bluuebunny/crossref_metadata_2025 huggingface dataset:
Placental Hofbauer cells and complications of pregnancy
Authors: Tang Zhonghua , Abrahams Vikki M. , Mor Gil , Guller Seth
Published: March 2011

Hofbauer cells (HBCs) are placental macrophages that are present in the villus across gestation. Despite their identification more than 100 years ago, their specific role in placental function remains largely unelucidated. We initially review aspects of their history and biology as well as evidence for putative sites of origin. To gain insight into their potential function, we then describe complications of pregnancy including villitis of unknown etiology (VUE) and histological chorioamnionitis (HCA), in which alterations in numbers, gene expression, or other characteristics of HBCs have been documented to occur. We further review methods for isolation of HBCs and in vitro studies that explore their role in relation to other major cell types in the placenta and examine their actions in cytokine‐mediated inflammation. We conclude that HBCs play a key role in placental pathophysiology, and future advances in their isolation and culture would enable mechanistic insight into their villus function.
Oocyte donation programs: strategy for improving results
Authors: Borini Andrea , Suriano Rosanna , Barberi Marzia , Dal Prato Luca , Bulletti Carlo
Published: March 2011

Oocyte donation is now a useful option for women who cannot start a spontaneous pregnancy for reasons related to advanced age, iatrogenic factors, early depletion of ovarian reserve, or genetic disorders. Embryo implantation rates, pregnancy rates, and pregnancy outcomes among women included in oocyte donation programs were shown to be comparable to those of spontaneous or in vitro fertilization (IVF) pregnancies. With oocyte freezing and cryobanks, recipients may have a successful response to oocyte donation, with no need to be on waiting lists, access to a larger number of oocytes from the same donor, and a lesser risk of infectious disease transmission.
Epigenetic regulatory mechanisms during preimplantation embryo development
Authors: Palini Simone , De Stefani Silvia , Scala Valentina , Dusi Ludovica , Bulletti Carlo
Published: March 2011

Within the past few years, developmental scientists have switched their attention from the study of DNA sequencing to the epigenetic state of the genome. Studying epigenetics could present the key to understanding gene expression changes without altering the basic structure of DNA. For example, the blastocyst, trophectoderm, and inner cell mass grow within the same environment, having the same genome, but differentiate differently. Each stage of embryo development is characterized by a specific epigenetic pattern. These modifications give the embryos the ability to interact with the uterus. Gene expression profiles change dramatically, and chromatin remodeling allows for core histone elements to undergo significant modifications. In the past, epigenetic mechanisms were studied using less accurate technologies such as PCR techniques and gel electrophoresis. Today microarray, DNA analyzers, and other in silico approaches give us the capability to understand the epigenetic impact on differentiation and cell fate.
Isolation, characterization, and function of EBAF/LEFTY B: role in infertility
Authors: Tabibzadeh Siamak
Published: March 2011

Human endometrium exhibits cyclic stromal and glandular remodeling in preparation of embryo implantation. We identified EBAF/LEFTY B as a soluble cytokine of the TGF‐β superfamily that is expressed at a low level in human endometrium during the receptivity period, while it is maximally expressed during perimenstrual and menstrual phases. Transfection of cells with EBAF/LEFTY B resulted in expression of a 42 kD protein that was proteolytically processed to release two polypeptides of 34 and 28 kD. EBAF/LEFTY B causes tissue remodeling by induction of collagenolysis by matrix metalloproteases. In a subset of infertile patients, the expression EBAF/LEFTY B was prematurely increased during the implantation window. We showed that induced overexpression of EBAF/LEFTY B in transgenic mice impairs implantation. EBAF/LEFTY B inhibits the expression of key decidual proteins, IGFBP‐1 and PRL, through regulation of transcription factors FOXO1 and ETS1. Together, these findings show that during embryonic development, EBAF/LEFTY B plays important roles in decidualization and embryo implantation.
Ultrasonographic staging: a new staging system for deep endometriosis
Authors: Coccia Maria Elisabetta , Rizzello Francesca
Published: March 2011

Modern imaging techniques allow for the noninvasive diagnosis of endometriosis. Preoperative staging of pelvic endometriosis helps the gynecologist plan therapy and offer a prognosis to patients. The challenge of creating a satisfactory classification of endometriosis remains. The ability of the current classification schemes to predict pregnancy outcome, or aid in the management of pelvic pain, is recognized to be inadequate. The study of deeply infiltrating endometriosis and adenomyosis is greatly hampered by a lack of clear terminology and the absence of a consensus classification of the lesions. A reviewed consensus classification of endometriosis in general, with a more detailed consideration on deep endometriosis, is urgently required. We suggest a new staging system for deep, infiltrating endometriosis based on ultrasonographic findings. Prospective data collection and review in large centers may provide a larger clinical base from which to derive empirical point scores and breakpoints in the classification scheme.
Ovarian cryopreservation strategies and the fine control of ovarian follicle development in vitro
Authors: Johnson Joshua , Patrizio Pasquale
Published: March 2011

Preserving the fertility of women whose reproductive tract function is threatened by disease or disease treatment is a challenge for the assisted reproduction clinic. In particular, the ovary is susceptible to damage by chemo‐ and radiotherapeutic treatments, as the reserve of primordial follicles is susceptible to damage and death. To overcome this challenge, experimental strategies are being developed where ovarian tissue or an entire ovary is removed and cryopreserved during the patient's treatment. After recovery, the tissue is thawed and reintroduced orthotopically to the patient, in hopes that ovarian function will recover. In complementary approaches, attempts are being made to foster ovarian follicle development in vitro so that mature eggs can be produced. Here, we consider the state of the art of these strategies, and consider the question of whether whole ovary freezing has a place in fertility preservation in the foreseeable future.
Intensive care treatment of ovarian hyperstimulation syndrome (OHSS)
Authors: Sansone Pasquale , Aurilio Caterina , Pace Maria Caterina , Esposito Raffaella , Passavanti Maria Beatrice , Pota Vincenzo , Pace Leonardo , Pezzullo Martina Gilda , Bulletti Carlo , Palagiano Antonio
Published: March 2011

Ovarian hyperstimulation syndrome (OHSS) is a morbid, iatrogenic, and potentially lethal condition caused by the pharmacological induction of ovulation. In OHSS, the ovaries are the site of significant stromal edema with bilateral cystic ovarian enlargement. It has been suggested that ascitic fluid accumulation may be due to increased capillary permeability, which results in fluid shift from the intravascular to the extravascular compartment. While the exact etiology is not clear, several biochemical markers are considered to be possible mediators. There is an incidence of OHSS in about 3% to 8% of all therapy cycles. Severe OHSS is the rarest form, with an incidence characterized by a degree of variability ranging from 0.1% to 2.5% of women subjected to induced polyovulation.
The artificial womb
Authors: Bulletti Carlo , Palagiano Antonio , Pace Caterina , Cerni Angelica , Borini Andrea , de Ziegler Dominique
Published: March 2011

The availability of computer‐controlled artificial hearts, kidneys, and lungs, as well as the possibility of implanting human embryos in ex vivo uterus models or an artificial endometrium, presents new perspectives for creating an artificial uterus. Survival rates have also improved, with fetuses surviving from as early as 24 weeks of gestation. These advances bring new opportunities for complete or partial ectogenesis through the creation of an artificial womb, one that could sustain the growth and development of fetuses outside of the human body.
Modification of Buffered Peptone Water for Improved Recovery of Heat‐Injured Salmonella Typhimurium
Authors: Taskila Sanna , Osmekhina Ekaterina , Tuomola Mika , Ruuska Jari , Neubauer Peter
Published: March 2011

Abstract:  Rapid detection of Salmonella in foods is often limited by the high demand for the sensitivity of detection, poor physiological conditions of the target cells, and high concentration of background flora. In this study, the conditions of nonselective enrichment cultivation were modified in order to improve the quantitative detection of heat‐injured Salmonella in minced meat. The effect of the modifications on the recovery was observed by means of RNA‐based sandwich hybridization, which was adjusted for the quantification of Salmonella enterica 23S rRNA in crude cell extracts. The supplementation of buffered peptone water with the enzyme‐controlled substrate delivery system EnBase‐Flo® and ferrioxamine E was shown to improve the recovery of cells in both single strain cultures and in the presence of minced meat. The presented results can be used for the development of more efficient enrichment cultivation media for faster detection of food borne Salmonella.
Vitamin C and Sugar Levels as Simple Markers for Discriminating Spanish Honey Sources
Authors: León‐Ruiz Virginia , Vera Soledad , González‐Porto Amelia V. , Andrés María Paz San
Published: March 2011

Abstract:  In this work, 7 Spanish honeys with different botanical origins were studied. The honey origins were rosemary, chestnut, lavender, echium, thyme, multifloral, and honeydew. The chemical compounds determined were ascorbic acid (vitamin C), hydroxymethylfurfural, and major sugar contents (glucose and fructose). The physicochemical parameters, pH, conductivity, moisture, free acidity, and color, were also measured. Vitamin C is an important antioxidant in food, and the possibility to use it as discriminate parameter among different honeys was studied. The determination of vitamin C in honey samples was carried out by 2 different methods, volumetric and chromatographic comparing the results by both statistically. Vitamin C content was higher in thyme honeys than in the other types; however a wide dispersion in the values was found. Through a linear discriminant analysis (LDA), conductivity, glucose, fructose, and vitamin C content were the most important discriminant parameters.

Practical Application:  Vitamin C content in different honey sources has been determined by a simple and rapid chromatographic method (less than 3 min) in honeys from 6 botanical origins. The results together with glucose and fructose content and some physicochemical parameters have been studied in order to discriminate the botanical origin of honeys and in the future certified their quality. A statistical LDA was applied to the data, and differentiation of honey sources was possible with very good agreement. The vitamin C content found in thymus honeys was significantly higher than in other types. This fact makes vitamin C a special marker for thymus honeys that have a higher antioxidant effect than the others giving it special properties. The identification of honey sources is essential for beekeepers in order to certify honeys for consumers.
Preparation and Characterization of Agar/Clay Nanocomposite Films: The Effect of Clay Type
Authors: Rhim Jong‐Whan , Lee Soo‐Bin , Hong Seok‐In
Published: March 2011

Abstract:  Agar‐based nanocomposite films with different types of nanoclays, such as Cloisite Na+, Cloisite 30B, and Cloisite 20A, were prepared using a solvent casting method, and their tensile, water vapor barrier, and antimicrobial properties were tested. Tensile strength (TS), elongation at break (E), and water vapor permeability (WVP) of control agar film were 29.7 ± 1.7 MPa, 45.3 ± 9.6%, and (2.22 ± 0.19) × 10−9 g·m/m2·s·Pa, respectively. All the film properties tested, including transmittance, tensile properties, WVP, and X‐ray diffraction patterns, indicated that Cloisite Na+ was the most compatible with agar matrix. TS of the nanocomposite films prepared with 5% Cloisite Na+ increased by 18%, while WVP of the nanocomposite films decreased by 24% through nanoclay compounding. Among the agar/clay nanocomposite films tested, only agar/Cloisite 30B nanocomposite film showed a bacteriostatic function against Listeria monocytogenes.
Effect of Modified Atmosphere Packaging on Quality Index Method (QIM) Scores of Farmed Gilthead Seabream (Sparus aurata L.) at Low and Abused Temperatures
Authors: Campus Marco , Bonaglini Elia , Cappuccinelli Roberto , Porcu Maria Cristina , Tonelli Roberto , Roggio Tonina
Published: March 2011

Abstract: A Quality Index Method (QIM) scheme was developed for modified atmosphere packaging (MAP) packed gilthead seabream, and the effect of MAP gas mixtures (60% CO2and 40% N2; 60% CO2, 30% O2, and 10% N2), temperature (2, 4, and 8 °C), and time of storage on QI scores was assessed. QI scores were crossed with sensory evaluation of cooked fish according to a modified Torry scheme to establish the rejection point. In order to reduce redundant parameters, a principal component analysis was applied on preliminary QIM parameters scores coming from the best performing MAP among those tested. The final QIM scheme consists of 13 parameters and a maximum demerit score of 25. The maximum storage time was found to be 13 d at 4 °C for MAP 60% CO2and 40% N2. Storage at 2 °C do not substantially improved sensory parameters scores, while storage under temperature abuse (8 °C) accelerated drastically the rate of increase of QI scores and reduced the maximum storage time to 6 d.
Toward Better Control of Salmonella Contamination by Taking Advantage of the Egg's Self‐Defense System: A Review
Authors: Zhang Wei , Zheng Jiang‐Xia , Xu Gui‐Yun
Published: March 2011

Abstract:  Egg‐associated salmonellosis is a major problem for food safety. It can be caused by vertical transmission (transovarian transmission) in hens and horizontal transmission though penetration. Despite a series of physical and chemical defense mechanisms naturally found in eggs, they cannot provide complete protection for them. Environmental hygiene, bacteria vectors such as birds, rodent, flies, and beetles along with feed and water contamination are the most frequently reported causes of Salmonella colonization in hens, and finally to eggs. In addition, inappropriate egg handling will cause eggs to lose their self‐protection ability, thus resulting in the survival and multiplication of Salmonella in an egg's contents, which contributes to the horizontal dissemination. The routes of Salmonella contamination were discussed, and the effectiveness and shortcomings of different decontamination methods were evaluated in this review. Various studies on egg storage indicated that the low‐temperature storage without temperature fluctuation was beneficial for the control of Salmonella. This review, based on an understanding of the stages of Salmonella transmission and an egg's self‐protection mechanisms, highlights a comprehensive strategy toward Salmonella control in a process from egg production and handling to human consumption.
Antifungal Effectiveness of Potassium Sorbate Incorporated in Edible Coatings Against Spoilage Molds of Apples, Cucumbers, and Tomatoes during Refrigerated Storage
Authors: Mehyar Ghadeer F. , Al‐Qadiri Hamzah M. , Abu‐Blan Hifzi A. , Swanson Barry G.
Published: March 2011

Abstract:  Predominant spoilage molds of fresh apples, cucumbers, and tomatoes stored at 4 °C were isolated and examined for resistance to potassium sorbate (PS) incorporated in polysaccharide edible coatings. The isolated molds were Penicillium expansum, Cladosporium herbarum, and Aspergillus niger from apples. P. oxalicum and C. cucumerinum were isolated from cucumbers and P. expansium and C. fulvum from tomatoes. Guar gum edible coating incorporated with PS was the most effective mold inhibitor, significantly (P < 0.05) reducing the isolated spoilage molds for 20, 15, and 20 d of storage at 4 °C on apples, cucumbers, and tomatoes, respectively. PS incorporated into pea starch edible coating was less effective and selectively inhibited the isolated mold species, causing significant (P < 0.05) reduction in mold on apples, cucumbers, and tomatoes counts for 20, 10 to 15, and 15 to 20 d of storage at 4 °C, respectively. The isolated mold species exhibited different resistances to PS incorporated in the edible coatings. The greatest inhibition (2.9 log CFU/g) was obtained with C. herbarum on apples and the smallest (1.1 log CFU/g) was with P. oxalicum on cucumbers and the other isolated mold species exhibited intermediate resistance. The coatings tested, in general, inhibited molds more effectively on apples than on tomatoes and cucumbers. Addition of PS to pea starch and guar gum, edible coatings improved the antifungal activity of PS against isolated spoilage molds on apples, cucumbers, and tomatoes. PS inhibition was most effective against C. herbarum on apples and least effective against P. oxalicum on cucumbers.
Believing in a secular age: Anthropology, sociology and religious experience1
Authors: Baldacchino Jean‐Paul , Kahn Joel S.
Published: March 2011

Charles Taylor’sA Secular Agegenerated a great deal of attention—and has stimulated important debates—among a diverse range of scholars in sociology, history, politics, religious studies and to a lesser extent, anthropologists. Much of the debate has focused on the implications of Taylor’s work for the so‐called secularisation thesis and the place (or non‐place) of religion in the so‐called public sphere. The essays in this volume arise less out of such concerns and more from Taylor’s discussion of secularism in a third, ‘experiential’ sense. Each paper addresses the question of what it is like to ‘believe’ (or not ‘believe’) in the modern world. Among other things the authors of the essays published in this Special Issue are concerned to develop better understandings of the conditions under which belief and unbelief may be experienced as open, rather than closed, to the possibility of other ontological construals, thereby building on Taylor’s insights into the phenomenology of modern secularism.
Credentialing of pharmacy technicians in the USA
Authors: Alkhateeb Fadi M , Shields Kelly M , Broedel-Zaugg Kimberly , Bryan Amy , Snell Jennifer
Published: March 2011

Objective

This review will compare the USA and UK regarding pharmacy technicians' roles, it will summarize the current roles and responsibilities of pharmacy technicians in the USA, public perception of pharmacy technicians, pharmacy organizations' perspectives on pharmacy technician credentialing, academic programmes for pharmacy technicians, accreditation of pharmacy technician programmes, pharmacy technician certification exams and differing perspectives on the push for standardized technician training. It will conclude with observations regarding the importance of standardized pharmacy technician training.

Methods

Articles were identified via searches of PubMed and IPA from inception to November 2010 related to credentialing of pharmacy technicians. Search terms included pharmacy technician, pharmacy technician certification, pharmacy registration, technician education and technician requirements. Articles describing the roles and responsibilities of a technician, public perception of technicians, demographics, certification processes and the future of technician roles were included. An Internet search was also performed to identify articles in the lay press related to this topic.

Key findings

Providing a pharmacy technician with proper training and education is necessary for operating a successful pharmacy. In the USA, mandating a national standardized training programme is the source of the debate. Current rules and regulations regarding the training and education needed for a pharmacy technician vary from state to state in the USA. Attitudes of technicians towards standardized training may be difficult to change.

Conclusion

This paper provides a review of pharmacy technicians' roles and responsibilities across different pharmacy settings. Although the literature provides some insight, more studies are needed to assess the value and impact of the knowledge and skills possessed by certified pharmacy technicians with standardized training compared with technicians with site-specific or limited training.
Characterization of the Highly Active Polyhydroxyalkanoate Synthase of Chromobacterium sp. Strain USM2
Authors: Bhubalan Kesaven , Chuah Jo-Ann , Shozui Fumi , Brigham Christopher J. , Taguchi Seiichi , Sinskey Anthony J. , Rha ChoKyun , Sudesh Kumar
Published: March 2011

The synthesis of bacterial polyhydroxyalkanoates (PHA) is very much dependent on the expression and activity of a key enzyme, PHA synthase (PhaC). Many efforts are being pursued to enhance the activity and broaden the substrate specificity of PhaC. Here, we report the identification of a highly active wild-type PhaC belonging to the recently isolated
Chromobacterium
sp. USM2 (PhaC

Cs

). PhaC

Cs

showed the ability to utilize 3-hydroxybutyrate (3HB), 3-hydroxyvalerate (3HV), and 3-hydroxyhexanoate (3HHx) monomers in PHA biosynthesis. An
in vitro
assay of recombinant PhaC

Cs

expressed in
Escherichia coli
showed that its polymerization of 3-hydroxybutyryl-coenzyme A activity was nearly 8-fold higher (2,462 ± 80 U/g) than that of the synthase from the model strain
C. necator
(307 ± 24 U/g). Specific activity using a Strep2-tagged, purified PhaC

Cs

was 238 ± 98 U/mg, almost 5-fold higher than findings of previous studies using purified PhaC from
C. necator
. Efficient poly(3-hydroxybutyrate) [P(3HB)] accumulation in
Escherichia coli
expressing PhaC

Cs

of up to 76 ± 2 weight percent was observed within 24 h of cultivation. To date, this is the highest activity reported for a purified PHA synthase. PhaC

Cs

is a naturally occurring, highly active PHA synthase with superior polymerizing ability.
Plasmid Typing and Resistance Profiling of Escherichia fergusonii and Other Enterobacteriaceae Isolates from South Korean Farm Animals
Authors: Rayamajhi Nabin , Cha Seung Bin , Shin Seung Won , Jung Byeong Yeal , Lim Suk-Kyung , Yoo Han Sang
Published: March 2011

In this study, we focused on determining the distribution and prevalence of major plasmid replicons in β-lactam-resistant
Escherichia fergusonii
and
Enterobacteriaceae
of animal and human origin. A high degree of plasmid variability and multiple plasmid replicons were observed among the isolates. The IncF and IncI1 replicons were the most prevalent in
E. fergusonii
and
Salmonella enterica
serovar Indiana isolated from swine and poultry in South Korea, respectively. The presence of broad-host-range plasmid replicons such as IncN, IncA/C, IncHI1, and IncHI2 that are associated with important virulence genes and toxins as well as antimicrobial resistance determinants indicates that
E. fergusonii
has the potential to become an important pig pathogen and possible emerging opportunistic zoonotic pathogen.
Variability in Protist Grazing and Growth on Different Marine Synechococcus Isolates
Authors: Apple Jude K. , Strom Suzanne L. , Palenik Brian , Brahamsha Bianca
Published: March 2011

Grazing mortality of the marine phytoplanktonSynechococcusis dominated by planktonic protists, yet rates of consumption and factors regulating grazer-Synechococcusinteractions are poorly understood. One aspect of predator-prey interactions for which little is known are the mechanisms by whichSynechococcusavoids or resists predation and, in turn, how this relates to the ability ofSynechococcusto support growth of protist grazer populations. Grazing experiments conducted with the raptorial dinoflagellateOxyrrhis marinaand phylogenetically diverseSynechococcusisolates (strains WH8102, CC9605, CC9311, and CC9902) revealed marked differences in grazing rates—specifically that WH8102 was grazed at significantly lower rates than all other isolates. Additional experiments using the heterotrophic nanoflagellateGoniomonas pacificaand the filter-feeding tintinnid ciliateEutintinnissp. revealed that this pattern in grazing susceptibility among the isolates transcended feeding guilds and grazer taxon.Synechococcuscell size, elemental ratios, and motility were not able to explain differences in grazing rates, indicating that other features play a primary role in grazing resistance. Growth of heterotrophic protists was poorly coupled to prey ingestion and was influenced by the strain ofSynechococcusbeing consumed. AlthoughSynechococcuswas generally a poor-quality food source, it tended to support higher growth and survival ofG. pacificaandO. marinarelative toEutintinnissp., indicating that suitability ofSynechococcusvaries among grazer taxa and may be a more suitable food source for the smaller protist grazers. This work has developed tractable model systems for further studies of grazer-Synechococcusinteractions in marine microbial food webs.
Persistence of Mycobacterium avium subsp. paratuberculosis at a Farm-Scale Biogas Plant Supplied with Manure from Paratuberculosis-Affected Dairy Cattle
Authors: Slana I. , Pribylova R. , Kralova A. , Pavlik I.
Published: March 2011

In this study, products from all steps of anaerobic digestion at a farm-scale biogas plant supplied with manure from paratuberculosis-affected dairy cattle were examined and quantified for the presence of the causal agent of paratuberculosis,
Mycobacterium avium
subsp.
paratuberculosis
, using culture and quantitative real-time PCR (qPCR). Viable
M. avium
subsp.
paratuberculosis
cells were detected using culture in fermentors for up to 2 months; the presence of
M. avium
subsp.
paratuberculosis
DNA (10
1
cells/g) was demonstrated in all anaerobic fermentors and digestate 16 months after initiation of work at a biogas plant, using IS
900
qPCR.
F57
qPCR was able to detect
M. avium
subsp.
paratuberculosis
DNA (10
2
cells/g) at up to 12 months. According to these results, a fermentation process that extended beyond 2 months removed all viable
M. avium
subsp.
paratuberculosis
cells and therefore rendered its product
M. avium
subsp.
paratuberculosis
free. However,
M. avium
subsp.
paratuberculosis
DNA was found during all the examined periods (more than 1 year), which could be explained by either residual DNA being released from dead cells or by the presence of viable cells whose amount was under the limit of cultivability. As the latter hypothesis cannot be excluded, the safety of the final products of digestion used for fertilization or animal bedding cannot be defined, and further investigation is necessary to confirm or refute this risk.
Production of Poly(3-Hydroxybutyrate-
co
-3-Hydroxyhexanoate) from Plant Oil by Engineered Ralstonia eutropha Strains
Authors: Budde Charles F. , Riedel Sebastian L. , Willis Laura B. , Rha ChoKyun , Sinskey Anthony J.
Published: March 2011

The polyhydroxyalkanoate (PHA) copolymer poly(3-hydroxybutyrate-
co
-3-hydroxyhexanoate) [P(HB-
co
-HHx)] has been shown to have potential to serve as a commercial bioplastic. Synthesis of P(HB-
co
-HHx) from plant oil has been demonstrated with recombinant
Ralstonia eutropha
strains expressing heterologous PHA synthases capable of incorporating HB and HHx into the polymer. With these strains, however, short-chain-length fatty acids had to be included in the medium to generate PHA with high HHx content. Our group has engineered two
R. eutropha
strains that accumulate high levels of P(HB-
co
-HHx) with significant HHx content directly from palm oil, one of the world's most abundant plant oils. The strains express a newly characterized PHA synthase gene from the bacterium
Rhodococcus aetherivorans
I24. Expression of an enoyl coenzyme A (enoyl-CoA) hydratase gene (
phaJ
) from
Pseudomonas aeruginosa
was shown to increase PHA accumulation. Furthermore, varying the activity of acetoacetyl-CoA reductase (encoded by
phaB
) altered the level of HHx in the polymer. The strains with the highest PHA titers utilized plasmids for recombinant gene expression, so an
R. eutropha
plasmid stability system was developed. In this system, the essential pyrroline-5-carboxylate reductase gene
proC
was deleted from strain genomes and expressed from a plasmid, making the plasmid necessary for growth in minimal media. This study resulted in two engineered strains for production of P(HB-
co
-HHx) from palm oil. In palm oil fermentations, one strain accumulated 71% of its cell dry weight as PHA with 17 mol% HHx, while the other strain accumulated 66% of its cell dry weight as PHA with 30 mol% HHx.
Importance of Porins for Biocide Efficacy against Mycobacterium smegmatis
Authors: Frenzel Elrike , Schmidt Stefan , Niederweis Michael , Steinhauer Katrin
Published: March 2011

Mycobacteria are among the microorganisms least susceptible to biocides but cause devastating diseases, such as tuberculosis, and increasingly opportunistic infections. The exceptional resistance of mycobacteria to toxic solutes is due to an unusual outer membrane, which acts as an efficient permeability barrier, in synergy with other resistance mechanisms. Porins are channel-forming proteins in the outer membrane of mycobacteria. In this study we used the alamarBlue assay to show that the deletion of Msp porins in isogenic mutants increased the resistance of
Mycobacterium smegmatis
to isothiazolinones (methylchloroisothiazolinone [MCI]/methylisothiazolinone [MI] and octylisothiazolinone [2-
n
-octyl-4-isothiazolin-3-one; OIT]), formaldehyde-releasing biocides {hexahydrotriazine [1,3,5-tris (2-hydroxyethyl)-hexahydrotriazine; HHT] and methylenbisoxazolidine [
N
,
N
′-methylene-bis-5-(methyloxazolidine); MBO]}, and the lipophilic biocides polyhexamethylene biguanide and octenidine dihydrochloride 2- to 16-fold. Furthermore, the susceptibility of the porin triple mutant against a complex disinfectant was decreased 8-fold compared to wild-type (wt)
M. smegmatis
. Efficacy testing in the quantitative suspension test EN 14348 revealed 100-fold improved survival of the porin mutant in the presence of this biocide. These findings underline the importance of porins for the susceptibility of
M. smegmatis
to biocides.
Genetic Control of Amadori Product Degradation in Bacillus subtilis via Regulation of
frlBONMD
Expression by FrlR
Authors: Deppe Veronika Maria , Klatte Stephanie , Bongaerts Johannes , Maurer Karl-Heinz , O'Connell Timothy , Meinhardt Friedhelm
Published: March 2011

Bacillus subtilis
is capable of degrading fructosamines. The phosphorylation and the cleavage of the resulting fructosamine 6-phosphates is catalyzed by the
frlD
and
frlB
gene products, respectively. This study addresses the physiological importance of the
frlBONMD
genes (formerly
yurPONML
), revealing the necessity of their expression for growth on fructosamines and focusing on the complex regulation of the corresponding transcription unit. In addition to the known regulation by the global transcriptional regulator CodY, the
frl
genes are repressed by the convergently transcribed FrlR (formerly YurK). The latter causes repression during growth on substrates other than fructosamines. Additionally, we identified in the first intergenic region of the operon an FrlR binding site which is centrally located within a 38-bp perfect palindromic sequence. There is genetic evidence that this sequence, in combination with FrlR, contributes to the remarkable decrease in the transcription downstream of the first gene of the
frl
operon.
Involvement of the SmeAB Multidrug Efflux Pump in Resistance to Plant Antimicrobials and Contribution to Nodulation Competitiveness in Sinorhizobium meliloti
Authors: Eda Shima , Mitsui Hisayuki , Minamisawa Kiwamu
Published: March 2011

The contributions of multicomponent-type multidrug efflux pumps to antimicrobial resistance and nodulation ability in
Sinorhizobium meliloti
were comprehensively analyzed. Computational searches identified genes in the
S. meliloti
strain 1021 genome encoding 1 pump from the ATP-binding cassette family, 3 pumps from the major facilitator superfamily, and 10 pumps from the resistance-nodulation-cell division family, and subsequently, these genes were deleted either individually or simultaneously. Antimicrobial susceptibility tests demonstrated that deletion of the
smeAB
pump genes resulted in increased susceptibility to a range of antibiotics, dyes, detergents, and plant-derived compounds and, further, that specific deletion of the
smeCD
or
smeEF
genes in a Δ
smeAB
background caused a further increase in susceptibility to certain antibiotics. Competitive nodulation experiments revealed that the
smeAB
mutant was defective in competing with the wild-type strain for nodulation. The introduction of a plasmid carrying
smeAB
into the
smeAB
mutant restored antimicrobial resistance and nodulation competitiveness. These findings suggest that the SmeAB pump, which is a major multidrug efflux system of
S. meliloti
, plays an important role in nodulation competitiveness by mediating resistance toward antimicrobial compounds produced by the host plant.
Enzymatic Dysfunction of Mitochondrial Complex I of the Candida albicansgoa1Mutant Is Associated with Increased Reactive Oxidants and Cell Death
Authors: Li Dongmei , Chen Hui , Florentino Abigail , Alex Deepu , Sikorski Patricia , Fonzi William A. , Calderone Richard
Published: March 2011

We have previously shown that deletion ofGOA1(growth andoxidantadaptation) ofCandida albicansresults in a loss of mitochondrial membrane potential, ATP synthesis, increased sensitivity to oxidants and killing by human neutrophils, and avirulence in a systemic model of candidiasis. We established that translocation of Goa1p to mitochondria occurred during peroxide stress. In this report, we show that thegoa1Δ (GOA31), compared to the wild type (WT) and a gene-reconstituted (GOA32) strain, exhibits sensitivity to inhibitors of the classical respiratory chain (CRC), including especially rotenone (complex I [CI]) and salicylhydroxamic acid (SHAM), an inhibitor of the alternative oxidase pathway (AOX), while potassium cyanide (KCN; CIV) causes a partial inhibition of respiration. In the presence of SHAM, however, GOA31 has an enhanced respiration, which we attribute to the parallel respiratory (PAR) pathway and alternative NADH dehydrogenases. Interestingly, deletion ofGOA1also results in a decrease in transcription of the alternative oxidase geneAOX1in untreated cells as well as negligibleAOX1andAOX2transcription in peroxide-treated cells. To explain the rotenone sensitivity, we measured enzyme activities of complexes I to IV (CI to CIV) and observed a major loss of CI activity in GOA31 but not in control strains. Enzymatic data of CI were supported by blue native polyacrylamide gel electrophoresis (BN-PAGE) experiments which demonstrated less CI protein and reduced enzyme activity. The consequence of a defective CI in GOA31 is an increase in reactive oxidant species (ROS), loss of chronological aging, and programmed cell death ([PCD] apoptosis)in vitrocompared to control strains. The increase in PCD was indicated by an increase in caspase activity and DNA fragmentation in GOA31. Thus,GOA1is required for a functional CI and partially for the AOX pathway; loss ofGOA1compromises cell survival. Further, the loss of chronological aging is new to studies ofCandidaspecies and may offer an insight into therapies to control these pathogens. Our observation of increased ROS production associated with a defective CI and PCD is reminiscent of mitochondrial studies of patients with some types of neurodegenerative diseases where CI and/or CIII dysfunctions lead to increased ROS and apoptosis.
Two Major Inositol Transporters and Their Role in Cryptococcal Virulence
Authors: Wang Yina , Liu Tong-bao , Delmas Guillaume , Park Steven , Perlin David , Xue Chaoyang
Published: March 2011

Cryptococcus neoformans
is an AIDS-associated human fungal pathogen and the most common cause of fungal meningitis, with a mortality rate over 40% in AIDS patients. Significant advances have been achieved in understanding its disease mechanisms. Yet the underlying mechanism of a high frequency of cryptococcal meningitis remains unclear. The existence of high inositol concentrations in brain and our earlier discovery of a large inositol transporter (
ITR
) gene family in
C. neoformans
led us to investigate the potential role of inositol in
Cryptococcus
-host interactions. In this study, we focus on functional analyses of two major
ITR
genes to understand their role in virulence of
C. neoformans
. Our results show that
ITR1A
and
ITR3C
are the only two
ITR
genes among 10 candidates that can complement the growth defect of a
Saccharomyces cerevisiae
strain lacking inositol transporters. Both
S. cerevisiae
strains heterologously expressing
ITR1A
or
ITR3C
showed high inositol uptake activity, an indication that they are major inositol transporters. Significantly,
itr1a itr3c
double mutants showed significant virulence attenuation in murine infection models. Mutating both
ITR1A
and
ITR3C
in an
ino1
mutant background activates the expression of several remaining
ITR
candidates and does not show more severe virulence attenuation, suggesting that both inositol uptake and biosynthetic pathways are important for inositol acquisition. Overall, our study provides evidence that host inositol and fungal inositol transporters are important for
Cryptococcus
pathogenicity.
Catabolite Repression of the Bacillus subtilis FadR Regulon, Which Is Involved in Fatty Acid Catabolism
Authors: Tojo Shigeo , Satomura Takenori , Matsuoka Hiroshi , Hirooka Kazutake , Fujita Yasutaro
Published: March 2011

The
Bacillus subtilis fadR
regulon involved in fatty acid degradation comprises five operons,
lcfA-fadR-fadB-etfB-etfA
,
lcfB
,
fadN-fadA-fadE
,
fadH-fadG
, and
fadF-acdA-rpoE
. Since the
lcfA-fadRB-etfBA
,
lcfB
, and
fadNAE
operons, whose gene products directly participate in the β-oxidation cycle, had been found to be probably catabolite repressed upon genome-wide transcript analysis, we performed Northern blotting, which indicated that they are clearly under CcpA-dependent catabolite repression. So, we searched for catabolite-responsive elements (
cre
's) to which the complex of CcpA and P-Ser-HPr binds to exert catabolite repression by means of a web-based
cis
-element search in the
B. subtilis
genome using known
cre
sequences, which revealed the respective candidate
cre
sequences in the
lcfA
,
lcfB
, and
fadN
genes. DNA footprinting indicated that the complex actually interacted with these
cre
's
in vitro
. Deletion analysis of each
cre
using the
lacZ
fusions with the respective promoter regions of the three operons with and without it, indicated that these
cre
's are involved in the CcpA-dependent catabolite repression of the operons
in vivo
.
More than One Way To Control Hair Growth: Regulatory Mechanisms in Enterobacteria That Affect Fimbriae Assembled by the Chaperone/Usher Pathway
Authors: Clegg Steven , Wilson Janet , Johnson Jeremiah
Published: March 2011

Many Gram-negative enterobacteria produce surface-associated fimbriae that facilitate attachment and adherence to eucaryotic cells and tissues. These organelles are believed to play an important role during infection by enabling bacteria to colonize specific niches within their hosts. One class of these fimbriae is assembled using a periplasmic chaperone and membrane-associated scaffolding protein that has been referred to as an usher because of its function in fimbrial biogenesis. The presence of multiple types of fimbriae assembled by the chaperone/usher pathway can be found both within a single bacterial species and also among different genera. One way of controlling fimbrial assembly in these bacteria is at the genetic level by positively or negatively regulating fimbrial gene expression. This minireview considers the mechanisms that have been described to control fimbrial gene expression and uses specific examples to demonstrate both unique and shared properties of such regulatory mechanisms.
Complete Genome Sequence of Streptococcus suis Serotype 14 Strain JS14
Authors: Hu Pan , Yang Ming , Zhang Anding , Wu Jiayan , Chen Bo , Hua Yafeng , Yu Jun , Xiao Jingfa , Jin Meilin
Published: March 2011

Streptococcus suis
is an important zoonotic agent leading to a variety of diseases in swine and can be transmitted to human beings upon close contact. Here, we report the complete genome sequence of
S. suis
serotype 14 strain JS14 which was isolated from a diseased pig in Jiangsu Province, China.
Biosynthesis of the RNA Polymerase Inhibitor Streptolydigin in Streptomyces lydicus: Tailoring Modification of 3-Methyl-Aspartate
Authors: Horna Dina H. , Gómez Cristina , Olano Carlos , Palomino-Schätzlein Martina , Pineda-Lucena Antonio , Carbajo Rodrigo J. , Braña Alfredo F. , Méndez Carmen , Salas José A.
Published: March 2011

The asparaginyl-tRNA synthetase-like SlgZ and methyltransferase SlgM enzymes are involved in the biosynthesis of the tetramic acid streptolydigin in
Streptomyces lydicus
. Inactivation of
slgZ
led to a novel streptolydigin derivative. Overexpression of
slgZ
,
slgM
, or both in
S. lydicus
led to a considerable increase in streptolydigin production.
Complete Genome Sequence of the Hemotrophic Mycoplasma suis Strain KI3806
Authors: Oehlerking Joern , Kube Michael , Felder Kathrin M. , Matter Denise , Wittenbrink Max M. , Schwarzenbach Sarah , Kramer Manuela M. , Hoelzle Katharina , Hoelzle Ludwig E.
Published: March 2011

Mycoplasma suis
, a member of the hemotrophic mycoplasma (HM) group, parasitize erythrocytes of pigs. Increasing evidence suggests that
M. suis
is also a zoonotic agent. Highly pathogenic strains of
M. suis
(e.g.,
M. suis
KI3806) have been demonstrated to invade erythrocytes. This complete sequenced and manually annotated genome of
M. suis
KI3806 is the first available from this species and from the HM group. The DNA was isolated from blood samples of experimentally infected pigs due to the lack of an
in vitro
cultivation system. The small circular chromosome of 709,270 bp, encoding an unexpectedly high number of hypothetical proteins and limited transport and metabolic capacities, could reflect the unique lifestyle of HM on the surface of erythrocytes.
Genome Sequence of Acinetobacter baumannii MDR-TJ
Authors: Gao Feng , Wang Yue , Liu Yan-Jie , Wu Xiao-Meng , Lv Xing , Gan Yi-Ru , Song Shi-Duo , Huang He
Published: March 2011

Acinetobacter baumannii
is a pathogenic species of bacteria, identified as an aerobic Gram-negative bacterium, that is resistant to most antibiotics. In this study, the MDR-TJ strain was isolated at the Second Hospital of Tianjin Medical University, China, and was found to be resistant to penicillin, cephalosporins, aminoglycosides, quinolones, and also imipenem. The genome sequence of
Acinetobacter baumannii
strain MDR-TJ was determined by using a combination of 454 pyrosequencing and paired-end sequencing performed with the Roche Genome Sequencer FLX system to generate a scaffolded assembly.
Genome Sequence of the Verrucomicrobium Opitutus terrae PB90-1, an Abundant Inhabitant of Rice Paddy Soil Ecosystems
Authors: van Passel Mark W. J. , Kant Ravi , Palva Airi , Copeland Alex , Lucas Susan , Lapidus Alla , Glavina del Rio Tijana , Pitluck Sam , Goltsman Eugene , Clum Alicia , Sun Hui , Schmutz Jeremy , Larimer Frank W. , Land Miriam L. , Hauser Loren , Kyrpides Nikolaos , Mikhailova Natalia , Richardson P. Paul , Janssen Peter H. , de Vos Willem M. , Smidt Hauke
Published: March 2011

Bacteria of the deeply branching phylum
Verrucomicrobia
are rarely cultured yet commonly detected in metagenomic libraries from aquatic, terrestrial, and intestinal environments. We have sequenced the genome of
Opitutus terrae
PB90-1, a fermentative anaerobe within this phylum, isolated from rice paddy soil and capable of propionate production from plant-derived polysaccharides.
Complete Genome Sequence of the Cellulose-Degrading Bacterium Cellulosilyticum lentocellum
Authors: Miller David A. , Suen Garret , Bruce David , Copeland Alex , Cheng Jan-Feng , Detter Chris , Goodwin Lynne A. , Han Cliff S. , Hauser Loren J. , Land Miriam L. , Lapidus Alla , Lucas Susan , Meincke Linda , Pitluck Sam , Tapia Roxanne , Teshima Hazuki , Woyke Tanja , Fox Brian G. , Angert Esther R. , Currie Cameron R.
Published: March 2011

Cellulosilyticum lentocellum
DSM 5427 is an anaerobic, endospore-forming member of the
Firmicutes
. We describe the complete genome sequence of this cellulose-degrading bacterium, which was originally isolated from estuarine sediment of a river that received both domestic and paper mill waste. Comparative genomics of cellulolytic clostridia will provide insight into factors that influence degradation rates.
Genome Sequence of a Dominant, Multidrug-Resistant Acinetobacter baumannii Strain, TCDC-AB0715
Authors: Chen Chun-Chen , Lin Yu-Chi , Sheng Wang-Huei , Chen Yee-Chun , Chang Shan-Chwen , Hsia Ko-Chiang , Liao Mei-Hui , Li Shu-Ying
Published: March 2011

Acinetobacter baumannii
has emerged as a significant nosocomial pathogen worldwide. The increasing trend of carbapenem and fluoroquinolone resistance in
A. baumannii
severely limits the usage of therapeutic antimicrobial agents. Here we report the genome sequence of a multidrug-resistant
A. baumannii
strain, TCDC-AB0715, harboring both
bla
OXA-23
and
bla
OXA-66
.
Regulation of Cytotoxicity by Quorum-Sensing Signaling in Vibrio vulnificus Is Mediated by SmcR, a Repressor of hlyU
Authors: Shao Chung-Ping , Lo Horng-Ren , Lin Jen-Hsing , Hor Lien-I
Published: March 2011

Cytotoxicity is an important virulence determinant in the pathogenesis of
Vibrio vulnificus
, and two cytotoxins, RTX (encoded by
rtxA1
) and cytolysin/hemolysin (encoded by
vvhA
), have been identified in this organism. We showed that the quorum-sensing regulator LuxO controlled the cytotoxicity of this organism: a Δ
luxO
mutant exhibited low cytotoxicity, whereas a constitutively activated
luxO
mutant,
luxO(D47E)
, remained highly cytotoxic. The cytotoxicity of the Δ
luxO
mutant was restored when
smcR
, a
Vibrio harveyi luxR
homologue repressed by
luxO
, was further deleted. SmcR then was shown to repress the expression of both
rtxA1
and
vvhA
. A DNA library of
V. vulnificus
was screened in
Escherichia coli
for clones that upregulated
vvhA
in the presence of SmcR, and
hlyU
, which has been shown to positively regulate
rtxA1
and
vvhA
, was identified. We demonstrated that SmcR repressed the expression of
hlyU
and bound to a region upstream of
hlyU
in
V. vulnificus
. The deletion of
hlyU
resulted in the loss of cytotoxicity and reduced cytolysin/hemolysin production in the Δ
smcR
mutant. The Δ
smcR
Δ
hlyU
mutant regained cytotoxicity and cytolysin/hemolysin activity when
hns
, which has been shown to repress the transcription of
rtxA1
and interfere with
hlyU
, was further removed. Collectively, our data suggest that SmcR mediates the regulation of cytotoxicity by quorum-sensing signaling in
V. vulnificus
by repressing
hlyU
, an activator of
rtxA1
and
vvhA
.