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Background: Relapsed or refractory peripheral T-cell lymphomas (r/r PTCLs) are a group of rare and aggressive diseases that lack effective therapies. Constitutive activation of the Janus kinase (JAK)/signal transducer and activator of transcription (STAT) pathway is reported to be associated with PTCLs. Golidocitinib is an oral, potent JAK1 selective inhibitor evaluated in a phase I/II multinational study in patients with r/r PTCLs., Patients and Methods: Patients with r/r PTCLs were eligible. The primary objectives were to assess safety and tolerability of golidocitinib and to define its recommended phase II dose (RP2D). The secondary objectives were to evaluate its antitumor activity and pharmacokinetics (PK)., Results: A total of 51 patients were enrolled and received golidocitinib treatment at 150 or 250 mg once daily (QD). The median prior lines of therapies were 2 (range: 1-8). Golidocitinib was tolerated at both doses tested, while a higher incidence of serious adverse events and dose modifications at 250 mg were observed. The most common grade ≥3 drug-related treatment-emergent adverse events were neutropenia (27.5%) and thrombocytopenia (11.8%). An objective response rate of 39.2% and a complete response rate of 21.6% were observed. With median follow-up time of 14.7 and 15.9 months, the median duration of response (DoR) and progression-free survival were 8.0 and 3.3 months, respectively. Based on these data, 150 mg QD was defined as the RP2D. Golidocitinib demonstrated a favorable PK profile as an oral agent. Biomarker analysis suggested a potential correlation between JAK/STAT pathway aberrations and clinical activity of golidocitinib., Conclusions: In this phase I study, golidocitinib demonstrated an acceptable safety profile and encouraging antitumor efficacy in heavily pretreated patients with r/r PTCLs. These results support the initiation of the multinational pivotal study in patients with r/r PTCLs., Competing Interests: Disclosure This study was sponsored by Dizal Pharmaceutical. MW and ZY reported employment by Dizal Pharmaceutical. All remaining authors have declared no conflicts of interest., (Copyright © 2023 European Society for Medical Oncology. Published by Elsevier Ltd. All rights reserved.)

Background: Suvemcitug (BD0801), a novel humanized rabbit monoclonal antibody against vascular endothelial growth factor, has demonstrated promising antitumor activities in preclinical studies., Patients and Methods: The phase Ia/b trials investigated the safety and tolerability and antitumor activities of suvemcitug for pretreated advanced solid tumors and in combination with FOLFIRI (leucovorin and fluorouracil plus irinotecan) in second-line treatment of metastatic colorectal cancer using a 3 + 3 dose-escalation design. Patients received escalating doses of suvemcitug (phase Ia: 2, 4, 5, 6, and 7.5 mg/kg; phase Ib: 1, 2, 3, 4, and 5 mg/kg plus FOLFIRI). The primary endpoint was safety and tolerability in both trials., Results: All patients in the phase Ia trial had at least one adverse event (AE). Dose-limiting toxicities included grade 3 hyperbilirubinemia (one patient), hypertension and proteinuria (one patient), and proteinuria (one patient). The maximum tolerated dose was 5 mg/kg. The most common grade 3 and above AEs were proteinuria (9/25, 36%) and hypertension (8/25, 32%). Forty-eight patients (85.7%) in phase Ib had grade 3 and above AEs, including neutropenia (25/56, 44.6%), reduced leucocyte count (12/56, 21.4%), proteinuria (10/56, 17.9%), and elevated blood pressure (9/56, 16.1%). Only 1 patient in the phase Ia trial showed partial response, [objective response rate 4.0%, 95% confidence interval (CI) 0.1% to 20.4%] whereas 18/53 patients in the phase Ib trial exhibited partial response (objective response rate 34.0%, 95% CI 21.5% to 48.3%). The median progression-free survival was 7.2 months (95% CI 5.1-8.7 months)., Conclusions: Suvemcitug has an acceptable toxicity profile and exhibits antitumor activities in pretreated patients with advanced solid tumors or metastatic colorectal cancer., Competing Interests: Disclosure Jiangsu Simcere Pharmaceuticals Co. Ltd., the sponsor of this trial, provided all trial costs, including investigational drugs, laboratory tests, and associated operational cost. WS, LL, YW, LS, XF, and JZ are full-time employees of Shandong Simcere Bio-Pharmaceutical Co., Ltd. Both Jiangsu Simcere Pharmaceuticals Co. Ltd. and Shandong Simcere Bio-Pharmaceutical Co., Ltd are subsidiaries of Simcere Pharmaceuticals Co., Ltd. All other authors have declared no conflicts of interest., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

As a pituitary hormone, prolactin (PRL) is also synthesized by immune system cells, and exerts its effects on the immune system by binding to its receptor (PRLR) via endocrine and paracrine/autocrine pathways. The immune organs adapt to the presence of fetal alloantigens during pregnancy, and the immune system is composed of primary organs and secondary organs. The objective of this study is to analyze the effects of early pregnancy on expression of PRL and PRLR in maternal immune organs in sheep. In this study, the thymus, lymph node, the spleen and the liver were sampled at day 16 of the estrous cycle, and at days 13, 16, and 25 of pregnancy in ewes. Expression of PRL and PRLR was analyzed through quantitative real-time PCR, Western blot and immunohistochemistry. Our data showed that there were an upregulation of PRL and PRLR in the thymus, lymph node and the spleen, and a downregulation in the liver during early pregnancy in ewes. In conclusion, it is reported for the first time that early pregnancy has tissue specific effects on expression of PRL isoform and PRLR isoform in the thymus, lymph node, the spleen and the liver, which may be owing to these organs exerting different functions during early pregnancy, and necessary for the successful pregnancy in sheep., (Copyright © 2022. Published by Elsevier Inc.)

Objectives: The aims of this study were to assess trends in maternal age in central China and to examine the relationship between maternal age and adverse pregnancy outcomes., Study Design: This was a retrospective and observational study., Methods: Data were analysed from the Wuhan Maternal and Child Health Management Information System, which included all pregnant women who were at 24-42 weeks' gestation, with singleton pregnancies and who lived in Wuhan between 2010 and 2017. Joinpoint regression was used to analyse the trends in mean maternal age and the proportion of women with advanced maternal age. Adverse pregnancy outcomes, including pregnancy-induced hypertension (PIH) disorder, gestational diabetes mellitus (GDM), caesarean delivery, postpartum haemorrhage, preterm birth, small for gestational age, large for gestational age (LGA) and 5-min Apgar score <7 among women aged <20, 20-24, 30-35 and ≥40 years were compared with women aged 25-29 years using multivariate binary logistic regression analysis and stratified analysis., Results: Among the 583,571 women included in this study, 1.2% were aged <20 years, 20.8% were aged 20-24 years, 47.9% were aged 25-29 years, 22.0% were aged 30-34 years, 6.9% were aged 35-39 years and 1.2% were aged ≥40 years. Between 2010 and 2017, the mean maternal age increased from 27.1 years to 29.7 years, and the proportion of women aged ≥35 years increased from 4.3% to 13.9%. Relative to women aged 25-29 years, women aged ≥30 years carried higher risks of PIH, GDM, caesarean delivery, preterm birth, LGA and 5-min Apgar score <7. Relative to older multiparous women, older nulliparous women were more likely to experience caesarean delivery, preterm birth and 5-min Apgar score <7., Conclusions: The average maternal age and the proportion of advanced maternal age showed increasing trends between 2010 and 2017, which may be related to the relaxation of the one-child policy in China. Older maternal age (≥30 years) is independently associated with various adverse pregnancy outcomes. The risks of adverse pregnancy outcomes may occur earlier than the commonly used definition of advanced maternal age (≥35 years) and may also differ by parity. Ensuring age- and parity-specific clinical counselling, antenatal surveillance and health interventions may significantly improve pregnancy outcomes in older mothers., (Copyright © 2022 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Purpose: This study aimed to compare the absolute numbers of various types of lymphocytes in the peripheral blood before and after chemotherapy following radio-chemotherapy in patients with cervical cancer, so as to explore the correlation between the changes in the absolute numbers of peripheral various types of lymphocytes and the overall survival rate of patients., Methods: Data of 205 patients with cervical cancer admitted to the hospital from June 2014 to August 2016 were retrospectively analyzed. These patients underwent concurrent radio-chemotherapy, followed by chemotherapy. The absolute numbers of peripheral blood lymphocytes and subtypes were compared before and after re-chemotherapy., Results: For patients with a good prognosis, the number of lymphocytes, T cells, and cytotoxic T cells (Tc) significantly decreased (P<0.05) after re-chemotherapy, while this phenomenon was not observed in patients with poor prognosis. Kaplan-Meier univariate analysis showed that patients with cervical cancer who had an advanced FIGO stage before treatment, presence of lymph node metastasis, and increased numbers of Tc and T helper cells after re-chemotherapy showed a low 30-month survival rate; the overall log-rank analysis showed significant differences (P<0.05). Multivariate Cox proportional hazards regression analysis, presence of lymph node metastasis: HR=9.718, P=0.002, 95% CI=0.183-0.679; Tc grouping: HR=3.239, P=0.0.072, 95% CI=0.950-03.347; Th group: HR=3.197, P=0.074, 95% CI=0.943-3.564., Conclusions: After radio-chemotherapy, in patients with advanced cervical cancer, the change in the numbers of peripheral blood Tc and Th before and after re-chemotherapy was associated with the 30-month overall survival rate of patients. It can be considered as a predictor of the survival of patients with advanced cervical cancer., (Copyright © 2020. Published by Elsevier Masson SAS.)

Objectives: In December 2019, a novel coronavirus disease (COVID-19) emerged in Wuhan city, China, which has subsequently led to a global pandemic. At the time of writing, COVID-19 in Wuhan appears to be in the final phase and under control. However, many other countries, especially the US, Italy and Spain, are still in the early phases and dealing with increasing cases every day. Therefore, this article aims to summarise and share the experience of controlling the spread of COVID-19 in Wuhan and provide effective suggestions to enable other countries to save lives., Study Design: Data from the National Health Commission of China are used to investigate the evolution trajectory of COVID-19 in Wuhan and discuss the impacts of the intervention strategies., Methods: A four-stage modified Susceptible-Exposed-Infectious-Removed (SEIR) model is presented. This model considers many influencing factors, including chunyun (the Spring festival), sealing off the city and constructing the Fangcang shelter hospitals. In addition, a novel method is proposed to address the abnormal data on 12-13 February as a result of changing diagnostic criteria. Four different scenarios are considered to capture different intervention measures in practice. The exposed population in Wuhan who moved out before sealing off the city have also been identified, and an analysis on where they had gone was performed using the Baidu Migration Index., Results: The results demonstrate that the four-stage model was effective in forecasting the peak, size and duration of COVID-19. We found that the combined intervention measures are the only effective way to control the spread and not a single one of them can be omitted. We estimate that England will be another epicentre owing to its incorrect response at the initial stages of COVID-19. Fortunately, big data technology can help provide early warnings to new areas of the pandemic., Conclusions: The four-stage SEIR model was effective in capturing the evolution trajectory of COVID-19. Based on the model analysis, several effective suggestions are proposed to prevent and control the pandemic for countries that are still in the initial phases., (Copyright © 2020 The Royal Society for Public Health. Published by Elsevier Ltd. All rights reserved.)

Background and Aims: Elevated resting heart rate (RHR) is associated with risk of type 2 diabetes mellitus (T2DM). However, the association of change in RHR (ΔRHR) and incident T2DM is not fully elucidated. We aimed to assess the dose-response association between 6-year ΔRHR and T2DM., Methods and Results: A total of 12155 non-T2DM participants ≥18 years old were enrolled during 2007-2008 and followed up during 2013-2014. ΔRHR was calculated by subtracting the baseline RHR from the RHR value at 6-year follow-up. Age-, sex-, and RHR-specific relative risks (RRs) and 95% confidence intervals (CIs) for the effect of ΔRHR on incident T2DM were calculated by using modified Poisson regression models. As compared with ΔRHR of 0 beats/min, the adjusted risk of T2DM was significantly increased with RHR increment and reduced with RHR reduction. ΔRHR was positively associated with future risk of T2DM [RR per unit increase: 1.03 (1.03-1.04)]. As compared with stable change in RHR group (-5<ΔRHR<5 beats/min), for ΔRHR ≤ -10 beats/min, -10<ΔRHR ≤ -5 beats/min, 5≤ΔRHR<10 beats/min, and ΔRHR ≥10 beats/min groups, the pooled adjusted RR (95% CI) of T2DM was 0.69 (0.55-0.86), 0.90 (0.73-1.11), 1.31 (1.07-1.61), and 1.90 (1.59-2.26), respectively. This significant association still existed on subgroup analyses based on age, sex, and baseline RHR and sensitivity analyses., Conclusions: Dynamic RHR change was significantly associated with incident T2DM. Our study suggests that RHR may be a non-invasive clinical indicator for interventions aiming to reduce incident T2DM in the general population., (Copyright © 2018 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Background: CBCSG006 trial reported the superior efficacy of cisplatin plus gemcitabine (GP) regimen than paclitaxel plus gemcitabine (GT) regimen as first-line treatment of metastatic triple-negative breast cancer (mTNBC). This study focused on the updated survival data and the explorations of potential biomarkers for efficacy., Patients and Methods: Germ-line mutations of homologous recombination (HR) panel, BRCA1/2 included, were evaluated in 55.9% (132/236) patients. PD-L1 expression was evaluated in 48.3% (114/236) patients. A nonparametric sliding-window subpopulation treatment effect pattern plot (STEPP) methodology was used to analyze the absolute survival benefits. All statistical tests were two-sided., Results: Median progression-free survival (PFS) was 7.73 [95% confidence interval (CI) 6.46-9.00] months for GP arm and 6.07 (95% CI 5.32-6.83) months for GT arm (P = 0.005). No significant difference in overall survival (OS) was observed. There was significant interaction between HR status and treatment for PFS and status of HR deficient significantly correlated with higher objective response rate (ORR) and longer PFS in GP arm than in GT arm (71.9% versus 38.7%, P = 0.008; 10.37 versus 4.30 months, P = 0.011). There was no significant interaction between germ-line BRCA1/2 (gBRCA1/2) status and treatment for PFS. Patients with gBRCA1/2 mutation had numerically higher ORR and prolonged PFS in GP arm than in GT arm (83.3% versus 37.5%, P = 0.086; 8.90 versus 3.20 months, P = 0.459). There was no significant interaction between PD-L1 status and treatment for PFS, and no significant differences in ORR, PFS or OS between two arms regardless of PD-L1 status. In STEPP analysis, patients with lower composite risks had more absolute benefits in PFS than those with higher composite risks., Conclusions: GP regimen has superior efficacy than GT regimen as first-line chemotherapy for mTNBC patients. Germ-line mutations of BRCA1/2 and HR panel are possible biomarkers for better performance of cisplatin-based regimens. A composite risk model was developed to guide patient selection for GP treatment in TNBC patients., Trial Registration: ClinicalTrials.gov, NCT01287624.

New-onset diabetes after liver transplantation (NODALT) is a frequent complication with an unfavorable outcome. We previously demonstrated a crucial link between donor graft genetics and the risk of NODALT. We selected 15 matched pairs of NODALT and non-NODALT liver recipients using propensity score matching analysis. The donor liver tissues were tested for the expression of 10 microRNAs (miRNAs) regulating human hepatic glucose homeostasis. The biological functions of potential target genes were predicted using gene ontology and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis. Both miR-103 and miR-181a were significantly highly expressed in the NODALT group as compared to the non-NODALT group. The predicted target genes (e.g. Irs2, Pik3r1, Akt2, and Gsk3b) were involved in glucose import and the insulin signaling pathway. We also observed dysregulation of miRNAs (e.g. let-7, miR-26b, miR-145, and miR-183) in cultured human hepatocytes treated with tacrolimus or high glucose, the two independent risk factors of NODALT identified in this cohort. The hepatic miRNA profiles altered by tacrolimus or hyperglycemia were associated with insulin resistance and glucose homeostatic imbalance as revealed by enrichment analysis. The disease susceptibility miRNA expressive pattern could be imported directly from the donor and consolidated by the transplant factors., (© 2016 The Authors. American Journal of Transplantation published by Wiley Periodicals, Inc. on behalf of American Society of Transplant Surgeons.)

Aim: The objective of this meta-analysis was to evaluate the feasibility, safety, and short-term clinical outcomes of transanal total mesorectal excision (TaTME) comparing with laparoscopy total mesorectal excision (LapTME) for mid and low rectal cancer., Methods: Relevant studies were searched from the databases of Pubmed, Embase, and the Cochrane Library. The qualities of all of the included studies were evaluated using Newcastle-Ottawa Scale (NOS). The synthesized outcomes were pooled using fixed-effects models or random-effects models, which weighted the odds ratio (OR) or mean difference (MD) with 95% confidence intervals (95% CI). A funnel plot was used to evaluate the publication bias., Results: Seven original studies including 209 TaTME patients and 257 LapTME patients with rectal cancer met the inclusion criteria in this meta-analysis. Compared with LapTME, TaTME showed a longer CRM, lower rate of positive CRM, complete TME rate, and less operative time. There were no significant differences in the outcomes of the harvested lymph nodes, distal margin distance, hospital stay, intraoperative complications, anastomotic leakage, postoperative complications, reoperation, readmission, or conversion between the TaTME group and the LapTME group., Conclusions: Compared with LapTME, TaTME is a feasible and safe approach for patients with mid and low rectal cancer. In addition, TaTME showed a better short-term clinical outcomes, such as a longer CRM, lower risk of positive CRM, higher complete quality of TME rate, and shorter operative duration. Further prospective studies with long-term follow-up are required., (Copyright © 2016 Elsevier Ltd, BASO ~ The Association for Cancer Surgery, and the European Society of Surgical Oncology. All rights reserved.)

Islet transplantation represents a potentially curative approach for individuals with Type I Diabetes. The requirement for systemic immune suppression to control immune-mediated rejection of transplanted islets and the limited human islet supply represent significant roadblocks to progress for this approach. Islet microencapsulation in alginate offers limited protection in the absence of systemic immunosuppression, but does not support long-term islet survival. The chemokine, CXCL12, can repel effector T cells while recruiting immune-suppressive regulatory T cells (Tregs) to an anatomic site while providing a prosurvival signal for beta-cells. We proposed that coating or encapsulating donor islets with CXCL12 would induce local immune-isolation and protect and support the function of an allo- or xenograft without systemic immune suppression. This study investigated the effect of alginate microcapsules incorporating CXCL12 on islet function. Islet transplantation was performed in murine models of insulin-dependent diabetes. Coating of islets with CXCL12 or microencapsulation of islets with alginate incorporating the chemokine, resulted in long-term allo- and xenoislet survival and function, as well as a selective increase in intragraft Tregs. These data support the use of CXCL12 as a coating or a component of an alginate encapsulant to induce sustained local immune-isolation for allo- or xenoislet transplantation without systemic immunosuppression., (© Copyright 2015 The American Society of Transplantation and the American Society of Transplant Surgeons.)

The effects of monochromatic light on the ontogeny, differentiation, and Muc2 expression level in goblet cells were studied in the small intestines of late-stage broiler embryos. The embryos were exposed to blue light (B group), green light (G group), red light (R group), or darkness (D group) throughout the incubation period. On d 15 of incubation (E15), a few acidic goblet cells (only the sulfated subtype) were observed, and Muc2 mRNA expression was detected. On E18, however, neutral, acidic, and intermediate types, as well as the sulfated subtype, were observed in the small intestine, and a decreasing gradient of goblet cell density was found along the duodenum to ileum axis. Up to E21, 3 types of goblet cells and 3 acidic cell subtypes were found in all the small intestines. The goblet cell density increased along the duodenum to ileum axis. Monochromatic light stimulation resulted in no significant differences in the density and types of goblet cells between the different treatment groups on E15 and E18, but an increased Muc2 mRNA expression level was detected on E18 in the G group compared with the other treatment groups. On E21, the goblet cell density, proportion of acidic goblet cells, and Muc2 mRNA expression level increased in the G group compared with other treatment groups. These results suggest that the ontogeny and differentiation of goblet cells in broiler embryos display temporal and spatial differences. Green monochromatic light may have the potential to promote the proliferation and maturation of as well as the expression of Muc2 mRNA in goblet cells of broiler embryos., (© 2014 Poultry Science Association Inc.)

To study the activation of caspase-9 and its potential influence in conditioning, longissimus thoracis (LT), semitendinosus (STN) and psoas minor (PMi) muscles were used to analyze the ratio of pro-apoptotic bax to anti-apoptotic bcl-2 in fresh tissues and observe the changes in ATP, cytosolic cytochrome c and caspase-9 activity levels during storage at 4°C. Caspase-9 activity at 5 h is higher than the activity at 0 and 24 h in the muscles (P<0.001). The ATP content decreased between 0 and 3 h, between 8 and 14 h in the PMi and LT muscles (P<0.0001), whereas between 0 and 5 h, between 8 and 14 h in the STN muscle (P<0.0001). There is 60.2%, 55.3% and 43.1% available ATP in the STN, LT and PMi muscles at 5 h, respectively. The cytosolic cytochrome c level increased during 5 and 24 h storage in the LT and PMi muscles (P<0.0001), during 5 and 96 h in the STN muscle (P<0.0001). The cytosolic cytochrome c at 24 h (P<0.001) and ratio of bax to bcl-2 (P<0.05) was higher in the PMi than in other muscles. We concluded that the increase in cytosolic cytochrome c and available intracellular ATP should be responsible for the increase in caspase-9 activity; the activation of caspase-9 could be limited by the subsequent depletion of ATP; the postmortem release level of cytochrome c could be determined by the ratio of bax to bcl-2 in fresh tissues.

Background: Resistance to non-depolarizing neuromuscular blocking agents induced by sepsis is associated with the qualitative change in the nicotinic acetylcholine receptor (nAChR). This study aims to investigate the effects of sepsis on the neuromuscular block properties of vecuronium in relation to the expression of fetal and neuronal α7 type nAChR., Methods: Male Sprague-Dawley rats were randomly divided into sham and sepsis groups. Sepsis was induced by caecal ligation and puncture (CLP). The rats were injected i.v. with ulinastatin or normal saline on Day 10. Neuromuscular block properties of vecuronium were evaluated and neuromuscular function was assessed by electromyography on Days 1, 3, 7, and 14 after CLP. Expression of fetal and neuronal type α7-nAChR on the tibialis anterior muscle was assessed using immunohistochemistry and western blot. The mRNA encoding for γ- and α7 subunits was evaluated by real-time polymerase chain reaction., Results: The half maximal inhibitory response of vecuronium in the sepsis group significantly increased, peaked on Day 7, and then declined on Day 14 (P<0.05). The neuromuscular function decreased with increasing postoperation time in the sepsis group (P<0.05). Sepsis significantly increased the expression of γ- and α7-nAchR along with expression of γ- and α7 subunits mRNA, peaked on Day 7, and declined on Day 14 (P<0.05). Ulinastatin suppressed the expression of receptor protein and mRNA encoding for γ- and α7 subunits (P<0.05)., Conclusions: Pharmacodynamic changes with vecuronium seem to be associated with the expression of γ- and α7-nAChR in the skeletal muscle. Ulinastatin can improve this effect by inhibiting the expression of these receptors.

This study determined the effects of melatonin (MEL) and its receptors on monochromatic light-induced bursal B-lymphocyte proliferation in broiler chickens. In vivo, green light (GL) enhanced the proliferation of B lymphocytes in bursas by 16.49% to 30.83% and the expression of MEL receptor subtypes 1a (Mel1a), Mel1b, and Mel1c receptors in bursas by 6.91% to 366.98% than other light colors. However, pinealectomy reduced these parameters and eliminated the differences between GL and other light groups. In vitro, the MEL-induced bursal B-lymphocyte proliferation was most suppressed by prazosin (P = 0.001, selective Mel1c antagonist), followed by luzindole (P = 0.022, nonselective Mel1a/Mel1b antagonist), but not by 4-phenyl-2-propionamideotetralin (P = 0.144, selective Mel1b antagonist). Similarly, dibutyryl-cyclic adenosine monophosphate (cAMP; analog of cAMP; P = 0.017) but not 8-(4-chloro-phenylthio)-2'-O-methyladenosine-3',5'-cyclic monophosphate (P = 0.736; activator of exchange protein directly activated by cAMP) significantly inhibited bursal B-lymphocyte proliferation. These results suggest that MEL mediates GL-induced bursal B-lymphocyte proliferation through Mel1c and Mel1a receptors but not Mel1b receptors by activating the cAMP/protein kinase A pathway., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Our previous study suggested that green light promotes broiler growth during the early stage [posthatching day (P) 0 to P26], and blue light enhances growth during the later stage (P27 to P49). The purpose of this study was to improve broiler growth and productive performance by using a combination of monochromatic lights at critical points between the early and later stages of growth. A total of 512 male Arbor Acres broilers on P0 were reared under white light (W), red light (R), green light (G), and blue light (B) by using light-emitting diode lamps at 15 ± 0.2 lx from P0 to P26 (16 replicate pens/group, 8 birds/pen), and then switching to another color of light until P49 (4 replicate pens/group, 8 birds/pen). As compared with single monochromatic lights, broilers reared in environments under combinations of monochromatic lights, W→G, R→B, G→B, and B→G, attained heavier BW than those reared in environments under W→W (3.18 to 12.00%), R→R (1.96 to 18.14%), G→G (0.85 to 5.08%), and B→B (0.39 to 4.70%), respectively. In addition, feed conversion ratios in the W→B, R→B, and G→B combinations were lower than feed conversion ratios for W→W (15.86%, P < 0.05), R→R (18.41%, P < 0.05), and G→G (3.37%), respectively. Moreover, the eviscerated carcass weight and breast, thigh, and crus muscle weights under G→B were greater by 0.40 to 56.23% than were those for the other light groups except W→B (eviscerated carcass) and B→G (breast muscle). The results suggest that the application of the G→B and B→G exchanges can be used successfully to improve growth and productive performance in broilers.

Tenderness is considered as the most important quality determinant of meat. Calcium chloride application has been shown to improve tenderness by regulating endogenous proteinases. This study was designed to determine the effect of 300 mM calcium chloride injection on myofibrillar structures, caspase activities and shear force in longissimus muscles of bulls during postmortem storage of 7 days. Myofibrillar fragmentation index was determined as an index of proteolysis occurring in muscle fibers and associated proteins. Maximum tenderness was observed at days 4 and 7 in both treated and control samples. The injection of calcium chloride significantly increased myofibrillar proteolysis and improved tenderness at postmortem days 4 and 7. The treatment reduced caspase-9 activity at 4 h and day 4, whereas those of caspase-8 and -3 activities at days 1 and 4 with respect to control. The improved tenderness and increased myofibril fragmentation with decreased caspase activities suggested that the proteolytic systems activated with calcium chloride injection possibly behave independent of the caspase system.

Our previous studies demonstrated that green and blue monochromatic lights were effective to stimulate immune response of the spleen in broilers. This study was designed to investigate the effects of monochromatic light on both gut mucosal mechanical and immunological barriers. A total of 120 Arbor Acre male broilers on post-hatching day (P) 0 were exposed to red light, green light (GL), blue light (BL), and white light (WL) for 49 d, respectively. As compared with broilers exposed to WL, the broilers exposed to GL showed that the villus height of small intestine was increased by 19.5% (P = 0.0205) and 38.8% (P = 0.0149), the crypt depth of small intestine was decreased by 15.1% (P = 0.0049) and 10.1% (P = 0.0005), and the ratios of villus height to crypt depth were increased by 39.3% (P < 0.0001) and 52.5% (P < 0.0001) at P7 and P21, respectively. Until P49, an increased villus height (33.6%, P = 0.0076), a decreased crypt depth (15.4%, P = 0.0201), and an increased villus height-to-crypt depth ratio (58.5%, P < 0.0001) were observed in the BL group as compared with the WL group. On the other hand, the numbers of intestinal intraepithelial lymphocytes (27.9%, P < 0.0001 and 37.0%, P < 0.0001), goblet cells (GC, 22.1%, P < 0.0001 and 18.1%, P < 0.0001), and IgA(+) cells (14.8%, P = 0.0543 and 47.9%, P = 0.0377) in the small intestine were significantly increased in the GL group as compared with the WL group at P7 and P21, respectively. The numbers of intestinal intraepithelial lymphocytes (36.2%, P < 0.0001), GC (26.5%, P < 0.0001), and IgA(+) cells (68.0%, P = 0.0177) in the BL group were also higher than those in the WL group at P49. These results suggest that both mucosal mechanical and immunological barriers of the small intestine may be improved by rearing broilers under GL at an early age and under BL at an older age.

A cDNA library from white alpaca (Vicugna pacos) skin was constructed using SMART technology to investigate the global gene expression profile in alpaca skin and identify genes associated with physiology of alpaca skin and pigmentation. A total of 5359 high-quality EST (expressed sequence tag) sequences were generated by sequencing random cDNA clones from the library. Clustering analysis of sequences revealed a total of 3504 unique sequences including 739 contigs (assembled from 2594 ESTs) and 2765 singletons. BLAST analysis against GenBank nr database resulted in 1287 significant hits (E-value < 10(-10)), of which 863 were annotated through gene ontology analysis. Transcripts for genes related to fleece quality, growth and coat color (e.g. collagen types I and III, troponin C2 and secreted protein acidic and rich in cysteine) were abundantly present in the library. Other genes, such as keratin family genes known to be involved in melanosome protein production, were also identified in the library. Members (KRT10, 14 and 15) of this gene family are evolutionarily conserved as revealed by a cross-species comparative analysis. This collection of ESTs provides a valuable resource for future research to understand the network of gene expression linked to physiology of alpaca skin and development of pigmentation.

Carvacrol and eugenol were encapsulated in micellar nonionic surfactant solutions to increase active component concentrations in the aqueous phase and used to treat two strains of Listeria monocytogenes (Scott A and 101) and two strains of Escherichia coli O157:H7 (4388 and 43895) grown as biofilms in a Centers for Disease Control and Prevention reactor. L. monocytogenes biofilms were grown in two different growth media, 1:20 TSB and Modified Welshimer's broth (MWB), while E. coli O157:H7 was grown in M9. In general, L. monocytogenes strains were more resistant to both micelle-encapsulated antimicrobials than E. coli O157:H7 strains. The two antimicrobials were equally effective against both strains of E. coli O157:H7, decreasing viable counts by 3.5 to 4.8 log CFU/cm(2) within 20 min. For both bacteria, most of the bactericidal activity took place in the first 10 min of antimicrobial exposure. Biofilm morphology and viability were assessed by the BacLight RedoxSensor CTC Vitality kit and confocal scanning laser microscopy, revealing an increasing number of dead cells when biofilms were treated with sufficiently high concentrations of carvacrol- or eugenol-loaded micelles. This study demonstrates the effectiveness of the application of surfactant-encapsulated essential oil components on two pathogen biofilm formers such as E. coli O157:H7 and L. monocytogenes grown on stainless steel coupons.

Interspecific facilitation on phosphorus uptake was observed in faba bean/maize intercropping systems in previous studies. The mechanism behind this, however, remained unknown. Under nitrate supply, the difference in rhizosphere acidification potential was studied by directly measuring pH of the solution and by visualizing and quantifying proton efflux of roots between faba bean (Vicia faba L. cv. Lincan No.5), soybean (Glycine max L. cv. Zhonghuang No. 17) and maize (Zea mays L. cv. Zhongdan No.2) in monoculture and intercrop, supplied without or with 0.2 mmol L(-1) P as KH(2)PO(4). The pH of the nutrient solution grown faba bean was lower than initial pH of 6.0 from day 1 to day 22 under P deficiency, whereas the pH of the solution with maize was declined from day 13 after treatment. Growing soybean increased solution pH irrespective of P supply. Under P deficiency, the proton efflux of faba bean both total (315.25 nmol h(-1) plant(-1)) and specific proton efflux (0.47 nmol h(-1) cm(-1)) was greater than that those of soybean (21.80 nmol h(-1) plant(-1) and 0.05 nmol h(-1) cm(-1), respectively). Faba bean had much more ability of rhizosphere acidification than soybean and maize. The result can explain partly why faba bean utilizes sparingly soluble P more effectively than soybean and maize do, and has an important implication in understanding the mechanism behind interspecific facilitation on P uptake by intercropped species.

A total of 260 one-day-old Arbor Acres male broilers were exposed to red light (RL), green light (GL), blue light (BL), and white light (WL), respectively, by using a light-emitting diode system for 7 wk. There were 5 replicate pens for each light treatment and 13 birds per pen. The effects of monochromatic light on the immune response were studied. The results indicated that proliferation of peripheral blood T lymphocytes in the GL group was significantly increased (by 80.8 and 54.8%) compared with those in the RL and BL groups, respectively, at 21 d of age (P < 0.05). At 49 d of age, however, the proliferation response was significantly increased in the BL group compared with the RL group (26.9%, P< 0.05). Moreover, the GL group showed a significant elevation in the serum anti-Newcastle disease virus level as compared with that of the RL group at 28 d of age (32.9%, P < 0.05). In contrast, no significant difference in serum anti-Newcastle disease virus level was observed among the BL, RL, and WL groups at this age (P > 0.05). By 49 d of age, the antibody titer was higher in the BL group than in the RL group (62.8%, P < 0.05). However, no significant difference in antibody titer was seen among the BL, GL, and WL groups at this age. Interestingly, the BL group showed a 44.0% reduction in the level of serum interleukin-1beta as compared with that in the RL group at 49 d of age (P < 0.05). These results suggest that GL and BL enhance the immune response better than RL, and that BL may play a role in alleviating the stress response in broilers.

Concentrations of environmental microflora and Listeria monocytogenes were monitored at multiple environmental locations within a seafood-processing facility over the course of 6 months. Concentrations of L. monocytogenes were determined using a most-probable-number (MPN) enrichment procedure. Two floor drains had persistent low concentrations of L. monocytogenes (0.03 to >1,100 MPN/cm2). In comparison, concentrations of the other organisms in the drain were much higher (heterotrophic plate count range of 10(5) to 10(8) CFU/cm2). Concentrations of environmental organisms (heterotrophic aerobic plate counts and counts of pseudomonads, Shewanella spp., Aeromonas hydrophila, and coliforms) were not correlated with concentrations of L. monocytogenes. The 178 confirmed L. monocytogenes isolates from the MPN procedure were further characterized by randomly amplified polymorphic DNA analysis. Sixteen different banding patterns were identified, and nine of the patterns were identified from samples collected on two or more collection dates. From all locations, banding type A was observed in 98 confirmed isolates (55%). Although present, L. monocytogenes was a relatively minor component in the ecosystem of the floor drains in this seafood-processing facility.

Atmospheric dustfall samples from 23 locations in Tianjin, China, were collected and analyzed for 16 polycyclic aromatic hydrocarbons (PAHs) classified by the Environmental Protection Agency as priority pollutants from March 2002 to March 2003. SigmaPAH16 (sum of 16 PAH compounds) concentrations in the dustfall collected during heating season ranged from 2.5 to 85.5 mug/g, while that during the non-heating season varied from 1.0 to 48.2 microg/g dry weight. The dominant components in the heating season included naphthalene, phenanthrene, fluoranthene, and chrysene, while naphthalene, fluorene, phenanthrene, and fluoranthene were dominant during the non-heating season. Compared with the non-heating season, the heating season was characterized by a higher fraction of high-molecular-weight PAHs with four to six rings with exception of the samples from the east industrial area. The east industrial area had more significant correlations between individual PAH compounds, and more discrete triangular components of three-, four-, five- and six-ring PAHs. No significant correlations were observed between the PAHs concentrations and total organic carbon (TOC) in the dustfall samples. The deposition fluxes of sigmaPAH15 (sum of 15 PAHs except naphthalene), sigmaPAH6 (sum of 6 carcinogenic PAHs recommended by IARC) and benzo[a]pyrene (BaP) from atmospheric deposition to the whole area were estimated as 1911, 196, and 53 microg/m2/year, respectively. The deposition rates for PAH compounds in the east industrial area were higher than those in the urban and rural areas. Furthermore, the deposition contribution of PAHs during domestic heating season in winter was not significant relative to the annual inputs.

To decipher the molecular mechanism of EGCG induced HL-60 cell apoptosis, alterations of gene expression spectra in HL-60 cell line cells after treatment with EGCG were screened by cDNA chip, and analyzed with the GenePix 3.0 twice; and the cDNA chip results further identified by RT-PCR. Ninety-seven genes among the total 8398 (1.15%) showed consistent significant differential expression in the duplicated cDNA chip assessments. Thirty-nine genes (40.2%) were up-regulated and 58 genes (59.8%) were down-regulated; and the randomly selected four performed RT-PCR results agreed with the cDNA chip data. The results suggest that the apoptosis of HL-60 cells induced by EGCG is a progressive transformation process regulated by a variety of genes.

Several types of vegetables were collected from two contaminated sites in Tianjin, China. The bulk soil and the rhizosphere soil samples were also collected from the same plots. Sixteen PAHs in the samples were measured. The total concentrations of PAH16 in the bulk soil from the two sites were 1.08 and 6.25 microg/g, respectively, with similar pattern. The concentrations of PAH16 and individual compounds in the rhizosphere were significantly higher than those in the bulk soil with mean values of 2.25 and 7.82 microg/g for the two sites, respectively. The contents of both total and dissolved organic matter in the rhizosphere were also higher than those in the bulk soil. Almost all PAH compounds studied were detected in both roots and aerial parts of the vegetables studied. Abundance of higher molecular weight PAHs in vegetable, however, was lower than that in soil. Concentrations of PAH16 in vegetable were higher than those reported in the literature for other areas. It appears that agricultural soils and vegetables in Tianjin, especially those from the site located immediately next to an urban district and irrigated with wastewater for several decades, are severely contaminated by PAHs. Among the eight types of vegetable studied, the highest concentration of PAHs was found in cauliflower. By average, the concentration of PAH16 in the aerial part of vegetables was 6.5 times higher as that in vegetable root, suggesting that foliar uptake is the primary transfer pathway of PAHs from environment to vegetables.

An accelerated solvent extraction (ASE) procedure using water, n-hexane and a mixture of n-hexane and acetone as solvents in sequence was developed and tested to evaluate the bioavailability of DDT and its metabolites including p,p'-DDT, o,p'-DDT, p,p'-DDE, and p,p'-DDD (SigmaDDTs) to wheat uptake from soils characterized by varied organic carbon contents. Results indicated that the extractability of SigmaDDTs with water was enhanced considerably in the presence of water soluble organic carbon (WSOC), while the amount of SigmaDDTs extracted with n-hexane was negatively correlated to the content of water insoluble organic carbon (WIOC). The interaction between SigmaDDTs and WIOC also reduced the bioavailability of the pesticides to wheat roots during uptake. There was a good positive correlation between the amount of SigmaDDTs extracted by n-hexane and the amount of SigmaDDTs accumulated in wheat roots, suggesting some potential for the use of the n-hexane ASE-extracted fraction as an indicator of SigmaDDTs' bioavailability to plant uptake. As such, the three sequentially extracted fractions may be viewed as representing the mobile, bioavailable, and fixed pools of SigmaDDTs in the soil.

Extracellular matrix metalloproteinase inducer (EMMPRIN), a glycoprotein present on the cancer cell plasma membrane, enhances fibroblast synthesis of matrix metalloproteinases (MMPs). The demonstration that peritumoral fibroblasts synthesize most of the MMPs in human tumors rather than the cancer cells themselves has ignited interest in the role of EMMPRIN in tumor dissemination. In this report we have demonstrated a role for EMMPRIN in cancer progression. Human MDA-MB-436 breast cancer cells, which are tumorigenic but slow growing in vivo, were transfected with EMMPRIN cDNA and injected orthotopically into mammary tissue of female NCr nu/nu mice. Green fluorescent protein was used to visualize metastases. In three experiments, breast cancer cell clones transfected with EMMPRIN cDNA were considerably more tumorigenic and invasive than plasmid-transfected cancer cells. Increased gelatinase A and gelatinase B expression (demonstrated by in situ hybridization and gelatin substrate zymography) was demonstrated in EMMPRIN-enhanced tumors. In contrast to de novo breast cancers in humans, human tumors transplanted into mice elicited minimal stromal or inflammatory cell reactions. Based on these experimental studies and our previous demonstration that EMMPRIN is prominently displayed in human cancer tissue, we propose that EMMPRIN plays an important role in cancer progression by increasing synthesis of MMPs.

The CED4/Apaf-1 family of proteins functions as critical regulators of apoptosis and NF-kappaB signaling pathways. A novel human member of this family, called CARD12, was identified that induces apoptosis when expressed in cells. CARD12 is most similar in structure to the CED4/Apaf-1 family member CARD4, and is comprised of an N-terminal caspase recruitment domain (CARD), a central nucleotide-binding site (NBS), and a C-terminal domain of leucine-rich repeats (LRR). The CARD domain of CARD12 interacts selectively with the CARD domain of ASC, a recently identified proapoptotic protein. In addition, CARD12 coprecipitates caspase-1, a caspase that participates in both apoptotic signaling and cytokine processing. CARD12 may assemble with proapoptotic CARD proteins to coordinate the activation of downstream apoptotic and inflammatory signaling pathways., (Copyright 2001 Academic Press.)

To determine the extent of brick tea consumption fluorosis in children living at elevations of 2000 and 4300 m, 519 children aged 8-15 years living in Xiege'er Town at 4300 m and Zhangmu Town at 2000 m were examined for dental fluorosis, their urinary fluoride concentration was determined, their dietary structure investigated, and the fluoride concentrations of various foods, freshwaters, soils, and fuels determined. Fifteen Tibetan families living in these two areas of the nature reserve of Mount Qomolangma were studied according to UNEP, FAO, and WHO guidelines for the study of dietary intake of chemical contaminants, Horowitz's classification and examination of dental fluorosis, and Dean's dental fluorosis index. The results demonstrated that dental fluorosis in Tibetan children living at an elevation of 2000 m was significantly lower than that of children at 4300 m (P<0.01). Higher elevation can worsen the extent of fluorosis, leading to retention of fluoride in tissues as a result of hypoxia, but fluorosis can also be associated with the deterioration of the ecological environment at high elevation and a low-level economy. Beverages and foods mixed with brick tea water are responsible for the dental fluorosis in the children.

A specific, sensitive and reliable index for assessment of human zinc status has not been developed, and continues to present a considerable challenge for nutritionists in the trace element field. We have focused on metallothionein (MT) expression as a potential index. A protocol involving 16 men and a 10-d supplementation period plus a 4-d postsupplementation period was used to examine the relative response of MT expression in erythrocytes, monocytes, peripheral blood mononuclear cells (PBMC) and cells from a dried blood spot (DBS). Zinc was supplemented at the current adult male recommended dietary allowance (RDA) of 15 mg. Erythrocyte MT protein, as measured by ELISA, increased gradually to about twofold over the placebo group during zinc supplementation and remained elevated for 4 d postsupplementation. Competitive reverse transcriptase-polymerase chain reaction showed that MT mRNA levels in both monocytes and PBMC increased (up to 4.7- and 2.7-fold, respectively) after 2 d of supplementation, with greater expression in monocytes compared with PBMC. Total RNA extracted from dried blood spots, representing cells from 50 microL of blood, showed a comparable change in MT mRNA upon zinc supplementation. In each leukocyte population isolated, when zinc supplementation was withdrawn, MT mRNA levels decreased. Collectively, these experiments show that, in men, MT gene expression increases during supplementation at the RDA, and that the DBS sampling method will be of value in measuring MT expression in a variety of clinical and survey situations.

Paracheirodon innesi was exposed (simultaneously or sequentially) to copper and lead in varying concentrations in a synthetic water solution. Metal accumulations in fish gills and the fish (without gills) were measured at the end of the exposure period. Results of the exposure experiment indicated that lead seems to facilitate the uptake of copper and vice versa when the exposure process is simultaneous. In the case of sequential exposure first to copper for 7 days then to lead for 20 days, lead accumulations in the fish and the gills were enhanced by copper, indicating a possible alteration of the physiological absorption process. The synergistic effects observed during either simultaneous or sequential exposure experiments can be described using logistic equations.

The effects of potassium phosphate buffer and its concentration upon the capillary zone electrophoretic separation of 23 sulphonamides and a neutral marker were examined at pH 7. The resolution between the pairs was improved with the increased concentration of the buffer from 65 mM to 174 mM. Nineteen sulphonamides, a hydrolysis product and several unidentified minor components were baseline resolved in both 101 and 138 mM phosphate buffers. In 174 mM buffer all 21 ionised sulphonamides and the other compounds were separated. A simple relationship between the resolution of analyte pairs (Rs) and the square root of the mean analysis time for the pair (square root of tapp) was derived, but few of the pairs displayed this behaviour. For the majority of pairs of compounds, Joule heating appeared to cause a maximum in the Rs versus square root of tapp relationship, while non-ideality and shifts in ionisation with increasing salt concentration appeared dominant in other cases.

The effects of histamine, cimetidine, and diphenhydramine on picryl chloride (PCl)-induced ear contact sensitivity, as well as liver injury, were examined in mice. Histamine was found to produce less response in mice to PCl. In contrast, cimetidine, a selective antagonist of histamine type 2 receptor, significantly enhanced the response, while diphenhydramine, a selective antagonist of histamine type 1 receptor showed no effect. The pre-treatment of 2,4, 6-trinitrobenzene sulphonic acid (TNBS) significantly caused a tolerance to the formation of the liver injury induced by delayed-type hypersensitivity (DTH) to PCl. Against the tolerance, the single intravenous administration of 150 mg kg-1 cyclophosphamide (Cy) at 3 days before the TNBS-treatment recovered the response and induced a remarkable elevation of serum transaminases. On the other hand, cyclosporin A protected the liver injury. These observations revealed that the development of acute PCl-DTH liver injury was regulated by the functional state of suppressor and helper T cells., (Copyright 1999 The Italian Pharmacological Society.)

A reporter gene system that allows in situ detection of cells that have suffered a specific frameshift mutation was developed. To construct the reporter gene, the open reading frame of a human placental alkaline phosphatase (PLAP) gene was disrupted by insertion of either 5 or 7 G:C basepairs, which formed mutant alleles carrying 9 and 11 consecutive G residues, respectively. The mutant PLAP genes did not produce alkaline phosphatase activity in cultured mouse cells in transient transfection assays. Several cell lines that contained integrated copies of the mutant PLAP genes were made. Histochemical staining of fixed cells showed that these cell lines contained a small number of cells that expressed PLAP activity and bound antibodies directed against PLAP. Cells carrying the allele with 11 consecutive G residues (G11 allele) acquired PLAP activity at a rate between 2 x 10(-3) and 2 x 10(-4) events per cell per generation, depending on the cell line. Cells carrying the allele with 9 consecutive G residues (G09 allele) acquired PLAP activity at a rate between 2 x 10(-5) and 2 x 10(-6) events per cell per generation, depending on the cell line. Cultures of PLAP+ cells were derived from cell lines carrying PLAP mutant genes. All the cells in these cultures had PLAP activity and bound anti-PLAP antibody. PLAP mRNA levels were the same in cultures where all cells were PLAP+ and in cultures where less than 1% of the cells expressed PLAP activity. DNA sequence analysis of PLAP+ cells showed that the G11 allele reverted by losing one basepair, and the G09 allele reverted by gaining one basepair.

The phosphorylation state of phosphofructokinase from the mantle tissue of the facultative anaerobe mollusk Mytilus galloprovincialis was determined by a back-phosphorylation technique. The incubation of intact mantle tissue with 8-bromoadenosine 3':5'-cyclic monophosphate increased significantly the phosphate content of phosphofructokinase, which indicates that the enzyme can be phosphorylated in vivo by endogenous cAMP-dependent protein kinase. The phosphate content of mussel phosphofructokinase changes significantly during the year, in agreement with the kinetic data that show a more active enzyme form in earlier autumn. These results suggest that cAMP-dependent phosphorylation of phosphofructokinase can be partially responsible for the observed glycolytic changes associated with the annual gametogenic cycle that takes place in the mantle tissue of the mollusk. On the contrary, no differences were observed between aerobic and 24-h hypoxic mussels with regard to the phosphorylation state and the kinetic constants of phosphofructokinase. This result is inconsistent with the hypothesis that phosphorylation of phosphofructokinase is involved in the glycolytic depression that takes place during the long-term environmental hypoxia that the mollusk can undergo.

Background: The chemical mechanism of the final step of ethylene biosynthesis (the conversion of 1-aminocyclopropanecarboxylic acid, ACC, to ethylene by ACC oxidase, the ethylene-forming enzyme, EFE) is poorly understood. Two possibilities have been suggested: a radical mechanism and an N-hydroxylation mechanism. We investigated reaction pathways available to radical intermediates in this reaction using an ACC analog, 1-aminocyclobutanecarboxylic acid (ACBC) as a substrate., Results: ACBC was converted to dehydroproline (delta 1-pyrroline-2-carboxylic acid) by the EFE via a ring expansion process. The possibility that an N-hydroxy-aminoacid (produced during two-electron oxidation) acts as an intermediate in this process was eliminated by control experiments. Chemical model reactions involving two-electron oxidants, such as a positive halogen (X+), which presumably generate N-halo derivatives, produce only decarboxylation products. Radical-based oxidants, in contrast, generate dehydroproline. Model reactions involving sequential single-electron transfer mechanisms also produce dehydroproline; thus our results support the proposal that the EFE-catalyzed step of ethylene biosynthesis proceeds using a radical-based mechanism., Conclusions: Our results provide support for a radical mechanism in the final step of ethylene biosynthesis and refute an alternative N-hydroxylation mechanism. This work extends the idea that the intrinsic chemical reactivity of a high energy iron-oxo intermediate can account for the observed products in ethylene biosynthesis.

The kinetics of lymphocyte function associated antigen 1 (LFA-1) expression on spleen cells (SPC) and liver non-parenchymal cells (NPC), and intercellular adhesion molecule 1 (ICAM-1) expression on hepatocytes (HC) was examined in acute liver injury mice induced by a DTH reaction to picryl chloride (PCl). The peak expression of LFA-1 on SPC was seen at 6 hr after eliciting liver injury, and then that of LFA-1 on NPC and ICAM-1 on HC appeared at 12 hr. Thereafter, the serum ALT elevation reached to a peak at 18 hr. A splenectomy before the PCl elicitation significantly reduced the ALT elevation. Both SPC and NPC from liver injury mice induced a remarkable release of ALT from HC in vitro, in parallel with their LFA-1 expression. The pre-treatment of NPC or SPC with anti-LFA-1 mAb, irrespective of the presence of complement, completely blocked the ALT release. Also, when HC was prebound with anti-ICAM-1 mAb, neither NPC nor SPC showed a cytotoxicity against the HC. Furthermore, the treatment of NPC with either anti-Thy1.2 or anti-CD4 mAb in the presence but not absence of complement, showed a complete abolishment of ALT release. Anti-CD8 mAb plus complement also tended to inhibit ALT release. The twofold increase in CD4+ LFA-1+ and mild increase in CD8+ LFA-1+ populations were also confirmed in NPC at 12 hr. These results suggest that PCl elicitation in liver may trigger an increased expression of LFA-1 on SPC and NPC and ICAM-1 on HC. LFA-1/ICAM-1 interaction between liver-infiltrating NPC, mainly including CD4+ and CD8+ T cells, and HC may be an essential step for the hepatocyte damage in PCl-DTH liver injury.

Helicobacter pylori is the major cause of gastritis. The aim of this investigation was to develop a specific antibody, which recognizes both coccoid and spiral forms of Helicobacter pylori and to test this antibody on gastric biopsy sections known to harbour coccoid bacteria. Murine monoclonal antibodies against glycine-acid extracts of five strains of Helicobacter pylori were raised. Immunofluorescence and immunoelectron microscopy showed that one antibody of the IgG1 subclass was specific for both the spiral and coccoid forms. It reacted with a 28 kDa protein that was present in all the five strains tested. Using this antibody in an indirect immunofluorescence assay of formalin-fixed antral and corpus biopsy specimens from Helicobacter pylori-associated gastritis patients showed that nine of the nine antral and five of six corpus specimens harboured the coccoid form of Helicobacter pylori. This technique thus provides a rapid and specific detection of both the spiral and coccoid forms.

Capillary dropout is an initial event in diabetic retinopathy, but the etiology is unknown. Recent evidence suggests that similar events may occur in the diabetic choroid. We have developed a method to evaluate the relationship between the polymorphonuclear leukocytes (PMNs) and capillary dropout in the human diabetic choroid using alkaline phosphatase (APase) histochemistry to label blood vessels and nonspecific esterase activity to identify PMNs. The number and distribution of PMNs in diabetic and nondiabetic choroidal capillaries (choriocapillaris) were analyzed in the flat perspective and the tissue then flat embedded in glycol methacrylate for histological sectioning. The total number of PMNs was increased within the choriocapillaris in five diabetic eyes (170.9 +/- 12.9 PMNs/mm2 of choroid) compared with five nondiabetic eyes (84.2 +/- 16.9 PMNs/mm2; P < 0.001). PMNs were almost always within blood vessel lumens and not in interstitial tissue. In the diabetic choroid, increased numbers of PMNs were present in areas of choriocapillaris with pathological changes (loss in APase activity and choroidal neovascularization) compared with nonpathological choriocapillaris (205.1 +/- 46.9 PMNs/mm2 in pathological versus 152.3 +/- 23.4 PMNs/mm2 in nonpathological areas; P < 0.001). PMNs were often queued up within the lumens of capillaries, demonstrating loss in APase activity. We have observed an increased number of PMNs in diabetic choroid compared with control nondiabetic choroids, and PMNs in diabetic choroid were associated with loss in APase activity, which was related to loss in viable endothelial cells. The results suggest that PMNs contribute to vaso-occlusive processes and endothelial cell injury in the diabetic choroid.

Various drugs for clinical hepatitis were applied to a new model of liver injury induced in mice by delayed-type hypersensitivity to picryl chloride (PCI-DTH). The hepatoprotective agent, biphenyl dimethyl dicarboxylate showed a remarkable improvement against the elevation of serum transaminase levels as well as the histopathological changes when given during the induction phase but not during the effector phase of DTH reaction. Cyclophosphamide (Cy), an immunosuppressive agent, significantly inhibited the enzymatic elevation given in both induction and effector phases. However, Cy did not affect the sustaining of liver injury 4 weeks after the liver injury eliciting. Moreover, the consecutive administration of prednisolone (Pred), in both induction phase and sustaining process of liver injury, conversely caused a more severe liver damage. Such exacerbation by Pred might be resulted from its toxic action to hepatocytes. As an immunomodulatory and antiinflammatory agent, glycyrrhizin remarkably improved the sustaining process but not the acute phase of the liver injury. Krestin and malotilate also showed an improving effect on the sustaining development of liver injury. These findings that most of above drugs showed an improving action in their respective manner suggest that this model may be useful for the pharmacological evaluation of drugs especially immunomodulating agents for hepatitis.

A lacZ transgene recombination system that reports homologous recombination events involving duplicated lacZ segments was used to study recombination in monkey cells exposed to ionizing radiation at different points in the cell cycle. With this system, recombination events can be detected in single cells by histochemical staining soon after exposure of cells to DNA-damaging treatment. Ionizing radiation rapidly induced recombination 5-10-fold in cells that were at the mitosis stage of the cell cycle. Irradiation either of cells at other points in the cell cycle or of nonsynchronized cells had less of an effect on recombination between lacZ segments.

The developmental characteristics of liver injury induced by a delayed-type hypersensitivity (DTH) mechanism against picryl chloride were examined for 9 consecutive weeks in 3 mouse strains, BALB/c, Kunming and ICR mice. The changes of most biochemical parameters were similar in these three strains, namely, the activities of serum transaminases, lactic dehydrogenase, and prolidase were elevated significantly on day 1, during the first several weeks, and almost throughout the duration, respectively, of liver injury. The content of liver hydroxyproline was also increased after 1-9 weeks of liver injury. In addition, a significant decrease of liver weight, serum alkaline phosphatase and albumin level was observed in BALB/c and Kunming mice. Similar changes in liver histology were also found in the three strains. The hepatocellular necrosis and inflammatory infiltration into the portal area were the predominant features on day 1 and were still distinct during the subsequent several weeks. The mild or moderate hepatocellular degeneration, regeneration and connective tissue hyperplasia were observed after 1 or 3 weeks. A bridging necrosis between portal and portal was observed in several BALB/c and ICR mice, reflecting the possibility of exacerbation of liver injury. These results suggest that the liver injury could be caused and sustained by a one-shot DTH reaction to picryl chloride. The chronicity of the biochemical and histopathological characteristics may be helpful in elucidating the mechanisms of chronic development of liver injury.

Three experiments were conducted with day-old chicks to study the effects of dietary Fe concentration and age on Fe accumulation in tissues as an estimate of supplemental Fe bioavailability, and of delaying the time of initial high Fe supplementation up to 7 d of age on feed intake to 3 wk of age. In Experiment 1, chicks were fed a basal corn-soybean meal diet (188 mg/kg Fe, DM basis) or the basal supplemented with 400, 600, or 800 mg/kg added Fe as reagent grade FeSO4.7H2O for either 1, 2, or 3 wk. Dietary Fe depressed (P < 0.001) feed intake and body weight gain, especially at 3 wk. Kidney Fe concentrations increased linearly (P < 0.001) with increasing dietary Fe. Liver Fe concentration also increased linearly, but reached a plateau in birds fed 600 mg/kg Fe. Bone Fe increased linearly (P < 0.05) at 1 wk, but not at 2 or 3 wk. Liver and kidney Fe regressed on daily Fe intake had the best fit to a linear model at 2 wk. In Experiment 2, chicks were fed either a basal diet (320 mg/kg Fe, DM basis) continuously, the basal supplemented with 800 mg/kg added Fe as FeSO4.7H2O continuously, or were started on the control diet and switched to the high Fe diet on Day 3, 5, or 7. Feed intake was lower (P < 0.05) in birds started on Fe on Days 1 or 3, but delaying feeding of high Fe diets until Day 5 resulted in intake at 3 wk similar to that of birds fed the basal diet. In Experiment 3, the basal diet (123 mg/kg Fe) was fed to chicks for 6 d, then experimental diets were fed for 14 d. Diets were the basal or basal supplemented with 400, 600, or 800 mg/kg added Fe as reagent grade or feed grade Fe sulfate or an Fe methionine complex. When estimated from regression of log10 liver Fe concentration on total analyzed dietary Fe concentration, relative bioavailability was set at 100% for reagent grade Fe sulfate, and the feed grade sulfate was 92.3% and Fe methionine was 88.3%. Liver Fe concentrations may be useful criteria for determining Fe bioavailability and 2 wk of feeding was the optimal time required for such a bioassay. Delaying feeding high Fe diets until 5 d of age alleviated the decreased feed intake associated with high Fe diets.

The immunoglobin heavy chain variable region (VH) gene usage in multiple myeloma (MM) has not been reported, although a few studies have incidentally identified the VH gene rearranged in small cohorts of MM patients. We used a reverse transcriptase-polymerase chain reaction based technique to analyze the VH gene usage in MM. The VH sequences were obtained after amplification of bone marrow cDNA using the seven VH family-specific and constant region primers. The VH sequences of 72 patients were successfully identified. The frequency of VH family usage in decreasing order was VH3>VH4>VH1>VH5>VH2>VH6>VH7 and corresponded to the functional germline complexity of the VH families. Individual VH genes (VH1-69, VH3-9, VH3-23, and VH3-30) were overrepresented in our cohort of MM patients; some VH genes [VH3-49, VH3-53, and VH4.21 (VH4-34)], which are rearranged with increased frequency in normal circulating B cells, autoimmune diseases, and other B-cell malignancies, were not detected in any MM patient. Compared with germline sequences, an average of 8.8% (range, 2.7% to 16.5%) of the nucleotides had evidence of mutation within each VH sequence. Based on these results, we conclude that (1) the VH gene usage in MM is unique compared with other malignant and nonmalignant B-cell populations, (2) the physiologic process of clonal deletion functions to remove clones that have rearranged VH genes (VH4.21) capable of expressing antibodies, which recognize self-antigens, and (3) the complete lack of VH4.21 gene rearrangement may help to partially explain the paucity of autoimmune phenomena in MM.

A major potential problem of autologous transplantation in the treatment of advanced malignancy is the infusion of tumor cells. A multi-institutional study of purified CD34-selected peripheral blood progenitor cell (PBPC) transplantation was conducted in 37 patients with advanced multiple myeloma receiving myeloablative chemotherapy. Fourteen days after intermediate-dose cyclophosphamide, prednisone, and granulocyte colony-stimulating factor (G-CSF), a median of 3 (range, 2 to 5) 10-L leukaphereses yielded 9.8 x 10(8)/kg (range, 3.7 to 28.3) mononuclear cells. The adsorbed (column-bound) fraction contained 5.9 x 10(6) cells/kg (range, 1.6 to 25.5) with 4.65 x 10(6) CD34 cells/kg (range, 1.2 to 23.3). Using Poisson distribution analysis of positive polymerase chain reactions with patient-specific complementarity-determining region 1 (CDR1) and CDR3 Ig-gene primers, tumor was detected in leukapheresis products from 8 to 14 unselected patients and ranged from 1.13 x 10(4) to 2.14 x 10(6) malignant cells/kg. After CD34 selection, residual tumor was detected in only three patients' products. Overall, a greater than 2.7- to 4.5-log reduction in contaminating multiple myeloma cells was achieved. CD34 PBPCs were infused 1 day after busulfan (14 mg/kg) and cyclophosphamide (120 mg/kg), and granulocyte-macrophage colony-stimulating factor was used until hematologic recovery. The median time to both neutrophil and platelet recovery was 12 days (range, 11 to 16 days and 9 to 52 days, respectively). The median number of erythrocyte and platelet transfusions was 7 (range, 2 to 37) and 3 (range, 0 to 85), respectively. Patients receiving fewer than 2 x 10(6) CD34 cells/kg had significantly prolonged neutropenia, thrombocytopenia, and an increased red blood cell and platelet transfusion requirement. Thus, CD34 selection of PBPCs markedly reduces tumor contamination in multiple myeloma and provides effective hematopoietic support for patients receiving myeloablative therapy.

Autologous stem cell transplantation has become an important therapy in multiple myeloma (MM). To develop adequate autograft purging methods, it is necessary to determine whether antigens expressed on early hematopoietic progenitors exist on malignant cells. The Ig heavy chain produced by the MM cells shows evidence of prior somatic mutation without intraclonal diversity. As a result, this sequence can be used as a specific marker to detect all members of the malignant clone. The Ig heavy chain sequence expressed by the MM cells was obtained in five patients with advanced disease. Patient specific oligonucleotide primers were designed based on the complementarity determining regions (CDR) of each MM Ig sequence and used to amplify DNA by polymerase chain reaction for the detection of malignant cells. A highly purified collection of CD34+ cells was obtained after passage of the initial bone marrow cells through an immunoadsorption column and fluorescence-activated cell sorting. Despite an assay sensitivity of 1 tumor cell in 2,500 to 44,000 normal cells, none of the CD34+ samples showed product with the myeloma-specific CDR primers. Therefore, positive selection for cells bearing this antigen should yield a tumor-free autograft capable of providing hematopoietic recovery after myeloablative chemotherapy.

Inhalation exposure of mice to 50, 200, 500 or 1300 ppm of 1,3-butadiene for 6 h per day for 5 consecutive days caused micronuclei in mouse bone marrow and peripheral blood erythrocytes. The dose response was non-linear. The slope of the curve flattened with increasing exposure concentration. Coat color spots were found in the mouse spot test after exposure of pregnant females on pregnancy days 8-12 to 500 ppm of 1,3-butadiene. Dominant lethal mutations were induced in spermatozoa and late spermatids after exposure of male mice to 1300 ppm with the 5-day exposure regimen. Thus, in the mouse 1,3-butadiene is a somatic and germ cell mutagen.