Effect of cell cycle arrest on intermediate metabolism in the marine diatom Phaeodactylum tricornutum.

The inhibitor NU 2058 [6-(cyclohexylmethoxy)-9H-purin-2-amine] leads to G1-phase cell cycle arrest in the marine diatom, Phaeodactylum tricornutum, by binding to two cyclin-dependent kinases, CDKA1 and CDKA2. NU 2058 has no effect on photosynthetic attributes, such as Fv/Fm, chlorophyll a/cell, levels of D2 PSII subunits, or RbcL; however, cell cycle arrest leads to unbalanced growth whereby photosynthetic products that can no longer be used for cell division are redirected toward carbohydrates and triacylglycerols (TAGs). Arrested cells up-regulate most genes involved in fatty acid synthesis, including acetyl-CoA carboxylase, and three out of five putative type II diglyceride acyltransferases (DGATs), the enzymes that catalyze TAG production. Correlation of transcriptomes in arrested cells with a flux balance model for P. tricornutum predicts that reactions in the mitochondrion that supply glycerate may support TAG synthesis. Our results reveal that sources of intermediate metabolites and macromolecular sinks are tightly coupled to the cell cycle in a marine diatom, and that arresting cells in the G1 phase leads to remodeling of intermediate metabolism and unbalanced growth.

Investigation of Toxoplasma gondii and association with early pregnancy and abortion rates in New Zealand farmed red deer (Cervus elaphus).

This study tested for association between Toxoplasma gondii and pregnancy and abortion to investigate sub-optimal reproduction in farmed red deer (Cervus elaphus). Sera from a sub-sample (n = 2304) of pregnant and non-pregnant hinds in early gestation at first pregnancy scan (scan 1) and approximately at the end of second trimester at second pregnancy scan (scan 2) were tested for T. gondii antibodies using a validated ELISA. Foetuses and/or uteri from pregnant, non-pregnant, and aborting hinds at scan 1, scan 2, or weaning were tested for T. gondii DNA by nested PCR. At scan 1, 31.1% of 861 rising two-year-old (R2) and 28.3% of 357 mixed-aged (MA, ≥ 2 years) hinds were sero-positive. There was no association between scan 1 serology and non-pregnancy at animal (R2, p = 0.05 and MA, p = 0.43) or herd level (R2, p = 0.37). Toxoplasma gondii DNA was detected in 3/18 placenta and 4/18 foetal brains from aborting R2 hinds and 15/157 R2 and 3/21 MA uteri from non-pregnant hinds at scan 1. At scan 2, sero-prevalence was higher (odds ratio = 1.6, 95% CI = 1.04-2.48) in aborted (34.3% of 268) than in non-aborted (23.5% of 446) R2 hinds (p = 0.03) and 7.9% of abortions between scans were attributable to T. gondii exposure. Within-herd sero-prevalence at scan 2 was positively associated with daily abortion rate in R2 herds with aborted hinds (p < 0.001) but not in MA herds (p = 0.07). Toxoplasma gondii DNA was detected in 27/169 uteri, 2/20 cotyledons, and 1/5 foetal brains from aborted hinds at scan 2 and in uteri from 5/33 hinds not rearing a calf to weaning. Toxoplasma gondii RFLP genotyping of five loci revealed a unique type I/III genotype pattern, TgRDNZ1, in a foetal brain sample, not been previously reported in deer. These findings provide serological and molecular evidence that T. gondii infection is associated with abortion in red deer, possibly in all three trimesters.

First report of Toxoplasma gondii sporulated oocysts and Giardia duodenalis in commercial green-lipped mussels (Perna canaliculus) in New Zealand.

Pollution of marine ecosystems with the protozoan parasites Toxoplasma gondii, Cryptosporidium spp. and Giardia duodenalis can be studied using bivalve shellfish as biosentinels. Although evidence suggests that these parasites are present in New Zealand coastal waters, the extent of protozoal pollution has not been investigated. This study used optimised molecular methods to detect the presence of Cryptosporidium spp., G. duodenalis and T. gondii in commercially sourced green-lipped mussel (Perna canaliculus), an endemic species found throughout coastal New Zealand. A nested polymerase chain reaction was validated for detection of T. gondii DNA and applied to 104 commercially sourced mussels. Thirteen mussels were positive for T. gondii DNA with an estimated true prevalence of 16.4% using Bayesian statistics, and the presence of T. gondii in mussels was significantly associated with collection during the summer compared with that in the winter (P = 0.003). Consumption of contaminated shellfish may also pose a health risk for humans and marine wildlife. As only sporulated T. gondii oocysts can be infectious, a reverse transcriptase-polymerase chain reaction was used to confirm presence of a sporozoite-specific marker (SporoSAG), detected in four mussels. G. duodenalis assemblage B, known to be pathogenic in humans, was also discovered in 1% mussels, tested by polymerase chain reaction (n = 90). Cryptosporidium spp. was not detected in the sampled mussel haemolymph. Results suggest that New Zealand may have high levels of coastal contamination with T. gondii, particularly in summer months, and that naturally exposed mussels can ingest and retain sporulated oocysts, further establishing shellfish consumption as a health concern.

Four cases of fatal toxoplasmosis in three species of endemic New Zealand birds.

Four cases of fatal toxoplasmosis in three endemic New Zealand avian species are reported. Between 2009 and 2012, two kereru (Hemiphaga novaeseelandiae), one North Island brown kiwi (Apteryx mantelli), and one North Island kaka (Nestor meridionalis) were submitted for necropsy examination. On gross postmortem, the kiwi had marked hepatosplenomegaly while the kaka and two kereru had swollen, slightly firm, deep-red lungs. Histologically there was extensive hepatocellular necrosis in the liver of the kiwi while the kaka and kereru showed severe fibrinous bronchointerstitial pneumonia. In the kiwi, protozoal organisms were present within both hepatocytes and Kupffer cells of the liver and within the epithelial cells and macrophages of the interstitium of the lungs in the kaka and two kereru. The diagnosis of toxoplasmosis was confirmed with immunohistochemistry and PCR of paraffin-embedded formalin-fixed tissue of the liver, lungs, or both. Genotyping of up to seven markers revealed that an atypical Type II isolate of Toxoplasma gondii was present in at least three of the cases. This study provides evidence that T. gondii can cause mortality in these endemic species and suggests further research is needed to determine the full extent of morbidity and mortality caused by this parasite in New Zealand's unique avifauna.

Soil warming, carbon-nitrogen interactions, and forest carbon budgets.

Soil warming has the potential to alter both soil and plant processes that affect carbon storage in forest ecosystems. We have quantified these effects in a large, long-term (7-y) soil-warming study in a deciduous forest in New England. Soil warming has resulted in carbon losses from the soil and stimulated carbon gains in the woody tissue of trees. The warming-enhanced decay of soil organic matter also released enough additional inorganic nitrogen into the soil solution to support the observed increases in plant carbon storage. Although soil warming has resulted in a cumulative net loss of carbon from a New England forest relative to a control area over the 7-y study, the annual net losses generally decreased over time as plant carbon storage increased. In the seventh year, warming-induced soil carbon losses were almost totally compensated for by plant carbon gains in response to warming. We attribute the plant gains primarily to warming-induced increases in nitrogen availability. This study underscores the importance of incorporating carbon-nitrogen interactions in atmosphere-ocean-land earth system models to accurately simulate land feedbacks to the climate system.

Acupuncture can improve absorption of baicalin from extracts of Scutellaria baicalensis Georgi in rats.

The aim of this research was to explore whether acupuncture could improve the absorption of baicalin from extracts of Scutellaria baicalensis Georgi in normal rats. Twenty male Sprague-Dawley (SD) rats were divided randomly into two groups. Group A was treated with a combination of oral administration of Scutellaria baicalensis Georgi extracts and acupuncture. Group B was treated only with oral administration of Scutellaria baicalensis Georgi extracts, in the same dosage as Group A. After treatment, the plasma concentration of baicalin was detected and pharmacokinetic analysis was conducted using high performance liquid chromatography (HPLC). The results showed that the plasma concentration of baicalin in Group A was significantly higher than in Group B, and that acupuncture at the acupoints Jizhong (Du6), Dazhui (Du14) and Zhongwan (Ren12) made the plasma concentration-time course of baicalin produce a bimodal phenomenon, increased the area under the curve (AUC) and slowed the elimination of baicalin, thus significantly improving the absorption of baicalin from extracts of Scutellaria baicalensis Georgi in rats.

High-throughput screening assay for biological hydrogen production.

This paper describes a screening assay, compatible with high-throughput bioprospecting or molecular biology methods, for assessing biological hydrogen (H2) production. While the assay is adaptable to various physical configurations, we describe its use in a 96-well, microtiter plate format with a lower plate containing H2-producing cyanobacteria strains and controls and an upper, membrane-bottom plate containing a color indicator and a catalyst. H2 produced by cells in the lower plate diffuses through the membrane into the upper plate, causing a color change that can be quantified with a microplate reader. The assay is reproducible, semiquantitative, sensitive down to at least 20 nmol of H2, and largely unaffected by oxygen, carbon dioxide, or volatile fatty acids at levels appropriate to biological systems.

Auricular acupressure may improve absorption of flavanones in the extracts from Citrus aurantium L. in the human body.

OBJECTIVE: To explore whether auricular acupressure could improve the absorption of flavanones in the extracts from Citrus aurantium L. in the human body. METHODS: Ten (10) healthy male volunteers were randomly divided into two groups. Group A was treated with the combination of oral administration of C. aurantium L. extracts and auricular acupressure. Group B was treated only with the oral administration of C. aurantium L. extracts in the same dosage as Group A. After 7 days of treatment, the plasma concentration of naringenin and hesperetin of the two groups were determined using high-performance liquid chromatography. RESULTS: The plasma concentration of naringenin and hesperetin in Group A were significantly higher than Group B. CONCLUSIONS: Auricular acupressure at some specific acupoints can significantly improve the absorption of naringenin and hesperetin in the extracts form C. aurantium L. in the human body.

Stability of maternal preferences for pediatric health states in the perinatal period and 1 year later.

BACKGROUND: We have previously shown that parents of premature children provided relatively high valuation of their children's health state in adolescence. However, stability of parental preferences for future pediatric health states is unknown during the antenatal and neonatal periods and infancy. OBJECTIVE: To determine whether preference-based, health-related quality-of-life scores obtained serially from 2 cohorts (women with high-risk pregnancies [antenatal cohort] and mothers of very low-birth-weight newborns [VLBW cohort]) are stable during the first year after birth. DESIGN AND METHODS: Longitudinal cohort study. Participants included 80 high-risk pregnant women recruited at 24 +/- 2 weeks of gestation, and 75 mothers of VLBW infants recruited within 1 week of delivery. We conducted 2 to 3 standardized interviews (antenatally, at 1 week after delivery, and at the 12-month corrected age visit) using the Standard Gamble technique to elicit preferences for 5 pediatric hypothetical health states with varying disabilities. RESULTS: Seventy-three mothers with high-risk pregnancies (91%) and 72 mothers of VLBW infants (96%) completed all scheduled interviews. As expected, preference scores were affected by the level of severity of the hypothetical health states (antenatal cohort, F(4,288) = 87.0 [P<.001]; VLBW cohort, F(4,284) = 64.2 [P<.001]). At each assessment, at least 38% of mothers rated 1 or more health states as worse than death. Repeated-measures analysis showed no change in preference scores over time (antenatal cohort, F(2,144) = 1.3 [P =.29]; VLBW cohort, F(1,71) = 0.7 [P =.42]). Maternal socioemotional factors, infant severity of illness at birth, and global health at 12 months did not affect preference scores. CONCLUSION: In our population, maternal preference scores for disabling health states appear to be stable during the first year of life and are unaffected by key maternal and infant variables.

Use of antihypertensive medications in pregnancy and the risk of adverse perinatal outcomes: McMaster Outcome Study of Hypertension In Pregnancy 2 (MOS HIP 2).

BACKGROUND: Uncertainty remains about the potential harmful effects of antihypertensive therapy on the developing fetus, especially for beta-blockers (betab). METHODS: We prospectively enrolled all singleton women with a blood pressure >/= 140/90 mm Hg during pregnancy. The main analysis included 1948 women with all forms of hypertension and compared the use of betab drugs, non-betab drugs or a combination of both, to no treatment. The primary study outcome was a composite of the diseases of prematurity, need for assisted ventilation for greater than 1 day, or perinatal death. A sub-group analysis evaluated the four treatment options among 583 singleton women with chronic hypertension before 20 weeks gestation. RESULTS: In the main analysis, no association was observed between betab use and the primary composite outcome [adjusted odds ratio (OR) 1.4, 95% CI 0.9-2.2], while an association was seen with non-betab therapy (OR 5.0, 95% CI 2.6-9.6) and combination therapy (OR 2.9, 95% CI 1.8-4.7). In the sub-group of 583 women with hypertension before 20 weeks, use of a non-betab drug (OR 4.9, 95% CI 1.7-14.2) or combination therapy (OR 2.9. 95% CI 1.1-7.7) was significantly associated with the primary composite outcome, while betab monotherapy was not (OR 1.4, 95% CI 0.6-3.4). CONCLUSIONS: Maternal use of antihypertensive medications other than betabs was associated with both major perinatal morbidity and mortality, while betab monotherapy was not. The combined use of betab and non-betab medications demonstrated the strongest association. Before definitive conclusions can be drawn, a large multicentre randomized controlled trial is needed to address the issues of both maternal efficacy and fetal safety with the use of one or more antihypertensive agents in pregnancy.

Quantum requirements for growth and fatty acid biosynthesis in the marine diatom Phaeodactylum tricornutum (Bacillariophyceae) in nitrogen replete and limited conditions.

We determined the quantum requirements for growth (1/ϕµ ) and fatty acid (FA) biosynthesis (1/ϕFA ) in the marine diatom, Phaeodactylum tricornutum, grown in nutrient replete conditions with nitrate or ammonium as nitrogen sources, and under nitrogen limitation, achieved by transferring cells into nitrogen free medium or by inhibiting nitrate assimilation with tungstate. A treatment in which tungstate was supplemented to cells grown with ammonium was also included. In nutrient replete conditions, cells grew exponentially and possessed virtually identical 1/ϕµ of 40-44 mol photons · mol C(-1) . In parallel, 1/ϕFA varied between 380 and 409 mol photons · mol C(-1) in the presence of nitrate, but declined to 348 mol photons · mol C(-1) with ammonium and to 250 mol photons · mol C(-1) with ammonium plus tungstate, indicating an increase in the efficiency of FA biosynthesis relative to cells grown on nitrate of 8% and 35%, respectively. While the molecular mechanism for this effect remains poorly understood, the results unambiguously reveal that cells grown on ammonium are able to direct more reductant to lipids. This analysis suggests that when cells are grown with a reduced nitrogen source, fatty acid biosynthesis can effectively become a sink for excess absorbed light, compensating for the absence of energetically demanding nitrate assimilation reactions. Our data further suggest that optimal lipid production efficiency is achieved when cells are in exponential growth, when nitrate assimilation is inhibited, and ammonium is the sole nitrogen source.

Arcobacter species in diarrhoeal faeces from humans in New Zealand.

AIM: To determine the prevalence, genetic diversity and antimicrobial susceptibility of Arcobacter spp in faecal samples from humans with diarrhoea in New Zealand. METHODS: An enrichment method was used to isolate Arcobacter spp from diarrhoeal human faeces submitted to a community laboratory in Hawke's Bay. The identity of isolates was confirmed by PCR and their diversity was determined by pulsed field gel electrophoresis (PFGE). Antibiotic susceptibility was established with E test strips. RESULTS: Arcobacter spp were isolated from 12 of 1380 diarrhoeal faecal samples examined (0.9%), including 7 A. butzleri and 5 A. cryaerophilus. Additional enteric pathogens were detected in four of these diarrhoeal faecal samples. All the Arcobacter isolates were genetically distinct and susceptible to ciprofloxacin. Most were also susceptible to erythromycin (92%) but fewer to tetracycline (67%) and ampicillin(50%). CONCLUSION: A. butzleri and A. cryaerophilus cause a small proportion of cases of diarrhoea in humans resident in New Zealand.

Effects of selected electron transport chain inhibitors on 24-h hydrogen production by Synechocystis sp. PCC 6803.

One factor limiting biosolar hydrogen (H(2)) production from cyanobacteria is electron availability to the hydrogenase enzyme. In order to optimize 24-h H(2) production this study used Response Surface Methodology and Q2, an optimization algorithm, to investigate the effects of five inhibitors of the photosynthetic and respiratory electron transport chains of Synechocystis sp. PCC 6803. Over 3 days of diurnal light/dark cycling, with the optimized combination of 9.4 mM KCN (3.1 µmol 10(10) cells(-1)) and 1.5 mM malonate (0.5 µmol 10(10) cells(-1)) the H(2) production was 30-fold higher, in EHB-1 media previously optimized for nitrogen (N), sulfur (S), and carbon (C) concentrations (Burrows et al., 2008). In addition, glycogen concentration was measured over 24 h with two light/dark cycling regimes in both standard BG-11 and EHB-1 media. The results suggest that electron flow as well as glycogen accumulation should be optimized in systems engineered for maximal H(2) output.

Chinese medicinal herbs in relieving perimenopausal depression: a randomized, controlled trial.

OBJECTIVE: To explore the effects of GengNianLe (GNL, also called perimenopausal depression relieving formula), a defined formula of Chinese medicinal herbs in relieving perimenopausal depression in Chinese women. METHODS: Between September 2004 and April 2008, 47 Chinese women were randomized into a GNL group (n = 21) and a control group which received tibolone (n = 26) using a randomization chart. Depression was rated with the 24-item Hamilton Depression Scale (HAMD). The serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), and estradiol (E(2)) were detected before and after the treatment. RESULTS: After 12 weeks of treatment, HAMD scores in both groups decreased significantly (p < 0.05) with no significant difference between the groups (p > 0.05). The levels of FSH decreased significantly and the level of E(2) increased significantly in both groups, and they changed more in the control group. No side-effect of treatment was reported in either group during treatment. CONCLUSIONS: The Chinese medicinal formula GNL showed promise in relieving perimenopausal depression and merits further study.

Optimization of pH and nitrogen for enhanced hydrogen production by Synechocystis sp. PCC 6803 via statistical and machine learning methods.

The nitrogen (N) concentration and pH of culture media were optimized for increased fermentative hydrogen (H(2)) production from the cyanobacterium, Synechocystis sp. PCC 6803. The optimization was conducted using two procedures, response surface methodology (RSM), which is commonly used, and a memory-based machine learning algorithm, Q2, which has not been used previously in biotechnology applications. Both RSM and Q2 were successful in predicting optimum conditions that yielded higher H(2) than the media reported by Burrows et al., Int J Hydrogen Energy. 2008;33:6092-6099 optimized for N, S, and C (called EHB-1 media hereafter), which itself yielded almost 150 times more H(2) than Synechocystis sp. PCC 6803 grown on sulfur-free BG-11 media. RSM predicted an optimum N concentration of 0.63 mM and pH of 7.77, which yielded 1.70 times more H(2) than EHB-1 media when normalized to chlorophyll concentration (0.68 +/- 0.43 micromol H(2) mg Chl(-1) h(-1)) and 1.35 times more when normalized to optical density (1.62 +/- 0.09 nmol H(2) OD(730) (-1) h(-1)). Q2 predicted an optimum of 0.36 mM N and pH of 7.88, which yielded 1.94 and 1.27 times more H(2) than EHB-1 media when normalized to chlorophyll concentration (0.77 +/- 0.44 micromol H(2) mg Chl(-1) h(-1)) and optical density (1.53 +/- 0.07 nmol H(2) OD(730) (-1) h(-1)), respectively. Both optimization methods have unique benefits and drawbacks that are identified and discussed in this study.

Genetic analysis of Aralia cordata Thunb by RAPD.

In the research, genetic analysis of Aralia cordata Thunb. (Araliaceae) was conducted using randomly amplified polymorphism DNA (RAPD). 161 loci were detected with 12 RAPD primers. Percentage of Polymorphic Band (PPB) varied from 34.78% to 63.35%. All the samples were respectively collected from the eight provinces richest in Aralia cordata Thunb resources in China, including Hunan, Yunnan, Zhejiang, Sichuan, Jiangxi, Anhui, Shanxi and Gansu. The results showed that Hunan Province enjoyed the highest level of genetic differentiation and Gansu was the lowest. The total genetic diversity (H(T)) of RAPD, intraspecific genetic diversity (H(S)) and genetic diversity (D(ST)) of the various places was respectively 26.33%, 11.14%, and 49.36%. The differentiation among the species accounted for 98.76% of total genetic diversity (G(ST)). Based on the cluster results of genetic distance, the 8 samples were classified into three groups. It is concluded that Hunan Province enjoyed the highest level of genetic differentiation of Aralia cordata Thunb and Gansu was the lowest, which provides a basis for the taxonomic identification and germplasm resource research of Aralia cordata Thunb in the future.

Acupuncture may improve the absorption of baicalin from the extracts of Scutellaria baicalensis Georgi in rats with pelvic inflammation.

The aim of the present study was to explore whether acupuncture may improve the absorption of baicalin from the extracts of Scutellaria baicalensis Georgi in rats with pelvic inflammation. 24 female Sprague-Dawley (SD) rats were randomly divided into groups A (a combination of oral administration of Scutellaria baicalensis Georgi extracts and acupuncture), B (oral administration of Scutellaria baicalensis Georgi extracts), C (model group) and D (normal control group), with 6 rats in each group. After treatment, the plasma concentration of baicalin was determined using high performance liquid chromatography (HPLC), the serum levels of tumor necrosis factor-alpha (TNF-alpha) and Interleukin 6 (IL-6) were detected with enzyme-linked immunosorbent assay (ELISA). The results showed that the plasma concentration of baicalin in group A was significantly higher than in group B, and the serum levels of TNF-alpha and IL-6 in group A were significantly lower than in group B. This indicates that acupuncture at certain acupoints may significantly improve the curative effects of Scutellaria baicalensis Georgi in rats with pelvic inflammation by enhancing the absorption of baicalin from the Scutellaria baicalensis Georgi extracts.

Treating histiocytic necrotizing lymphadenitis with Chinese herbs: a case report.

We present the case of a young woman who suffered from right cervical lymphadenopathy with associated malaise for 8 weeks. The sonography showed multiple, enlarged lymph nodes in the cervical, submaxillary, and subclavicular chains on the right side. The biopsy of a right cervical lymph node disclosed necrosis, histiocytic accumulation, infiltration with lymphocytes and immunoblasts, and cellular debris, demonstrating histiocytic necrotizing lymphadenitis (HNL). Although the patient had received treatment with dexamethasone for 5 weeks and antibiotic drugs for 6 weeks, the lymphadenopathy and the associated malaise had not improved. For fear of the side-effects caused by dexamethasone and the antibiotic drugs, the patient and her parents asked for treatment with Chinese herbs instead of dexamethasone and antibiotics. With the patient's and her parents' consent, the dose of dexamethasone was tapered according to its specifications and Chinese herbs were applied to treat her disease. After 6 weeks of orally administered Chinese herbs both the lymphadenopathy and the associated malaise disappeared. Follow-up by phone 6 months later showed no abnormality. Chinese medicinal herbs, as a natural treatment, can be used as an alternative in treating HNL.

Assessment of diagnostic tests to inform policy decisions--visual electrodiagnosis.

OBJECTIVES: To conduct a systematic review of the evidence for the effectiveness of five visual electrodiagnostic tests to inform the Medical Services Advisory Committee (MSAC) of the Department of Health and Ageing (Australia) in its decision in allocating public funding for new technologies. METHODS: We searched the biomedical literature to identify English-language articles published from 1966 to September 2000. We assessed validity of methodology of included studies against the following criteria: investigators (i) compared test with an appropriate reference test; (ii) tested an appropriate spectrum of patients; (iii) masked assessment of study and reference tests; (iv) measured the study test independently of clinical information; and (v) measured the reference test before any interventions. RESULTS: Sixty-one articles met inclusion criteria for critical appraisal: nineteen were cross-sectional studies that compared a study test with another test, thirty-four were case-control studies that compared a test in a group of patients with an eye disease to a group of subjects without eye disease, and eight studies were case series. None of the included studies met all of the validity criteria. Only four studies provided enough information to calculate diagnostic characteristics but were flawed due to inclusion of patients already diagnosed with disease or lack of an appropriate reference test and, thus, overestimated test accuracy. CONCLUSIONS: Identified studies did not provide sufficient valid evidence of the clinical value of the five visual tests in diagnosing diseases of the retina or optic nerve. Thus, MSAC recommended that the tests not be supported by public funding.