and GLP5) highly responsive to Fe deficiency was identified, which included both known and unknown novel proteins involved in tolerating Fe deficiency. We propose that the correlation analysis with the cosine correlation coefficients is a powerful method for finding important proteins of interest to several biological processes through comprehensive data sets.

Predictive factors for the outcome of multidisciplinary treatments in chronic low back pain at the first multidisciplinary pain center of Japan.

[Purpose] Multidisciplinary treatments are recommended for treatment of chronic low back pain. The aim of this study was to show the associations among multidisciplinary treatment outcomes, pretreatment psychological factors, self-reported pain levels, and history of pain in chronic low back pain patients. [Subjects and Methods] A total of 221 chronic low back pain patients were chosen for the study. The pretreatment scores for the 10-cm Visual Analogue Scale, Hospital Anxiety and Depression Scale, Pain Catastrophizing Scale, Short-Form McGill Pain Questionnaire, Pain Disability Assessment Scale, pain drawings, and history of pain were collected. The patients were divided into two treatment outcome groups a year later: a good outcome group and a poor outcome group. [Results] One-hundred eighteen patients were allocated to the good outcome group. The scores for the Visual Analogue Scale, Pain Disability Assessment Scale, and affective subscale of the Short-Form McGill Pain Questionnaire and number of nonorganic pain drawings in the good outcome group were significantly lower than those in the poor outcome group. Duration of pain in the good outcome group was significantly shorter than in the poor outcome group. [Conclusion] These findings help better predict the efficacy of multidisciplinary treatments in chronic low back pain patients.

Peptide separation methodologies for in-depth proteomics in Arabidopsis.

In the post-genome era, several tools that have increased our global understanding of the molecular basis of several cell-based phenomena have been developed. However, proteomics has not been efficiently integrated with the other 'omics' (e.g. transcriptomics and metabolomics), because of the relatively low number of proteins identified by mass spectrometry (MS). Peptides from low-abundance proteins are often not detected by MS due to ionization suppression. To improve the number of peptide identifications in MS analyses, we propose three separation methodologies; namely, OFFGEL electrophoresis, 2D-liquid chromatography (LC) and the long monolithic silica-C18 capillary column method, with the common aim to decrease peptide complexity prior to MS analyses. Proteomics using the above three peptide separation methods were separately applied to protoplasts collected from the epidermal cell layer of Arabidopsis roots using fluorescence-activated cell sorting. In each method alone, 1,132, 836 and 795 proteins were specifically identified, respectively. This has allowed the identification of 1,493 proteins with no redundancy and with <1.0% false discovery rate. Moreover, approximately two-thirds of these proteins are identified here for the first time in the epidermal cell layer. These results show that use of different proteomic approaches can increase the total number of proteins identified. We propose that the integration of data from these methodologies represents a powerful tool for generation of proteome maps by enabling identification of low-abundance proteins in the various Arabidopsis root cell layers.

[Exploring for Drug Development Seeds for Children Using a Large Claims Data-Survey for Powderization or Decapsulation of Oral Tablets or Capsule Formula in Children].

Medical big data is accumulated numerous medical related data day by day. These data may have tips for new approach for drug development. Authors tried to find drug-development-needs in children using medical big data analysis with prescription survey. Medical big data were provided from JMDC (Japan Medical Data Centre) Inc. about 3 million participants between January 2005 and June 2017. In these, we identified randomly identified 22787 participants from 466701 participants who are aged ≤11 years. In these participants, 9644 were administered "capsule," "tablet," "orally distegrating tablet," "controlled release tablet/capsule" or "enteric coated tablet" formula drugs. In these, 514 were administered these as powderization or decapsulation. Sixty components administered in 145 participants (28.2%) are not marketed for pediatric formula. On the other hands, 92 components administered in 369 participants (71.8%) are decapsulation or powderization, though pediatric formulas are marketed. These 152 components may have a development seeds for children. In conclusion, prescription survey using medical big data may partially resolve the drug-development-need in pediatrics because by using medical big data will leads low biased data depending each institution.

In vivo kinetics of indoxyl sulfate in humans and its renal interaction with angiotensin-converting enzyme inhibitor quinapril in rats.

Indoxyl sulfate (IS) is an organic anion uremic toxin that accumulates in patients with chronic kidney disease (CKD). The aims of this study were to examine the kinetic profiles of IS in humans at a steady state after multiple doses of L-Trp, a precursor of IS, and the in vivo interaction of IS with the angiotensin-converting enzyme inhibitor quinapril, whose active metabolite is a substrate of organic anion transporter 3 (OAT3) in rats. First, 12-h kinetics after single doses of Trp (2, 4, and 8 g) were examined in two healthy volunteers. Second, 24-h kinetics after a single dose of 2 g of Trp was studied in six volunteers. Third, 35-h kinetics after single and multiple doses of 2 g of Trp were examined in five volunteers. In anesthetized rats, quinapril or probenecid, an inhibitor of OATs, was given intravenously before IS, and blood and urine samples were taken until 90 min. Trp and IS concentrations were determined by high-performance liquid chromatography. Ultrafiltration was used to measure serum unbound IS concentrations. Renal tubular secretion of IS accounted for more than 90% of its renal clearance in the steady state of serum IS levels after multiple doses in humans. In animals, the serum area under the curve of IS increased in conjunction with a decrease in renal clearances after coadministration of IS with quinapril or probenecid. It is concluded that quinapril may inhibit the urine excretion of IS via OAT3-mediated renal tubular transport in patients with CKD.

Characteristics of patients with subchorionic hematomas in the second trimester.

AIM: The aim of this study was to investigate the etiological characteristics of patients diagnosed with subchorionic hematoma. METHODS: A case-controlled study was performed to compare characteristics of patients and controls. Via ultrasound examination, 47 pregnant patients were found to have subchorionic hematomas and 1075 had no evidence of subchorionic hematomas (controls). In the second trimester, patients were compared with regard to endocervical Chlamydia trachomatis and other vaginal microorganisms. RESULTS: The overall incidence of subchorionic hematomas in this pregnant population was 4.2%. Maternal clinical characteristics did not differ between cases and controls. Evaluation of the vaginal flora revealed that the positive rates of coagulase-negative staphylococci (cases: 12.8%; controls: 4.1%; P<0.01) and Gardnerella vaginalis (cases: 12.8%; controls: 2.5%; P<0.001) in the cases were significantly higher than those of the controls. The negative rate of Lactobacillus in the cases was significantly higher than that of the controls (cases: 42.6%; controls: 27.6%; P<0.05). CONCLUSION: Pregnant women with subchorionic hematoma in the first trimester showed changes in vaginal flora in the second trimester, which suggests a possible association with subchorionic hematoma and vaginal flora change.

Idiopathic central precocious puberty with Prader-Willi syndrome: pubertal development with discontinuation of gonadotropin-releasing hormone analog.

Summary: Prader-Willi syndrome (PWS) is a genetic imprinting disorder that is characterized by obesity, short stature, and hypogonadism. Hypogonadism is characterized by normal luteinizing hormone (LH), high follicle-stimulating hormone (FSH), low testosterone, low inhibin B, and relatively low anti-Müllerian hormone (AMH). Only a few cases of central precocious puberty (CPP) have been reported in PWS, and follow-up for CPP with PWS is not established. Hence, we present a boy with PWS accompanied by CPP. Gonadotropin-releasing hormone analog (GnRHa) therapy was started at 7 years of age, CPP was adequately arrested, and GnRHa therapy was discontinued at 11.3 years of age. Growth hormone (GH) therapy was started at 12 years of age due to inadequate growth. He grew close to his final height, and his testes developed with normal LH, increased FSH, normal testosterone, and reduced AMH corresponding to puberty at 13.5 years of age. The features of 16 patients with PWS with CPP, including our patient, were summarized. Out of seven male patients, five were treated with GnRHa, as well as four out of nine female patients. Out of 16 patients, 6 were assessed with pubertal development over 13 years of age. Pubertal development was considered to be restored in four patients who had GnRHa therapy discontinuation. We should carefully follow-up on pubertal development in CPP. GnRHa therapy is useful for adequate puberty blockage, and pubertal development could be restored with GnRHa therapy discontinuation. Learning points: Pubertal development in Prader-Willi syndrome (PWS) varies from hypogonadism to precocious puberty. Pubertal development assessment based on clinical features and hormone levels is needed in central precocious puberty (CPP) treatment with PWS. Gonadotropin-releasing hormone analog (GnRHa) therapy is useful for CPP with PWS, and pubertal development can be restored with GnRHa therapy discontinuation.

C-reactive protein levels at pre-/post-indicated cervical cerclage predict very preterm birth.

AIMS: To investigate the relation between serum levels of C-reactive protein (CRP) at pre-/post-cerclage points and preterm birth at <33 weeks of gestation in women with indicated cervical cerclage (CC). METHODS: Fifty-eight women with CC indicated for a short or soft cervix, but no visible or protruding fetal membranes into the vagina, between 17 and 26 weeks of gestation, were reviewed. Serum CRP levels were examined three times: just before cerclage, and on day 1 and day 2 post-cerclage. RESULTS: Serum CRP levels on day 1 and day 2, but not just before cerclage, predicted the occurrence of very preterm birth. In women with cervical dilatation of <3.0 cm, serum CRP levels on post-cerclage day 1 were associated with the increase of very preterm birth [CRP ≥1.5 mg/dL vs. <1.5 mg/dL: 4/5 (80%) vs. 8/31 (26%), P=0.033]. In women with cervical dilatation of <3.0 cm, serum CRP ≥3.0 mg/dL on post-cerclage day 2 was also associated with the increase of very preterm birth. CONCLUSION: In women with indicated CC between 17 and 26 weeks of gestation, increased levels of serum CRP on post-cerclage day 1 or 2 might be ominous signs for very preterm birth.

Reproducible induction of depressive-like behavior in C57BL/6J mice exposed to chronic social defeat stress with a modified sensory contact protocol.

AIMS: C57BL/6J mice are well-known to exhibit resilience to chronic social defeat stress (CSDS) for induction of depressive-like behavior. Establishment of protocols for reproducible induction of depressive-like behavior in C57BL/6J mice would be useful to elucidate the underlying molecular mechanisms using target gene-knock-in and -out mice whose background is generally C57BL/6J. Here, we developed a modified CSDS protocol for reproducible induction of depressive-like behavior in C57BL/6J mice, and compared the profile of their gut microbiota with that with the standard CSDS protocol. MAIN METHODS: To prevent acclimation of defeated C57BL/6J mice to aggressive ICR mice, the sensory contact following a daily 10 min-defeat episode was performed by housing an individual defeated mouse in a cage set next to a cage for the aggressor one. KEY FINDINGS: The number of attacks by ICR mice on C57BL/6J ones was significantly increased with the modified CSDS protocol, and the susceptible mice exhibited greater hippocampal inflammation and an increased immobility time in the forced swim test, compared in the case of the standard CSDS protocol, and the reproducibility was confirmed in another set of experiments. Both the standard and modified CSDS protocols changed the diversity and relative composition of gut microbiota in the susceptible mice, but there was no apparent difference in them between the standard and modified CSDS-susceptible mice. SIGNIFICANCE: We established a CSDS protocol for reproducible induction of depressive-like behavior in C57BL/6J mice, and the features of the gut microbiota were similar in the susceptible mice with and without the depressive-like behavior.

Cytokine profiles of seventeen cytokines, growth factors and chemokines in cord blood and its relation to perinatal clinical findings.

Few papers have investigated the cytokine profiles of multiple cytokines in cord blood. We obtained cord blood samples from 224 infants admitted to our neonatal intensive care unit. Cytokine profiles of 17 cytokines were investigated using cytometric bead array technology. We found a wide variety of cytokines of various levels which ranged from 0.59pg/ml (in Interleukin (IL)-4) to 222.0pg/ml (in macrophage inflammatory protein-1beta. Pro-inflammatory cytokines were highly correlated with each other and with granulocyte-colony stimulating factor and IL-8. On the contrary, IL-5, IL-13, and IL-17 did not show any significant correlation with other cytokines. Several maternal factors were strongly related to several cytokines in cord blood. IL-6, IL-8 and monocyte chemotactic protein-1 were closely related to certain neonatal diseases in preterm neonates. Some cytokines may be regulated independently of each other, while others appear to work as a network affecting physiological and pathological conditions in the fetus.

Xanthogranuloma of the sellar region accompanied by growth hormone deficiency: case report and literature review.

Background Xanthogranuloma of the sellar region is a rare entity. Its pathology is controversial and it is difficult to strictly differentiate it from craniopharyngioma or Rathke's cyst. Case presentation We report a case of xanthogranuloma accompanied by growth hormone deficiency in an 11-year-old girl. She did not show any other pituitary hormone deficiency or neurological symptoms before operation. The preoperative diagnosis was craniopharyngioma, but histological findings showed small areas of epithelium. Thus, the final diagnosis was xanthogranuloma. Xanthogranuloma is an important cause of growth delay. We reviewed 16 cases reported after 2000, and included our case, of xanthogranuloma in children. Conclusions Endocrinological symptoms are often regarded as one of the few apparent symptoms in xanthogranuloma compared with craniopharyngioma. Therefore, we should follow up carefully and accumulate cases.

Factors Associated with possibly Inappropriate Histological Evaluation of Excised Specimens in Cold-snare Polypectomy for Small Colorectal Polyps.

BACKGROUND AND AIMS: Small colorectal polyps may be removed with cold snare polypectomy (CSP). Some of these polyps may contain unexpectedly advanced neoplasia. Thus, it is important to establish criteria for excision that will ensure that the CSP specimens are adequate for accurate histological assessment. We retrospectively investigated depth of excised small polyps and their vertical margins in patients who underwent CSP. METHOD: CSP-excised specimens of 376 small colorectal polyps were examined. We histologically evaluated negative tumor vertical margins and complete resection through the muscularis mucosae, which was defined as muscularis mucosae present under the tumor along more than 80% of its horizontal axis. We also evaluated the fragmentation of the retrieved specimens. RESULTS: The mean size of the 376 polyps was 4.9 +/-1.4 mm, and 25 polyps (6.6%) had unexpectedly advanced histology. Thirty-two lesions (8.5%) were fragmented. In 275 (79.9%) of the remaining 344 unfragmented polyps, muscularis mucosae resection was judged complete. Vertical margins were confirmed negative in 99.6% (274/275) of polyps that had complete muscularis mucosae resection, but in only 33.3% (23/69) of polyps with incomplete resection. In 79 polyps (21%) (32 fragmented specimens and 47 unfragmented specimens), including 5 polyps with advanced histology, negative vertical margins could not be confirmed. Sessile morphology and location in the cecum were independent risk factors for incomplete muscularis mucosae resection and fragmentation. CONCLUSION: Incomplete muscularis mucosae resection and fragmentation at retrieval are risk factors for inadequate histological evaluation of CSP-excised small colorectal polyps, especially for sessile polyps and polyps in the cecum.