bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of Depigmenting Chemical Agents on Hair and Skin Color in Yellow (Pheomelanic) and Black (Eumelanic) Mice
Authors: QUEVEDO WALTER C. , HOLSTEIN THOMAS J. , DYCKMAN JACOB , NORDLUND JAMES J.
Published: August 2006

Topical applications of monobenzylether of hydroquinone (MBEH) or intraperitoneal injections of phenol induced graying of hair in eumelanic mice but had little effect on hair color in pheomelanic mice. Amcinonide, an anti‐inflammatory agent, elicited whitening of a few hairs in both pheomelanic and eumelanic mice. In phenol‐treated eumelanic mice, damaged follicular melanocytes were uprooted from hair bulbs and incorporated into the developing hair. The fate of follicular melanocytes in MBEH‐ or amcinonide‐treated mice was not determined since hair growth and graying were more variable than in phenol‐treated mice.

In contrast to the susceptibility of eumelanic hair follicles to depigmentation by phenol or MBEH, the tail skin of eumelanic or pheomelanic mice was not depigmented by these agents. Overall, during the 3 week period of treatment that was sufficient for phenol or MBEH to elicit graying of hair, epidermal melanocytes of the tails of eumelanic or pheomelanic mice either failed to respond (phenol) or were stimulated in their “proliferative” and melanogenic activity (MBEH). In contrast, amcinonide brought about a marked reduction in the numbers of DOPA‐positive epidermal melanocytes inhabiting the tails of eumelanic or pheomelanic mice. Amcinonide exerted a deleterious influence on the structure and function of tail epidermis. Its actions were partly reversed by simultaneous treatment with MBEH but not with prostaglandin (PGE2).
Concentrations of estradiol in ewe cerebrospinal fluid are modulated by photoperiod through pineal‐dependent mechanisms
Authors: Thiery Jean‐Claude , Lomet Didier , Schumacher Michael , Liere Philippe , Tricoire Helene , Locatelli Alain , Delagrange Philippe , Malpaux Benoit
Published: August 2006

Abstract: In the ewe, seasonal anestrus results from an increased responsiveness of the hypothalamus to the negative feedback of estradiol (E2) on the gonadotropic axis under long‐day conditions. However, this seasonal effect could also depend upon variable uptake of steroids by the brain. The aim of the present experiment was to compare the concentration of E2 in the blood plasma and in the cerebrospinal fluid (CSF) from the third ventricle in groups of ovariectomized, estradiol treated ewes maintained under short day (SD) or long day (LD) conditions and to study the involvement of the pineal gland in this photoperiodic regulation. Pinealectomized and sham‐operated ewes were equipped with an intracerebral cannula to sample the CSF. The plasma E2 concentrations showed no difference between LD and SD in sham‐operated and pinealectomized animals. In contrast, in the CSF, E2 concentration was higher in the LD than the SD group, and pinealectomy suppressed this effect of photoperiod. Concomitantly, the stimulatory effect of SD on luteinizing hormone levels observed in sham‐operated ewes was abolished by pinealectomy. The results demonstrate that LD increases the E2 concentration in the CSF by a mechanism involving the pineal gland.
Evidence for Early Endosome‐like Fusion of Recently Endocytosed Synaptic Vesicles
Authors: Rizzoli Silvio O. , Bethani Ioanna , Zwilling Daniel , Wenzel Dirk , Siddiqui Tabrez J. , Brandhorst Dorothea , Jahn Reinhard
Published: August 2006

Early endosomes are well‐established acceptor compartments of endocytic vesicles in many cell types. Little evidence of their existence or function has been obtained in synapses, and it is generally believed that synaptic vesicles recycle without passing through an endosomal intermediate. We show here that the early endosomal SNARE proteins are enriched in synaptic vesicles. To investigate their function in the synapse, we isolated synaptic nerve terminals (synaptosomes), stimulated them in presence of different fluorescent markers to label the recycling vesicles and used these vesicles in in vitro fusion assays. The recently endocytosed vesicles underwent homotypic fusion. They also fused with endosomes from PC12 and BHK cells. The fusion process was dependent upon NSF activity. Moreover, fusion was dependent upon the early endosomal SNAREs but not upon the SNAREs involved in exocytosis. Our results thus show that at least a fraction of the vesicles endocytosed during synaptic activity are capable of fusing with early endosomes and lend support to an involvement of endosomal intermediates during recycling of synaptic vesicles.
Which Thrombophilic Gene Mutations are Risk Factors for Recurrent Pregnancy Loss?
Authors: Goodman Cyle S. , Coulam Carolyn B. , Jeyendran Rajasingam S. , Acosta Vida A. , Roussev Roumen
Published: August 2006

Problem  Thrombophilia has been associated with poor obstetrical outcomes. To determine the association of specific inherited thrombophilias and recurrent pregnancy loss, 10 thrombophilic genes were investigated.

Method of study  A total of 550 women with a history of recurrent pregnancy loss had buccal swabs taken for DNA analyses of the following gene mutations: factor V G1691A, factor V H1299R (R2), factor V Y1702C, factor II prothrombin G20210A, factor XIII V34L, β‐fibrinogen ‐455G>A, PAI‐1 4G/5G, HPA1 a/b(L33P), methylenetetrahydrofolate reductase (MTHFR) C677T, MTHFR A1298C. The frequencies of these mutations were compared with controls published in the literature.

Results  When examined individually, PAI‐1 4G/5G (P = 0.009), factor XIII V34L (P < 0.0001), and homozygous MTHFR C667T (P < 0.0001) correlated significantly with recurrent pregnancy loss compared with controls. The frequency of the factor V Y1702C mutation was extremely low in patients and controls; thus, this gene was removed from further calculations. The remaining six mutated genes, when analyzed cumulatively, also corresponded with recurrent pregnancy loss (P < 0.0001).

Conclusion  A panel of thrombogenic gene mutations consisting of factor V G1691A, factor V H1299R (R2), factor II prothrombin G20210A, factor XIII V34L, β‐fibrinogen ‐455G>A, PAI‐1 4G/5G, HPA1 a/b(L33P), MTHFR C677T, and MTHFR A1298C can identify individuals at risk for recurrent pregnancy loss.
Utilization of Dental Services by the Elderly1
Authors: Asuman Kiyak H.
Published: August 2006

Older persons utilize dental services at a significantly lower rate than they use other health services, and lower than the rates for younger age groups. This presentation will focus on utilization behavior within the framework of a person‐environment congruence model. Much of the previous research in this area has been concerned with the person component, specifically predisposing (demographic) and enabling (financial resources) variables. But these variables have rarely been found to predict a significant proportion of the variance in utilization behavior. More recent studies have examined perceived and objective need for dental care, and have found low perceived need among non‐utilizers. The author's own research on psychosocial factors in dental behaviors points to the role of attitudes, personality, and perceived oral health status in determining the older person's oral self‐care and utilization behaviors. Our research with elderly Caucasians and Pacific Asians suggests that reducing the traditional barriers to dental care delivery and educating older persons about oral health will not significantly improve their health care behaviors. Attitudinal barriers must first be overcome. Research on other psychological and environmental barriers to dental service utilization, and alternative methods of eliminating these barriers is needed. Areas of potentially valuable research are discussed. Only by moving from descriptive to more experimental studies such as these can we enhance the oral health behaviors of the elderly population.
Inhibitory properties and solution structure of a potent Bowman–Birk protease inhibitor from lentil (Lens culinaris, L) seeds
Authors: Ragg Enzio M. , Galbusera Valerio , Scarafoni Alessio , Negri Armando , Tedeschi Gabriella , Consonni Alessandro , Sessa Fabio , Duranti Marcello
Published: August 2006

Bowman–Birk serine protease inhibitors are a family of small plant proteins, whose physiological role has not been ascertained as yet, while chemopreventive anticarcinogenic properties have repeatedly been claimed. In this work we present data on the isolation of a lentil (Lens culinaris, L., var. Macrosperma) seed trypsin inhibitor (LCTI) and its functional and structural characterization. LCTI is a 7448 Da double‐headed trypsin/chymotrypsin inhibitor with dissociation constants equal to 0.54 nm and 7.25 nm for the two proteases, respectively. The inhibitor is, however, hydrolysed by trypsin in a few minutes timescale, leading to a dramatic loss of its affinity for the enzyme. This is due to a substantial difference in the kon and k*on values (1.1 µm−1·s−1 vs. 0.002 µm−1·s−1), respectively, for the intact and modified inhibitor. A similar behaviour was not observed with chymotrypsin. The twenty best NMR structures concurrently showed a canonical Bowman–Birk inhibitor (BBI) conformation with two antipodal β‐hairpins containing the inhibitory domains. The tertiary structure is stabilized by ion pairs and hydrogen bonds involving the side chain and backbone of Asp10‐Asp26‐Arg28 and Asp36‐Asp52 residues. At physiological pH, the final structure results in an asymmetric distribution of opposite charges with a negative electrostatic potential, centred on the C‐terminus, and a highly positive potential, surrounding the antitryptic domain. The segment 53–55 lacks the anchoring capacity found in analogous BBIs, thus rendering the protein susceptible to hydrolysis. The inhibitory properties of LCTI, related to the simultaneous presence of two key amino acids (Gln18 and His54), render the molecule unusual within the natural Bowman–Birk inhibitor family.
Palaeophytochemical Constituents of Cretaceous Ginkgo coriacea Florin Leaves
Authors: Zhao You‐Xing , Li Cheng‐Sen , Luo Xiao‐Dong , Wang Yu‐Fei , Zhou Jun
Published: August 2006

Chemical investigation of the organic solvent extract of Cretaceous Ginkgo coriacea Florin leaves by liquid chromatography‐mass spectroscopy (LC‐MS) and gas chromatography‐mass spectrometry (GC‐MS), analogous to those from extant leaves of Ginkgo biloba L., led to the detection of a group of natural flavonoids and other volatiles. The similarity of the chemical constituents in these two species of Ginkgo suggest that the secondary metabolism of extant G. biloba is close to that of the Cretaceous species. The remaining natural products may be one explanation why the leaves of the Cretaceous G. coriacea have been preserved morphologically in fossilization. The detection of flavonoids suggests that the leaves of G. coriacea experienced a mild post‐depositional environment during their fossilization. This appears to be the oldest occurrence of flavonoids in plant fossils.

(Managing editor: Wei Wang)
Enhanced Cadmium Accumulation in Transgenic Tobacco Expressing the Phytochelatin Synthase Gene of Cynodon dactylon L.
Authors: Li Jiangchuan , Guo Jiangbo , Xu Wenzhong , Ma Mi
Published: August 2006

Bermudagrass (Cynodon dactylon L. cv. Goldensun) is highly resistant to and accumulates large amounts of cadmium (Cd). A phytochelatin synthase (PCS) cDNA (CdPCS1) was isolated from this grass by rapid amplification of cDNA ends. The putative CdPCS1 protein shared a high homology with PCS from other plants, with 79% homology at the N‐terminal and 47% homology at the C‐terminal. However, 16 Cys residues were found at the C‐terminal of CdPCS1, and among these residues, three positions were different from other PCS proteins. Semiquantitative reverse transcription‐polymerase chain reaction analysis showed that Cd stress induced CdPCS1 expression in both roots and leaves in Bermudagrass. We verified that CdPCS1 plays an important role in Cd tolerance in yeast cells by expressing the gene in ABDE1, a Cd‐sensitive mutant. CdPCS1 was then introduced into tobacco plants. The phytochelatin level in some transgenic tobacco lines increased 3.88‐fold more than in wild type plants and Cd accumulation in these transgenic plants was enhanced 3.21‐fold accordingly. The results suggested that CdPCS1 could be used as a gene element for phytoremediation in the future.

(Managing editor: Ya‐Qin Han)
Who Wants Informal Courts? Paradoxical Evidence from a Yugoslav Attempt to Create Workers' Courts for Labor Cases
Authors: Hayden Robert M.
Published: August 2006

The comrades' courts of the East European socialist countries are considered by those who favor alternative means of dispute resolution to be admirable examples of informal courts in modern industrial societies. However, these courts have not been extensively investigated. This article presents the results of an intensive observational study of one kind of socialist alternative court, the Yugoslav Courts of Associated Labor, comparing them with an ideal model of informal courts and with the available data on comrades' courts in other East European socialist countries. We find that, in contrast with the latter, the Yugoslav courts are indeed workers' courts, in the sense that they are used by workers—over 90% of their cases are brought by individual workers. On the other hand, they are not workers' courts in the sense of being controlled by workers—they are instead dominated by legal professionals. We conclude that these Yugoslav courts are attractive to individual workers precisely because they are not informal, social courts, but rather are independent legal agencies from which workers may receive unprejudiced decisions and substantial remedies.
Cerebral Microbleeds: Old Leaks and New Haemorrhages
Authors: Fiehler Jens
Published: August 2006

Black dots in the brain parenchyma diagnosed in T2*-weighted magnetic resonance imaging should be interpreted in light of the patient's history as well as the location, number and distribution of the lesions and associated imaging findings. These dots will correspond to haemoglobin degradation products in most of the cases referred to as cerebral microbleeds in patients with small vessel disease. Cerebral microbleeds have a prevalence of 5.7 % (range 3.7–7.7%) and are observed more frequently with increasing age. cerebral microbleeds have been observed in 47–80% of the primary intracerebral haemorrhage patients and in 0–78% of the patients with ischaemic cerebrovascular disease and appear to be a general marker of various types of bleeding-prone, small vessel disease and a predictor of recurrent vascular events. The occurrence and the number of cerebral microbleeds are associated with the degree of cerebral white matter abnormalities. Current data does not support the hypothesis that cerebral microbleeds are associated with a higher risk for a clinically relevant intracerebral haemorrhage, after anticoagulation/antiaggregation therapy, or after thrombolytic therapy in stroke patients. Therefore, the current data do not support the general exclusion of patients from therapy based on the presence of cerebral microbleeds. In the future, cerebral microbleeds may be incorporated into the design of clinical trials of anticoagulation/antiaggregation drugs in stroke patients for potential individual stratification for both the risk of recurrent ischaemic stroke and for the risk of intracerebral haemorrhage.
Burden of Stroke: Indian Perspective
Authors: Dalal Praful M.
Published: August 2006

India will face an enormous socio-economic burden to meet the costs of stroke. Stroke prevention planning, reliable epidemiological infromation on pattern of disease and exposure to major risk factors and morbidity or mortality trends for CVD in defined populations is imperative. There is a great need to monitor these trends in a simple and reproducible way. The World Health Organization (WHO) STEPs program does this and is now being implemented in India through the Indian Collaborative Acute Stroke Study (ICASS). The initial data supports the high prevalence of vascular risk factors and the relatively young age of stroke cases in India. A number of stroke prevalence studies have been performed in recent years, further confirming the high burden of stroke in this country. Recognition of the trend in the burden of stroke in India is essential to assist target prevention and management strategies.
Antioxidant Properties of Genistein in a Model Edible Oil System
Authors: Russin T.A. , Boye J.I. , Pham H.M. , Arcand Y.
Published: August 2006

ABSTRACT:  Antioxidant properties of genistein were tested during the low temperature oxidation of linseed oil. Four linseed oil solutions were prepared: linseed oil, linseed oil containing 2 μmol added genistein/g oil, linseed oil containing 4 μmol added genistein/g oil, and linseed oil containing 200 ppm butylated hydroxyanisole (BHA). Dimethyl sulfoxide (DMSO) (1.7%) was added to all solutions to allow for the solubilization of genistein. Antioxidant activities were analyzed using 2 techniques: (i) the oven stability test carried out at 60 °C with periodic determination of peroxide values (PV); and (ii) oxidation on a heated (60 °C) FTIR‐ATR crystal with periodic scanning and collection of infrared spectra. Induction points (IP) for each solution were determined. A strong correlation (r= 0.91) was noted between the 2 analytical methods used. Three significantly (P < 0.10) different levels of IP were generally observed. The shortest IP (17 h for PV, 36.9 h for FTIR) was found for the linseed oil. Intermediate IP's were obtained for the 2 μmol added genistein/g oil (32.4 h for PV, 43.5 h for FTIR) and 4 μmol added genistein/g oil (33.9 h for PV, 44.2 h for FTIR) samples. The longest IP (39.9 h for PV, 54.2 h for FTIR) was obtained for the 200 ppm BHA containing solution. Although genestein was not as effective as BHT in retarding oxidation, the results clearly demonstrate that it is able to act as an antioxidant in a bulk oil system.
Functional Characterization of Steam Jet‐Cooked β‐Glucan‐Rich Barley Flour as an Oil Barrier in Frying Batters
Authors: Lee Suyong , Inglett George E.
Published: August 2006

ABSTRACT:  The effect of steam jet‐cooking on the hydration, pasting, and rheological properties of barley flour was investigated. The thermo‐mechanical shear during steam jet‐cooking led to significant increases in the water absorption, water solubility, and swelling power of the barley flour. Also, the pasting profile showed elevated initial viscosity and reduced final viscosity. In addition, the suspensions of the steam jet‐cooked barley flour demonstrated typical shear thinning and dynamic viscoelastic responses of random coil polysaccharides with entanglements. The steam jet‐cooked barley flour was also evaluated as an oil barrier in fried foods. Its incorporation into frying batters increased batter pickup and viscosity while the moisture loss of fried batters was reduced. These combined effects significantly lowered the oil uptake of batters.
Potassium Lactate and Fresh‐Pork‐Sausage Formulation Effects on Shelf Life in Lighted and Unlighted Display
Authors: Seyfert Mark , Hunt Melvin C. , Grobbel Jeannine P. , Ryan Suzanne M. , Johnson Dallas E. , Monderen Rene A.
Published: August 2006

ABSTRACT:  The objective was to examine the effects of potassium lactate (KL) and raw‐meat blend on the shelf‐life of fresh‐pork‐sausage patties in lighted and unlighted display. Four fresh‐pork‐sausage blends (70% prerigor meat, 30% fresh‐pork trim; 80%/20%; 90%/10%; and 100%/0%) were formulated (n= 3) with 0% and 3% KL, and patties were displayed for 10 d at 4 °C in lighted display or for 7 d in unlighted display followed by 3 d in lighted display. Patties with KL had lower aerobic plate counts (APC), more stable color (greater a* and chroma, less visual discoloration), and less lipid oxidation (P < 0.05) during display than did patties without KL. Patties in the light for 10 d were more discolored and had more lipid oxidation (P < 0.05) than those in the dark for 7 d followed by the light for 3 d. Patties with KL in the light for 10 d had similar color (P > 0.05) to patties without KL that were in the dark for the first 7 d. Adding KL resulted in similar color stability (P > 0.05) among patties made from 80, 90, and 100% prerigor meat and greater color stability (P < 0.05) than similarly formulated patties without KL. By using 3% KL, fresh‐pork‐sausage patty color stability was maximized, APC and lipid oxidation were minimized, and formulation costs can be decreased by using less‐expensive fresh‐pork trim for up to 20% of the blend. Adding KL also minimized the effects of display lighting on myoglobin and lipid oxidation.
Kinetic Analysis of Bifidobacterial Metabolism Reveals a Minor Role for Succinic Acid in the Regeneration of NAD
+
through Its Growth-Associated Production
Authors: Van der Meulen Roel , Adriany Tom , Verbrugghe Kristof , De Vuyst Luc
Published: August 2006

Several strains belonging to the genus
Bifidobacterium
were tested to determine their abilities to produce succinic acid.
Bifidobacterium longum
strain BB536 and
Bifidobacterium animalis
subsp.
lactis
strain Bb 12 were kinetically analyzed in detail using in vitro fermentations to obtain more insight into the metabolism and production of succinic acid by bifidobacteria. Changes in end product formation in strains of
Bifidobacterium
could be related to the specific rate of sugar consumption. When the specific sugar consumption rate increased, relatively more lactic acid and less acetic acid, formic acid, and ethanol were produced, and vice versa. All
Bifidobacterium
strains tested produced small amounts of succinic acid; the concentrations were not more than a few millimolar. Succinic acid production was found to be associated with growth and stopped when the energy source was depleted. The production of succinic acid contributed to regeneration of a small part of the NAD
+
, in addition to the regeneration through the production of lactic acid and ethanol.
Multiple-Subunit Genes of the Aromatic-Ring-Hydroxylating Dioxygenase Play an Active Role in Biphenyl and Polychlorinated Biphenyl Degradation in
Rhodococcus
sp. Strain RHA1
Authors: Iwasaki Takumi , Miyauchi Keisuke , Masai Eiji , Fukuda Masao
Published: August 2006

A gram-positive strong polychlorinated biphenyl (PCB) degrader,
Rhodococcus
sp. strain RHA1, can degrade PCBs by cometabolism with biphenyl or ethylbenzene. In RHA1, three sets of aromatic-ring-hydroxylating dioxygenase genes are induced by biphenyl. The large and small subunits of their terminal dioxygenase components are encoded by
bphA1
and
bphA2
,
etbA1
and
etbA2
, and
ebdA1
and
ebdA2
, respectively, and the deduced amino acid sequences of
etbA1
and
etbA2
are identical to those of
ebdA1
and
ebdA2
, respectively. In this study, we examined the involvement of the respective subunit genes in biphenyl/PCB degradation by RHA1. Reverse transcription-PCR and two-dimensional polyacrylamide gel electrophoresis analyses indicated the induction of RNA and protein products of
etbA1
and
ebdA1
by biphenyl. Single- and double-disruption mutants of
etbA1
,
ebdA1
, and
bphA1
were constructed by insertional inactivation. The 4-chlorobiphenyl (4-CB) degradation activities of all the mutants were lower than that of RHA1. The results indicated that all of these genes are involved in biphenyl/PCB degradation. Furthermore, we constructed disruption mutants of
ebdA3
and
bphA3
, encoding ferredoxin, and
etbA4
, encoding ferredoxin reductase components. The 4-CB degradation activities of these mutants were also lower than that of RHA1, suggesting that all of these genes play a role in biphenyl/PCB degradation. The substrate preferences of
etbA1A2
/
ebdA1A2
- and
bphA1A2
-encoded dioxygenases for PCB congeners were examined using the corresponding mutants. The results indicated that these dioxygenase isozymes have different substrate preferences and that the
etbA1A2
/
ebdA1A2
-encoded isozyme is more active on highly chlorinated congeners than the
bphA1A2
-encoded one.
Rickettsia felisfrom Cat Fleas: Isolation and Culture in a Tick-Derived Cell Line
Authors: Pornwiroon Walairat , Pourciau Susan S. , Foil Lane D. , Macaluso Kevin R.
Published: August 2006

Rickettsia felis, the etiologic agent of spotted fever, is maintained in cat fleas by vertical transmission and resembles other tick-borne spotted fever group rickettsiae. In the present study, we utilized anIxodes scapularis-derived tick cell line, ISE6, to achieve isolation and propagation ofR. felis. A cytopathic effect of increased vacuolization was commonly observed inR. felis-infected cells, while lysis of host cells was not evident despite large numbers of rickettsiae. Electron microscopy identified rickettsia-like organisms in ISE6 cells, and sequence analyses of portions of the citrate synthase (gltA), 16S rRNA,Rickettsiagenus-specific 17-kDa antigen, and spotted fever group-specific outer membrane protein A (ompA) genes and, notably,R. felisconjugative plasmids indicate that this cultivatable strain (LSU) wasR. felis. Establishment ofR. felis(LSU) in a tick-derived cell line provides an alternative and promising system for the expansion of studies investigating the interactions betweenR. felisand arthropod hosts.
Peptide aMptD-Mediated Capture PCR for Detection ofMycobacterium aviumsubsp.paratuberculosisin Bulk Milk Samples
Authors: Stratmann Janin , Dohmann Karen , Heinzmann Julia , Gerlach Gerald-F.
Published: August 2006

A peptide-mediated capture PCR for the detection ofMycobacterium aviumsubsp.paratuberculosisin bulk milk samples was developed and characterized. Capture of the organism was performed using peptide aMptD, which had been shown to bind to theM. aviumsubsp.paratuberculosisMptD protein (J. Stratmann, B. Strommenger, R. Goethe, K. Dohmann, G. F. Gerlach, K. Stevenson, L. L. Li, Q. Zhang, V. Kapur, and T. J. Bull, Infect. Immun.72:1265-1274, 2004). Consistent expression of the MptD receptor protein and binding of the aMptD ligand were demonstrated by capturing differentMycobacterium aviumsubsp.paratuberculosistype I and type II strains and subsequent PCR analysis using ISMav2-based primers. The analytical sensitivity of the method was determined to be 5 × 102CFU ml−1for artificially contaminated milk. The specificity of aMptD binding was confirmed by culture and competitive capture assays, showing selective enrichment ofM. aviumsubsp.paratuberculosis(at a concentration of 5 × 102CFU ml−1) from samples containing 100- and 1,000-fold excesses of other mycobacterial species, includingM. aviumsubsp.aviumandM. aviumsubsp.hominissuis. The aMptD-mediated capture ofM. aviumsubsp.paratuberculosisusing paramagnetic beads, followed by culture, demonstrated the ability of this approach to capture viable target cells present in artificially contaminated milk. Surface plasmon resonance experiments revealed that the aMptD peptide is a high-affinity ligand with a calculated association rate constant of 9.28 × 103and an association constant of 1.33 × 109. The potential use of the method on untreated raw milk in the field was investigated by testing 423 bulk milk samples obtained from different dairy farms in Germany, 23 of which tested positive. Taken together, the results imply that the peptide-mediated capture PCR might present a suitable test for paratuberculosis screening of dairy herds, as it has an analytical sensitivity sufficient for detection ofM. aviumsubsp.paratuberculosisin bulk milk samples under field conditions, relies on a defined and validated ligand-receptor interaction, and is adaptable to routine diagnostic laboratory automation.
Dominant Microbial Populations in Limestone-Corroding Stream Biofilms, Frasassi Cave System, Italy
Authors: Macalady Jennifer L. , Lyon Ezra H. , Koffman Bess , Albertson Lindsey K. , Meyer Katja , Galdenzi Sandro , Mariani Sandro
Published: August 2006

Waters from an extensive sulfide-rich aquifer emerge in the Frasassi cave system, where they mix with oxygen-rich percolating water and cave air over a large surface area. The actively forming cave complex hosts a microbial community, including conspicuous white biofilms coating surfaces in cave streams, that is isolated from surface sources of C and N. Two distinct biofilm morphologies were observed in the streams over a 4-year period. Bacterial 16S rDNA libraries were constructed from samples of each biofilm type collected from Grotta Sulfurea in 2002. β-, γ-, δ-, and ε-proteobacteria in sulfur-cycling clades accounted for ≥75% of clones in both biofilms. Sulfate-reducing and sulfur-disproportionating δ-proteobacterial sequences in the clone libraries were abundant and diverse (34% of phylotypes). Biofilm samples of both types were later collected at the same location and at an additional sample site in Ramo Sulfureo and examined, using fluorescence in situ hybridization (FISH). The biomass of all six stream biofilms was dominated by filamentous γ-proteobacteria with
Beggiatoa
-like and/or
Thiothrix
-like cells containing abundant sulfur inclusions. The biomass of ε-proteobacteria detected using FISH was consistently small, ranging from 0 to less than 15% of the total biomass. Our results suggest that S cycling within the stream biofilms is an important feature of the cave biogeochemistry. Such cycling represents positive biological feedback to sulfuric acid speleogenesis and related processes that create subsurface porosity in carbonate rocks.
Metabolic Profiles and Genetic Diversity of Denitrifying Communities in Activated Sludge after Addition of Methanol or Ethanol
Authors: Hallin Sara , Throbäck Ingela Noredal , Dicksved Johan , Pell Mikael
Published: August 2006

External carbon sources can enhance denitrification rates and thus improve nitrogen removal in wastewater treatment plants. The effects of adding methanol and ethanol on the genetic and metabolic diversity of denitrifying communities in activated sludge were compared using a pilot-scale plant with two parallel lines. A full-scale plant receiving the same municipal wastewater, but without external carbon source addition, was the reference. Metabolic profiles obtained from potential denitrification rates with 10 electron donors showed that the denitrifying communities altered their preferences for certain compounds after supplementation with methanol or ethanol and that methanol had the greater impact. Clone libraries of
nirK
and
nirS
genes, encoding the two different nitrite reductases in denitrifiers, revealed that methanol also increased the diversity of denitrifiers of the
nirS
type, which indicates that denitrifiers favored by methanol were on the rise in the community. This suggests that there might be a niche differentiation between
nirS
and
nirK
genotypes during activated sludge processes. The composition of
nirS
genotypes also varied greatly among all samples, whereas the
nirK
communities were more stable. The latter was confirmed by denaturing gradient gel electrophoresis of
nirK
communities on all sampling occasions. Our results support earlier hypotheses that the compositions of denitrifier communities change during predenitrification processes when external carbon sources are added, although no severe effect could be observed from an operational point of view.
Cloning and Expression of Two Crystal Protein Genes,
cry30Ba1
and
cry44Aa1
, Obtained from a Highly Mosquitocidal Strain,
Bacillus thuringiensis
subsp.
entomocidus
INA288
Authors: Ito Takeshi , Ikeya Tomonori , Sahara Ken , Bando Hisanori , Asano Shin-ichiro
Published: August 2006

Two novel crystal protein genes,
cry30Ba
and
cry44Aa
, were cloned from
Bacillus thuringiensis
subsp.
entomocidus
INA288 and expressed in an acrystalliferous strain. Cry44Aa crystals were highly toxic to second-instar
Culex pipiens pallens
(50% mortality concentration [LC
50
] = 6 ng/ml) and
Aedes aegypti
(LC
50
= 12 ng/ml); however, Cry30Ba crystals were not toxic.
Molecular Fingerprinting of
Cryptosporidium
Oocysts Isolated during Water Monitoring
Authors: Nichols Rosely A. B. , Campbell Brian M. , Smith Huw V.
Published: August 2006

We developed and validated a PCR-based method for identifying
Cryptosporidium
species and/or genotypes present on oocyst-positive microscope slides. The method involves removing coverslips and oocysts from previously examined slides followed by DNA extraction. We tested four loci, the 18S rRNA gene (N18SDIAG and N18SXIAO), the
Cryptosporidium
oocyst wall protein (COWP) gene (STN-COWP), and the dihydrofolate reductase (
dhfr
) gene (by multiplex allele-specific PCR), for amplifying DNA from low densities of
Cryptosporidium parvum
oocysts experimentally seeded onto microscope slides. The N18SDIAG locus performed consistently better than the other three tested. Purified oocysts from humans infected with
C. felis
,
C. hominis
, and
C. parvum
and commercially purchased
C. muris
were used to determine the sensitivities of three loci (N18SDIAG, STN-COWP, and N18SXIAO) to detect low oocyst densities. The N18SDIAG primers provided the greatest number of positive results, followed by the N18SXIAO primers and then the STN-COWP primers. Some oocyst-positive slides failed to generate a PCR product at any of the loci tested, but the limit of sensitivity is not entirely based on oocyst number. Sixteen of 33 environmental water monitoring
Cryptosporidium
slides tested (oocyst numbers ranging from 1 to 130) contained mixed
Cryptosporidium
species. The species/genotypes most commonly found were
C. muris
or
C. andersoni
,
C. hominis
or
C. parvum
, and
C. meleagridis
or
Cryptosporidium
sp. cervine, ferret, and mouse genotypes. Oocysts on one slide contained
Cryptosporidium
muskrat genotype II DNA.
Peptidoglycan from
Bacillus cereus
Mediates Commensalism with Rhizosphere Bacteria from the
Cytophaga-Flavobacterium
Group
Authors: Peterson Snow Brook , Dunn Anne K. , Klimowicz Amy K. , Handelsman Jo
Published: August 2006

Previous research in our laboratory revealed that the introduction of
Bacillus cereus
UW85 can increase the populations of bacteria from the
Cytophaga-Flavobacterium
(CF) group of the
Bacteroidetes
phylum in the soybean rhizosphere, suggesting that these rhizosphere microorganisms have a beneficial relationship (G. S. Gilbert, J. L. Parke, M. K. Clayton, and J. Handelsman, Ecology 74:840-854, 1993). In the present study, we determined the frequency at which CF bacteria coisolated with
B. cereus
strains from the soybean rhizosphere and the mechanism by which
B. cereus
stimulates the growth of CF rhizosphere strains in root exudate media. In three consecutive years of sampling, CF strains predominated among coisolates obtained with
B. cereus
isolates from field-grown soybean roots. In root exudate media, the presence of
B. cereus
was required for CF coisolate strains to reach high population density. However, rhizosphere isolates from the phylum
Proteobacteria
grew equally well in the presence and absence of
B. cereus
, and the presence of CF coisolates did not affect the growth of
B. cereus
. Peptidoglycan isolated from
B. cereus
cultures stimulated growth of the CF rhizosphere bacterium
Flavobacterium johnsoniae
, although culture supernatant from
B. cereus
grown in root exudate media did not. These results suggest
B. cereus
and CF rhizosphere bacteria have a commensal relationship in which peptidoglycan produced by
B. cereus
stimulates the growth of CF bacteria.
σBActivation under Environmental and Energy Stress Conditions inListeria monocytogenes
Authors: Chaturongakul Soraya , Boor Kathryn J.
Published: August 2006

To measure σBactivation inListeria monocytogenesunder environmental or energy stress conditions, quantitative reverse transcriptase PCR (TaqMan) was used to determine the levels of transcripts for the σB-dependentopuCAandclpCgenes in strains having null mutations in genes encodingregulator ofsigmaBproteins (rsbTandrsbV) and sigma B (sigB) and in theL. monocytogeneswild-type 10403S strain under different stress conditions. The ΔsigB, ΔrsbT, and ΔrsbVstrains previously exhibited increased hemolytic activities compared to the hemolytic activity of the wild-type strain; therefore, transcript levels forhlywere also determined. RsbT, RsbV, and σBwere all required foropuCAexpression during growth under carbon-limiting conditions or following exposure to pH 4.5, salt, ethanol, or the protonophore carbonyl cyanidem-chlorophenylhydrazone (CCCP). Expression ofclpCwas RsbT, RsbV, and σBdependent in the presence of CCCP but not under the other conditions.hlyexpression was not RsbT, RsbV, or σBdependent in the presence of either CCCP or salt.opuCAtranscript levels did not increase in the presence of rapidly lethal stresses (i.e., pH 2.5 or 13 mM cumene hydroperoxide) despite the enhanced survival of the wild type compared with the survival of the mutant strains under these conditions. These findings highlight the importance of complementing phenotypic characterizations with gene expression studies to identify direct and indirect effects of null mutations in regulatory genes, such assigB. Overall, our data show that while σBactivation occurs through a single pathway under both environmental and energy stress conditions, regulation of expression of some stress response and virulence genes in the σBregulon (e.g.,clpC) appears to require networks involving multiple transcriptional regulators.
Examination of breast needle core biopsy specimens performed for screen-detected microcalcification
Authors: Kumaraswamy V , Carder P J
Published: August 2006

Aims:To establish the number of histological levels necessary for the evaluation of breast needle core biopsy (NCB) specimens taken from areas of mammographic calcification in patients presenting via the UK National Health Service Breast Screening Programme.

Methods:Retrospective review of a series of breast NCB specimens initially examined routinely at nine levels. The presence of calcification within the histological sections in each of three sets of levels (levels 1–3, 4–6 and 7–9) and the (cumulative) diagnostic B category that would have pertained after each were assessed.

Results:Accurate diagnostic classification was possible after examination of three levels in 89% cases. Examination of a further three levels permitted accurate diagnosis in a further eight cases (total 97% cases). In only three cases were nine levels necessary for accurate classification. In only a single case (1%) was it likely that routine examination of six levels could have led to significant misclassification. In a significant group of patients (18%), nine levels were considered to provide additional useful information, although this information did not alter the diagnosis.

Conclusions:NCBs for screen-detected mammographic calcification should be routinely examined at six levels. Further levels may be needed in occasional cases to identify more conclusively the associated pathological abnormality. Further levels may be of particular value when assessing atypical intraductal proliferative epithelial lesions.
Squamous differentiation in primary urothelial carcinoma of the urinary tract as seen by MAC387 immunohistochemistry
Authors: Lopez-Beltran Antonio , Requena Maria J , Alvarez-Kindelan Jose , Quintero Ana , Blanca Ana , Montironi Rodolfo
Published: August 2006

Squamous differentiation (SqD) is variably present in urinary tract tumours, but its significance remains unclear. In this study, SqD was assessed by immunohistochemistry using the monoclonal antibody Mac387 in 145 urothelial tumours (bladder, n = 115; renal pelvis, n = 30). Mac387 detects the myelomonocytic L1 antigen; a member of the calgranulin family shared by epithelial cells and keratinocytes. L1 antigen was shown in SqD in urothelial carcinomas of the bladder or the renal pelvis, including 11 cases with focal SqD unrecognised by conventional analysis. SqD is more frequent in renal pelvic tumours (p = 0.027) and increases with grade/stage mainly in bladder carcinoma (grade, p = 0.05; stage, p = 0.005). Stage Ta/T1 bladder carcinomas with SqD recurred more (p = 0.021). In conclusion, Mac387 efficiently shows SqD in urothelial tumours.
Getting across the nuclear pore complex: new insights into nucleocytoplasmic transportThis paper is one of a selection of papers published in this Special Issue, entitled The Nucleus: A Cell Within A Cell.
Authors: Stoffler Daniel , Schwarz-Herion Kyrill , Aebi Ueli , Fahrenkrog Birthe
Published: August 2006

Small ions and molecules can traverse the nuclear pore complex (NPC) simply by diffusion, whereas larger proteins and RNAs require specific signals and factors that facilitate their passage through the NPC. Our understanding of the factors that participate and regulate nucleocytoplasmic transport has increased tremendously over the past years, whereas the actual translocation step through the NPC has remained largely unclear. Here, we present and discuss recent findings on the interaction between the NPC and transport receptors and provide new evidence that the NPC acts as a constrained diffusion pore for molecules and particles without retention signal and as an affinity gate for signal-bearing cargos.
Synergistic Effect of Ginger and Nifedipine on Human Platelet Aggregation: A Study in Hypertensive Patients and Normal Volunteers
Authors: Young Haw-Yaw , Liao Jung-Chun , Chang Yuan-Shiun , Luo Yen-Lin , Lu Ming-Chin , Peng Wen-Huang
Published: August 2006

In this study, we evaluated the synergistic effect of ginger and nifedipine on anti-platelet aggregation in normal human volunteers and hypertensive patients. The results showed that the percentage of platelet aggregation induced by collagen, ADP and epinephrine in hypertensive patients was larger than that in normal volunteers. Either aspirin or ginger could potentiate the anti-platelet aggregation effect of nifedipine in normal volunteer and hypertensive patients. These results suggested that ginger and nifedipine possessed synergistic effect on anti-platelet aggregation. A combination of 1 g ginger with 10 mg nifedipine per day could be valuable for cardiovascular and cerebrovascular complication due to platelet aggregation.
Effects of Safflower Seed Oil in Osteoporosis Induced-Ovariectomized Rats
Authors: Alam Md. Rafiqul , Kim Soo Mi , Lee Jong Il , Chon Seung Ki , Choi Sung Jin , Choi In Hyuk , Kim Nam Soo
Published: August 2006

The effects of Korean Safflower (Carthamus tinctorius L) seed oil (SSO) on osteoporosis induced-ovariectomized rats were investigated. A total of 90 female Sprague-Dawley rats, 4-month-old, weighing 200–230 g were randomly assigned into 3 groups (30 animals/group). The animals in group I were sham operated and those of group II and III were ovariectomized (Ovx). After eight weeks, the animals of group I and II received solvent vehicle daily, whereas those of group III were administered SSO orally (1 ml/kg) daily for 30 days. The changes in the serum levels of insulin-like growth factor-I (IGF-I), IGF-II, insulin-like growth factor binding protein-3 (IGBP-3), estrogen, total alkaline phosphatase (TALP), bone-specific alkaline phosphatase (BALP), calcium and phosphorous in serum, and also the histomorphology of the proximal tibia metaphysis and femur/body weight (F/B) ratio were examined in all the groups at every 10 days interval. Thirty days later, IGF-I, IGF-II, IGBP-3 and BALP levels were significantly increased ( p < 0.05) in group III as compared to groups I and II. There were no significant differences in serum levels of estrogen, TALP and F/B ratio between groups II and III, but estrogen levels were higher in group I. These results suggest that the safflower seeds have possible roles in the improvement of osteoporosis induced-ovariectomized rats.
Shen-Fu Attenuates Endotoxin-Induced Acute Lung Injury in Rats
Authors: Qian Yanning , Sun Jie , Wang Zhongyun , Yang Jianjun
Published: August 2006

Sepsis is associated with the highest risk of progression to acute lung injury or acute respiratory distress syndrome. Shen-Fu has been advocated to treat many severely ill patients. Our study was designed to investigate the effect of Shen-Fu on endotoxin-induced acute lung injury in vivo. Adult male Wistar rats were randomly divided into 6 groups: controls; those challenged with endotoxin (5 mg/kg) and treated with saline; those challenged with endotoxin (5 mg/kg) and treated with Shen-Fu (1 mg/kg); those challenged with endotoxin (5 mg/kg) and treated with Shen-Fu (10 mg/kg); increase challenged with endotoxin (5 mg/kg) and treated with Shen-Fu (100 mg/kg); saline injected and treated with Shen-Fu (100 mg/kg). TNF-α, IL-6, and NF-kappa B were investigated in the lung two hours later. Myeloperoxidase (MPO) activity and wet/dry weight ratio were investigated six hours later. Intravenous administration of endotoxin provoked significant lung injury, which was characterized by increment increase of MPO activity and wet/dry lung weight ratio, and TNF-α and IL-6 expression and NF-kappa B activation. Shen-Fu (10,100 mg/kg) decreased MPO activity and wet/dry weight ratio and inhibited TNF-α and IL-6 production, endotoxin-induced NF-kappa B activation. Our results indicated that Shen-Fu at a dose of higher than 10 mg/kg inhibited endotoxin-induced pulmonary inflammation in vivo.
EVOLUTION OF FRACTAL SHAPES FOR ARTISTS AND DESIGNERS
Authors: LUTTON EVELYNE
Published: August 2006

We analyse in this paper the way randomness is considered and used in ArtiE-Fract. ArtiE-Fract is an interactive software, that allows the user (artist or designer) to explore the space of fractal 2D shapes with help of an interactive genetic programming scheme. The basic components of ArtiE-Fract are first described, then we focus on its use by two artists, illustrated by samples of their works. These "real life" tests have led us to implement additional components in the software. It seems obvious for the people who use ArtiE-Fract that this system is a versatile tool for creation, especially regarding the specific use of controlled random components.
CONICAL EMISSION OF TI:SAPPHIRE FEMTOSECOND LASER PULSES PROPAGATING IN WATER
Authors: ZHANG SHIAN , SUN ZHENRONG , YANG XIHUA , WANG ZUGENG , XU ZHIZHAN , LI RUXIN
Published: August 2006

Conical emission is investigated for Ti :sapphire femtosecond laser pulses propagating in water. The colored rings can be observed in the forward direction due to the constructive and destructive interference of transverse wavevector, which are induced by the spatio-temporal gradient of the free-electron density. With increasing input laser energy, due to filamentation and pulse splitting induced by the plasma created by multiphoton excitation of electrons from the valence band to the conduction band, the on-axis spectrum of the conical emission is widely broadened and strongly modulated with respect to input laser spectrum, and finally remains fairly constant at higher laser energy due to intensity clamping in the filaments.