bluuebunny/crossref_metadata_2025_split huggingface dataset:
Controlling pH in shake flasks using polymer-based controlled-release discs with pre-determined release kinetics
Authors: Scheidle Marco , Dittrich Barbara , Klinger Johannes , Ikeda Hideo , Klee Doris , Büchs Jochen 
Published: March 2011

Abstract

Background
There are significant differences in the culture conditions between small-scale screenings and large-scale fermentation processes. Production processes are usually conducted in fed-batch cultivation mode with active pH-monitoring and control. In contrast, screening experiments in shake flasks are usually conducted in batch mode without active pH-control, but with high buffer concentrations to prevent excessive pH-drifts. These differences make it difficult to compare results from screening experiments and laboratory and technical scale cultivations and, thus, complicate rational process development. In particular, the pH-value plays an important role in fermentation processes due to the narrow physiological or optimal pH-range of microorganisms. To reduce the differences between the scales and to establish a pH-control in shake flasks, a newly developed easy to use polymer-based controlled-release system is presented in this paper. This system consists of bio-compatible silicone discs embedding the alkaline reagent Na2CO3. Since the sodium carbonate is gradually released from the discs in pre-determined kinetics, it will ultimately compensate the decrease in pH caused by the biological activity of microorganisms.

Results
The controlled-release discs presented here were successfully used to cultivate E. coli K12 and E. coli BL21 pRSET eYFP-IL6 in mineral media with glucose and glycerol as carbon (C) sources, respectively. With glucose as the C-source it was possible to reduce the required buffer concentration in shake flask cultures by 50%. Moreover, with glycerol as the C-source, no buffer was needed at all.

Conclusions
These novel polymer-based controlled-release discs allowed buffer concentrations in shake flask media to be substantially reduced or omitted, while the pH remains in the physiological range of the microorganisms during the whole cultivation time. Therefore, the controlled-release discs allow a better control of the pH, than merely using high buffer concentrations. The conditions applied here, i.e. with significantly reduced buffer concentrations, enhance the comparability of the culture conditions used in screening experiments and large-scale fermentation processes.
Methicillin resistant Staphylococcus aureus adhesion to human umbilical vein endothelial cells demonstrates wall shear stress dependent behaviour
Authors: Viegas Kayla D , Dol Sharul S , Salek M Mehdi , Shepherd Robert D , Martinuzzi Robert M , Rinker Kristina D 
Published: March 2011

AbstractBackgroundMethicillin-resistantStaphylococcus aureus(MRSA) is an increasingly prevalent pathogen capable of causing severe vascular infections. The goal of this work was to investigate the role of shear stress in early adhesion events.MethodsHuman umbilical vein endothelial cells (HUVEC) were exposed to MRSA for 15-60 minutes and shear stresses of 0-1.2 Pa in a parallel plate flow chamber system. Confocal microscopy stacks were captured and analyzed to assess the number of MRSA. Flow chamber parameters were validated using micro-particle image velocimetry (PIV) and computational fluid dynamics modelling (CFD).ResultsUnder static conditions, MRSA adhered to, and were internalized by, more than 80% of HUVEC at 15 minutes, and almost 100% of the cells at 1 hour. At 30 minutes, there was no change in the percent HUVEC infected between static and low flow (0.24 Pa), but a 15% decrease was seen at 1.2 Pa. The average number of MRSA per HUVEC decreased 22% between static and 0.24 Pa, and 37% between 0.24 Pa and 1.2 Pa. However, when corrected for changes in bacterial concentration near the surface due to flow, bacteria per area was shown to increase at 0.24 Pa compared to static, with a subsequent decline at 1.2 Pa.ConclusionsThis study demonstrates that MRSA adhesion to endothelial cells is strongly influenced by flow conditions and time, and that MSRA adhere in greater numbers to regions of low shear stress. These areas are common in arterial bifurcations, locations also susceptible to generation of atherosclerosis.
Cystic cavernous malformation of the cerebellopontine angle: Case report and literature review
Authors: Huang Haiyan , Xu Kan , Qu Limei , Li Ye , Yu Jinlu 
Published: March 2011

Abstract

Background
Cavernous malformations (CMs) in the cerebellopontine angle (CPA) are rare, and most of such CMs reported to date are solid and extend from the internal auditory canal into the CPA. In contrast, cystic CMs that arise in the CPA and do not involve the internal auditory canal and dura of the skull base are extremely rare.

Case presentation
A 50-year-old man presented with vertigo and progressive hearing loss in the right ear. MRI examination revealed a lesion in the CPA with solid and cystic components. Surgery was performed. Well-circumscribed adhesion to cranial nerves, the cerebellum, or the brain stem was noted during surgery. The lesion was totally resected. Pathological examination suggested the lesion to be a CM. At 1-year follow-up, the symptoms at presentation had resolved and no complications had occurred.

Conclusion
Although cystic CMs of the CPA have no established imaging features, a diagnosis of CMs may be suspected when a cystic lesion is present in the CPA and does not involve internal acoustic meatus or dura mater of the skull base. Skillful microsurgical techniques and monitoring of cranial nerves will secure good outcomes for patients with cystic CMs in the CPA.
Severe teratozoospermia and its influence on pronuclear morphology, embryonic cleavage and compaction
Authors: Berger Dara S , AbdelHafez Faten , Russell Helena , Goldfarb James , Desai Nina 
Published: March 2011

AbstractBackgroundFertilization, cell division and embryo development depend on genomic contributions from male and female gametes. We hypothesize that teratozoospermic sperm influences early embryo development and embryo compaction.MethodsWe conducted a retrospective analysis of embryos derived from intracytoplasmic sperm injection (ICSI) cycles. Two hundred thirty-five consecutive ICSI cycles were included in the study; all treatment was provided at the Cleveland Clinic Fertility Center. Patient cycles were divided by sperm morphology based on Kruger's strict criteria: Group A, embryos where teratozoospermic sperm (0-2% normal) were used for ICSI and Group B, embryos where dysmorphic sperm (5-13% normal) were used for ICSI. All cycles analyzed were of patients doing day 3 embryo transfers. Outcome measures assessed included pronuclear (PN) pattern, syngamy, early cleavage, cell number, rate of compaction and blastulation of embryos left in culture and not transferred on day 3.ResultsA total of 1762 embryos were analyzed. PN patterns were similar in Group A and Group B embryos. No differences were noted in syngamy, cleavage, cell number or blastulation rate. Studying the development of embryos in culture after day 3 transfer revealed a difference in the timeline for compaction. By day 4, 25% of Group A embryos had compacted compared to 36% in Group B (P = 0.0007). There was no difference found between Group A and Group B embryos in regards to blastulation.ConclusionsWe did not find an association between sperm morphology and clinical outcomes. The impact of teratozoospermia may be masked in ICSI cycles where fertilization, implantation rate and clinical pregnancy rate are the primary outcome measures. However, by examining the timeline of development, we were better able to discern a potential paternal effect at critical transition points from fertilization through activation.
SERPINE2, an inhibitor of plasminogen activators, is highly expressed in the human endometrium during the secretory phase
Authors: Lee Robert Kuo-Kuang , Fan Chi-Chen , Hwu Yuh-Ming , Lu Chung-Hao , Lin Ming-Huei , Chen Ying-Jie , Li Sheng-Hsiang 
Published: March 2011

Abstract

Background
SERPINE2, also known as protease nexin-1, belongs to the serine protease inhibitor (SERPIN) superfamily. It is one of the potent SERPINs that modulates the activity of plasminogen activators (PAs). PAs and their SERPIN inhibitors, such as SERPINB2 and SERPINE1, were expressed in the human endometrium and were implicated in implantation. However, expression data about SERPINE2 in the human endometrium is still unknown. Thus, we conducted an investigation to reveal the spatiotemporal and cellular expression of SERPINE2 in the human uterus during the menstrual cycle.

Methods
Seven patients who underwent a hysterectomy and samples of 120 archived patients' endometrial curettage or parts of the uterus that were formalin-fixed and embedded in paraffin. Western blotting was performed to evaluate the specificity and sensitivity of the antibody. Immunohistochemistry was conducted to localize the SERPINE2 expression site. Quantitative analysis was conducted to evaluate expression levels of SERPINE2 in various sub-phases of the menstrual cycle.

Results
The SERPINE2 protein was primarily detected in the uterine fluid during the mid- and late-secretory phases of the menstrual cycle. It was predominantly expressed in the luminal and glandular epithelium, less in the myometrium, and only dispersedly in certain stromal cells throughout the menstrual cycle. A quantitative analysis of expression levels of SERPINE2 in the glandular epithelium revealed that it was highly expressed in the endometrium during the secretory phase compared to the proliferative phase.

Conclusions
The SERPINE2 protein is highly expressed in the endometrium during the secretory phase, indicating that it may participate in tissue remodeling involved in implantation.
High ERCC1 expression predicts cisplatin-based chemotherapy resistance and poor outcome in unresectable squamous cell carcinoma of head and neck in a betel-chewing area
Authors: Chiu Tai-Jan , Chen Chang-Han , Chien Chih-Yen , Li Shau-Hsuan , Tsai Hsin-Ting , Chen Yi-Ju 
Published: March 2011

Abstract

Background
This study was to evaluate the effect of excision repair cross-complementation group 1(ERCC1) expression on response to cisplatin-based induction chemotherapy (IC) followed by concurrent chemoradiation (CCRT) in locally advanced unresectable head and neck squamous cell carcinoma (HNSCC) patients.

Methods
Fifty-seven patients with locally advanced unresectable HNSCC who received cisplatin-based IC followed by CCRT from January 1, 2006 through January 1, 2008. Eligibility criteria included presence of biopsy-proven HNSCC without a prior history of chemotherapy or radiotherapy. Immunohistochemistry was used to assess ERCC1 expression in pretreatment biopsy specimens from paraffin blocks. Clinical parameters, including smoking, alcohol consumption and betel nuts chewing, were obtained from the medical records.

Results
The 12-month progression-free survival (PFS) and 2-year overall survival (OS) rates of fifty-seven patients were 61.1% and 61.0%, respectively. Among these patients, thirty-one patients had low ERCC1 expression and forty-one patients responded to IC followed by CCRT. Univariate analyses showed that patients with low expression of ERCC1 had a significantly higher 12-month PFS rates (73.3% vs. 42.3%, p < 0.001) and 2-year OS (74.2 vs. 44.4%, p = 0.023) rates. Multivariate analysis showed that for patients who did not chew betel nuts and had low expression of ERCC1 were independent predictors for prolonged survival.

Conclusions
Our study suggest that a high expression of ERCC1 predict a poor response and survival to cisplatin-based IC followed by CCRT in patients with locally advanced unresectable HNSCC in betel nut chewing area.
Can graphene make better HgCdTe infrared detectors?
Authors: Xu Wen , Gong Youpin , Liu Liwei , Qin Hua , Shi Yanli 
Published: March 2011

AbstractWe develop a simple and low-cost technique based on chemical vapor deposition from which large-size graphene films with 5-10 graphene layers can be produced reliably and the graphene films can be transferred easily onto HgCdTe (MCT) thin wafers at room temperature. The proposed technique does not cause any thermal and mechanical damages to the MCT wafers. It is found that the averaged light transmittance of the graphene film on MCT thin wafer is about 80% in the mid-infrared bandwidth at room temperature and 77 K. Moreover, we find that the electrical conductance of the graphene film on the MCT substrate is about 25 times larger than that of the MCT substrate at room temperature and 77 K. These experimental findings suggest that, from a physics point of view, graphene can be utilized as transparent electrodes as a replacement for metal electrodes while producing better and cheaper MCT infrared detectors.
Impairment of brain endothelial glucose transporter by methamphetamine causes blood-brain barrier dysfunction
Authors: Muneer P M Abdul , Alikunju Saleena , Szlachetka Adam M , Murrin L Charles , Haorah James 
Published: March 2011

Abstract

Background
Methamphetamine (METH), an addictive psycho-stimulant drug with euphoric effect is known to cause neurotoxicity due to oxidative stress, dopamine accumulation and glial cell activation. Here we hypothesized that METH-induced interference of glucose uptake and transport at the endothelium can disrupt the energy requirement of the blood-brain barrier (BBB) function and integrity. We undertake this study because there is no report of METH effects on glucose uptake and transport across the blood-brain barrier (BBB) to date.

Results
In this study, we demonstrate that METH-induced disruption of glucose uptake by endothelium lead to BBB dysfunction. Our data indicate that a low concentration of METH (20 μM) increased the expression of glucose transporter protein-1 (GLUT1) in primary human brain endothelial cell (hBEC, main component of BBB) without affecting the glucose uptake. A high concentration of 200 μM of METH decreased both the glucose uptake and GLUT1 protein levels in hBEC culture. Transcription process appeared to regulate the changes in METH-induced GLUT1 expression. METH-induced decrease in GLUT1 protein level was associated with reduction in BBB tight junction protein occludin and zonula occludens-1. Functional assessment of the trans-endothelial electrical resistance of the cell monolayers and permeability of dye tracers in animal model validated the pharmacokinetics and molecular findings that inhibition of glucose uptake by GLUT1 inhibitor cytochalasin B (CB) aggravated the METH-induced disruption of the BBB integrity. Application of acetyl-L-carnitine suppressed the effects of METH on glucose uptake and BBB function.

Conclusion
Our findings suggest that impairment of GLUT1 at the brain endothelium by METH may contribute to energy-associated disruption of tight junction assembly and loss of BBB integrity.
Full genome re-sequencing reveals a novel circadian clock mutation in Arabidopsis
Authors: Ashelford Kevin , Eriksson Maria E , Allen Christopher M , D'Amore Rosalinda , Johansson Mikael , Gould Peter , Kay Suzanne , Millar Andrew J , Hall Neil , Hall Anthony 
Published: March 2011

AbstractMap based cloning in Arabidopsis thaliana can be a difficult and time-consuming process, specifically if the phenotype is subtle and scoring labour intensive. Here, we have re-sequenced the 120-Mb genome of a novel Arabidopsis clock mutant early bird (ebi-1) in Wassilewskija (Ws-2). We demonstrate the utility of sequencing a backcrossed line in limiting the number of SNPs considered. We identify a SNP in the gene AtNFXL-2 as the likely cause of the ebi-1 phenotype.
Flight restriction prevents associative learning deficits but not changes in brain protein-adduct formation during honeybee ageing
Authors: Tolfsen Christina C. , Baker Nicholas , Kreibich Claus , Amdam Gro V. 
Published: March 2011

SUMMARYHoneybees (Apis mellifera) senesce within 2 weeks after they discontinue nest tasks in favour of foraging. Foraging involves metabolically demanding flight, which in houseflies (Musca domestica) and fruit flies (Drosophila melanogaster) is associated with markers of ageing such as increased mortality and accumulation of oxidative damage. The role of flight in honeybee ageing is incompletely understood. We assessed relationships between honeybee flight activity and ageing by simulating rain that confined foragers to their colonies most of the day. After 15 days on average, flight-restricted foragers were compared with bees with normal (free) flight: one group that foraged for ∼15 days and two additional control groups, for flight duration and chronological age, that foraged for ∼5 days. Free flight over 15 days on average resulted in impaired associative learning ability. In contrast, flight-restricted foragers did as well in learning as bees that foraged for 5 days on average. This negative effect of flight activity was not influenced by chronological age or gustatory responsiveness, a measure of the bees' motivation to learn. Contrasting their intact learning ability, flight-restricted bees accrued the most oxidative brain damage as indicated by malondialdehyde protein adduct levels in crude cytosolic fractions. Concentrations of mono- and poly-ubiquitinated brain proteins were equal between the groups, whereas differences in total protein amounts suggested changes in brain protein metabolism connected to forager age, but not flight. We propose that intense flight is causal to brain deficits in aged bees, and that oxidative protein damage is unlikely to be the underlying mechanism.
A simple technique to manipulate foraging costs in seed-eating birds
Authors: Koetsier Egbert , Verhulst Simon 
Published: March 2011

SUMMARY
Food availability is a key factor in ecology and evolution, but available techniques to manipulate the effort to acquire food in vertebrates are technically challenging and/or labour intensive. We present a simple technique to increase foraging costs in seed-eating birds that can be applied with little effort and at low monetary cost for prolonged periods (years) to solitary or group-housed animals. The essence of the technique is that food is offered in a container above ground level, with holes in the sides from which the food can be taken, forcing birds into energetically demanding hovering flight to forage. As a control treatment we offered a similar container but with perches mounted beneath the holes, allowing birds to eat without extra flights. Increasing foraging costs in this way induced zebra finches to double the time spent foraging, and to decrease their basal metabolic rate, in agreement with results obtained using more laborious techniques to increase foraging costs. The technique was not too severe because mortality was low during a winter with sub-zero temperatures. As foraging costs under natural conditions are generally higher than those under standard laboratory conditions, we suggest that measuring behaviour and physiology when animals have to work for food may better reflect their natural state.
Ocellar adaptations for dim light vision in a nocturnal bee
Authors: Berry Richard P. , Wcislo William T. , Warrant Eric J. 
Published: March 2011

SUMMARYGrowing evidence indicates that insect ocelli are strongly adapted to meet the specific functional requirements in the environment in which that insect lives. We investigated how the ocelli of the nocturnal bee Megalopta genalis are adapted to life in the dim understory of a tropical rainforest. Using a combination of light microscopy and three-dimensional reconstruction, we found that the retinae contain bar-shaped rhabdoms loosely arranged in a radial pattern around multi-layered lenses, and that both lenses and retinae form complex non-spherical shapes reminiscent of those described in other ocelli. Intracellular electrophysiology revealed that the photoreceptors have high absolute sensitivity, but that the threshold location varied widely between 109 and 1011 photons cm–2 s–1. Higher sensitivity and greater visual reliability may be obtained at the expense of temporal resolution: the corner frequencies of dark-adapted ocellar photoreceptors were just 4–11 Hz. Spectral sensitivity profiles consistently peaked at 500 nm. Unlike the ocelli of other flying insects, we did not detect UV-sensitive visual pigments in M. genalis, which may be attributable to a scarcity of UV photons under the rainforest canopy at night. In contrast to earlier predictions based on anatomy, the photoreceptors are not sensitive to the e-vector of polarised light. Megalopta genalis ocellar photoreceptors possess a number of unusual properties, including inherently high response variability and the ability to produce spike-like potentials. These properties bear similarities to photoreceptors in the compound eye of the cockroach, and we suggest that the two insects share physiological characteristics optimised for vision in dim light.
Regulatory function of hyperosmotic stress-induced signaling cascades in the expression of transcription factors and osmolyte transporters in freshwater Japanese eel primary gill cell culture
Authors: Chow S. C. , Wong Chris K. C. 
Published: March 2011

SUMMARY
In the present study, we investigated the early activation of osmotic stress-related protein kinases, with the aim of characterizing their functional links with downstream effectors (i.e. transcription factors and osmolyte transporters). Freshwater eel primary gill cells were cultured in hypertonic medium (500 mosmol l–1) for 6 h. Protein lysates and total RNA were collected for western blotting and quantitative real-time PCR assays. In this study, the osmotic challenge stimulated histone H3 phosphorylation, various signaling pathways (i.e. ERK1/2, p38 MAPK, JNK, CREB, MARCKS and MLCK) and expression of some downstream effectors (i.e. Na+/K+-ATPase, TauT and Ostf). Increased phosphorylation of acetylated histone is known to promote chromatin relaxation for global gene transcription, probably leading to the activation of downstream effectors for osmotic responses. In addition, the importance of the p38 MAPK and MLCK pathways in the regulation of the expression of Na+/K+-ATPase and TauT was demonstrated. Inhibition of the p38 MAPK pathway by SB202190 reduced histone H3 phosphorylation and TauT mRNA expression. Moreover, inhibition of the MLCK pathway by ML-7 decreased the expression level of Na+/K+-ATPase but increased the transcript level of TauT. Collectively, the present study reveals possible functional links of osmosensing signaling cascades to the regulation of downstream effectors.