IL-11 inhibits Clostridium difficile toxin A enterotoxicity in rat ileum.
Interleukin-11 (IL-11) is a stromal cell-derived cytokine with several biological activities against hematopoietic cells. Recent results indicated that IL-11 reduced mucosal damage in animal models of colitis. This study aimed to explore the action of recombinant human IL-11 (rhIL-11) on the intestinal effects of Clostridium difficile toxin A, an inflammatory enterotoxin, and cholera toxin, a noninflammatory enterotoxin in rat ileum. We administered rhIL-11 subcutaneously to rats before injection of toxin A into ileal loops and measured fluid secretion, epithelial permeability to mannitol, histopathological damage, and release of rat mast cell protease II (RMCP II) from intestinal mast cells and of tumor necrosis factor-alpha (TNF-alpha) and macrophage inflammatory protein-2 (MIP-2) from lamina propria macrophages. rhIL-11 (50-1,000 micrograms/kg) inhibited toxin A but not cholera toxin-mediated secretion and permeability in a dose-dependent fashion and reduced toxin A-induced epithelial cell damage. Rats treated with rhIL-11 also showed reduced RMCP II, TNF-alpha, and MIP-2 release in response to toxin A. Exposure of rat peripheral monocytes in vitro to rhIL-11 (1 microgram/ml) inhibited lipopolysaccharide and toxin A-mediated increases in TNF-alpha mRNA and protein levels. We conclude that rhIL-11 blocks the intestinal effects of C. difficile toxin A, possibly by inhibiting release of inflammatory mediators from mucosal mast cells and intestinal macrophages.


Feasibility study on direct fermentation of soybean meal by Bacillus stearothermophilus under non-sterile conditions.
To evaluate the feasibility of high-temperature solid-state fermentation (SSF) using soybean meal (SBM) during the non-sterile process, Bacillus stearothermophilus was employed to assess the nutritional quality and bioactivity of SBM after fermentation. The fermented SBM (FSBM) without autoclaving showed significant improvements in nutritional quality and bioactivity. The contents of peptides and crude and soluble proteins increased by 131.21%, 5.3% and 15.52%, respectively. Meanwhile, DPPH (2,2-diphenyl-1-picrylhydrazyl) scavenging ability, reducing ability and hydroxyl free radical-scavenging activity rose by 57.07%, 238.92% and 368.26%, respectively. The inhibitory activity of angiotensin I-converting enzyme increased from 1.43 ± 0.83% to 26.89 ± 1.03%, while the trypsin inhibitor activity decreased by 74.05%. The contents of neutral and alkaline proteases and the growth of microorganisms in FSBM without autoclaving were higher and better than in steam-treated FSBM. After steam treatment, the water-holding capacity of SBM decreased, and a high crosslink density was observed on the surface of SBM particles. It is feasible to ferment SBM by high-temperature SSF using B. stearothermophilus under non-sterile conditions. Adverse effects of SSF using sterile SBM might be owing to the low water-holding capacity caused by autoclaving. © 2018 Society of Chemical Industry.


Reactive species variation in cold atmospheric pressure plasma jet discharge under the influence of intrinsic parameters and its effect on E. coli inactivation.
Cold atmospheric pressure plasma jet (CAPJ) has piqued the interest of researchers for various antimicrobial applications such as disinfection, wound decontamination, etc. In the current context, a deeper understanding of the correlation between CAPJ's intrinsic parameters, discharge characteristics, species composition, and antimicrobial activity is required for any successful application. This research evaluated the effect of intrinsic operational parameters such as voltage, frequency, gas flow rate, and operating gas on the reactive species composition of an in-house-developed CAPJ discharge along with the antimicrobial activity. It was observed that the identified excited atoms (Ar I, He I, N2, and O I), ions (Ar+, N2+, N+, H2O+, H3O+, etc.), radical reactive oxygen and nitrogen species (RONS) (OH•), and nonradical RONS (O I, O+, OH+, NO+, O2+, O2-, NO2-, N2O2-, NO3-, N2O3-, etc.) might play a synergistic role in bacterial inactivation via oxidative and electrostatic stress. The variation in voltage, frequency, gas flow rate, and operating gas influenced the discharge chemistry, leading to variation in bacterial inactivation. The reactive species in the discharge responsible for such variation was evaluated extensively. This investigation into various operational parameters would aid in determining the most effective settings for a developed CAPJ to achieve high productivity.


Synthesis, photodynamic activities, and cytotoxicity of new water-soluble cationic gallium(III) and zinc(II) phthalocyanines.
The cationic Ga(III) and Zn(II) phthalocyanines carrying N-methyl-pyridinium groups at eight peripheral β-positionshave been synthesized. These complexes are highly soluble in dimethyl sulfoxide (DMSO) and moderately soluble in water and phosphate buffered saline (PBS); both Ga(III)Cl and Zn(II) complexes have shown no aggregation in water up to 1.2 × 10-4 and 1.5 × 10-5 M, respectively. A higher water-solubility of Ga(III)Cl complex as compared to Zn(II) complex is ascribed to the presence of an axially coordinated chloride. The spectroscopic properties, photogeneration of singlet oxygen (1O2), and cytotoxicity of these complexes have been investigated. The absolute quantum yields (ΦΔabsolute) for the photogeneration of singlet oxygen using Ga(III)Cl and Zn(II) complexes have been determined to be 4.4 and 5.3%, respectively, in DMSO solution. The cytotoxicity and intracellular sites of localization of Ga(III)Cl and Zn(II) complexes have been evaluated in human HEp2 cells. Both complexes, localized intracellularly in multiple organelles, have shown no cytotoxicity in the dark. Upon exposure to a low light dose (1.5 J/cm2), however, Zn(II) complex has exhibited a high photocytotoxicity. The result suggests that Zn(II) complex can be considered as a potential photosensitizer for Photodynamic therapy (PDT).


High-throughput anaerobic screening for identifying compounds acting against gut bacteria in monocultures or communities.
The human gut microbiome is a key contributor to health, and its perturbations are linked to many diseases. Small-molecule xenobiotics such as drugs, chemical pollutants and food additives can alter the microbiota composition and are now recognized as one of the main factors underlying microbiome diversity. Mapping the effects of such compounds on the gut microbiome is challenging because of the complexity of the community, anaerobic growth requirements of individual species and the large number of interactions that need to be quantitatively assessed. High-throughput screening setups offer a promising solution for probing the direct inhibitory effects of hundreds of xenobiotics on tens of anaerobic gut bacteria. When automated, such assays enable the cost-effective investigation of a wide range of compound-microbe combinations. We have developed an experimental setup and protocol that enables testing of up to 5,000 compounds on a target gut species under strict anaerobic conditions within 5 d. In addition, with minor modifications to the protocol, drug effects can be tested on microbial communities either assembled from isolates or obtained from stool samples. Experience in working in an anaerobic chamber, especially in performing delicate work with thick chamber gloves, is required for implementing this protocol. We anticipate that this protocol will accelerate the study of interactions between small molecules and the gut microbiome and provide a deeper understanding of this microbial ecosystem, which is intimately intertwined with human health.


E-Health interventions for anxiety and depression in children and adolescents with long-term physical conditions.
Long-term physical conditions affect 10% to 12% of children and adolescents worldwide; these individuals are at greater risk of developing psychological problems, particularly anxiety and depression. Access to face-to-face treatment for such problems is often limited, and available interventions usually have not been tested with this population. As technology improves, e-health interventions (delivered via digital means, such as computers and smart phones and ranging from simple text-based programmes through to multimedia and interactive programmes, serious games, virtual reality and biofeedback programmes) offer a potential solution to address the psychological needs of this group of young people. To assess the effectiveness of e-health interventions in comparison with attention placebos, psychological placebos, treatment as usual, waiting-list controls, or non-psychological treatments for treating anxiety and depression in children and adolescents with long-term physical conditions. We searched the Cochrane Common Mental Disorders Group's Controlled Trials Register (CCMDTR to May 2016), the Cochrane Central Register of Controlled Trials (CENTRAL) (Issue 8, 2017), Web of Science (1900 - 18 August 2016, updated 31 August 2017) and Ovid MEDLINE, Embase, PsycINFO (cross-search 2016 to 18 Aug 2017). We hand-searched relevant conference proceedings, reference lists of included articles, and the grey literature to May 2016. We also searched international trial registries to identify unpublished or ongoing trials. We included randomised controlled trials (RCTs), cluster-randomised trials, and cross-over trials of e-health interventions for treating any type of long-term physical condition in children and adolescents (aged 0 to 18 years), and that measured changes in symptoms or diagnoses of anxiety, depression, or subthreshold depression. We defined long-term physical conditions as those that were more than three-months' duration. We assessed symptoms of anxiety and depression using patient- or clinician-administered validated rating scales based on DSM III, IV or 5 (American Psychological Association 2013), or ICD 9 or 10 criteria (World Health Organization 1992). Formal depressive and anxiety disorders were diagnosed using structured clinical interviews. Attention placebo, treatment as usual, waiting list, psychological placebo, and other non-psychological therapies were eligible comparators. Two review authors independently reviewed titles, abstracts, and full-text articles; discrepancies were resolved through discussion or addressed by a third author. When available, we used odds ratio (OR) to compare dichotomous data and standardised mean differences (SMD) to analyse continuous data, both with 95% confidence intervals (CI). We undertook meta-analysis when treatments, participants, and the underlying clinical question were adequately similar. Otherwise, we undertook a narrative analysis. We included five trials of three interventions (Breathe Easier Online, Web-MAP, and multimodal cognitive behavioural therapy (CBT)), which included 463 participants aged 10 to 18 years. Each trial contributed to at least one meta-analysis. Trials involved children and adolescents with long-term physical conditions, such as chronic headache (migraine, tension headache, and others), chronic pain conditions (abdominal, musculoskeletal, and others), chronic respiratory illness (asthma, cystic fibrosis, and others), and symptoms of anxiety or depression. Participants were recruited from community settings and hospital clinics in high income countries.For the primary outcome of change in depression symptoms versus any control, there was very low-quality evidence meaning that it could not be determined whether e-health interventions were clearly better than any comparator (SMD -0.06, 95% CI -0.35 to 0.23; five RCTs, 441 participants). For the primary outcome of change in anxiety symptoms versus any comparator, there was very low-quality evidence meaning that it could not be determined whether e-health interventions were clearly better than any comparator (SMD -0.07, 95% CI -0.29 to 0.14; two RCTs, 324 participants). For the primary outcome of treatment acceptability, there was very low-quality evidence that e-health interventions were less acceptable than any comparator (SMD 0.46, 95% CI 0.23 to 0.69; two RCTs, 304 participants).For the secondary outcome of quality of life, there was very low-quality evidence meaning that it could not be determined whether e-health interventions were clearly better than any comparator (SMD -0.83, 95% CI -1.53 to -0.12; one RCT, 34 participants). For the secondary outcome of functioning, there was very low-quality evidence meaning that it could not be determined whether e-health interventions were clearly better than any comparator (SMD -0.08, 95% CI -0.33 to 0.18; three RCTs, 368 participants). For the secondary outcome of status of long-term physical condition, there was very low-quality evidence meaning that it could not be determined whether e-health interventions were clearly better than any comparator (SMD 0.06, 95% CI -0.12 to 0.24; five RCTs, 463 participants).The risk of selection bias was considered low in most trials. However, the risk of bias due to inadequate blinding of participants or outcome assessors was considered unclear or high in all trials. Only one study had a published protocol; two trials had incomplete outcome data. All trials were conducted by the intervention developers, introducing another possible bias. No adverse effects were reported by any authors. At present, the field of e-health interventions for the treatment of anxiety or depression in children and adolescents with long-term physical conditions is limited to five low quality trials. The very low-quality of the evidence means the effects of e-health interventions are uncertain at this time, especially in children aged under 10 years.Although it is too early to recommend e-health interventions for this clinical population, given their growing number, and the global improvement in access to technology, there appears to be room for the development and evaluation of acceptable and effective technologically-based treatments to suit children and adolescents with long-term physical conditions.


The human fetal retinal nerve fiber layer and optic nerve head: a DiI and DiA tracing study.
The organization of the primate nerve fiber layer and optic nerve head with respect to the positioning of central and peripheral axons remains controversial. Data were obtained from 32 human fetal retinae aged between 15 and 21 weeks of gestation. Crystals of the carbocyanine dyes, DiI or DiA, and fluorescence microscopy were used to identify axonal populations from peripheral retinal ganglion cells. Peripheral ganglion cell axons were scattered throughout the vitreal-scleral depth of the nerve fiber layer. Such a scattered distribution was maintained as the fibers passed through the optic nerve head and along the optic nerve. There was a rough topographic representation within the optic nerve head according to retinal quadrant such that both peripheral and central fibers were mixed within a wedge extending from the periphery to the center of the nerve. There was no indication that the fibers were reorganized in any way as they passed through the optic disc and into the nerve. The present results suggest that any degree of order present within the fiber layer and optic nerve is not an active process but a passive consequence of combining the fascicles of the retinal nerve fiber layer. Optic axons are not instructed to establish a retinotopic order and the effect of guidance cues in reordering fibers, particularly evident prechiasmatically and postchiasmatically, does not appear to be present within the nerve fiber layer or optic nerve head in humans.


Effect of dexamethasone on immunoreactive corticotropin-releasing factor in the rat median eminence and intermediate-posterior pituitary.
Immunoreactive ACTH (I-ACTH) concentrations in the anterior pituitary, intermediate-posterior pituitary (IP), and plasma and immunoreactive corticotropin-releasing factor (I-CRF) concentrations in the median eminence and IP were determined in rats receiving dexamethasone for various periods from 16 h to 10 days. Plasma I-ACTH concentrations were decreased 16 h after a single injection of dexamethasone. Anterior pituitary I-ACTH concentrations did not decrease until 4 days after the start of dexamethasone medication. IP I-ACTH concentrations did not change throughout these periods. I-CRF concentrations in median eminence and IP rapidly decreased after dexamethasone administration. These results raise the possibility that the source of I-CRF in the IP is hypothalamic.


Development of the Leapfrog methodology for evaluating hospital implemented inpatient computerized physician order entry systems.
The 1999 Institute of Medicine report raised public awareness of the frequency and cost of adverse drug events in medicine. In response, in November 2000 a coalition of healthcare purchasers announced the formation of the Leapfrog Group, an organization dedicated to making "great leaps forward" in the safety and quality of health care in America. Their first target-computerized physician order entry (CPOE)-was selected specifically for its potential to reduce harm to patients from medications. The Leapfrog inpatient CPOE standard included a requirement that the organization operating CPOE should demonstrate via a test that their inpatient CPOE system can alert physicians to at least 50% of common serious prescribing errors. This paper outlines the development of this test which evaluates the ability of implemented CPOE systems to prevent the occurrence of medication errors that have a high likelihood of leading to adverse drug events. A framework was developed to include 12 different categories of CPOE based decision support that could prevent prescribing errors leading to adverse drug events. A scoring system was developed based on the known frequency and severity of adverse drug events. Simulated test patients and accompanying simulated test medication orders were developed to evaluate the ability of a CPOE system to intercept prescribing errors in all 12 decision support categories. The test was validated at a number of inpatient sites using both commercially available and custom developed CPOE systems. A web based application was developed to allow hospitals to self-administer the evaluation.


STD management by private pharmacies in Hanoi: practice and knowledge of drug sellers.
Prompt treatment of sexually transmitted infections may reduce the incidence of HIV/AIDS infections. With health sector reforms private pharmacies are increasingly the first and only contact with health delivery services. To find out how patients with STDs are treated at private pharmacies in Hanoi, and what drug sellers know about STD management. Five simulated clients were taught to adopt a scenario stating that their friend had a urethral discharge. They visited 60 randomly selected private pharmacies in urban Hanoi and noted all questions asked, advice offered, and treatment given. Afterwards interviewers administered a semistructured questionnaire to all people working in the 60 pharmacies. Drug treatment was given in 84% of the 297 encounters averaging 1.5 drugs and 1.2 antibiotics per encounter. Quinolones were given 188 times. No dispensing was adequate for chlamydia or was in accordance with the national guidelines. No questions were asked in 55% of encounters and no advice was given in 61%. Questions on sexual activity were asked in 23% (69) of cases and about the health of the partner twice (1%). Advice to practise safe sex was given in 1% of encounters and for the partner to seek treatment only once. Of 69 questionnaires administered 51% said they would refer to a doctor, 16% said they would ask about the sexual activity 1% said they would ask about the health of the partner, 7% said they would advise using a condom, and 1% advised telling the partner to seek treatment. Even after prompting, 61% would ask no questions and 80% would give no advice. Even though 74% of pharmacists and drug sellers know that they should not treat STD patients, 84% actually did. None gave syndromically correct treatment. In both the questionnaire and during the simulated client methods, numbers advising on partner notification and condom use were very poor. Educational or peer awareness interventions are urgently needed among private pharmacists in Vietnam.


Effective inhibition of colon cancer cell growth with MgAl-layered double hydroxide (LDH) loaded 5-FU and PI3K/mTOR dual inhibitor BEZ-235 through apoptotic pathways.
Colon cancer is the third most common cancer and the third largest cause of cancer-related death. Fluorouracil (5-FU) is the front-line chemotherapeutic agent for colon cancer. However, its response rate is less than 60%, even in combination with other chemotherapeutic agents. The side effects of 5-FU also limit its application. Nanoparticles have been used to deliver 5-FU, to increase its effectiveness and reduce side effects. Another common approach for colon cancer treatment is targeted therapy against the phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt) pathway. A recently-invented inhibitor of this pathway, BEZ-235, has been tested in several clinical trials and has shown effectiveness and low side effects. Thus, it is a very promising drug for colon cancer treatment. The combination of these two drugs, especially nanoparticle-packed 5-FU and BEZ-235, has not been studied. In the present study, we demonstrated that nanoparticles of layered double hydroxide (LDH) loaded with 5-FU were more effective than a free drug at inhibiting colon cancer cell growth, and that a combination treatment with BEZ-235 further increased the sensitivity of colon cancer cells to the treatment of LDH-packed 5-FU (LDH-5-FU). BEZ-235 alone can decrease colon cancer HCT-116 cell viability to 46% of the control, and the addition of LDH-5-FU produced a greater effect, reducing cell survival to 8% of the control. Our data indicate that the combination therapy of nanodelivered 5-FU with a PI3K/Akt inhibitor, BEZ-235, may promise a more effective approach for colon cancer treatment.


Nephrin in experimental glomerular disease.
The recently identified gene NPHS1 with its mutations causing congenital nephrotic syndrome of the Finnish type (CNF) is highly promising in providing new understanding of pathophysiology of proteinuria. Earlier we cloned a rat NPHS1 homologue, as well as characterized and raised antibodies to the respective protein product nephrin. Changes in the expression levels of nephrin-specific mRNA in commonly used experimental models of proteinuria were examined using semiquantitative reverse transcription-polymerase chain reaction, immunofluorescence, and immunoelectron microscopy (IEM) of nephrin. Notably, a 40% down-regulation of the nephrin-specific mRNA of cortical kidney was seen already at day 3 after induction of the puromycin aminonucleoside nephrosis (PAN), while no major elevation of urinary protein secretion was seen at this stage. A further decrease of 80% of nephrin message was seen at the peak of proteinuria at day 10. A similar decrease of up to 70% from the basal levels was seen in mercuric chloride-treated rats. Changes in the protein expression paralleled those of the mRNA in indirect immunofluorescence. Interestingly, a remarkable plasmalemmal dislocation from the normal expression site at the interpodocyte filtration slits could be observed in IEM. Nephrin appears to be an important causative molecule of proteinuria and shows a remarkable redistribution from the filtration slits to the podocyte plasma membrane, especially in PAN.


Bone marrow-derived cells are required for the induction of a pulmonary inflammatory response mediated by CD40 ligation.
The expression of inflammatory mediators by various cells following in vitro CD40 ligation is well known. However, knowledge of the role and interaction with these cells in the establishment and maintenance of in vivo immune-mediated inflammation is limited. In this report, a chimeric mouse model based on CD40 knockout and wild-type mice was used to assess the role of bone marrow (BM)-derived and non-BM-derived cells in a CD40-mediated pulmonary inflammation response. CD40+ BM-derived cells were required for initial cell recruitment, pulmonary edema, and weight loss associated with this response. The structural CD40+ non-BM-derived cells of the lung, such as fibroblasts, epithelial cells, and endothelial cells, could not by themselves establish any level of pulmonary inflammation. However, both the CD40+ BM-derived cells and the structural CD40+ non-BM-derived cells of the lung were required to maximize the level of pulmonary inflammation. Both B cells and T cells played a contributing role in macrophage recruitment and pulmonary edema but neither contributed to the inflammation-associated weight loss. These experiments indicate that CD40+ BM-derived cells were critical to the induction of pulmonary inflammation and that alveolar macrophages, B cells, and T cells contributed to selective aspects of the response.


Diagnosis and management costs of suspicious skin lesions from a population-based melanoma screening programme.
Recently, massive increases in health-care costs for the diagnosis and management of skin lesions have been observed (2000-05). The aim of this study was to describe the health system costs attributed to the diagnosis and management of suspicious skin lesions detected during a trial of a population melanoma screening programme (1998-2001). Queensland, Australia. Data from the trial and Medicare Australia were used to categorize and cost all suspicious skin lesions arising from the trial, which included general practitioner consultations, diagnosis/management and pathology. Comparisons were made with other screened and unscreened populations. Overall, 2982 lesions were treated within the trial producing a mean cost of Aus$118 per lesion. Excisions for benign lesions contributed the greatest proportion of total costs (45%). The total cost burden was approximately 10% higher for men than women, and 63% of overall costs were for persons aged >or=50 years. For diagnosis and management procedures, the estimated average cost per 1000 individuals was Aus$23,560 for men aged >or=50 years from the skin cancer screening trial, compared with Aus$26,967 for BreastScreen Australia and Aus$3042 for the National Cervical Screening Program. The proportion of costs for benign skin lesions and biopsies arising from the screening programme were no higher than in the two-year period outside the trial. While comparisons are difficult, it appears that diagnostic and management costs for skin cancer as a result of screening may be comparable with those for BreastScreen Australia, if screening is targeted at men aged >or=50 years.


Morphology and distribution of pit organs and canal neuromasts in non-teleost bony fishes.
The morphology of the pit organs and canal neuromasts of the actinopterygian Polypterus and the lungfishes Lepidosiren, Neoceratodus and Protopterus are described using light microscopy, scanning electron microscopy and transmission electron microscopy. Pit organs resemble canal neuromasts in their hair cell polarization, but they differ in size, shape, hair cell density and in the length of the kinocilia. Pit line neuromasts in Lepidosiren, Polypterus and Protopterus are arranged in dense lines, with numerous neuromasts between pores in the canal, and are an order of magnitude smaller than canal neuromasts in Polypterus. We suggest that the evolutionary transformation of canal neuromasts into superficial neuromasts occurred through the evolutionary reduction of the lateral line canals, either as the result of selection for changes in neuromast function, or as the result of non-adaptive reduction of dermal bone around the lateral line canals and their neuromasts. Protopterus (a lepidosirenid lungfish) is apparently unique among bony fishes in that it has multiple neuromasts between adjacent pores in lateral line canals on the head. An analysis of the distribution of this character among major gnathostome clades indicates that multiple canal neuromasts between pore positions is a primitive gnathostome character. Latimeria, Neoceratodus and actinopterygian fishes are characterized by an alternative character state, the presence of one canal neuromast between adjacent pores.


Psychosocial assessment of the family in the clinical setting.
Children develop in the context of the family. Family functioning prominently shapes the psychosocial adaptation and mental health of the child. Several family psychosocial risk factors have been shown to increase the risk of behavioral problems in children. Early identification of families with psychosocial profiles associated with a higher risk of having children with behavioral problems may be valuable for targeting these children for prevention and early intervention services. We developed the Family Health Questionnaire (FHQ) for the purpose of evaluating families' psychosocial risk profiles in the primary care setting. The questionnaire included 10 formative indicators that have been shown to influence children's behavioral health. We aimed to establish a correlation between the family risk factors on the FHQ and child behavioral health. In addition, we examined the properties of the questionnaire as a screening tool for use in primary care. Families of 313 of children 4-6 years of age presenting for well child examinations at two primary care clinics completed both the FHQ and the Pediatric Symptom Checklist 17 (PSC-17), a validated screening instrument for pediatric behavioral problems. We found that the FHQ was positively and significantly correlated with the PSC score (r = .50, p < .05). The FHQ may be a valuable screening tool for identifying families with psychosocial risk profiles associated with increased risk of childhood behavioral problems.


Compensation-free, all-fiber-optic, two-photon endomicroscopy at 1.55 μm.
We present an all-fiber-optic scanning multiphoton endomicroscope with 1.55 μm excitation without the need for prechirping femtosecond pulses before the endomicroscope. The system consists of a 1.55 μm femtosecond fiber laser, a customized double-clad fiber for light delivery and fluorescence collection, and a piezoelectric scan head. We demonstrate two-photon imaging of cultured cells and mouse tissue, both labeled with indocyanine green. Free-space multiphoton imaging with near-IR emission has previously shown benefits in reduced background fluorescence and lower attenuation for the fluorescence emission. For fiber-optic multiphoton imaging there is the additional advantage of using the soliton effect at the telecommunication wavelengths (1.3-1.6 μm) in fibers, permitting dispersion-compensation-free, small-footprint systems. We expect these advantages will help transition multiphoton endomicroscopy to the clinic.


A propensity score matching analysis of gasless endoscopic transaxillary thyroidectomy with five-settlement technique versus conventional open thyroidectomy in patients with papillary thyroid microcarcinoma.
In a previous study, we proposed a novel anatomy-based five-settlement method for transaxillary endoscopic thyroidectomy (fs-TAT) for patients with papillary thyroid carcinoma. The safety of this new method has been reported in a retrospective study of a single cohort. The safety and short-term oncological outcome of this method was confirmed by comparing it with conventional open surgery (COT) in patients with papillary thyroid microcarcinoma. The medical records of patients who underwent fs-TAT or COT by a single surgeon from February 2019 to December 2021 were reviewed retrospectively. All patients were diagnosed with papillary thyroid microcarcinoma and underwent lobectomy and ipsilateral central compartment neck dissection. Propensity score matching was used to compare the technical safety and short-term oncologic outcomes of fs-TAT and COT for the purpose of reducing potential selection bias. Reporting was consistent with the STROCSS 2021 guidelines. After propensity score matching, 460 (fs-TAT: 230; COT: 230) patients remained in the study population. There were no significant differences in sex, age, tumor size, Hashimoto's thyroiditis, or tumor multifocality between the groups. The operative time was longer [104.5 (90.3, 120.0) vs. 62.0 (52.0, 76.0), P < 0.001] and the total postoperative drainage volume [135(90, 210) vs. 75 (55, 115), P < 0.001] was greater in the fs-TAT group than in the COT group. However, intraoperative bleeding [3.0 (2.0, 5.0) vs. 5.0 (5.0, 7.5), P < 0.001] was greater, and the median number of lymph nodes yielded [5.0 (2.3, 8.0) vs. 7.0 (5.0, 11.0), P < 0.001] was greater in the COT group than in the fs-TAT group. The groups exhibited no significant difference in the rate of complications (fs-TAT: 2.2% vs. COT: 2.6%, P = 0.856), rate of positive lymph nodes (fs-TAT: 32.2% vs. COT: 36.5%, P = 0.377), length of postoperative hospital stay (3 days vs. 3 days, P = 0.305) or total medical costs (26,936 vs. 26,549, P = 0.144). Compared to conventional open surgery, fs-TAT offered excellent safety and acceptable short-term oncological outcomes in a selected cohort of patients with papillary thyroid microcarcinoma.


A Rare Coexistence of Multiple Myeloma and Polycythemia Vera.
Multiple myeloma (MM) is classically associated with organ dysfunction leading to hypercalcemia, renal insufficiency, anemia and bone disease, known as the CRAB criteria. More than 70% of patients with MM present with anemia. Few rare case reports, however, have demonstrated the presentation of MM associated with polycythemia. We present an interesting case of a 65-year-old female who was initially diagnosed with monoclonal gammopathy of undetermined significance (MGUS) which progressed to smoldering myeloma and later developed into MM. The patient also had coexisting polycythemia vera (PCV). We discuss the typical patient presentations as well as the expanded diagnostic criteria for MM. The pathophysiology explaining the coexistence of polycythemia and MM will be explored as well.


Prevention of group B streptococcal disease in the newborn.
Group B streptococcus (GBS) is a leading cause of morbidity and mortality among newborns. Universal screening for GBS among women at 35 to 37 weeks of gestation is more effective than administration of intrapartum antibiotics based on risk factors. Lower vaginal and rectal cultures for GBS are collected at 35 to 37 weeks of gestation, and routine dindamycin and erythromycin susceptibility testing is performed in women allergic to penicillin. Women with GBS bacteriuria in the current pregnancy and those who previously delivered a GBS-septic newborn are not screened but automatically receive intrapartum antibiotics. Intrapartum chemoprophylaxis is selected based on maternal allergy history and susceptibility of GBS isolates. Intravenous penicillin G is the preferred antibiotic, with ampicillin as an alternative. Penicillin G should be administered at least four hours before delivery for maximum effectiveness. Cefazolin is recommended in women allergic to penicillin who are at low risk of anaphylaxis. Clindamycin and erythromycin are options for women at high risk for anaphylaxis, and vancomycin should be used in women allergic to penicillin and whose cultures indicate resistance to clindamycin and erytbromycin or when susceptibility is unknown. Asymptomatic neonates born to GBS-colonized mothers should be observed for at least 24 hours for signs of sepsis. Newborns who appear septic should have diagnostic work-up including blood culture followed by initiation of ampicillin and gentamicin. Studies indicate that intrapartum prophylaxis of GBS carriers and selective administration of antibiotics to newborns reduce neonatal GBS sepsis by as much as 80 to 95 percent.


[Methods of making the plaster copies of skulls for the medical and criminalistic identification of individuals].
A possibility was studied to use the plaster copies of skulls in the forensic medical identification of personality by applying the craniofacial method. No dependence was found between the plaster casts and the moulding types. Differences between the same craniometric sizes in the skull and in its cast are of an incidental nature. Such incidental deviation is related with an accuracy of determining the localization of craniometric points and with an instrumental precision of measuring devices. The necessity is substantiated that, while making a craniofacial examination, it is necessary to mark all anthropometric points in the skull cast, with the original being a control. The use of a plaster skull cast is possible in those cases, when the burial of the remains must be done in full.


A conserved arginine residue in the terminal protein domain of hepatitis B virus polymerase is critical for RNA pre-genome encapsidation.
Hepadnaviruses, including human hepatitis B virus (HBV) and duck hepatitis B virus (DHBV), replicate their DNA genome through reverse transcription. Although hepadnaviral polymerase (Pol) is distantly related to retroviral reverse transcriptases, some of its features are distinct. In particular, in addition to the reverse transcriptase and RNase H domains, which are commonly encoded by retroviral reverse transcriptases, the N-terminally extended terminal protein (TP) domain confers unique features such as protein-priming capability. Importantly, the TP domain is also essential for encapsidation of the viral RNA pre-genome. To gain further insight into the TP domain, this study used clustered charged residue-to-alanine mutagenesis of HBV Pol. Of the 20 charged residues examined, only one arginine (R105) was critical for RNA encapsidation. This result contrasts with previous findings for DHBV Pol regarding the critical residue of the TP domain required for RNA binding. Firstly, R128 of DHBV Pol, which corresponds to R105 of HBV Pol, was reportedly tolerable to alanine substitution for RNA binding. Secondly, the C-terminal arginine residue of the DHBV Pol TP domain (R183) was shown to be critical for RNA binding, whereas alanine substitution of the corresponding arginine residue of the HBV Pol TP domain (R160) remained able to support RNA encapsidation. Together, these data highlight the divergence between avian and mammalian hepadnaviral Pols with respect to an arginine residue critical for RNA encapsidation.


Investigating latitudinal clines for life history and stress resistance traits in Drosophila simulans from eastern Australia.
Latitudinal clines have been demonstrated for many quantitative traits in Drosophila and are assumed to be due to climatic selection. However, clinal studies are often performed in species of Drosophila that contain common cosmopolitan inversion polymorphisms that also show clinal patterns. These inversion polymorphisms may be responsible for much of the observed clinal variation. Here, we consider latitudinal clines for quantitative traits in Drosophila simulans from eastern Australia. Drosophila simulans does not contain cosmopolitan inversion polymorphisms, so allows the study of clinal selection on quantitative traits that are not confounded by associations with inversions. Body size showed a strong linear cline for both females and males. Starvation resistance exhibited a weak linear cline in females, whereas chill-coma recovery exhibited a significant nonlinear cline in females only. No clinal pattern was evident for development time, male chill-coma recovery, desiccation or heat resistance. We discuss these results with reference to the role inversion polymorphisms play in generating clines in quantitative traits of Drosophila.


Stress and coping in wildland firefighting dispatchers.
To gain a better understanding of the stressors faced by wildland firefighting dispatchers and how they cope with it. Qualitative method of phenomenology. Dispatch centers around the western United States. Subjects were recruited via e-mail solicitation. Only currently employed wildland firefighting dispatchers with extensive dispatching experience were selected. Dispatchers included in this study were employed at the local (3), geographic (4), or national level (4). Eleven dispatchers in total were interviewed, six females and five males. Average experience level as a dispatcher was 14.2 years. In-person interviews. Three broad categories of stressors were revealed: balancing personal and professional lives, contending with job-related issues, and dealing with issues related to control. Four coping strategies also emerged: taking time off, exercising, providing a service to firefighters, and receiving support from others. In general, a complex and at times even a paradoxical relationship between the dispatchers interviewed and stress was noted. In other words, while subjects felt that the stressors experienced as a dispatcher had the ability to negatively affect their performance, they also believed that stress was beneficial at times. Future research is recommended to further our understanding of workplace stressors for wildland fire dispatchers, and how they cope with them.


Estrogen opposes the apoptotic effects of bone morphogenetic protein 7 on tissue remodeling.
Interactions between estrogen and growth factor signaling pathways at the level of gene expression play important roles in the function of reproductive tissues. For example, estrogen regulates transforming growth factor beta (TGFbeta) in the uterus during the proliferative phase of the mammalian reproductive cycle. Bone morphogenetic protein 7 (BMP-7), a member of the TGFbeta superfamily, is also involved in the development and function of reproductive tissues. However, relatively few studies have addressed the expression of BMP-7 in reproductive tissues, and the role of BMP-7 remains unclear. As part of an ongoing effort to understand how estrogen represses gene expression and to study its interactions with other signaling pathways, chick BMP-7 (cBMP-7) was cloned. cBMP-7 mRNA levels are repressed threefold within 8 h following estrogen treatment in the chick oviduct, an extremely estrogen-responsive reproductive tissue. This regulation occurs at the transcriptional level. Estrogen has a protective role in many tissues, and withdrawal from estrogen often leads to tissue regression; however, the mechanisms mediating regression of the oviduct remain unknown. Terminal transferase-mediated end-labeling and DNA laddering assays demonstrated that regression of the oviduct during estrogen withdrawal involves apoptosis, which is a novel observation. cBMP-7 mRNA levels during estrogen withdrawal increase concurrently with the apoptotic index of the oviduct. Furthermore, addition of purified BMP-7 induces apoptosis in primary oviduct cells. This report demonstrates that the function of BMP-7 in the oviduct involves the induction of apoptosis and that estrogen plays an important role in opposing this function.


Haloperidol and azaperone in drive-net captured Southern chamois (Rupicapra pyrenaica).
We investigated the effect of haloperidol and azaperone in drive-net captured Southern chamois (Rupicapra pyrenaica). Both tranquilizers have been successfully used in a wide range of wild species for reducing postcapture stress response. During 2005, 39 free-ranging chamois were captured, randomly injected intramuscularly with haloperidol (0.29 +/- 0.12 mg/kg; n=24), azaperone (1.1 +/- 0.82 mg/kg; n=6), or saline (0.5 ml; n=9), and restrained for 3 hr. Heart rate was higher in the treated chamois; erythrocyte parameters and total protein concentration decreased over time owing to splenic sequestration, hemodilution, vasodilation, and reflex tachycardia. Creatinine, sodium, and chloride remained stable only in the haloperidol-treated group, suggesting an improvement in renal perfusion. Nevertheless, the azaperone-treated chamois displayed higher body temperature, and both treated groups had higher serum muscular enzymes than the control group, suggesting higher muscle stress. These results lead us not to recommend the use of these tranquilizers-especially azaperone-as first-choice neuroleptics in chamois.


Variable Cre Recombination Efficiency in Placentas of Cyp19-Cre ROSAmT/mG Transgenic Mice.
The aromatase-Cre recombinase (Cyp19-Cre) transgenic mouse model has been extensively used for placenta-specific gene inactivation. In a pilot study, we observed unexpected phenotypes using this mouse strain, which prompted an extensive characterization of Cyp19-Cre placental phenotypes using ROSAmT/mG transgenic reporter mice. The two strains were mated to generate bi-transgenic Cyp19-Cre;ROSAmT/mG mice following a standard transgenic breeding scheme, and placental and fetal tissues were analyzed on embryonic day 17.5. Both maternal and paternal Cre inheritance were analyzed by mating the respective Cyp19-Cre and ROSAmT/mG males and females. The genotype results showed the expected percentage of Cyp19-Cre;ROSAmT/mG fetuses (73%) and Cre mRNA was expressed in all of the Cyp19-Cre placentas. However, surprisingly, only about 50% of the Cyp19-Cre;ROSAmT/mG placentas showed Cre-mediated recombinase activity as demonstrated by placental enhanced green fluorescent protein (EGFP) expression. Further genetic excision analysis of the placentas revealed consistent results showing the absence of excision of the tdTomato in all of the Cyp19-Cre;ROSAmT/mG placentas lacking EGFP expression. Moreover, among the EGFP-expressing placentas, there was wide variability in recombination efficiency, even in placentas from the same litter, leading to a mosaic pattern of EGFP expression in different zones and cell types of the placentas. In addition, we observed a significantly higher percentage of Cre recombination activity in placentas with maternal Cre inheritance. Our results show frequent mosaicism, inconsistent recombination activity, and parent-of-origin effects in placentas from Cyp19-Cre;ROSAmT/mG mice, suggesting that tail-biopsy genotype results may not necessarily indicate the excision of floxed genes in Cyp19-Cre positive placentas. Thus, placenta-specific mutagenesis studies using the Cyp19-Cre model require extensive characterization and careful interpretation of the placental phenotypes for each floxed allele.


[Adenomyomatosis of the gall bladder in rabbits].
Possibility of the adenomiomatosis development of the gallbladder in rabbits of different age is studied. Adenomiomatosis appeared in one of 10 animals of older age animal group. The role of serotonin in the development of adenomiomatosis is discussed.


The economic burden of prostate cancer survivorship care.
Most men live with rather than die of prostate cancer. As a result, survivors have a protracted course, harboring considerable clinical and economic implications. Thus, we investigated the extent to which health service use and expenditures vary during continuing prostate cancer care. We identified 105,961 patients diagnosed with prostate cancer between 1992 and 2005 using Surveillance, Epidemiology and End Results-Medicare data. All Medicare payments for prostate cancer related care were assigned to a phase of care (initial, continuing care or end of life), price adjusted and standardized. Patients were sorted into 5 equal groups (quintiles) based on annual per capita continuing care expenditures. We then enumerated the use of common prostate cancer health services. Average annual per capita continuing care phase expenditures were $36 to $4,724 in the lowest to the highest expenditure group, respectively. Office visits (27.3%) and androgen deprivation (62.7%) comprised most physician related payments for prostate cancer survivorship care. The use of each common health service grew with increasing continuing care intensity (each p <0.001). However, the magnitude of variation in each prostate cancer risk stratum was substantially greater than that between risk strata, eg low risk and metastatic disease, in patients with similar spending. In Medicare beneficiaries the prostate cancer continuing care phase is associated with substantial variation, resulting in potentially unnecessary excess cost to the health care system. Variation was evident across the spectrum of disease severity and implies the need for better evidence to inform clinical practice.


Targeting T regulatory cells in autoimmune diseases.
Among the several mechanisms that play a role in maintaining peripheral self-tolerance is the existence of a unique CD4+CD25+ population of naturally occurring regulatory T cells, which actively prevent both the activation and the effector function of autoreactive T cells that have escaped different mechanisms of tolerance. Many studies have shown the benefit of targeting this cell population by restoring self-tolerance. Therapies that could possibly increase the suppressive ability of T regulatory cells were proven to improve the course of autoimmune diseases.


Calcitonin gene-related peptide receptor activation produces PKA- and PKC-dependent mechanical hyperalgesia and central sensitization.
Calcitonin gene-related peptide (CGRP), acting through CGRP receptors, produces behavioral signs of mechanical hyperalgesia in rats and sensitization of wide dynamic range (WDR) neurons in the spinal cord dorsal horn. Although involvement of CGRP receptors in central sensitization has been confirmed, the second-messenger systems activated by CGRP receptor stimulation and involved in pain transmission are not clear. This study tested whether the hyperalgesia and sensitizing effects of CGRP receptor activation on WDR neurons are mediated by protein kinase A or C (PKA or PKC) signaling. Intrathecal injection of CGRP in rats produced mechanical hyperalgesia, as shown by paw withdrawal threshold tests. CGRP-induced hyperalgesia was attenuated significantly by the CGRP1 receptor antagonist, CGRP8-37. The effect was also attenuated significantly by a PKA inhibitor (H89) or a PKC inhibitor (chelerythrine chloride). Electrophysiological experiments demonstrated that superfusion of the spinal cord with CGRP-induced sensitization of spinal dorsal horn neurons. The CGRP effect could be blocked by CGRP8-37. Either a PKA or PKC inhibitor (H89 or chelerythrine) also attenuated this effect of CGRP. These results are consistent with the hypothesis that CGRP produces hyperalgesia by a direct action on CGRP1 receptors in the spinal cord dorsal horn and suggest that the effects of CGRP are mediated by both PKA and PKC second-messenger pathways.


An Orbitrap/Time-of-Flight Mass Spectrometer for Photofragment Ion Imaging and High-Resolution Mass Analysis of Native Macromolecular Assemblies.
We discuss the design, development, and evaluation of an Orbitrap/time-of-flight (TOF) mass spectrometry (MS)-based instrument with integrated UV photodissociation (UVPD) and time/mass-to-charge ratio (m/z)-resolved imaging for the comprehensive study of the higher-order molecular structure of macromolecular assemblies (MMAs). A bespoke TOF analyzer has been coupled to the higher-energy collisional dissociation cell of an ultrahigh mass range hybrid quadrupole-Orbitrap MS. A 193 nm excimer laser was employed to photofragment MMA ions. A combination of microchannel plates (MCPs)-Timepix (TPX) quad and MCPs-phosphor screen-TPX3CAM assemblies have been used as axial and orthogonal imaging detectors, respectively. The instrument can operate in four different modes, where the UVPD-generated fragment ions from the native MMA ions can be measured with high-mass resolution or imaged in a mass-resolved manner to reveal the relative positions of the UVPD fragments postdissociation. This information is intended to be utilized for retrieving higher-order molecular structural details that include the conformation, subunit stoichiometry, and molecular interactions as well as to understand the dissociation dynamics of the MMAs in the gas phase.


Caspase-mediated suppression of glutamate (AMPA) receptor channel activity in hippocampal neurons in response to DNA damage promotes apoptosis and prevents necrosis: implications for neurological side effects of cancer therapy and neurodegenerative disorders.
DNA damage in neurons is implicated in the pathogenesis of several neurodegenerative disorders and may also contribute to the often severe neurological complications in cancer patients treated with chemotherapeutic agents. DNA damage can trigger apoptosis, a form of controlled cell death that involves activation of cysteine proteases called caspases. The excitatory neurotransmitter glutamate plays central roles in the activation of neurons and in processes such as learning and memory, but overactivation of ionotropic glutamate receptors can induce either apoptosis or necrosis. Glutamate receptors of the AMPA (alpha-amino-3-hydroxy-5-methylisoxazole-4-propionate) type mediate such physiological and pathological processes in most neurons. We now report that DNA damage can alter glutamate receptor channel activity by a mechanism involving activation of caspases. Whole-cell patch clamp analyses revealed a marked decrease in AMPA-induced currents after exposure of neurons to camptothecin, a topoisomerase inhibitor that induces DNA damage; N-methyl-d-aspartate (NMDA)-induced currents were unaffected by camptothecin. The decrease in AMPA-induced current was accompanied by a decreased calcium response to AMPA. Pharmacological inhibition of caspases abolished the effects of camptothecin on AMPA-induced current and calcium responses, and promoted excitotoxic necrosis. Combined treatment with glutamate receptor antagonists and a caspase inhibitor prevented camptothecin-induced neuronal death. Caspase-mediated suppression of AMPA currents may allow neurons with damaged DNA to withdraw their participation in excitatory circuits and undergo apoptosis, thereby avoiding widespread necrosis. These findings have important implications for treatment of patients with cancer and neurodegenerative disorders.


Juvenile polyposis syndrome: An unusual case report of anemia and gastrointestinal bleeding in young infant.
Juvenile polyposis syndrome, a rare disorder in children, is characterized with multiple hamartomatous polyps in alimentary tract. A variety of manifestations include bleeding, intussusception, or polyp prolapse. In this study, we present an 8-month-old male infant of juvenile polyposis syndrome initially presenting with chronic anemia. To the best of our knowledge, this is the youngest case reported in the literature. We report a rare case of an 8-month-old male infant who presented with chronic anemia and gastrointestinal bleeding initially. Panendoscopy and abdominal computed tomography showed multiple polyposis throughout the entire alimentary tract leading to intussusception. Technetium-99m-labeled red blood cell (RBC) bleeding scan revealed the possibility of gastrointestinal tract bleeding in the jejunum. Histopathological examination on biopsy samples showed Peutz-Jeghers syndrome was excluded, whereas the diagnosis of juvenile polyposis syndrome was established. Enteroscopic polypectomy is the mainstay of the treatment. However, polyps recurred and occupied the majority of the gastrointestinal tract in 6 months. Supportive management was given. The patient expired for severe sepsis at the age of 18 months. Juvenile polyposis syndrome is an inherited disease, so it is not possible to prevent it. Concerning of its poor outcome and high mortality rate, it is important that we should increase awareness and education of the parents at its earliest stages.


Use of troponins in the classification of myocardial infarction from electronic health records. The Atherosclerosis Risk in Communities (ARIC) Study.
Electronic health record (EHR) data are underutilized for abstracting classification criteria for heart disease. We compared extraction of EHR data on troponin I and T levels with human abstraction. Using EHR for hospitalizations identified through the Atherosclerosis Risk in Communities (ARIC) Study in four US hospitals, we compared blood levels of troponins I and T extracted from EHR structured data elements with levels obtained through data abstraction by human abstractors to 3 decimal places. Observations were divided randomly 50/50 into training and validation sets. Bayesian multilevel logistic regression models were used to estimate agreement by hospital in first and maximum troponin levels, troponin assessment date, troponin upper limit of normal (ULN), and classification of troponin levels as normal (< ULN), equivocal (1-2× ULN), abnormal (>2× ULN), or missing. Estimated overall agreement in first measured troponin level in the validation data was 88.2% (95% credible interval: 65.0%-97.5%) and 95.5% (91.2-98.2%) for the maximum troponin level observed during hospitalization. The largest variation in probability of agreement was for first troponin measured, which ranged from 66.4% to 95.8% among hospitals. Extraction of maximum troponin values during a hospitalization from EHR structured data is feasible and accurate.


Infectious Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) in Exhaled Aerosols and Efficacy of Masks During Early Mild Infection.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) epidemiology implicates airborne transmission; aerosol infectiousness and impacts of masks and variants on aerosol shedding are not well understood. We recruited coronavirus disease 2019 (COVID-19) cases to give blood, saliva, mid-turbinate and fomite (phone) swabs, and 30-minute breath samples while vocalizing into a Gesundheit-II, with and without masks at up to 2 visits 2 days apart. We quantified and sequenced viral RNA, cultured virus, and assayed serum samples for anti-spike and anti-receptor binding domain antibodies. We enrolled 49 seronegative cases (mean days post onset 3.8 ± 2.1), May 2020 through April 2021. We detected SARS-CoV-2 RNA in 36% of fine (≤5 µm), 26% of coarse (>5 µm) aerosols, and 52% of fomite samples overall and in all samples from 4 alpha variant cases. Masks reduced viral RNA by 48% (95% confidence interval [CI], 3 to 72%) in fine and by 77% (95% CI, 51 to 89%) in coarse aerosols; cloth and surgical masks were not significantly different. The alpha variant was associated with a 43-fold (95% CI, 6.6- to 280-fold) increase in fine aerosol viral RNA, compared with earlier viruses, that remained a significant 18-fold (95% CI, 3.4- to 92-fold) increase adjusting for viral RNA in saliva, swabs, and other potential confounders. Two fine aerosol samples, collected while participants wore masks, were culture-positive. SARS-CoV-2 is evolving toward more efficient aerosol generation and loose-fitting masks provide significant but only modest source control. Therefore, until vaccination rates are very high, continued layered controls and tight-fitting masks and respirators will be necessary.


Chemotherapy is linked to severe vitamin D deficiency in patients with colorectal cancer.
Preclinical and clinical evidence support an association between vitamin D deficiency and an increased risk of colorectal cancer. Normal vitamin D status has been linked to favorable health outcomes ranging from decreased risk of osteoporosis to improved cancer mortality. We performed a retrospective study to assess the impact of metastatic disease and chemotherapy treatment on vitamin D status in patients with colorectal cancer residing in Western New York. Patients, 315, with colorectal cancer treated in a single institute were assayed for 25-OH vitamin D. The association of age, gender, primary disease site and stage, body mass index, and chemotherapy with vitamin D status was investigated. Vitamin D deficiency was common among participants with a median 25-OH vitamin D level of 21.3 ng/ml (optimal range 32-100 ng/ml). Primary site of disease and chemotherapy status were associated with very low 25-OH vitamin D levels (< or =15 ng/ml) on multivariate analysis. Patients receiving chemotherapy and patients with a rectal primary were 3.7 and 2.6-fold more likely to have severe vitamin D deficiency on multivariate analysis than nonchemotherapy patients and colon cancer primary patients, respectively. Chemotherapy is associated with a significant increase in the risk of severe vitamin D deficiency. Patients with colorectal cancer, especially those receiving chemotherapy, should be considered for aggressive vitamin D replacement strategies.


Speeding rod recovery improves temporal resolution in the retina.
The temporal resolution of the visual system progressively increases with light intensity. Under scotopic conditions, temporal resolution is relatively poor, and may be limited by both retinal and cortical processes. Rod photoresponses themselves are quite slow because of the slowly deactivating biochemical cascade needed for light transduction. Here, we have used a transgenic mouse line with faster than normal rod phototransduction deactivation (RGS9-overexpressors) to test whether rod signaling to second-order retinal neurons is rate-limited by phototransduction or by other mechanisms. We compared electrical responses of individual wild-type and RGS9-overexpressing (RGS9-ox) rods to steady illumination and found that RGS9-ox rods required 2-fold brighter light for comparable activation, owing to faster G-protein deactivation. When presented with flickering stimuli, RGS9-ox rods showed greater magnitude fluctuations around a given steady-state current amplitude. Likewise, in vivo electroretinography (ERG) and whole-cell recording from OFF-bipolar, rod bipolar, and horizontal cells of RGS9-ox mice displayed larger than normal magnitude flicker responses, demonstrating an improved ability to transmit frequency information across the rod synapse. Slow phototransduction recovery therefore limits synaptic transmission of increments and decrements of light intensity across the first retinal synapse in normal retinas, apparently sacrificing temporal responsiveness for greater overall sensitivity in ambient light.


A novel method to assist the detection of the Cyclic Alternating Pattern (CAP).
This study proposes a novel method to assist the detection of the components that build up the Cyclic Alternating Pattern (CAP). CAP is a sleep phenomenon formed by consecutive sequences of activations (A1, A2, A3) and non-activations during nonREM sleep. The main importance of CAP evaluation is the possibility of defining the sleep process more accurately. Ten recordings from healthy and good sleepers were included in this study. The method is based on inferential statistics to define the initial and ending points of the CAP components based only on an initialization point given by the expert. The results show concordance up to 95% for A1, 85% for A2 and 60% for A3, together with an overestimation of 1.5 s in A1, 1.3 s in A2 and 0 s in A3. The total CAP rate presents a total underestimation of 7 min. Those results suggest that the method is able to accurately detect the initial and ending points of the activations, and may be helpful for the physicians by reducing the time dedicated to the manual inspection task.


Clinical prediction of deeply infiltrating endometriosis before surgery: is it feasible? A review of the literature.
Endometriosis is a chronic benign gynecologic disease that can cause pelvic pain and infertility affecting almost 10% of reproductive-age women. Deeply infiltrating endometriosis (DIE) is a specific entity responsible for painful symptoms which are related to the anatomic location of the lesions. Definitive diagnosis requires surgery, and histological confirmation is advisable. The aim of this paper is to review the current literature regarding the possibility of diagnosing DIE accurately before surgery. Despite its low sensitivity and specificity, vaginal examination and evaluation of specific symptoms should not be completely omitted as a basic diagnostic tool in detecting endometriosis and planning further therapeutic interventions. Recently, transvaginal ultrasound (TVUS) has been reported as an excellent tool to diagnose DIE lesions in different locations (rectovaginal septum, retrocervical and paracervical areas, rectum and sigmoid, and vesical wall) with good accuracy. There are neither sufficiently sensitive and specific signs and symptoms nor diagnostic tests for the clinical diagnosis of DIE, resulting in a great delay between onset of symptoms and diagnosis. Digital examination, in addition to TVS, may help to gain better understanding of the anatomical extent and dimension of DIE which is of crucial importance in defining the best surgical approach.


From Believing to Doing: The Association Between Leadership Self-Efficacy and the Developmental Leadership Model.
The current study examined the association between leadership self-efficacy and the developmental leadership model. The purpose was to better understand how leadership training transfers to facets of developmental leadership. This was tested in a cross-sectional design with military commanders in the Swedish Armed Forces. The results show that the sub-domain of leader self-control efficacy (the cognitive and emotional ability to remain composure) did not predict developmental leadership, but that leader assertiveness efficacy (the ability to make rational decisions) predicted the two dimensions of the exemplary model, and inspiration and motivation in the developmental leadership model. One possibility is that leader self-control efficacy can be what enables the individual to function within an extreme context, but leader assertiveness efficacy can be what most determines the leadership performance within that context. The possibility for mediatory analyses in further research is discussed.


Gene expressions of TRP channels in glioblastoma multiforme and relation with survival.
Glioblastoma multiforme (GBM) is one of the most lethal forms of cancer in humans, with a median survival of 10 to 12 months. Glioblastoma is highly malignant since the cells are supported by a great number of blood vessels. Although new treatments have been developed by increasing knowledge of molecular nature of the disease, surgical operation remains the standard of care. The TRP (transient receptor potential) superfamily consists of cation-selective channels that have roles in sensory physiology such as thermo- and osmosensation and in several complex diseases such as cancer, cardiovascular, and neuronal diseases. The aim of this study was to investigate the expression levels of TRP channel genes in patients with glioblastoma multiforme and to evaluate the relationship between TRP gene expressions and survival of the patients. Thirty-three patients diagnosed with glioblastoma were enrolled to the study. The expression levels of 21 TRP genes were quantified by using qRT-PCR with dynamic array 48 × 48 chip (BioMark HD System, Fluidigm, South San Francisco, CA, USA). TRPC1, TRPC6, TRPM2, TRPM3, TRPM7, TRPM8, TRPV1, and TRPV2 were found significantly higher in glioblastoma patients. Moreover, there was a significant relationship between the overexpression of TRP genes and the survival of the patients. These results demonstrate for the first time that TRP channels contribute to the progression and survival of the glioblastoma patients.


Exposure of Heligmosomoides polygyrus and Trichuris muris to albendazole, albendazole sulfoxide, mebendazole and oxantel pamoate in vitro and in vivo to elucidate the pathway of drug entry into these gastrointestinal nematodes.
Millions of people are treated with anthelmintics to control soil-transmitted helminth infections; yet, drug distribution in the plasma and gastrointestinal tract compartments and the pathway of drug uptake into gastrointestinal nematodes responsible for the pharmacological effect are unknown. We assessed the distribution and uptake of albendazole, albendazole sulfoxide, albendazole sulfone in the hookworm Heligmosomoides polygyrus in vitro and in vivo as well as the distribution and uptake of albendazole, mebendazole, and oxantel pamoate in the whipworm Trichuris muris in vitro and in vivo. Oral and intraperitoneal treatments (100 mg/kg) were studied. Drug quantities in helminths and host compartments (stomach, the contents and mucosa of the small and large intestine, and the plasma) were determined using HPLC-UV/vis and anthelmintic activities were recorded using phenotypic readout. The influence of 1-aminobenzotriazole (ABT), an irreversible and unspecific cytochrome P450 inhibitor, on albendazole disposition in mice harboring H. polygyrus was evaluated. In vivo, albendazole was found in quantities up to 10 nmol per ten H. polygyrus and up to 31 nmol per ten T. muris. ABT did not change the levels of albendazole or its metabolites in the plasma of mice harboring H. polygyrus or in H. polygyrus, whereas drug levels in the gastrointestinal tract of host mice doubled. Mebendazole and oxantel pamoate quantities per ten T. muris were as high as 21 nmol and 34 nmol, respectively. Albendazole revealed a very dynamic distribution and high rate of metabolism, hence, H. polygyrus and T. muris are exposed to albendazole and both metabolites via multiple pathways. Diffusion through the cuticle seems to be the crucial pathway of oxantel pamoate uptake into T. muris, and likely also for mebendazole. No relationship between concentrations measured in helminths and concentrations in plasma, intestinal content and mucosa of mice, or drug efficacy was noted for any of the drugs studied.


A psychoeducational intervention (SWEEP) for depressed women with diabetes.
Clinically significant depression is present in 25 % of individuals with type 2 diabetes, its risk being doubled in women. To examine the effectiveness of the Study of Women's Emotions and Evaluation of a Psychoeducational (SWEEP), a group therapy for depression treatment based on cognitive behavioral therapy principles that was developed for women with type 2 diabetes was conducted. Women with significantly elevated depression symptoms (Center for Epidemiologic Studies Depression Scale ≥16) were randomized to SWEEP (n = 38) or usual care (UC, n = 36). Multilevel modeling indicated that SWEEP was more effective than UC in reducing depression (mean difference of -15 vs. -7, p < .01), decreasing trait anxiety (mean difference of -15 vs. -5, p < .01), and improving anger expression (mean difference of -12 vs. -5, p < .05). Although SWEEP and UC had improvements in fasting glucose (mean difference of -24 vs. -1 mg/dl) and HbA1c (mean difference of -0.4 vs. -0.1 %), there were no statistically significant differences between groups. SWEEP was more effective than UC for treating depressed women with type 2 diabetes. Addition of group therapy for depression meaningfully expands the armamentarium of evidence-based treatment options for women with diabetes.


Abatement of the clostridial load in the teats of lactating cows with lysozyme derived from donkey milk.
The use of a sterilized product for washing cows' udders before milking may be useful to reduce or prevent Clostridium tyrobutyricum contamination, the main cause of the late-blowing defect in hard and semi-hard cheeses. The aim of this research was to evaluate the antibacterial efficacy of an experimental formula containing 15% condensed donkey milk (lysozyme content 825 mg/L). The antimicrobial activity of condensed milk was first evaluated in vitro, using the disk diffusion method, on the following microorganisms: Bacillus megaterium, Bacillus mojavensis, Clavibacter michiganensis, and Clostridium tyrobutyricum. These results were compared with the effects of 2 antibiotics, ampicillin (100 mg/mL) and kanamycin (50 mg/ mL), and a commercial pre-dipping formula. The results showed that the inhibitory activity of lysozyme from donkey milk on all the considered microorganisms was higher than that of the commercial product and similar to that of the 2 antibiotics. Next, the formula with lysozyme was compared with a commercial pre-dipping formula on 48 lactating cows (24 cows in each group). Skin tests were performed on teats before and after pre-dipping. Results showed that the formula with condensed milk significantly reduced the clostridial load detected on the skin of cows' teats before cleaning (-55.61% vs. -27.99%) and in the bulk milk of the experimental group compared with the control group with commercial product (-52.53% vs. -32.42%).


Achieving High Reliability Through Cultural Mindfulness.
Healthcare has enthusiastically embraced quality and safety improvement. Yet, more radical transformation is clearly needed to make a more significant impact on error reduction and to ensure consistent quality. This need for transformation is leading healthcare to examine how other industries, such as nuclear power and aviation, improve safety to achieve a high degree of reliability and avoid potential catastrophes. Research has shown that successful organizations in high-risk industries achieve high reliability by maintaining a cultural mindfulness that allows them to continually reinvent themselves in complex environments. Healthcare faces similar challenges and could greatly benefit from instilling high-reliability principles in its operations. The Medical University of South Carolina, an academic health system, has been on a quest to improve safety and quality by implementing a high-reliability culture.


alpha-Lipoic acid supplementation inhibits oxidative damage, accelerating chronic wound healing in patients undergoing hyperbaric oxygen therapy.
Hyperbaric oxygen (HBO) therapy is successfully used for the treatment of a variety of conditions. However, prolonged exposure to high concentrations of oxygen induces production of reactive oxygen species, causing damage to the cells. Thus, antioxidant supplementation has been proposed as an adjuvant to attenuate such deleterious secondary effects. We evaluated the effects of alpha-lipoic acid (LA) in patients affected by chronic wounds undergoing HBO treatment. LA supplementation efficiently reduces both the lipid and DNA oxidation induced by oxygen exposure. LA exerted its antioxidant activity by directly interacting with free radicals or by recycling vitamin E. An inhibitory effect of LA on the pro-inflammatory cytokine interleukin-6 was observed. Taken together, we demonstrated an adjuvant effect of LA in HBO therapy used for impaired wound healing treatment. We propose that LA may be used to further promote the beneficial effects of HBO therapy.


Exposure to airborne fungi during conservative dental treatment.
The aim of the study was a mycological assessment of bioaerosol forming during conservative dental treatment, taking into account concentration and type of fungal microflora, and evaluation of the influence of DUWL disinfecting protocol on the fungal contamination of the bioaerosol. The research was conducted on 25 operative sites located in public dental clinics. The air contained in the space between a patient and a dentist during conservative dental treatment with the use of a high-speed handpiece was examined. Air samples were taken using the portable RCS PLUS Air Sampler (BIOTEST AG, Dreieich, Germany) and ready-to-use agar YM Strips for yeast and mould fungi culture. The volume of the sampled air was 100 litres. Before disinfection, the concentration of fungi in the collected air samples at individual operative sites ranged from 4 x 10(1) cfu/m3 to 34 x 10(1) cfu/m3. The most common species was Penicillium herquei (62.17% of the total count), followed by other fungi: Alternaria alternata - 12.68%, Penicillium roseopurpureum - 9.41%, Rhizopus nigricans - 5.93%, Aspergillus terreus - 3.89%, Geotrichum candidum - 2.25%, Aspergillus glaucus group - 2.04%, Cladosporium cladosporoides - 1.23% and Penicillium diversum - 0.41%. The concentration of Penicillium herquei at individual operative sites ranged from 0 to 34 x 10(1) cfu/m3, mean 121.6 cfu/m3, Penicillium roseopurpureum - from 0 to 11 x 10(1) cfu/m3, mean 18.4 cfu/m3 and Alternaria alternata - from 0 to 18 x 10(1) cfu/m3, mean 24.8 cfu/m3. After disinfection, like before disinfection procedures, the prevailing species of fungi were: Penicillium herquei, Penicillium reseopurpureum and Alternaria alternata, which amounted to 62.6%, 18.28% and 11.36% of the isolated fungi, respectively. The recorded levels of total airborne fungi were lower after DUWL disinfection compared to those before disinfection.


Anterior-only approaches to scoliosis.
Scoliosis is a three-dimensional spinal deformity for which surgery may be indicated when patients experience severe pain, curve progression, or progressive disability. Operative treatment has conventionally involved extensive posterior stabilization, either alone or in combination with anterior release procedures. Anterior-only approaches have a more limited role, but they should be considered in the appropriate setting when addressing this disease. Some advantages of anterior-only approaches to scoliosis include greater corrective ability with a greater fusion rate while mandating fewer motion segments to be involved in the instrumented construct. Preventing denervation of the posterior paraspinal musculature is also desirable and is inherently provided for by the anterior approach. Disadvantages include pulmonary morbidity associated with violating the thoracic cavity, as well as post-thoracotomy chronic pain. There are also limitations in the indications for which anterior-only surgery can be performed, and these will be discussed along with treatment options. Technological advances, including minimally invasive thoracic access as well as evolving instrumentation, may minimize the stated disadvantages and expand the scope of scoliosis cases that may be treated by anterior-only approaches. Hence, the enhanced corrective ability and biomechanical advantages of the anterior column can be used in the treatment of these patients. Although the indications for anterior-only strategies in scoliosis remain limited, such operations should be considered in the appropriate setting when addressing spinal deformity.


1.4 Mb candidate gene region for X linked dyskeratosis congenita defined by combined haplotype and X chromosome inactivation analysis.
Dyskeratosis congenita (DC) is a rare inherited disorder characterised by the early onset of reticulate skin pigmentation, nail dystrophy, and mucosal leucoplakia. In over 80% of cases bone marrow failure develops and this is the main cause of early mortality. The DC1 gene responsible for the X linked form (MIM 305000) of dyskeratosis congenita has been mapped to Xq28. In order to narrow the candidate gene region, genetic linkage analysis was performed in eight X linked pedigrees using a set of markers spanning Xq28. A maximum lod score of 5.31 with no recombinations was achieved with marker DXS1073. Two recombination events were identified; one of these uses X chromosome inactivation pattern analysis to determine carrier status and haplotype analysis to fine map the recombination breakpoint. The fine mapping of these recombination events has enabled the candidate gene region for X linked dyskeratosis congenita to be defined as the 1.4 Mb interval between Xq3274 and DXS1108.


Apoptosis-inducing factor deficiency sensitizes dopaminergic neurons to parkinsonian neurotoxins.
Mitochondrial complex I deficits have long been associated with Parkinson disease (PD). However, it remains unknown whether such defects represent a primary event in dopaminergic neurodegeneration. Apoptosis-inducing factor (AIF) is a mitochondrial protein that, independently of its proapoptotic properties, plays an essential physiologic role in maintaining a fully functional complex I. We used AIF-deficient harlequin (Hq) mice, which exhibit structural deficits in assembled complex I, to determine whether primary complex I defects linked to AIF depletion may cause dopaminergic neurodegeneration. Despite marked reductions in mitochondrial complex I protein levels, Hq mice did not display apparent alterations in the dopaminergic nigrostriatal system. However, these animals were much more susceptible to exogenous parkinsonian complex I inhibitors, such as 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). Subtoxic doses of MPTP, unable to cause damage to wild-type animals, produced marked nigrostriatal dopaminergic degeneration in Hq mice. This effect was associated with exacerbated complex I inhibition and increased production of mitochondrial-derived reactive oxygen species (ROS) in Hq brain mitochondria. The antioxidant superoxide dismutase-mimetic compound tempol was able to reverse the increased susceptibility of Hq mice to MPTP. Supporting an instrumental role for mitochondrial-derived ROS in PD-related neurodegeneration, transgenic mice overexpressing mitochondrially targeted catalase exhibited an attenuation of MPTP-induced mitochondrial ROS and dopaminergic cell death. Structural complex I alterations linked to AIF deficiency do not cause dopaminergic neurodegeneration but increase the susceptibility of dopaminergic neurons to exogenous parkinsonian neurotoxins, reinforcing the concept that genetic and environmental factors may interact in a common molecular pathway to trigger PD.


Contrast of directional influence upon motor overflow between submaximal and maximal static exertions.
This study investigated exertion-dependent motor overflow among healthy adults when they performed isometric tasks with contralateral joints in different task directions. Twenty healthy adults (10 males and 10 females, mean age = 26.2 yrs) were instructed to complete a set of isometric contractions of various force vectors with the shoulder, elbow, and wrist joints, in a total of ten motor tasks at submaximal and maximal intensities (50%, 100% maximal voluntary contractions). The electromyographical activities from eight muscles of the unexercised upper limb were recorded to characterize intensity of motor overflow during sustained isometric contraction. Both occurrence frequency and magnitude of motor overflow in terms of standardized net excitation (SNE) increased with exertion level for all joint movements (P < 0.001). Additionally, the motor overflow magnitude depended strongly on the task direction of maximal isometric contraction (P < 0.05). Motor overflow was particularly augmented by the contralateral isometric contractions where task directions were opposed to gravity. However, such a directional effect upon SNE was not evident during submaximal contraction (P > 0.05). The difference of the net excitation between maximal and submaximal contraction (DNE(100%-50%MVC) data) indicated that the pectoralis major and triceps brachii consistently exhibited a marked recruitment in reaction to change in task direction of isometric contraction. Patterned motor overflow may be physiologically relevant to topological mapping of the ipsilateral pathways and altered effectiveness of use-dependent interhemispherical connectivity. The current observations provide better insight into gain in muscle strength due to contralateral exercise.


Furosemide 1-O-acyl glucuronide. In vitro biosynthesis and pH-dependent isomerization to beta-glucuronidase-resistant forms.
A biosynthetic acyl-type glucuronic acid conjugate of furosemide was isolated from in vitro incubation of pregnenolone-16 alpha-carbonitrile-induced rat liver microsomes containing UDP-glucuronyltransferase activity, furosemide, and UDP-glucuronic acid. Furosemide 1-O-acyl glucuronide (FG) was specifically hydrolyzed by beta-glucuronidase (BG) and was also labile to alkaline hydrolysis. FG concentration decreased at an apparent first order rate when incubated at 37 degrees C in buffer solution of pH values greater than 6.0 with only moderate hydrolysis of the conjugate at pH values less than 8.5. Formation of rearrangement forms of FG that were resistant to BG but labile to alkaline hydrolysis accounted for most of the disappearance of FG at this pH range. Radiochemical labeling of the conjugate with either 14C-furosemide or 14C-UDP-glucuronic acid was detected in the BG-resistant isomerization products of FG as they were separated by HPLC. The structure of FG and its isomerization products was further verified by negative ion thermospray liquid chromatography/mass spectrometry. The abundant (M - 1)-ion at mass 505, the aglycone fragment at m/z 329, and the characteristic sugar fragment ion of mass 175 were found in the spectra of FG and three additional isomers. An ion at m/z 221 was noted only in the case of the parent conjugate and thus may prove to be a characteristic ion for 1-O-acyl-linked glucuronides under negative ion thermospray. In vivo as well as in vitro rearrangement of FG to BG-resistant forms might affect the results of furosemide disposition studies which use BG hydrolysis to determine FG formation.


A comparison of selected nutrient intakes derived from three diet assessment methods used in a low-fat maintenance trial.
In the vast majority of surveys and research in humans, dietary data are obtained from self-reports: recalls; records; or historical methods, usually food frequency questionnaires (FFQ). This study provides a rare opportunity to compare data derived from all three methods. A crossover study of dietary fat in which data were collected using an average of 11.4 food records and 11.7 24-h diet recalls. Using simple subtraction and correlation, energy and nutrient intakes derived from the three methods were compared to each other and with those derived from a single FFQ. Analysis of variance was used to evaluate sources of variability in nutrient intakes estimated from the individual days of records and recalls. An independent, free-standing medical research institute. 13 men who were compliant with study procedures. FFQ-derived estimates of energy and nutrient intake were highest (e.g. 1967 kcal versus 1858 kcal and 1936 kcal for the records and recalls, respectively). Mean differences in energy and nutrient intakes and their variances were lowest and correlation coefficients highest in comparing the records and recalls (e.g. for fat the mean difference was 5.0 g, and r = 0.85). Analysis of variance of individual days of record- and recall-derived data (n = 300) revealed that there was no effect due to either method (record or recall) or the sequence of administration. Results of this study indicate that the FFQ overestimated dietary intake. Energy and nutrient results obtained from the records and recalls were interchangeable. However, based on smaller SDs around the means, it appears that the recalls may perform slightly better in estimating dietary intake in groups such as these well-educated, highly compliant men.


Epitaxial antiperovskite/perovskite heterostructures for materials design.
Engineered heterostructures formed by complex oxide materials are a rich source of emergent phenomena and technological applications. In the quest for new functionality, a vastly unexplored avenue is interfacing oxide perovskites with materials having dissimilar crystallochemical properties. Here, we propose a unique class of heterointerfaces based on nitride antiperovskite and oxide perovskite materials as a previously unidentified direction for materials design. We demonstrate the fabrication of atomically sharp interfaces between nitride antiperovskite Mn3GaN and oxide perovskites (La0.3Sr0.7)(Al0.65Ta0.35)O3 and SrTiO3. Using atomic-resolution imaging/spectroscopic techniques and first-principles calculations, we determine the atomic-scale structure, composition, and bonding at the interface. The epitaxial antiperovskite/perovskite heterointerface is mediated by a coherent interfacial monolayer that interpolates between the two antistructures. We anticipate our results to be an important step for the development of functional antiperovskite/perovskite heterostructures, combining their unique characteristics such as topological properties for ultralow-power applications.


Interference of BAD (Bcl-xL/Bcl-2-associated death promoter)-induced apoptosis in mammalian cells by 14-3-3 isoforms and P11.
Apoptosis and survival of diverse cell types are under hormonal control, but intracellular mechanisms regulating cell death are unclear. The Bcl-2/Ced-9 family of proteins contains conserved Bcl-2 homology regions that mediate the formation of homo- or heterodimers important for enhancing or suppressing apoptosis. Unlike most other members of the Bcl-2 family, BAD (Bcl-xL/Bcl-2 associated death promoter), a death enhancer, has no C-terminal transmembrane domain for targeting to the outer mitochondrial membrane and nuclear envelope. We hypothesized that BAD, in addition to binding Bcl-xL and Bcl-2, may interact with proteins outside the Bcl-2 family. Using the yeast two-hybrid system to search for BAD-binding proteins in an ovarian fusion cDNA library, we identified multiple cDNA clones encoding different isoforms of 14-3-3, a group of evolutionally conserved proteins essential for signal transduction and cell cycle progression. Point mutation of BAD in one (S137A), but not the other (S113A), putative binding site found in diverse 14-3-3 interacting proteins abolished the interaction between BAD and 14-3-3 without affecting interactions between BAD and Bcl-2. Because the S137A BAD mutant presumably resembles an underphosphorylated form of BAD, we used this mutant to screen for additional BAD-interacting proteins in the yeast two-hybrid system. P11, a nerve growth factor-induced neurite extension factor and member of the calcium-binding S-100 protein family, interacted strongly with the mutant BAD but less effectively with the wild type protein. In Chinese hamster ovary (CHO) cells, transient expression of wild type BAD or its mutants increased apoptotic cell death, which was blocked by cotransfection with the baculovirus-derived cysteine protease inhibitor, P35. Cotransfection with 14-3-3 suppressed apoptosis induced by wild type or the S113A mutant BAD but not by the S137A mutant incapable of binding 14-3-3. Furthermore, cotransfection with P11 attenuated the proapoptotic effect of both wild type BAD and the S137A mutant. For both 14-3-3 and P11, direct binding to BAD was also demonstrated in vitro. These results suggest that both 14-3-3 and P11 may function as BAD-binding proteins to dampen its apoptotic activity. Because the 14-3-3 family of proteins could interact with key signaling proteins including Raf-1 kinase, protein kinase C, and phosphatidyl inositol 3 kinase, whereas P11 is an early response gene induced by the neuronal survival factor, nerve growth factor, the present findings suggest that BAD plays an important role in mediating communication between different signal transduction pathways regulated by hormonal signals and the apoptotic mechanism controlled by Bcl-2 family members.


Castration of piglets under CO2-gas anaesthesia.
It has become common practice in pig fattening production systems to castrate young boar piglets without the use of anaesthesia. In this study, we examined whether or not CO2 gas is capable of inducing an acceptable anaesthetic state during which castration can be performed. The first step was to identify the most promising CO2/O2 mixture. Based on the results from this first experiment, a mixture of 70% CO2 + 30% O2 was chosen for further investigation as a potential anaesthetic during the castration of young piglets. Thereby, it was established whether the duration and depth of anaesthesia were acceptable for castration where the animal has to be insensible and unconscious. Physiological effects were assessed based on electroencephalogram (EEG) and electrocardiogram (ECG) measurements, blood gas values and behavioural responses. During the induction phase, the only typical behaviour the piglets exhibited when exposed to the 70/30 gas mixture was heavy breathing. All piglets (n = 25) lost consciousness after approximately 30 s according to the EEG. Heart rate decreased slowly during the induction phase, a serious drop occurred when piglets lost their posture. Immediately after this drop, the heart rate neared zero or showed a very irregular pattern. Shortly after loss of posture, most animals showed a few convulsions. None of the animals showed any reaction to castration in behaviour and/or on the EEG and ECG. On average, the piglets recovered within 59 s, i.e. EEG returned to its pre-induction pattern and piglets were able to regain a standing position. After 120 s, heart rate returned to pre-induction levels. In order to explore the usage range of CO2 concentration, 24 piglets were exposed to 60% CO2 + 20% O2 + 20% N2 for up to 30 s after loss of consciousness (as registered on EEG), and castrated after removal from the chamber. Sixteen of the 24 animals showed a reaction to the castration on the EEG. To establish the maximum time piglets survive in 70% CO2 + 30% O2, five piglets were placed in this mixture for 3 min. Two of them died. After that, four piglets were placed in this mixture for 2 min after unconsciousness, one died after 2 min. It was concluded from this study that it is possible to anaesthetise piglets with a mixture of 70% CO2 + 30% O2, but that there are limits to its safety in terms of CO2 concentration and duration of exposure. Before implementation for practical use, further research is essential to assess the limits of gas concentration and exposure times.


Organizer-specific homeobox genes in Xenopus laevis embryos.
The dorsal blastopore lip of the early Xenopus laevis gastrula can organize a complete secondary body axis when transplanted to another embryo. A search for potential gene regulatory components specifically expressed in the organizer was undertaken that resulted in the identification of four types of complementary DNAs from homeobox-containing genes that fulfill this criterion. The most abundant of these encodes a DNA-binding specificity similar to that of the Drosophila melanogaster anterior morphogen bicoid. The other three are also homologous to developmentally significant Drosophila genes. These four genes may participate in the regulation of the developmental potential of the organizer.


Aging reduces EEG markers of recognition despite intact performance: Implications for forensic memory detection.
ERP-based forensic memory detection is based on the logic that guilty suspects will hold incriminating knowledge about crimes they have committed, and therefore should show parietal ERP positivities related to recognition when presented with reminders of their crimes. We predicted that such forensic memory detection might however be inaccurate in older adults, because of changes to recognition-related brain activity that occurs with aging. We measured both ERPs and EEG oscillations associated with episodic old/new recognition and forensic memory detection in 30 younger (age < 30) and 30 older (age > 65) adults. EEG oscillations were included as a complementary measure which is less sensitive to temporal variability and component overlap than ERPs. In line with predictions, recognition-related parietal ERP positivities were significantly reduced in the older compared to younger group in both tasks, despite highly similar behavioural performance. We also observed aging-related reductions in oscillatory markers of recognition in the forensic memory detection test, while the oscillatory effects associated with episodic recognition were similar across age groups. This pattern of results suggests that while both forensic memory detection and episodic recognition are accompanied by aging-induced reductions in parietal ERP positivities, these reductions may be caused by non-overlapping mechanisms across the two tasks. Our findings suggest that EEG-based forensic memory detection tests are less valid in older than younger populations, limiting their practical applications.


Proteome-wide profiling and mapping of post translational modifications in human hearts.
Post translational modifications (PTMs) are covalent modifications of proteins that can range from small chemical modifications to addition of entire proteins. PTMs contribute to regulation of protein function and thereby greatly increase the functional diversity of the proteome. In the heart, a few well-studied PTMs, such as phosphorylation and glycosylation, are known to play essential roles for cardiac function. Yet, only a fraction of the ~ 300 known PTMs have been studied in a cardiac context. Here we investigated the proteome-wide map of PTMs present in human hearts by utilizing high-resolution mass spectrometry measurements and a suite of PTM identification algorithms. Our approach led to identification of more than 150 different PTMs across three of the chambers in human hearts. This finding underscores that decoration of cardiac proteins by PTMs is much more diverse than hitherto appreciated and provides insights in cardiac protein PTMs not yet studied. The results presented serve as a catalogue of which PTMs are present in human hearts and outlines the particular protein and the specific amino acid modified, and thereby provides a detail-rich resource for exploring protein modifications in human hearts beyond the most studied PTMs.


Soy protein peptides regulate cholesterol homeostasis in Hep G2 cells.
The activation of LDL receptors was described recently in a human hepatoma cell line (Hep G2) exposed both to alpha + alpha' subunits from 7S soy globulin and to Croksoy(R)70, a commercial isoflavone-poor soy concentrate. To assess the final identity of the peptide(s) putatively responsible for the biochemical effect, experiments were performed in Hep G2 cells, exposed either to synthetic peptides corresponding to specific sequences of 7S soy globulin or to peptides from the in vitro digestion of Croksoy(R)70. Moreover, the ability of the whole 7S globulin, its subunits and whole Croksoy(R)70 to interfere in the apolipoprotein B (apo B) secretion in the medium as well as in sterol biosynthesis was evaluated in the same model. Increased (125)I-LDL uptake and degradation vs. controls were shown after Hep G2 incubation with a synthetic peptide (10(-)(4) mol/L, MW 2271 Da) corresponding to positions 127-150 of the 7S globulin. Cells exposed to Croksoy(R)70 enzyme digestion products showed a more marked up-regulation of LDL receptors vs. controls, compared with vs. Hep G2 cells incubated with undigested Croksoy(R)70. Among soy-derived products, only the 7S globulin inhibited apo B secretion and (14)C-acetate incorporation when tested in Hep G2 cells at a concentration of 1.0 g/L. These findings support the hypothesis that if one or more peptides can reach the liver after intestinal digestion, they may elicit a cholesterol-lowering effect. Moreover, the protein moiety, devoid of isoflavone components, is likely to be responsible for this major biochemical effect of soy protein.