[Early German research on virology and establishment of the specialised research institution].

Aftosa spread rapidly in Europe at the end of the 19th century.The German imperial government was concerned about it and appointed some scientists, such as Friedrich Loeffler and Paul Frosch, to do the research on it and they identified the virus of Aftosa in 1897. An experimental study was planned to be conducted by Friedrich Loeffler at the Greifswald Institute of Hygiene at a farm outside the city in 1899, but cancelled by the Government concerned about potential safety hazards in the research sites. After three years of preparation, the first virology research institution in the world was established by Friedrich Loeffler and others on Riems Island in Germany on October 10, 1910. This old virology research institution has now become the Federal Animal Health Research Center, making significant contributions to human research on zoonotic diseases.

Effect of Lonicera japonica extract on lactation performance, antioxidant status, and endocrine and immune function in heat-stressed mid-lactation dairy cows.

Here, we examined the effects of Lonicera japonica extract (LJE) on lactation performance, antioxidant status, and endocrine and immune function in heat-stressed mid-lactation dairy cows. Twenty-four healthy Chinese Holstein mid-lactation dairy cows, all with similar milk yield (30.0 ± 1.0 kg/d), parity (2.5 ± 0.3), and days in milk (105 ± 5 d) were allocated to 4 groups using a randomized complete block design: a negative control group (without LJE supplementation; CON) and groups that received LJE at 14, 28, and 56 g/d. The experiment lasted 10 wk over a hot summer, with a pre-feeding period of 2 wk. Cows were exposed to heat stress, as the average temperature-humidity index was greater than 72. The results showed that LJE had no effect on respiration rate; however, it reduced the rectal temperature of dairy cows experiencing heat stress in both a linear and quadratic manner; the lowest (39.03°C) was recorded for the LJE-28 group, lower than the CON group. Supplementation with LJE did not affect dry matter intake, milk yield, or milk composition. The majority of biochemical parameters in serum were unaffected by supplementation with different amounts of LJE; the exception was creatinine, which was reduced quadratically. Compared with the CON group, serum triiodothyronine concentrations increased significantly in the LJE-28 group. Addition of LJE to the diet increased thyroxine concentrations quadratically; values peaked at 18.62 ng/mL in the LJE-28 group. Furthermore, supplementation with increasing amounts of LJE quadratically increased the activity of glutathione peroxidase and total antioxidant capacity in serum but decreased concentration of malondialdehyde. Although we detected no differences in the concentrations of IgA, IgM, or cytokines, dairy cows in the LJE-28 group had higher IgG and IL-4 concentrations than did cows in the CON group. Supplementation with LJE increased concentrations of IgG and IL-4 in the serum quadratically but decreased that of IL-2. Finally, heat shock protein 72 concentrations in the serum tended to fall quadratically as the amount of LJE increased. In summary, LJE had no negative effects on lactation performance but helped to alleviate heat stress by improving antioxidant status and promoting endocrine and immune functions. Supplementation with LJE at 28 g/d is recommended for lactating dairy cows experiencing heat stress during hot summers.

Effects of different types of zinc supplement on the growth, incidence of diarrhea, immune function, and rectal microbiota of newborn dairy calves.

Neonatal diarrhea in dairy calves causes huge economic and productivity losses in the dairy industry. Zinc is an effective anti-diarrheal agent, but high doses may pose a threat to the environment. Therefore, we aimed to evaluate the effects of low-dose zinc supplementation on the growth, incidence of diarrhea, immune function, and rectal microbiota of newborn Holstein dairy calves. Thirty newborn calves were allocated to either a control group (without extra zinc supplementation), or groups supplemented with either 104 mg of zinc oxide (ZnO, equivalent to 80 mg of zinc/d) or 457 mg of zinc methionine (Zn-Met, equivalent to 80 mg of zinc/d) and studied them for 14 d. The rectal contents were sampled on d 1, 3, 7, and 14, and blood samples were collected at the end of the study. Supplementation with ZnO reduced the incidence of diarrhea during the first 3 d of life, and increased serum IgG and IgM concentrations. The Zn-Met supplementation increased growth performance and reduced the incidence of diarrhea during the first 14 d after birth. The results of fecal microbiota analysis showed that Firmicutes and Proteobacteria were the predominant phyla, and Escherichia and Bacteroides were the dominant genera in the recta of the calves. As the calves grew older, rectal microbial diversity and composition significantly evolved. In addition, dietary supplementation with ZnO reduced the relative abundance of Proteobacteria in 1-d-old calves, and increased that of Bacteroidetes, Lactobacillus, and Faecalibacterium in 7-d-old calves, compared with the control group. Supplementation with Zn-Met increased the relative abundance of the phylum Actinobacteria and the genera Faecalibacterium and Collinsella on d 7, and that of the genus Ruminococcus after 2 wk, compared with the control group. Thus, the rectal microbial composition was not affected by zinc supplementation but significantly evolved during the calves' early life. Zinc supplementation reduced the incidence of diarrhea in young calves. In view of their differing effects, we recommend ZnO supplementation for dairy calves during their first 3 d of life and Zn-Met supplementation for the subsequent period. These findings suggest that zinc supplementation may be an alternative to antibacterial agents for the treatment of newborn calf diarrhea.

Serum fibroblast growth factor 23 and mineral metabolism in patients with euthyroid Graves' diseases: a case-control study.

This study investigated the alterations of mineral metabolism in patients with Graves' disease (GD) who achieved euthyroidism. They had higher fibroblast growth factor 23 (FGF23) and phosphorus as compared with healthy subjects. Serum FGF23 was negatively correlated with serum phosphorus. These indicated abnormal mineral metabolism even after 1.6 years of euthyroid status. INTRODUCTION: FGF23 is involved in the mineral homeostasis, especially the regulation of serum phosphorus. Graves' disease (GD) is associated with accelerated bone turnover, hyperphosphatemia, and elevated serum FGF23. Evidence suggested that serum FGF23 decreased after a 3-month treatment of GD. However, it remains unclear whether serum FGF23, serum phosphorus, and other markers of mineral metabolism will be normalized after euthyroid status achieved. METHODS: A total of 62 patients with euthyroid GD and 62 healthy control subjects were enrolled, and the median duration of euthyroid status was 1.6 years. Endocrine profiles including thyroid function test, autoantibodies, serum FGF23, and bone turnover markers were obtained and compared between the two groups. RESULTS: Euthyroid GD patients had significantly higher serum FGF23 and phosphorus, and lower 25-hydroxyvitamin D (25(OH)D) and intact parathyroid hormone (iPTH) levels as compared with the control group. Serum FGF23 was significantly and negatively correlated with phosphorus level after adjusted for age, gender, calcium, iPTH, and 25(OH)D in the euthyroid GD group. CONCLUSION: Serum phosphorus and FGF23 levels remain higher in GD patients even after euthyroid status has been achieved for a median of 1.6 years. Serum FGF23 was negatively correlated with serum phosphorus in euthyroid GD patients. Underlying mechanisms warrant further investigations. TRIAL REGISTRATION: Registration number: NCT01660308 and NCT02620085.

[Outcome of postoperative adjuvant chemotherapy and surgery alone for patients with stage pT1b-3N0M0 squamous cell carcinoma of the thoracic esophagus].

Objective: To compare the outcome of postoperative adjuvant chemotherapy and surgery alone for stage pT1b-3N0M0 squamous cell carcinoma (SCC) of the thoracic esophagus. Methods: Two hundred and thirty-eight patients who underwent esophagectomy for stage pT1b-3N0M0 SCC of the thoracic esophagus in Department of Thoracic Surgery, Fourth Hospital, Hebei Medical University between January 2008 and February 2011 were analyzed retrospectively. These included postoperative adjuvant chemotherapy for 118 patients and surgery alone for 120 patients. In postoperative adjuvant chemotherapy arm, 3 or 4 cycles of taxol+ cisplatin (TP, n=33) or fluorouracil+ cisplatin (FP, n=85) regimens were given commencing from 4 to 6 weeks after surgery. Survival analysis was performed using Kaplan-Meier method. Univariate analysis for prognostic factors was performed by Log-rank test and multivariate by Cox regression model. Results: The 3- and 5-year disease-free survival (DFS) rates were 57.9% and 53.8%, respectively for postoperative adjuvant chemotherapy patients, compared to 40.4% and 32.5% for patients who underwent surgery alone (χ2=11.973, P=0.001). The 3- and 5-year overall survival (OS) rates were 64.4% and 55.9% in postoperative adjuvant chemotherapy patients, and 46.7% and 33.5% in surgery alone patients (χ2=13.110, P=0.000). Stratification analysis showed that there was significant difference in stage T3 patients (χ2=7.895, P=0.006), but not in stage T1b (χ2=0.762, P=0.383) and T2 (χ2=1.259, P=0.262) patients between adjuvant chemotherapy and surgery alone. Both DFS rate (χ2=1.748, P=0.186) and OS rate (χ2=2.200, P=0.138) of TP group were similar with FP group. Conclusion: In lymph node negative esophageal SCC patients, postoperative adjuvant chemotherapy shows survival benefits in stage T3 patients.

[Analysis of compiling date of Huang di nei jing (Huangdi's Internal Classic)].

There are Seven Medical Classics recorded in the Han shu Yi wen zhi(Bibliography in Annals of the Han Dynasty), among which the 18-volume Huang di nei jing (Huangdi's Internal Classic) is not the current edition of Ling shu (Miraculous Pivot) and Su wen(Plain Questions). The present edition of Huang di nei jing containing these two works might be merged in the Eastern Han Dynasty, benefited from the invention of Cai Lun's papermaking technique and the development of Nine needles, especially filiform needle manufacturing. Huang di nei jing was compiled on the basis of two major integrations: The first was joined by some medical experts such as Bian Que and tai yi ling (minister of imperial physicians) of the Qin State by using the language in the Warring States, contributing mainly to the theoretical system and forming its basic academic framework; The second was the second Royal book-collating activity in the Eastern Han Dynasty (100-110), which had revised the seven volumes of Chinese Medical Classics and incorporated them into two parts, Ling Shu and Su Wen, together with Bian Que's medical works and the new contents from the introductory Chapter on Nine Needles and Twelve Yuan(origin) Acupoints.

Growth inhibitive effect of betulinic acid combined with tripterine on MSB-1 cells and its mechanism.

Marek's disease (MD), a highly infectious lymphoproliferative disease in chickens, is caused by a cell-associated oncogenic herpesvirus, Marek's disease virus (MDV). MSB-1 is a MD-derived lymphoblastoid cell line and can induce tumors when inoculated into susceptible chickens. Betulinic acid, which is present as one of the major effective components in many traditional Chinese medicines, has recently been reported to inhibit growth of cancer cells and employed as a potential anticancer agent. Tripterine, a major active compound extracted from the Chinese herb Tripterygium wilfordii Hook F, has now also shown anti-tumor activities in various cancers. The aim of this study was to investigate the synergistic growth-inhibitive effect of betulinic acid combined with tripterine on MSB-1 cells and its mechanism. Viability of MSB-1 cells was assessed by 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) method. Cell apoptotic analysis was performed by fluorescence detection. NF-κB transcription activity was detected by measuring luciferase activity. Western blotting was used to analyze the expression of p65, IκB and Meq. Our results showed that the proliferation in the combination group was significantly decreased as compared with that of monotherapy using betulinic acid or tripterine, accompanied by an induction of apoptosis, inhibition of NF-κB transcriptional activity and its targeting oncogenic gene Meq. The results suggest that the combination of betulinic acid and tripterine at lower concentration may produce a synergistic inhibitive effect on MSB-1 cells that warrants further investigation for its potential clinical applications.

Effect of sample preparation on components and liver toxicity of Polygonum multiflorum.

It was shown that different extracts had significant differences in the toxicity of Polygonum multiflorum. In this study, the effect of sample preparation on components and liver toxicity of different extracts from P. multiflorum were determined. Hepatoxic components were discovered based on biomembrane extraction. Comparative chemistry and toxicology between ethanol and water extracts were also performed. The results showed that ethanol extract had much stronger hepatotoxicity, the content of emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion was significantly higher in ethanol extract than in water extract, while the human hepatocytes extraction showed that 2,3,5,4'-tetrahydroxystilbene-2-O-ß-d-glucopyranoside, emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion had interaction with human hepatocytes. The hepatotoxic effect of these components was investigated on human hepatocytes LO2 cells and emodin-8-O-ß-d-glucopyranoside, physcion-8-O-ß-d-glucopyranoside, emodin and physcion were finally confirmed to be, at least partial, hepatotoxic components. The results showed that sample preparation has significant effect on components in extracts of P. multiflorum especially the components related to hepatotoxicity. Water extract, the conventional administration form of Chinese herbs, is prefer for phytotherapy before well understanding their chemistry and biological activities.

Effects of dietary supplementation with green tea polyphenols on digestion and meat quality in lambs infected with Haemonchus contortus.

Ujumqin sheep are susceptible to infection by the gastrointestinal nematode Haemonchus contortus, which reduces productivity and total meat yield in sheep. Thus, the effects of green tea polyphenol (GTP) supplements (0, 2, 4, or 6g of GTP/kg feed) on dietary nutrient digestibility and meat quality in lambs infected with H. contortus were examined; control lambs were not infected. H. contortus infections did not affect digestion but the apparent digestibilities of nutrients were decreased by dietary 2g of GTP/kg feed supplementation. There was an interaction between treatment and sampling time on plasma total protein, urea nitrogen, and amino acid concentrations. The antioxidant activity and meat color of INFGTP0 lambs decreased. In conclusion, H. contortus infections in lambs decreased meat quality, but appropriate levels of dietary GTP supplementation diminished these negative effects though lower dose of GTP supplement showed negative effects on digestion.

Celastrol induces apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells: an in vitro and in vivo study.

Osteosarcoma is the most common primary malignant tumor of bone, the long-term survival of which has stagnated in the past several decades. Celastrol, a triterpene from traditional Chinese medicine, has been proved to possess potent anti-tumor effect on various cancers. However, the effect of celastrol on human osteosarcoma and the underlying mechanisms remains to be elucidated. We reported here that celastrol could inhibit cell proliferation by causing G2/M phase arrest. Exposure to celastrol resulted in the activation of caspase-3, -8, and -9, indicating that celastrol induced apoptosis through both extrinsic and intrinsic pathways. Autophagy occurred in celastrol-treated cells as evidenced by formation of autophagosome and accumulation of LC3B-II. The celastrol-induced cell death was remarkably restored by the combination of autophagy and apoptosis inhibitors. Furthermore, inhibition of apoptosis enhanced autophagy while suppression of autophagy diminished apoptosis. Celastrol also induced JNK activation and ROS generation. The JNK inhibitor significantly attenuated celastrol-triggered apoptosis and autophagy while ROS scavenger could completely reverse them. The ROS scavenger also prevented G2/M phase arrest and phosphorylation of JNK. Importantly, we found that celastrol had the similar effects on primary osteosarcoma cells. Finally, in vivo, celastrol suppressed tumor growth in the mouse xenograft model. Taken together, our results revealed that celastrol caused G2/M phase arrest, induced apoptosis and autophagy via the ROS/JNK signaling pathway in human osteosarcoma cells. Celastrol is therefore a promising candidate for development of antitumor drugs targeting osteosarcoma.

Early third trimester maternal response to glucose challenge and pregnancy outcome in Chinese women-relationship between upper distribution level and recommended diagnostic criteria.

BACKGROUND/OBJECTIVES: The objective of this study was to examine the relationship between upper distribution levels of glucose values in the 75-g oral glucose tolerance test (OGTT) and recommended diagnostic criteria for gestational diabetes mellitus (GDM) and adverse pregnancy outcomes. SUBJECTS/METHODS: The distribution of the OGTT 2-h values of 13,501 pregnant women, which were below the World Health Organization (WHO) threshold for overt diabetes mellitus (DM), and managed in one teaching hospital in China, was reviewed and related to maternal characteristics and pregnancy outcomes. RESULTS: For the entire group, the 90th and 95th percentile values of the OGTT 2-h glucose level, respectively, were close to the diagnostic cutoff values of the WHO and International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria. For adverse maternal outcomes, glucose level above the 90th percentile value was associated with increased hypertensive disorders, whereas no difference was seen with cutoff using the 95th percentile value. For perinatal outcomes, the 90th percentile was associated with increased neonatal intensive care unit admission and hypoglycemia, whereas the 95th percentile showed in addition association with phototherapy for jaundice and 5th-minute Apgar score <7. Although no differences in the incidence of adverse pregnancy outcomes were found using the different cutoffs, the >95th percentile cutoff value would have missed out 33.3-56.7% of the cases of adverse outcomes that would otherwise have been attributed to GDM. CONCLUSIONS: Further studies are warranted to clarify which diagnostic criterion is most appropriate universally to identify adverse pregnancy outcomes attributed to GDM, and which could be mitigated with treatment specific for GDM.

Advanced phosphorus removal for secondary effluent using a natural treatment system.

Mechanisms for low concentrations phosphorus removal in secondary effluent were studied, and a process was developed using limestone filters (LF), submerged macrophyte oxidation ponds (SMOPs) and a subsurface vertical flow wetland (SVFW). Pilot scale experimental models were applied in series to investigate the advanced purification of total phosphorus (TP) in secondary effluent at the Chengjiang sewage treatment plant. With a total hydraulic residence time (HRT) of 82.52 h, the average effluent TP dropped to 0.17 mg L(-1), meeting the standard for Class III surface waters. The major functions of the LF were adsorption and forced precipitation, with a particulate phosphorus (PP) removal of 82.93% and a total dissolved phosphorus (TDP) removal of 41.07%. Oxygen-releasing submerged macrophytes in the SMOPs resulted in maximum dissolved oxygen (DO) and pH values of 11.55 mg L(-1) and 8.10, respectively. This regime provided suitable conditions for chemical precipitation of TDP, which was reduced by a further 39.29%. In the SVFW, TDP was further reduced, and the TP removal in the final effluent reached 85.08%.

Type 2 diabetes prevalence and incidence among adults in Taiwan during 1999-2004: a national health insurance data set study.

AIM: To evaluate annual prevalence and incidence of Type 2 diabetes and to examine possible trends among adults in Taiwan. METHODS: A retrospective nationwide longitudinal study using the Taiwan National Health Insurance Research Database collected during 1999-2004. Adult patients aged > or = 20 years old with prevalent and incident Type 2 diabetes were identified using ICD-9-CM diagnostic codes. Age-specific and age-direct-standardized annual incidence and prevalence were calculated to describe their trends in different gender and age group and compared using Poisson regression. RESULTS: During the study years, the age-standardized prevalence of Type 2 diabetes increased from 4.7 to 6.5% for men and from 5.3 to 6.6% for women. The increasing trends in prevalence were significant and higher among people aged < 40 and > or = 80 years. The age-standardized incidence rates of Type 2 diabetes per 1000 person-years were approximately 7.6 and remain stable for men, but decreasing from 7.7 to 6.9 for women. However, the incidence increased significantly in younger adults aged < 40 years whose relative incidence (RI with 95% confidence interval) was 1.31 (1.20-1.42) for men and 1.04 (1.01-1.08) for women. The incidence trends for people aged > or = 40 years were decreased for men and women. The differences in incidence trends between age groups and between genders were all statistically significant (all P < 0.001). CONCLUSIONS: This study demonstrated a substantial increasing trend in Type 2 diabetes prevalence during 1999-2004 among adults in Taiwan. Despite the incidence decreased in older people, young men aged 20-40 years were most susceptible to higher incidence of Type 2 diabetes.

Antifungal activity of thyme oil against Geotrichum citri-aurantii in vitro and in vivo.

AIMS: To investigate antifungal effect of thyme oil on Geotrichum citri-aurantii arthroconidia germination and germ tube elongation, to reveal effects of thyme oil on morphological structures on fungal hyphae and arthroconidia and to assess potential bio-control capacities of thyme oil against disease suppression in vivo conditions. METHODS AND RESULTS: Thyme oil controlled the growth of G. citri-aurantii effectively. Arthroconidia germination and germ tube elongation in potato dextrose broth was greatly inhibited by thyme oil. At 600 microl l(-1), it inhibited the germination of about 94% of the arthroconidia and the germ tube length was only 4.32 +/- 0.28 microm. Observations using light microscope, scanning electron microscope and transmission electron microscope revealed ultrastructural modifications caused by thyme oil that included markedly shrivelled and crinkled hyphae and arthroconidia, plasma membrane disruption and mitochondrial disorganization. Thyme oil applied to 'Satsuma' mandarin oranges that had been artificially wounded and inoculated with G. citri-aurantii reduced sour rot from 78.1% among untreated control fruit to 14.1% after 5 days at 26 degrees C. Thyme oil applied to intact fruits reduced the decay from 76% among untreated control fruit to 35% after 30 days at 20 degrees C. Thyme oil treatment did not harm 'Satsuma' mandarin oranges when they were examined after treatment and storage at 20 degrees C for 30 days. CONCLUSIONS: Thyme oil may provide an alternative means of controlling postharvest sour rot on citrus fruit. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of such essential oil may constitute an important alternative to synthetic fungicides. They can be exploited in commercial production and applied under storage and greenhouse conditions.

A combination of grape seed-derived procyanidins and gypenosides alleviates insulin resistance in mice and HepG2 cells.

This study investigated the effects of grape seed-derived procyanidins (GSP), gypenosides (GPE), and combination procyanidins/gypenosides on insulin resistance in mice and HepG2 cells. ICR mice were randomly divided into 2 control and 4 treatment groups. The control mice were to receive either normal diet (ND) or high-fat diet (HFD), and the treatment groups were fed high-fat diet with either 80 mg/kg of GSP (GSP80), GPE (GPE80), GSP + GPE (1: 1, GSP40 + GPE40), or 500 mg/kg of metformin for a 6-wk period. All the groups of mice except the normal control were on high-fat diet along with fructose (15%) administered in drinking water throughout the period of treatment. An insulin-resistant HepG2 cell model was developed after 24 h of 5 x 10(-7) mol/L insulin incubation. The treatment of GPE80 could significantly reduce the index of insulin resistance (HOMA-IR) and increase hepatic glycogen concentration, compared with HFD group (P < 0.05). When GSP and GPE were administered simultaneously, synergic effects were observed in decreasing the HOMA-IR index and serum total cholesterol (TC) level and enhancing glucose tolerance. All treatment groups showed considerable raise of hepatic glucokinase activity (P < 0.05 compared with HFD group). GSP application increased the consumption of extracellular glucose in HepG2 cells. Our data suggest that the combination of GSP and GPE may have functional efficacy in consumers with insulin resistance.

First Report of Sooty Blotch and Flyspeck Caused by Species of Dissoconium, Mycosphaerella, and Peltaster on Hawthorn Fruit in China.

Sooty blotch and flyspeck (SBFS), a disease complex comprised of as many as 30 putative species of fungi, occurs on the cuticle of pome fruits in moist production regions worldwide, inciting cosmetic damage that causes significant economic losses (1). Chinese hawthorn (Crataegus pinnatifida Bge.) is an economically important tree species in China. Its fruit are sold fresh or dried and are used as a culinary spice as well as an ingredient in Chinese traditional medicine. In October of 2007, Chinese hawthorn fruit exhibiting SBFS signs were sampled from supermarkets in Yangling, Shaanxi Province and Luoyang, Henan Province, China. Thalli directly from the hawthorn fruit were transferred onto potato dextrose agar (PDA) slants under a dissecting microscope and cultured at 22 ± 1°C in darkness. DNA was extracted from pure isolates and the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA (nrDNA) was amplified and sequenced using primers ITS-1F and ITS4 (3). Phylogenetic analysis of the ITS sequences revealed that the 35 isolates generated in this study included five species in three genera: Dissoconium sp. (18 isolates), Mycosphaerella sp. (5 isolates), and Peltaster sp. 1 (4 isolates), Peltaster sp. 2 (4 isolates), and Peltaster sp. 3 (4 isolates). To fulfill Koch's postulates and verify that these fungi could also infest apple fruit, two representative isolates of each putative species were inoculated onto mature intact hawthorn and apple (cv. Fuji) fruit that had been surface disinfested with 75% ethanol and allowed to dry. Inoculum was prepared by comminuting 1-month-old cultures growing on PDA into a suspension of mycelial fragments and conidia (105 to ~106 CFU/ml) in a blender with sterile deionized water (SDW). Each isolate was inoculated on three hawthorn and three apple fruit by using cotton swabs. As controls, two surface-disinfested hawthorn and apple fruit were swabbed with SDW. After the inoculated hawthorn and apple fruit had been incubated in a moist chamber at 22 ± 1°C for 1 month, all inoculated hawthorn and apple fruit exhibited SBFS signs similar to those of the original colonies on hawthorn fruit, but the controls did not. Reservoir hosts have been inferred to play an important role in SBFS by providing the fungi with overwintering habitat and inoculum for infestations on apple. Many reservoir hosts have been reported in the United States and Japan (2). To our knowledge, this is the first report of fungi in the SBFS complex on hawthorn fruit and the first confirmation that fungi growing on hawthorn fruit can produce SBFS signs on apple fruit. These results identify hawthorn as a potential inoculum source for SBFS in apple orchards. References: (1) J. C. Batzer et al. Mycologia 97:1283, 2005. (2) K. Hemnani et al. Phytopathology 98(suppl):S66, 2008. (3) T. J. White et al. PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

High throughput crystallography of TB drug targets.

Tuberculosis (TB) infects one-third of the world population. Despite 50 years of available drug treatments, TB continues to increase at a significant rate. The failure to control TB stems in part from the expense of delivering treatment to infected individuals and from complex treatment regimens. Incomplete treatment has fueled the emergence of multi-drug resistant (MDR) strains of Mycobacterium tuberculosis (Mtb). Reducing non-compliance by reducing the duration of chemotherapy will have a great impact on TB control. The development of new drugs that either kill persisting organisms, inhibit bacilli from entering the persistent phase, or convert the persistent bacilli into actively growing cells susceptible to our current drugs will have a positive effect. We are taking a multidisciplinary approach that will identify and characterize new drug targets that are essential for persistent Mtb. Targets are exposed to a battery of analyses including microarray experiments, bioinformatics, and genetic techniques to prioritize potential drug targets from Mtb for structural analysis. Our core structural genomics pipeline works with the individual laboratories to produce diffraction quality crystals of targeted proteins, and structural analysis will be completed by the individual laboratories. We also have capabilities for functional analysis and the virtual ligand screening to identify novel inhibitors for target validation. Our overarching goals are to increase the knowledge of Mtb pathogenesis using the TB research community to drive structural genomics, particularly related to persistence, develop a central repository for TB research reagents, and discover chemical inhibitors of drug targets for future development of lead compounds.

Therapeutic effects of Sophora moorcroftiana alkaloids in combination with albendazole in mice experimentally infected with protoscolices of Echinococcus granulosus

The objective of the present study was to determine if the combination of alkaloids from Sophora moorcroftiana seeds and albendazole might be effective in the treatment of experimental echinococcosisin female NIH mice (6 weeks old and weighing 18-20 g, N = 8 in each group) infected withprotoscolices of Echinococcus granulosus. Viable protoscolices (N = 6 x 103) were cultured in vitro in 1640 medium and mortality was calculated daily. To determine the in vivo efficacy, mice were inoculated intraperitoneally with viable protoscolices and then treated once daily by gavage for three months with the alkaloids (50 mg kg-1 day-1) and albendazole (50 mg kg-1 day-1), separately and in combination (both alkaloids at 25 mg kg-1 day-1 and albendazole at 25 mg kg-1 day-1). Next, the hydatid cysts collected from the peritoneal cavity of the animals were weighed and serum IL-4, IL-2, and IgE levels were analyzed. Administration of alkaloids to cultured protoscolices showed significant dose- and time-dependent killing effects. The weight of hydatid cysts was significantly decreased upon treatment with each drug (P < 0.01), but the decrease was more prominent and the rate of hydatid cyst growth inhibition was much higher (76.1 percent) in the group receiving the combined treatments (18.3 ± 4.6 mg). IL-4 and total IgE were decreased (939 ± 447 pg/mL and 2.03 ± 0.42 IU/mL, respectively) in serum from mice treated with alkaloids and albendazole compared with the untreated control (1481 ± 619 pg/mL and 3.31 ± 0.37 IU/mL; P < 0.01). These results indicate that S. moorcroftiana alkaloids have protoscolicidal effects and the combination of alkaloids and albendazole has significant additive effects.

Relapses in leprosy patients treated with rifampicin plus dapsone after varying periods of dapsone monotherapy.

Leprosy patients treated formerly with dapsone monotherapy followed by combined therapy with rifampicin plus dapsone were surveyed for relapse and rifampicin resistance. The relapse rate was significantly low for the 482 multibacillary (MB) patients receiving > 12 months combined therapy compared with the 49 MB cases receiving < 12 months of combined therapy. The relapse rate was related to the duration of dapsone monotherapy prior to combined therapy. The difference in relapse rate in 247 paucibacillary (PB) patients following > 12 months combined therapy was also of significance, compared with the 66 PB cases who had received < 12 months combined therapy. Five strains of M. leprae isolated from relapsed patients were sensitive to rifampicin by mouse foot-pad test and all relapsed patients responded favourably to fixed duration MDT regimen for MB cases.

Circuits and mechanisms governing hypothalamic responses to stress: a tale of two paradigms.

The results of recent studies support a partitioning of stress models into at least two basic classes. While these have been referred to as 'systemic' and 'neurogenic', we would suggest that the terms interoceptive and exteroceptive, respectively, are more apt descriptors. This is based on the similarities in the overall patterns of activational responses seen as a consequence of exposure to a range of perturbations in the internal versus external environments. While stressors of each class may share in common such fundamental features as a capacity to enlist certain PVH effector populations and medullary catecholamine-containing neurons, both the capacity to involve specific output neuron classes and the dependence of hypothalamic effects on the integrity of aminergic afferents in at least some interoceptive and exteroceptive models, are clearly differential. The available evidence suggests that interoceptive stress effects on PVH effector populations may be conceived essentially as simple reflex responses, mediated at a subcortical level by cell groups and associated circumventricular organs that comprise the core of a system involved in the processing of visceral sensory information. Based on the general pattern of acute footshock-induced Fos expression and commonalities of cellular activation profiles seen in this and other acute exteroceptive paradigms, it seems a reasonable assumption that pathways that convey somatosensory/nociceptive information to the PVH are apt to mediate adaptive visceromotor responses in these models. Multiple candidates for such roles have been identified at various levels of what may be viewed as the ascent of the spinothalamic pathway through the brainstem and thalamus, and on through the limbic forebrain and hypothalamus. Dissecting the relative contributions of these in determining PVH output will speak to important conceptual issues concerning the extent to which the affective and visceromotor responses to exteroceptive stressors are organized, and the level(s) at which these different avenues of emotional expression may be integrated.