The repeating disaccharide unit of the mucopeptide of the cell wall of Mycobacterium smegmatis contains N-acetyl-glucosamine linked to N-glycolyl-muramic acid. This first identification of N-glycolyl-muramic acid as a natural product has been performed by mass spectrometry of the permethylated disaccharide. N-glycolyl muramic acid has been synthesized: the natural and synthetic products are identical.


Glass bead sterilisation of surgical dental burs.
Since burs used in OMF surgery are a source of microbial contamination, sterilisation of the burs is an important aspect of infection control. A two-part study was performed to evaluate the efficacy of using the glass bead steriliser to sterilise long-shank burs. The laboratory and clinical studies indicated that the glass bead steriliser was effective in sterilising long-shank burs in 60 s.


Hyperthermo-chemo-radiotherapy for carcinoma of the esophagus.
Hyperthermo-chemo-radiotherapy was prescribed preoperatively for eight patients with carcinoma of the esophagus. Histological study of the resected specimens showed marked effect in two, moderate effect in four and slight effect in two. In those with marked effect, there were no viable neoplastic cells, and severe necrosis and complete destruction were evident on the mucosal surface. These two patients have had no recurrence, now at 27th and 31st postoperative months. Those findings suggest that hyperthermo-chemo-radiotherapy is effective for selected patients with carcinoma of the esophagus.


Astrocytes resolve ER stress through mitochondrial fusion facilitated by biotin availability.
Structures of cellular organelles are intertwined with their functions that undergo alterations once the organelles are stressed. Since organelle functions are dependent on each other, an organelle-specific stress possibly influences the structure and function of its associated organelles. In this perspective, our study demonstrated that endoplasmic reticulum (ER)-specific stress induced by tunicamycin in primary astroglial culture is associated with altered mitochondrial dynamics and matched with the changes as observed in the aging rat brain. However, the exogenous addition of biotin, a highly lipogenic and mitochondrial vitamin, ameliorates ER stress even though its direct targets are not known within ER. Alternatively, the increased biotinylation of mitochondrial carboxylases preserves its basal respiratory capacity by upregulating mitofusin 2 (Mfn2) and, possibly, its associated role on mitochondrial fusion. Furthermore, the Mfn2 increase by biotin augments physical interaction between ER and functional mitochondria to exchange biomolecules as a part of ER stress resolution. This suggests an increased demand for micronutrient biotin under ER stress resolves the same by undergoing appropriate structural and metabolic contacts between ER and mitochondria. These findings provide a paradigm to resolve stress in one organelle by sustaining the metabolic commitments of another interdependent organelle. The findings also highlight a novel role of biotin in inducing Mfn2 expression and localization under ER stress in addition to its known role as a co-enzyme.


High-dose chemotherapy with autologous stem cell rescue in breast cancer.
Because metastatic breast cancer is a lethal disease despite some responsiveness to systemic chemotherapy, high-dose chemotherapy with autologous stem cell rescue is being utilized with increasing frequency. This analysis was undertaken to determine the outcome for such patients treated with intensive chemotherapy between 1989-1994, at the Hoag Cancer Center in Newport Beach, CA. During 1989, only patients with metastatic disease who had failed more than two standard breast cancer chemotherapy regimens were considered eligible for such treatment. They received high-dose BCNU/cyclophosphamide/cisplatinum chemotherapy with autologous bone marrow rescue. After January 1990, patients with metastatic disease were eligible only if they had received limited prior chemotherapy and demonstrated responsiveness to induction chemotherapy. Beginning June 1990, patients with metastatic disease were to receive mitoxantrone and thiotepa (MiTepa) followed by peripheral blood stem cell rescue, then ifosfamide, carboplatin and etoposide (ICE) chemotherapy followed by peripheral blood stem cell rescue. High-risk adjuvant patients were to receive one course of ICE followed by rescue. Between 1/89-12/94, 48 breast cancer patients underwent 65 intensive chemotherapy treatments followed by autologous stem cell rescue. During 1989, three of the eight patients with metastatic disease died within 60 days because of therapy-related complications. The longest failure-free survival (FFS) of these eight was 12.2 months, and the longest overall survival (OS) 20.5 months. Since 1/90, one physician has treated 24 patients with metastatic breast cancer, 17 of whom actually underwent two successive transplants with MiTepa/ICE. For the latter group, median FFS is 23.2 months; median OS is 39.7 months. There were no acute deaths, but two patients died > 60 days after initial transplant from therapy-related complications, veno-occlusive disease (5.2 months) and myelodysplastic syndrome (30.5 months), while five died of progressive disease at 22.5, 32.8, 39.4, 46.3, and 51.3 months. For the 24 metastatic patients treated 1990-1994, 1-, 2-, and 3-year FFS rates are 86%, 40%, and 17%, respectively, while OS rates are 91%, 80%, and 65%. Of 11 patients treated in the adjuvant setting, only one has relapsed (9.8 months) with follow-up from 3-61 months. Modifications made in the program, including selection of patients responsive to induction chemotherapy, transfusion of peripheral blood stem cells, implementation of hematopoietic colony stimulating factors, and use of tandem intensive treatments has been associated with a low rate of acute morbidity and encouraging survival rates.


Ball fracture with the 6120-model Starr-Edwards mitral valve prosthesis occurring late after implantation.
The Starr-Edwards (S-E) mitral valve, Model 6120, has been widely used in the last three decades. Fatty infiltration and subsequent damage of the silastic ball, a phenomenon termed ball variance, has been described only in the first generation aortic ball-valve prostheses, used until 1965. A case of partial ball fracture in a 6120-Model S-E mitral valve prosthesis, occurring 27 years after implantation and presenting with thromboembolism and progressive valve dysfunction, is described.


Chronic sun exposure-related fusion oncogenes EGFR-PPARGC1A in cutaneous squamous cell carcinoma.
Cutaneous squamous cell carcinoma (cSCC) differs from SCC of other organs in its strong association with chronic sun exposure. However, the specific driver mutations in cSCC remain unknown. Fusion genes in established cSCC cell lines (A431 and DJM-1) were predicted by transcriptome sequence, and validated by Sanger sequence, fluorescence in situ hybridization and G-banding. By transcriptome sequencing, we identified fusion gene EGFR-PPARGC1A in A431, which were expressed in 31 of 102 cSCCs. The lesions harboring the fusion gene tended to be located in sun-exposed areas. In vivo cutaneous implantation of EGFR-PPARGC1A-expressing NIH3T3 induced tumors resembling human cSCC, indicating its potent tumorigenicity. NIH3T3 transfected with EGFR-PPARGC1A as well as A431 showed increased cell proliferation activity. With regard to underlying mechanism, EGFR-PPARGC1A protein causes constitutive tyrosine phosphorylation, and induces the phosphorylation of wild-type full-length epidermal growth factor receptor (EGFR) by dimerization. Conversely, the RNAi-mediated attenuation of EGFR or CRISPR/Cas9-mediated knockdown of the fusion gene in A431 led to a decrease in the cell number, and may have therapeutic value. Our findings advance the knowledge concerning genetic causes of cSCC and the function of EGFR, with potential implications for new diagnostic and therapeutic approaches.


Optimizing the deformation behavior of stent with nonuniform Poisson's ratio distribution for curved artery.
Stent implantation at a highly curved artery has always been a challenge, considering the relatively high chance of in-stent restenosis (ISR) caused by severe straightening effect and high strain energy over the vessel wall. In this paper, a novel optimization based design method was proposed to manipulate the deformation behavior of the common ring-and-link stent. By changing the location of the connection point between rings and links, traditional ring-and-link structure was modified to achiever tunable Poisson's ratio (PR). With the nonuniform cellular structure design method proposed in a previous study, PR distribution of the stent structure was optimized to achieve the desired curvature. As a result, the obtained stent structure with nonuniform PR could perfectly fit into the curved artery after expansion, without causing any obvious vessel straightening. To validate the proposed method, two different vessel models were introduced. Firstly, a short vessel with a constant curvature was set as the design objective, and both numerical and experimental tests were conducted. Further, a patient-specific vessel was applied. Both test results showed that optimized stents would cause much smaller vessel straightening. Moreover, vessels stented by the optimized structures had much lower stress concentration and strain energy. All those properties will decrease the possibility of ISR significantly.


Temporal changes in vegetation around a shale gas development area in a subtropical karst region in southwestern China.
Over the past decade, various aspects of China's fragile karst environments, including net primary productivity (NPP), have been changed or threatened by shale gas development. This industry is still developing, so it is important to understand what drives environmental changes, particularly in NPP, when shale gas pads are constructed in sensitive areas. Few previous studies have addressed this issue, so we quantified how the NPP changed, and what drove the changes, when a large shale gas area was developed at the end of 2012 in a mountainous karst area in Sichuan Province. We calculated the trend in the normalized difference vegetation index (NDVI) from 2012 to 2017 and used the Carnegie-Ames-Stanford Approach (CASA) model to calculate the changes in NPP at different distances from the pads using remote sensing images for July 2012 and July 2017 and field survey data from July 2017. We then identified the factors that drove the changes with Geodetector. The results showed that the NDVI increased across 64.2% of the shale gas development area from 2012 to 2017 because of climate change, and only showed a significant decrease across 0.3% of the area, mainly because of the shale gas development. The NPP decreased by 110.1 t because of the shale gas development in July 2017, or by about 0.35% of the total NPP. Of this, 93.8 t were associated with the pad construction areas, and 16.3 t were associated with the area around the pads. The changes in NPP around the shale gas pads were mainly confined to within 150 m during the construction phase and 90 m once the construction was completed. The NPP at different distances from the pads during the construction period was related to the distance from the pad, slope, and land use. Once completed, the NPP mainly varied with distance, land use, and the distance from the pad to rural settlements. The NPP was most strongly influenced by the distance from the pad and the area of the pad. We suggest that, when planning the construction of shale gas pads, the pads should be sited on gently sloping areas, the number of wells on each pad should be optimized, land use type changes outside the pad should be limited, and the land beyond the pads should be reclaimed in a timely manner to allow the NPP to recover.


Synthesis of Eu(III) and Tb(III) complexes with novel pyridine dicarboxamide derivatives and their luminescence properties.
A novel ligand, N2,N6-bis[2-(3-methylpyridyl)]pyridine-2,6-dicarboxamide (L2) and the corresponding Eu(III) and Tb(III) hydrochlorate complexes have been synthesized and characterized in detail based on elemental analysis, IR and NMR. The crystal and molecular structure of the complexes was determined by X-ray crystallography. The Eu(III) and Tb(III) ions were found to coordinate to the amido nitrogen atoms and pyridine nitrogen atoms. The luminescence properties of lanthanide complexes in solid state, in different solutions and in different pH value were investigated. The result shows that Tb(III) complexes exhibit more efficient luminescence than Eu(III) complexes, and the ligand (L2) is an excellent sensitizer to Tb(III) ion.


Ingeniously designed Silica nanostructures as an exceptional support: Opportunities, potential challenges and future prospects for viable degradation of pesticides.
Despite significant advancements in modern agricultural practices, efficient handling of pesticides is a must as they are continuously defiling our terrestrial as well as aquatic life. During the last couple of decades, substantial efforts by various research groups have been devoted to find innovative solutions to remove pesticides from our environment in a greener way. In this regard, functionalized silica nanoparticles (NPs) have gained considerable attention of scientific community due to their notable properties such as amenable design, large surface area as well as fine-tunable and uniform pore structures which make them an ideal material for pesticides removal. The present review aims to proffer current scientific progress attained by silica-based nanostructures as an excellent material for effective removal of noxious agrochemicals. Further, a brief discussion on the synthetic strategies as well as intrinsic benefits associated with different morphologies of silica have also been highlighted in this article. It also summarizes the recent reports on silica assisted degradation of pesticides via enzymatic, chemical as well as advanced oxidation protocols. Additionally, it presents a critical analysis of different support materials for decontamination of our ecosystem. The review concludes with potential challenges, their possible solutions along with key knowledge gaps and future research directions for successful deployment of silica supported materials in degradation of pesticides at commercial scale.


The Tubulin Detyrosination Cycle: Function and Enzymes.
Microtubules are subjected to a variety of post-translational modifications (PTMs). The combination of different α- and β-tubulin isoforms and PTMs are referred to as the tubulin code. PTMs are generated by a suite of enzymes thought to affect tubulin-interacting proteins. One PTM is the cyclic removal and ligation of the C-terminal tyrosine of α-tubulin. This has been implicated in cellular processes such as mitosis, cardiomyocyte contraction, and neuronal function. Recently, vasohibins (VASHs) were identified as the first tubulin-detyrosinating enzymes, A cell-autonomous role for VASHs in regulating the cytoskeleton was unexpected due to their previous association with angiogenesis. This review discusses the functionality of the tubulin detyrosination cycle, the biology of VASHs, and highlights the emerging questions accompanying this link.


Corticosteroids and circadian rhythms in the cardiovascular system.
The mineralocorticoid receptor (MR) plays a central role in cardiac physiological function and disease and is thus an attractive therapeutic target for patients with heart failure. However, the incidence of significant side effects from mineralocorticoid receptor antagonist (MRA) treatment has led to investigation of new mechanisms that may enhance MR targeted therapies. Recent studies have identified the circadian clock as a novel, reciprocal interacting partner of the MR in the heart. While the closely related glucocorticoid receptor (GR) and its ligand, cortisol (corticosterone in rodents), are established regulators of the circadian clock, new data suggest that the MR can also regulate circadian clock gene expression and timing. This review will discuss the role of the MR and its ligands in the regulation of the circadian clock in the heart and the implications of dysregulation of these systems for cardiac disease progression, and for MR activation.


Early evening ranitidine administration promotes faster duodenal ulcer healing.
In a prospective double-blind clinical trial, 141 patients with endoscopically diagnosed duodenal ulcer were randomly assigned to treatment with ranitidine 300 mg, taken either at 6 PM or at 10 PM. After 2 wk of treatment, 52 of 70 patients (74%) in the 6 PM treatment group had healed, compared with 32 of 64 patients (50%) taking ranitidine at 10 PM (p less than 0.01). After 4 wk, the cumulative healing rates were 100% and 94%, respectively, for the 6 PM and 10 PM treatment regimens. These results suggest that ranitidine, taken as a single daily 300-mg dose at 6 PM after dinner, provides more rapid duodenal ulcer healing than the same dose of the drug taken at 10 PM.


The ACA's 65th Birthday Challenge: Moving from Medicaid to Medicare.
The Affordable Care Act (ACA) expanded Medicaid to millions of low-income near-elderly Americans, facilitating access to health care services, but did not change income eligibility for Medicaid for those 65 years and older. Therefore, following the ACA's coverage expansion, many newly-insured older enrollees will face a complex insurance transition on their 65th birthday: they will lose Medicaid coverage and transition from Medicaid to Medicare as their primary insurer. This transition in primary health insurance coverage includes changes to benefits, patient cost-sharing, and provider reimbursement, which could have profound consequences on the use of health services and associated health outcomes for low-income seniors. Using data from 2012, we estimate that 1.6 million current Medicaid beneficiaries and an additional 1.6 to 2.9 million low-income individuals who will gain Medicaid coverage under the ACA will be likely to make this transition in the next decade. Primary care physicians and policymakers can help mitigate the potential consequences of this insurance transition by preparing patients for Medicare's more restrictive insurance coverage, encouraging patients to sign up for available low-income subsidies, and understanding how the loss of Medicaid coverage impacts out-of-pocket costs.


The psychosocial impact of serological herpes simplex type 2 testing in an urban HIV clinic.
Herpes simplex virus type 2 (HSV-2) is a common infection among HIV infected people. HSV type specific serologies permit the diagnosis of previously unrecognised HSV-2 infection. While substantial psychosocial morbidity has been associated with a clinical diagnosis of genital herpes, the burden associated with a serological diagnosis of HSV-2 is unclear. This study prospectively measured the psychosocial response to a new serological HSV-2 diagnosis in patients receiving care at an urban HIV clinic. At entry, sera were tested for HSV-1 and HSV-2 antibodies by western blot. Participants completed a 90 item psychosocial and life quality questionnaire at enrollment, and at 2 weeks, 3 months, and 6 months after receiving test results. Of 248 HIV infected participants, 172 (69.4%) were HSV-2 seropositive and 116 (67.4%) seropositive people did not have a previous history of genital herpes. After correction for multiple comparisons, no statistically significant differences were detected on the psychosocial and life quality scales between those who received a new HSV-2 serological diagnosis compared with those who were HSV-2 seropositive with a history of genital herpes, or those who tested HSV-2 seronegative. Additionally, no significant changes in scores were observed during follow up. HSV-2 was a common but often unrecognised infection in this urban HIV clinic and participants coped well with a positive HSV-2 result. Concerns about psychosocial burden should not deter serological testing for HSV-2. Given the epidemiological and clinical interaction between HSV-2 and HIV, these data support routine HSV-2 testing of HIV infected people.


No signs of subclinical atherosclerosis after risk-reducing salpingo-oophorectomy in BRCA1/2 mutation carriers.
BRCA1/2 mutation carriers are generally exposed to early menopause due to risk-reducing salpingo-oophorectomy (RRSO) around the age of 40 years. This risk-reducing intervention is based on a 10-40% life-time risk of ovarian cancer in this population. Although effective, premature and acute menopause induces non-cancer related morbidity in both the short and long term. Little is known about the impact of RRSO on the cardiovascular system. This cross-sectional study explored the relationship between time since RRSO and signs of subclinical atherosclerosis, as measured by carotid intima-media thickness (CIMT) and pulse wave velocity (PWV), in 165 BRCA1/2 mutation carriers. All participants, aged 40 to 63 years, underwent RRSO before the age of 45 years, and at least 5 years ago. Cardiovascular risk factors were assessed by questionnaires and a single screening visit. Data were analyzed using linear regression models. Mean CIMT was 692.7 μm (SD 87.0), and mean central PWV 6.40 m/s (SD 1.42). After adjustment for age and several relevant cardiovascular risk factors, time since RRSO was not associated with CIMT (β=0.68 μm; 95% CI -4.02, 5.38) and PWV (β=44 mm/s; 95% CI -32, 120). Compared to women of a reference group from the general population, lower systolic blood pressure [mean difference 12 mmHg; 95% confidence interval (CI) 10, 14] was found in BRCA1/2 mutation carriers. We found that, in BRCA1/2 mutation carriers, at 5 to 24 years follow-up, time since RRSO is not related to development of subclinical atherosclerosis. However, the follow-up period in these relatively young women might have been too short.


Quantification of rRNA in Escherichia coli using capillary gel electrophoresis with laser-induced fluorescence detection.
Over the past 10 years, sophisticated powerful techniques have been developed for the quantification of messenger RNA (mRNA) and ribosomal RNA (rRNA), enabling researchers in science, industry, and molecular medicine to explore gene expression. These techniques require the (reverse) transcription of analyte RNA, hybridization with synthetic oligonucleotides, and other additional steps that make them costly, time-consuming, and quantitatively difficult to perform. The current work demonstrates how 16S and 23S rRNA can be quantified precisely using capillary gel electrophoresis with laser-induced fluorescence detection (CGE-LIF) directly after the extraction of total RNA without requiring further reactions or calibration. CGE-LIF normally is used for the qualitative examination of RNA preparations. Its quantitative performance could be improved significantly using MS2 bacteriophage RNA as an internal standard. The entire analytical procedure was validated for linearity, repeatability, reproducibility, and recovery. This validation also included total RNA extraction from bacterial cells, an aspect examined for the first time in absolute RNA quantification. Recovery is close to 100%, and the analytical precision was increased 10-fold (CV<3%), as compared with similar approaches. The demonstrated method is simple and opens up new possibilities for the absolute quantification of not only rRNA but also individual mRNAs.


Topical antimicrobial agents and antibiotics for the eye.
Antibiotics for eye infection can be given topically, by subconjunctival or intravitreous injection, or systemically. The bacteria responsible are reviewed and pharmacokinetics influencing topical concentration are discussed. An approach to treatment by the primary care specialist is suggested.


Chemical eye injury: pathophysiology, assessment and management.
Chemical eye injury (CEI) is an acute emergency which can threaten sight and life. These commonly occur at home or the workplace with the former being generally mild and the latter more severe and bilateral. Major workplace accidents involve other parts of the body and can be associated with inhalation or ingestion of the chemical. Alkali injuries cause damage by saponification of tissue and deeper penetration as a consequence. Acid injuries cause rapid coagulation of tissue, which impedes penetration and limits damage. Irritants such as alcohols, cause superficial epithelial denudation. Severe chemical insult can affect all anterior segment structures causing iris, pupil and lens abnormalities. Eye pressure is variably affected and can be low or high or start as one and rapidly change to the other. Chorioretinal changes in the form of vasculopathy are seen and ascribed to be secondary to anterior segment inflammation rather than due to the direct effect of CEI. Final outcome related to structure and function is determined by the injurious agent, duration of exposure, nature of treatment and the rapidity with which it is instituted. Prevention of further damage by profuse and prolonged eye wash, after ascertaining pH of both eyes, together with exploration and removal of all particulate matter, is the key. Other management principles include a complete and thorough assessment, control of inflammation, facilitation of healing and prevention and management of sequelae and complications. Intraocular pressure is often forgotten and must be assessed and managed. Management often requires a multidisciplinary approach.


Mapping lines of non-extension in persons with lower limb amputation to aid comfort-driven prosthetic socket design.
This study aimed to develop a new technique to map the strain field for persons with lower-limb amputations to use for the design of comfortable prostheses. Using a DSLR camera with stenciled 2D markers, we demonstrated a technique to measure skin strain around the residual limb of persons with lower limb amputations. We used open-source software programs to reconstruct a series of cloud points derived from the pictures of the marked residual limb into 3D models, then calculated the minimum, maximum, and non-extension lines from directional strain fields. A DSLR camera was successful in capturing 2D markers. The maximum mean principal strain was 68% ± 14%, observed around the patella. The minimum compressive mean principal strain of -31% ± 4% was observed posteriorly in the popliteal region of the knee. Although lines of non-extension (LoNE) appear separate in different participants, they are anatomically located in regions that could be generalized for the design of prostheses. Marker locations extracted from the video of different poses can be compared to calculate strains from which the position of LoNE can be generated. The use of LoNE could be valuable in reducing discomfort at the socket interface and informing future socket design.


Impact of endoscopic therapy on outcome of operation for bleeding peptic ulcers.
Since elective surgery for bleeding peptic ulcer disease has declined, the surgical opinion is that patients who undergo emergency operation have more advanced disease and possibly a poorer outcome. We examined current mortality for surgical correction of upper gastrointestinal (UGI) bleeding from peptic ulcer disease. Between July 1, 1986, and December 31, 1990, 1,213 patients had esophagogastroduodenoscopy for UGI bleeding (659 with peptic ulcer disease, 219 with gastroesophageal varices, 152 with esophagitis, 83 with other causes, and 100 with no source found). Of 110 patients with peptic ulcer disease treated by endoscopic methods, bleeding was controlled in 90, and 20 required operation for failed endoscopic control. Another 22 patients had primary operation for exigent bleeding with diagnostic endoscopy only. The overall results in our series compared favorably with two pre-therapeutic endoscopy index series (Nottingham, 1982; University of Pittsburgh, 1982). Our operation rate was 6%, with a mortality rate of 7%, compared with operation rates of 15% to 27%, with mortality rates of 21% to 22% in the historical control series. In conclusion, we found that: (1) endoscopic control of UGI bleeding from peptic ulcer disease has decreased the incidence of operation compared with historical series; (2) overall operative mortality is decreasing; and (3) the major postoperative complication is rebleeding.


Metastatic follicular carcinoma of the thyroid to the skin: a case confirmed by immunohistochemistry.
A 78-year-old woman with a history of follicular carcinoma of the thyroid developed a painless nodule on her face. A skin biopsy showed a dermal tumor composed of cells with granular, eosinophilic cytoplasm. Immunoperoxidase staining demonstrated thyroglobulin in the tumor cells, confirming the diagnosis of metastatic thyroid carcinoma.


Human liver RNA-programmed in vitro synthesis of a polypeptide related to human apolipoprotein B.
In an in vitro synthesizing system programmed with RNA from human liver a polypeptide with an estimated Mr of 80 000 (80 kDa) +/- 1400 (mean +/- SD, n = 5) was synthesized. This polypeptide could be precipitated with antiserum to a narrow density cut of LDL (d = 1.030-1.055) or antiserum against the high-Mr form of apoB (apoB 100 [4]). The synthesized protein is immunologically related to a 75 kDa protein isolated from LDL. We suggest that the 80 kDa protein represents a primary translation product of apoB synthesized in human liver.


Stimulation with human menopausal gonadotropin versus follicle-stimulating hormone after pituitary suppression in polycystic ovarian syndrome.
Stimulation with human menopausal gonadotropin (hMG) or follicle-stimulating hormone (FSH) was compared in 34 patients with polycystic ovarian syndrome after pituitary gonadotrope suppression with buserelin acetate. No differences were seen in the hormone parameters observed. Also, the duration of the stimulation period and the dose of gonadotropin used were the same. In both groups a multifollicular response was seen. Oocyte retrieval and in vitro fertilization resulted in identical ratios of mature to total oocytes and cleavage rates. Nine pregnancies occurred, four in the hMG group and five in the FSH group. Of the nine pregnancies, two were the result of transfer of frozen-thawed embryos in estradiol and progesterone substituted cycles.


Regional health care planning: a methodology to cluster facilities using community utilization patterns.
Community-based health care planning and regulation necessitates grouping facilities and areal units into regions of similar health care use. Limited research has explored the methodologies used in creating these regions. We offer a new methodology that clusters facilities based on similarities in patient utilization patterns and geographic location. Our case study focused on Hospital Groups in Michigan, the allocation units used for predicting future inpatient hospital bed demand in the state's Bed Need Methodology. The scientific, practical, and political concerns that were considered throughout the formulation and development of the methodology are detailed. The clustering methodology employs a 2-step K-means + Ward's clustering algorithm to group hospitals. The final number of clusters is selected using a heuristic that integrates both a statistical-based measure of cluster fit and characteristics of the resulting Hospital Groups. Using recent hospital utilization data, the clustering methodology identified 33 Hospital Groups in Michigan. Despite being developed within the politically charged climate of Certificate of Need regulation, we have provided an objective, replicable, and sustainable methodology to create Hospital Groups. Because the methodology is built upon theoretically sound principles of clustering analysis and health care service utilization, it is highly transferable across applications and suitable for grouping facilities or areal units.


Control of ventilation in the hypercapnic skate Raja ocellata: II. Cerebrospinal fluid and intracellular pH in the brain and other tissues.
This study examined the possible role(s) of central acid-base stimuli in the increase in ventilation induced by hypercapnia in the skate, a response that is not due to an O2 signal (Graham et al., Respir. Physiol., 1990, 80: 251-270). Skate were sampled for cerebrospinal fluid (CSF) acid-base status, intracellular pH of the brain (14C-DMO method), and pHi in other tissues throughout 24 h of exposure to PICO2 = 7.5 Torr. CSF PCO2 rapidly equilibrated with the elevated PaCO2. Despite the much lower non-HCO3- buffer capacity in the CSF, CSF pH was not depressed to the same extent as blood pHa. CSF pH was also regulated rapidly, returning to control levels by 8-10 h, whereas pHa remained significantly depressed at 24 h. Similarly, the pHis of the weakly buffered brain and heart ventricle were initially compensated more rapidly than those of more strongly buffered white muscle and red blood cells. However, brain pHi adjustment slowed markedly after 4 h and stabilized at only 70% compensation by 20-24 h, suggesting that brain intracellular acidosis may play a role in the long-term increase in ventilation. CSF and brain were the only compartments which did not exhibit an apparent compounding metabolic acidosis during the initial stages of hypercapnic exposure. While these results illustrate the primacy of central acid-base regulation, they do not support a role for CSF pH in the long-term elevation of ventilation in response to hypercapnia. Depressions in pHa and brain pHi appear the two most likely candidates for proximate stimuli.


Measurement of Residual Dipolar Couplings of Organic Molecules in Multiple Solvent Systems Using a Liquid-Crystalline-Based Medium.
Residual dipolar coupling (RDC), a robust anisotropic NMR parameter for structural elucidation of organic molecules, is only accessible in an anisotropic environment. Herein, we introduce a novel alignment medium based on the molecular self-assembly of oligopeptide amphiphile (OPA). This medium is compatible with different intermediate and polar solvent systems, such as CD3 OD, [D6 ]DMSO, and D2 O. The preparation of the OPA-based medium is simple and rapid, while only very weak background signals were observed from OPAs. Furthermore, we show that the purity of OPA has only a minor influence on the quality of the RDC data. These advantages allow RDC measurements of organic molecules with different polarities and solubilities with high efficiency and accuracy.


[Toward the appropriate use of teleradiology].
Teleradiology involves much more than merely transmitting images and information between two points: teleradiology consists of sharing knowledge and working together in a network. It facilitates rapid access to radiological reports and second opinions, remote consulting among physicians, improved patient care, access to complex tools for postprocessing and computer-aided diagnosis, support for research and training projects, ties between isolated healthcare providers and busier or more experienced providers, 24-hour coverage, and competition among radiology departments. A close relation with the radiologist leads to better care. However, teleradiology should not have negative effects on the efficacy of the clinical radiology service that is closest to the patient. This article focuses on the legal requirements of teleradiology services and on the clinical problems that can arise in teleradiology settings, with the ultimate aim of ensuring the appropriate use of teleradiology to improve healthcare.


A Novel Way of Measuring Dual-Task Interference: The Reliability and Construct Validity of the Dual-Task Effect Battery in Neurodegenerative Disease.
Decreased automaticity is common among individuals with neurodegenerative disease and is often assessed using dual-task (DT) paradigms. However, the best methods for assessing performance changes related to DT demands remain inconclusive. To investigate the reliability and validity of a novel battery of DT measures (DT Effect-Battery (DTE-B)) encompassing three domains: task-specific interference, task prioritization, and automaticity. Data for this retrospective cross-sectional study included 125 participants with Parkinson's disease (PD), 127 participants with Alzheimer's disease (AD), and 84 healthy older adults. Reliability analyses were conducted using a subset of each population. DTE-B measures were calculated from single and DT performance on the Timed Up and Go test and a serial subtraction task. Construct validity was evaluated via associations within the DTE-B and with theoretically supported measures as well as known-groups validity analyses. Good to excellent reliability was found for DTE-B measures of task interference (motor and cognitive DT effects) (ICCs≥.658) and automaticity (combined DT effect (cDTE)) (ICCs≥.938). Evidence for convergent validity was found with associations within the hypothesized constructs. Known-groups validity analyses revealed differences in the DTE-B among the healthy group and PD and AD groups (ps≤.001), excepting task prioritization (ps≥.061). This study provides evidence to support the DTE-B as a reliable measure of multiple constructs pertinent to DT performance. The cDTE demonstrated evidence to support its validity as a measure of automaticity. Further investigation of the utility of the DTE-B in both PD and AD, as well as other populations, is warranted.


Strategies for Development of a Next-Generation Protein Sequencing Platform.
Proteomic analysis can be a critical bottleneck in cellular characterization. The current paradigm relies primarily on mass spectrometry of peptides and affinity reagents (i.e., antibodies), both of which require a priori knowledge of the sample. An unbiased protein sequencing method, with a dynamic range that covers the full range of protein concentrations in proteomes, would revolutionize the field of proteomics, allowing a more facile characterization of novel gene products and subcellular complexes. To this end, several new platforms based on single-molecule protein-sequencing approaches have been proposed. This review summarizes four of these approaches, highlighting advantages, limitations, and challenges for each method towards advancing as a core technology for next-generation protein sequencing.


On-Site and Quantitative Detection of Trace Methamphetamine in Urine/Serum Samples with a Surface-Enhanced Raman Scattering-Active Microcavity and Rapid Pretreatment Device.
Here, it reports a high-throughput detection method for reliably quantitative analysis of illegal drugs in complex biological samples by means of a surface-enhanced Raman scattering (SERS) active microcavity and rapid pretreatment device. Based on the well-made hemispherical microcavities that regularly distributed on a glass array, the quality-controllable microcavity device is fabricated by the compact self-assembly of core-shell nanopeanuts (CSNPs) onto the inside surface. Both the CSNPs with a quantifiable internal standard signal of crystal violet acetate anchored inside their gap and the well-made microcavity referred to the physical amplification of the microscale groove surface will do well in trace analysis, which will allow us to realize the accurately quantitative SERS analysis of targeted analytes spread on the bottom area of the microcavity array. As an example, 0.8 nM malachite green and 160 ppb methamphetamine (MATM) have been successively detected in a wide range as standard, while even 0.01 ppm MATM mixed in the urine/serum samples has been efficiently tested by the microcavity device equipped with a rapid pretreatment device (manual monolithic column syringe needle). All of the above suggest that the SERS-active microcavity equipped with a rapid pretreatment device has potential in the on-site quick test of trace amounts of illegal drugs in bodily fluid samples or other field analysis of food sanitation, environmental safety, and public health.


Vagal Nerve Stimulation: Indications for Revision in Adult Refractory Epilepsy.
The goal of this study was to assess the indications of revision for vagal nerve stimulation at a single institution in an adult population with drug-resistant epilepsy. This was a retrospective review of a prospectively collected database who underwent vagal nerve stimulator implantation for drug-resistant epilepsy during 1992-2017. Patients receiving vagal nerve stimulation (VNS) implants were monitored throughout their perioperative and postoperative course and were classified according to type of seizure at the time of diagnosis and indications for VNS revision. In addition, response to dysfunctional VNS devices or adverse effects were noted. Most patients receiving VNS implants were given a diagnosis of complex partial seizures (CPSs) before implantation (95.1%). Other epileptic conditions identified requiring implantation included generalized seizures, generalized-atonic seizures, Lennox-Gastaut syndrome, CPS or generalized seizures, and tuberous sclerosis (with generalized characteristics). High lead impedance was the most common indication for revision (5.6%), whereas device ineffectiveness leading to continued seizures was the most common indication for removal (2.3%). Infection, lead fracture, and dual- to single-pin lead battery changes occurred at an incidence of 1.9%, requiring either implant removal or revision. Other events that occurred, albeit rarely, included skin extrusion (0.5%), postoperative hematoma (0.5%), and implant rejection (0.5%) necessitating removal. VNS implantation in adults was shown to be a well-tolerated procedure. In addition, indications for revision or removal of the VNS device was low in this population with lead fracture rates lower than the incidence reported in pediatric population literature.


Biodiversity: invasions by marine life on plastic debris.
Colonization by alien species poses one of the greatest threats to global biodiversity. Here I investigate the colonization by marine organisms of drift debris deposited on the shores of 30 remote islands from the Arctic to the Antarctic (across all oceans) and find that human litter more than doubles the rafting opportunities for biota, particularly at high latitudes. Although the poles may be protected from invasion by freezing sea surface temperatures, these may be under threat as the fastest-warming areas anywhere are at these latitudes.


Lie Markov models with purine/pyrimidine symmetry.
Continuous-time Markov chains are a standard tool in phylogenetic inference. If homogeneity is assumed, the chain is formulated by specifying time-independent rates of substitutions between states in the chain. In applications, there are usually extra constraints on the rates, depending on the situation. If a model is formulated in this way, it is possible to generalise it and allow for an inhomogeneous process, with time-dependent rates satisfying the same constraints. It is then useful to require that, under some time restrictions, there exists a homogeneous average of this inhomogeneous process within the same model. This leads to the definition of "Lie Markov models" which, as we will show, are precisely the class of models where such an average exists. These models form Lie algebras and hence concepts from Lie group theory are central to their derivation. In this paper, we concentrate on applications to phylogenetics and nucleotide evolution, and derive the complete hierarchy of Lie Markov models that respect the grouping of nucleotides into purines and pyrimidines-that is, models with purine/pyrimidine symmetry. We also discuss how to handle the subtleties of applying Lie group methods, most naturally defined over the complex field, to the stochastic case of a Markov process, where parameter values are restricted to be real and positive. In particular, we explore the geometric embedding of the cone of stochastic rate matrices within the ambient space of the associated complex Lie algebra.


Robust correlation analyses: false positive and power validation using a new open source matlab toolbox.
Pearson's correlation measures the strength of the association between two variables. The technique is, however, restricted to linear associations and is overly sensitive to outliers. Indeed, a single outlier can result in a highly inaccurate summary of the data. Yet, it remains the most commonly used measure of association in psychology research. Here we describe a free Matlab((R)) based toolbox (http://sourceforge.net/projects/robustcorrtool/) that computes robust measures of association between two or more random variables: the percentage-bend correlation and skipped-correlations. After illustrating how to use the toolbox, we show that robust methods, where outliers are down weighted or removed and accounted for in significance testing, provide better estimates of the true association with accurate false positive control and without loss of power. The different correlation methods were tested with normal data and normal data contaminated with marginal or bivariate outliers. We report estimates of effect size, false positive rate and power, and advise on which technique to use depending on the data at hand.


Autoimmunity in common variable immunodeficiency.
Autoimmunity has been increasingly recognized as a major issue in patients with common variable immunodeficiency (CVID), the most common symptomatic primary immunodeficiency in adulthood. Different authors report high prevalences of autoimmune diseases in CVID, and several mechanisms have been proposed to explain this apparent paradox. Genetic predisposition, under current surveillance, innate and adaptive immunity deficiencies leading to persistent/recurrent infections, variable degrees of immune dysregulation, and possible failure in central and peripheral mechanisms of tolerance induction or maintenance may all contribute to increased autoimmunity. Data on the clinical/immunological profile of affected patients and treatment are available mostly concerning autoimmune cytopenias, the most common autoimmune diseases in CVID. Treatment is based on conventional alternatives, in association with short experience with new agents, including rituximab and infliximab. Benefits of early immunoglobulin substitutive treatment and hypothetical premature predictors of autoimmunity are discussed as potential improvements to CVID patients' follow-up.


Activation of c-Jun N-terminal kinase 1 and caspase 3 in the tamoxifen-induced apoptosis of rat glioma cells.
The mechanisms of the antitumor effects of tamoxifen upon gliomas are still unclear. In this study, we investigated the role of c-Jun N-terminal kinase-1 (JNK1) and caspase 3 in the tamoxifen-induced apoptosis of rat glioma cells. Glioma cells were treated with tamoxifen, followed by a cytotoxicity assay to study its effects on the cells, and then a flow-activated cell sorter (FACS) analysis was performed to analyze the cellular apoptosis of the glioma cells. The expression of JNK1 and phospho-specific JNK1 in glioma cells treated with tamoxifen was investigated by Western blot analysis. The activity of caspase 3 in glioma cells was analyzed by caspase activity assay. Tamoxifen was demonstrated to exert cytotoxic effects upon and induced apoptosis of the glioma cells in a concentration- and time-dependent manner (P<0.05). Western blot analysis demonstrated that tamoxifen increased the expression of phospho-specific JNK1 in glioma cells, and an increasing concentration of tamoxifen induced an increasing expression of phospho-specific JNK1. Four-hour 50-microM tamoxifen treatment increased the expression of phospho-specific JNK1 to 3.2 times that of the control level in glioma cells. Tamoxifen also increased the activity of caspase 3 in glioma cells. Pretreatment of glioma cells with the antisense oligonucleotide (OGN) of JNK1 immediately prior to tamoxifen treatment suppressed the expression of phospho-specific JNK1 and the activity of caspase 3. The apoptosis fraction of glioma cells induced by 4-h treatment with 50 microM tamoxifen was decreased from 51% to 28% by pretreatment with the antisense OGN of JNK1 (P<0.003), and to 20% by pretreatment with caspase 3 inhibitor (DEVD-CHO) (P<0.0008). The results suggest that the tamoxifen-induced apoptosis of rat glioma cells is related to the activation of the JNK1/caspase 3 signaling pathway; however, the confirmation of the occurrence of such activation in vivo needs further investigation.


Endoscopic Ultrasound-Guided Laser Ablation Using a Diffusing Applicator for Locally Advanced Pancreatic Cancer Treatment.
Endoscopic ultrasound (EUS)-guided cylindrical interstitial laser ablation (CILA) procedures can be used to treat unresectable pancreatic cancer (PC). The aim of this study was to investigate the acute responses of pancreatic tissue after EUS-guided CILA in vivo in porcine models. Eight pigs were tested to compare the effects of different energy levels on pancreatic tissue ablation. A 1064 nm laser system was used to deliver 5 W through a diffusing applicator. The EUS-guided CILA was performed under four different energies: 200, 400, 600, and 800 J. Three days after the experiments, histological analysis was performed. The CILA consistently generated circular coagulated necrosis (CN) in the cross-sectioned pancreatic tissue. The ablation diameter was linearly dependent on the total energy delivery. The area of the CN initially increased with total energy delivery but became saturated at 600 J. The width of the degenerative parenchyma (DP) in the native tissue beyond the CN region increased with the total energy up to 600 J, and then decreased afterward. EUS-guided CILA can be a feasible approach for treating PC. Further animal studies will investigate the chronic responses of the pancreatic tissue to examine the efficacy and safety of the proposed method for clinical translation.


Relation of diet-induced thermogenesis to brown adipose tissue activity in healthy men.
Human brown adipose tissue (BAT) is a thermogenic tissue activated by the sympathetic nervous system in response to cold exposure. It contributes to energy expenditure (EE) and takes up glucose and lipids from the circulation. Studies in rodents suggest that BAT contributes to the transient rise in EE after food intake, so-called diet-induced thermogenesis (DIT). We investigated the relationship between human BAT activity and DIT in response to glucose intake in 17 healthy volunteers. We assessed DIT, cold-induced thermogenesis (CIT), and maximum BAT activity at three separate study visits within 2 wk. DIT was measured by indirect calorimetry during an oral glucose tolerance test. CIT was assessed as the difference in EE after cold exposure of 2-h duration as compared with warm conditions. Maximal activity of BAT was assessed by 18-F-fluoro-deoxyglucose (18F-FDG) 18F-FDG-PET/MRI after cold exposure and concomitant pharmacological stimulation with mirabegron. Seventeen healthy men (mean age = 23.4 yr, mean body mass index = 23.2 kg/m2) participated in the study. EE increased from 1,908 (±181) kcal/24 h to 2,128 (±277) kcal/24 h (P < 0.0001, +11.5%) after mild cold exposure. An oral glucose load increased EE from 1,911 (±165) kcal/24 h to 2,096 (±167) kcal/24 h at 60 min (P < 0.0001, +9.7%). The increase in EE in response to cold was significantly associated with BAT activity (R2 = 0.43, P = 0.004). However, DIT was not associated with BAT activity (R2 = 0.015, P = 0.64). DIT after an oral glucose load was not associated with stimulated 18F-FDG uptake into BAT, suggesting that DIT is independent from BAT activity in humans (Clinicaltrials.gov Registration No. NCT03189511).NEW & NOTEWORTHY Cold-induced thermogenesis (CIT) was related to BAT activity as determined by FDG-PET/MRI after stimulation of BAT. Diet-induced thermogenesis (DIT) was not related to stimulated BAT activity. Supraclavicular skin temperature was related to CIT but not to DIT. DIT in humans is probably not a function of BAT.


Novel dynamic measures of emetic behavior in musk shrews.
The emetic reflex occurs as a pattern of motor responses produced by a network of neurons in the hindbrain. Despite an understanding of the sequence of motor outputs that form an emetic episode (EE), the variability in the dynamics of multiple EEs across time remains a mystery. Many clinical investigations rely on once a day patient recall of total amount of vomiting, and preclinical studies frequently report only the total number of EE per unit time. The aim of the current study was to develop novel temporal measures of emetic activation in a preclinical model. Male and female musk shrews were tested with prototypical emetic stimuli: motion exposure (1 Hz), nicotine (5 mg/kg, sc), and copper sulfate (120 mg/kg, ig). New emetic measures included duration (time from first to last episode), rate, standard deviation of the inter-episode interval (SD-I), and a survival analysis of emetic latency (analyzed with Cox regression). Behavioral patterns associated with emesis were also assessed using statistical temporal pattern (T-pattern) analysis to measure nausea-like behaviors (e.g., immobility). The emetic stimuli produced different levels of total EE number, duration, rate, and SD-I. A typical antiemetic, the neurokinin 1 receptor antagonist CP-99,994, suppressed the number of EEs but was less effective for reducing the duration or prolonging the emetic latency. Overall, the current study shows the use of novel dynamic behavioral measures to more comprehensively assess emesis and the impact of therapies.


Antibody assay of rabbit sera for outer membrane vesicles of Haemophilus influenzae by enzyme-linked immunosorbent assay.
Enzyme-linked immunosorbent assay (ELISA) was applied to the detection of IgG and IgM antibodies against outer membrane vesicles (OMV) antigen of Haemophilus influenzae type b. In this ELISA system, IgG antibody titers were about 40 fold higher than those in indirect hemagglutination assay (IHA). The IgG antibody titers by this ELISA of rabbit sera obtained after immunization were comparable with those by radioimmunoassay (RIA) of the same sera. A significant correlation was established between these two assays (r = 0.973, P less than 0.001).


Nondiarrheal disorders of the gastrointestinal tract in neonatal foals.
Neonates can have a variety of gastrointestinal disorders, primary and secondary in nature. Important primary disorders include con-genital abnormalities and meconium retention. One of the most important secondary lesions is generalized ileus. Gastric ulceration can occur as a primary or secondary event. This article addresses the pathophysiology, diagnosis, and treatment of gastrointestinal problems commonly observed in neonatal foals.


Identification of new type 2B von Willebrand disease mutations: Arg543Gln, Arg545Pro and Arg578Leu.
We report the identification in five patients (three families) affected with type 2B von Willebrand disease (VWD) of three heterozygous nucleotide substitutions at the codon for arginine 543, 545 and 578 of the mature von Willebrand factor (VWF) subunit resulting in a glutamine, proline and leucine substitution, respectively. These mutations are located in the A1 loop where prevalent type 2B mutations (Arg543Trp, Arg545Cys and Arg578Gln) have been already identified at the same positions. By in vitro mutagenesis of full-length cDNA of VWF and transient expression in Cos-7 cells, we have shown that the six corresponding mutated recombinant VWFs (Gln543, Trp543, Cys545, Pro545, Leu578 and Gln578 rVWF) exhibited quantitatively normal expression and normal multimeric pattern but increased ristocetin- and botrocetin-induced binding to platelets as compared with that for wild-type rVWF. The two mutations at position 545 induced the greatest reactivity for GPIb of corresponding rVWFs as compared to the two mutations at positions 543 and 578.


[Paget's cancer of male breast].
The case of a male patient with Paget's cancer of the breast is reported. Radical mastectomy was performed. Two and a half years later the patient was free of symptoms.


Analysis of cell movement between skin and other anatomical sites in vivo using photoconvertible fluorescent protein "Kaede"-transgenic mice.
Clarification of the spatiotemporal regulation and function of immune cells within the skin is critical to the understanding of the role of immune cells and the skin in immune homeostasis. Here, we describe a novel assay system for monitoring cell movements in the entire body using the photoconvertible fluorescent protein "Kaede"-transgenic (Tg) mice. We can label immune cells by the change in color of Kaede from green to red in these cells following exposure to violet light and track these cells in the entire body. The Kaede-Tg system is an ideal tool for monitoring precise cellular movements between the skin and other anatomical sites in vivo at different stages of the immune response.


Autoimmune effector cells. V.A monoclonal antibody specific for rat helper T lymphocytes inhibits adoptive transfer of autoimmune encephalomyelitis.
The in vitro activation of spleen cells from donor rats immunized with myelin basic protein results in enhanced adoptive transfer of experimental allergic encephalomyelitis to syngeneic recipients. In this study, adoptive transfer was inhibited when monoclonal antibody W3/25, which defines an antigen on rat helper T lymphocytes, was included in the in vitro cultures. Partial inhibition was achieved with monoclonal antibody OX-3, which recognizes la antigen, whereas other anti-T cell monoclonal reagents were ineffective. These results support the conclusion that W3/25+T cells play an essential role in the induction of this autoimmune response.


Metformin, testosterone, or both in men with obesity and low testosterone: A double-blind, parallel-group, randomized controlled trial.
Men with obesity tend to be insulin resistant and often have low-normal testosterone concentrations. We conducted a clinical trial aimed to evaluate potential therapeutic strategies for low testosterone in men with obesity. We did a 1-year, parallel, randomized, double-blind, placebo-controlled trial, where we evaluated the independent and combined effects of metformin and testosterone in 106 men with obesity, aged 18-50 years, who had low levels of testosterone and no diabetes mellitus. The primary outcome was change in insulin resistance, measured as Homeostasis Model Assessment for Insulin Resistance (HOMA-IR) index. Secondary outcomes included changes in total and free serum testosterone, body composition, metabolic variables, erectile function, and health-related quality of life (HRQoL). In the intention-to-treat analysis, the HOMA-IR index decreased significantly in all active groups compared to placebo (metformin -2.4, 95 % CI -4.1 to -0.8, p = 0.004; testosterone -2.7, 95 % CI -4.3 to -1.1, p = 0.001; combination -3.4, 95 % CI -5.0 to -1.8, p < 0.001). Combination therapy was not superior to testosterone alone in decreasing insulin resistance (-0.7, 95 % CI -2.3 to 0.9, p = 0.383). Only the combination of metformin plus testosterone significantly increased total and free testosterone concentrations, compared to placebo. No significant changes in body composition (except for a higher decrease in fat mass in the metformin and combination group), metabolic variables, erectile function, or HRQoL were found with any treatment. Among men with obesity and low testosterone concentrations, the combination of metformin plus testosterone, metformin only, and testosterone only, compared to placebo, reduced insulin resistance with no evidence of additive benefit.


Obesity as malnutrition: the role of capitalism in the obesity global epidemic.
The global obesity epidemic remains poorly understood, partly because it has emerged alongside persisting under-nutrition in many populations. At an abstract level, obesity develops from exposure to the "obesogenic niche," comprising diverse factors predisposing to weight gain. This article first explores how susceptibility to the obesogenic niche is influenced by developmental and life-history experience. Human growth is sensitive to early-life ecological conditions, under the transducing effect of maternal phenotype. Such plasticity is associated with subsequent variability in body composition and metabolism, impacting susceptibility to the obesogenic niche, albeit with heterogeneity across populations. Both nutritional constraint and nutritional excess during early life are associated with variability in relevant molecular pathways. The article then considers the fundamental contribution of capitalist economics to population under-nutrition and over-nutrition. Historically, capitalism contributed to the under-nutrition of many populations through demand for cheap labor. As the limiting factor for economic growth switched to consumption, capitalism has increasingly driven consumer behavior inducing widespread over-nutrition. In populations undergoing nutritional transition, many individuals encounter both under- and over-nutrition within the life course, elevating both susceptibility and exposure to the obesogenic niche. The interactions between global economic forces and nutritional shifts are distributed across generations, and are strongly transduced by maternal effects. The structural connections between undernourished and overnourished worldwide and between under- and over-nutrition within individual life-courses highlight the central role of capitalist economics in the global obesity epidemic. Prevention policies targeting individual behavior have proved ineffective and economic policies are arguably the optimal target for intervention.


Case control study of prognostic markers and disease outcome in inflammatory carcinoma breast: a unique clinical experience.
Inflammatory breast carcinoma (IBC) is a rare but aggressive form of breast cancer. In this first-ever study, we investigated the role of nine prognostic markers' expression (estrogen receptor [ER], progesterone receptor [PR], p53, C-erbB-2, epidermal growth factor receptor [EGFR], cathepsin D [CD], proliferating cell nuclear antigen [PCNA], DNA ploidy, and S-phase fraction [SPF]) and disease outcome in IBC cases compared with the control group. A case control study of IBC was conducted on 40 test cases with two controls per case matching age, grade, and number of axillary lymph nodes sampled. During 7 years of this study, 10% of all patients with breast cancer had IBC. In this study, 84% of IBC cases showed positive axillary lymph nodes compared with 63% in control group. The expression of nine prognostic markers, that is, ER, PR, p53, C-erbB-2, EGFR, CD, PCNA, SPF, and DNA ploidy, was studied by immunohistochemistry and flow cytometry. Hormone receptor status showed an inverse correlation (p < 0.05). Among p53, C-erbB-2, EGFR, and CD in the IBC group, only p53 showed a significant correlation, with 70% positivity in IBC versus 48% positivity in the control group (p < 0.05). Much higher SPF and PCNA positivity was seen in the IBC group compared with the control group (p < 0.05). DNA ploidy also showed a significant correlation compared with the control group (p < 0.05). After a median follow up of 18 months, median overall survival in the IBC group was 1.8 years (range 0.6-5.8 years) compared with 3.0 years (range 2.5-7.0 years), with a p value of 0.0001.


Rescue of Alzheimer's disease phenotype in a mouse model by transplantation of wild-type hematopoietic stem and progenitor cells.
Alzheimer's disease (AD) is the most prevalent cause of dementia; microglia have been implicated in AD pathogenesis, but their role is still matter of debate. Our study showed that single systemic wild-type (WT) hematopoietic stem and progenitor cell (HSPC) transplantation rescued the AD phenotype in 5xFAD mice and that transplantation may prevent microglia activation. Indeed, complete prevention of memory loss and neurocognitive impairment and decrease of β-amyloid plaques in the hippocampus and cortex were observed in the WT HSPC-transplanted 5xFAD mice compared with untreated 5xFAD mice and with mice transplanted with 5xFAD HSPCs. Neuroinflammation was also significantly reduced. Transcriptomic analysis revealed a significant decrease in gene expression related to "disease-associated microglia" in the cortex and "neurodegeneration-associated endothelial cells" in the hippocampus of the WT HSPC-transplanted 5xFAD mice compared with diseased controls. This work shows that HSPC transplant has the potential to prevent AD-associated complications and represents a promising therapeutic avenue for this disease.


Changes of adenine nucleotides content and release reaction of human blood platelets following gamma irradiation.
The aim of this work was to examine the effect of gamma irradiation on the energy metabolism and physiological functions of blood platelets. Blood platelets were irradiated with a 60Co-source in the range of 0.5--8.0 krad. Before and after irradiation, the free nucleotide content of platelets and the ability of platelets to perform their hemostatic functions (the release reaction with thrombin) were determined. The obtained results demonstrated that: 1. ATP, ADP and AMP content reached minimum values at 1.5--2.0 krad. 2. The ability of platelets to perform the release reaction correlates with the ATP level at doses of 1.5--2.0 krad. At higher doses (6.0--8.0) disturbances of the release reaction, indicating damage to the platelet plasma membranes, were observed.


Antithrombotic and antiplatelet effects of plant-derived compounds: a great utility potential for primary, secondary, and tertiary care in the framework of 3P medicine.
Thromboembolism is the third leading vascular disease, with a high annual incidence of 1 to 2 cases per 1000 individuals within the general population. The broader term venous thromboembolism generally refers to deep vein thrombosis, pulmonary embolism, and/or a combination of both. Therefore, thromboembolism can affect both - the central and peripheral veins. Arterial thromboembolism causes systemic ischemia by disturbing blood flow and oxygen supply to organs, tissues, and cells causing, therefore, apoptosis and/or necrosis in the affected tissues. Currently applied antithrombotic drugs used, e.g. to protect affected individuals against ischemic stroke, demonstrate significant limitations. For example, platelet inhibitors possess only moderate efficacy. On the other hand, thrombolytics and anticoagulants significantly increase hemorrhage. Contextually, new approaches are extensively under consideration to develop next-generation antithrombotics with improved efficacy and more personalized and targeted application. To this end, phytochemicals show potent antithrombotic efficacy demonstrated in numerous in vitro, ex vivo, and in vivo models as well as in clinical evaluations conducted on healthy individuals and persons at high risk of thrombotic events, such as pregnant women (primary care), cancer, and COVID-19-affected patients (secondary and tertiary care). Here, we hypothesized that specific antithrombotic and antiplatelet effects of plant-derived compounds might be of great clinical utility in primary, secondary, and tertiary care. To increase the efficacy, precise patient stratification based on predictive diagnostics is essential for targeted protection and treatments tailored to the person in the framework of 3P medicine. Contextually, this paper aims at critical review toward the involvement of specific classes of phytochemicals in antiplatelet and anticoagulation adapted to clinical needs. The paper exemplifies selected plant-derived drugs, plant extracts, and whole plant foods/herbs demonstrating their specific antithrombotic, antiplatelet, and fibrinolytic activities relevant for primary, secondary, and tertiary care. One of the examples considered is antithrombotic and antiplatelet protection specifically relevant for COVID-19-affected patient groups.


Extracellular microvesicles and invadopodia mediate non-overlapping modes of tumor cell invasion.
Tumor cell invasion requires the molecular and physical adaptation of both the cell and its microenvironment. Here we show that tumor cells are able to switch between the use of microvesicles and invadopodia to facilitate invasion through the extracellular matrix. Invadopodia formation accompanies the mesenchymal mode of migration on firm matrices and is facilitated by Rac1 activation. On the other hand, during invasion through compliant and deformable environments, tumor cells adopt an amoeboid phenotype and release microvesicles. Notably, firm matrices do not support microvesicle release, whereas compliant matrices are not conducive to invadopodia biogenesis. Furthermore, Rac1 activation is required for invadopodia function, while its inactivation promotes RhoA activation and actomyosin contractility required for microvesicle shedding. Suppression of RhoA signaling blocks microvesicle formation but enhances the formation of invadopodia. Finally, we describe Rho-mediated pathways involved in microvesicle biogenesis through the regulation of myosin light chain phosphatase. Our findings suggest that the ability of tumor cells to switch between the aforementioned qualitatively distinct modes of invasion may allow for dissemination across different microenvironments.


Cardiac safety, efficacy, and correlation of serial serum HER2-extracellular domain shed antigen measurement with the outcome of the combined trastuzumab plus CMF in women with HER2-positive metastatic breast cancer: results from the EORTC 10995 phase II study.
Cardiotoxicity is a side effect of trastuzumab. We assessed efficacy and cardiac safety of CMF with trastuzumab (CMF+T) in HER2-positive metastatic breast cancer patients (MBC). In this phase II study, centrally confirmed, previously treated HER2-positive MBC patients with measurable disease (per RECIST v 1.0) were enrolled. Initially, patients were randomized between 8 CMF cycles alone or combined with trastuzumab during chemotherapy, followed by 3-weekly trastuzumab maintenance till progression. A protocol amendment dropped the CMF arm and thereafter all patients received CMF+T. Translational research for prediction of treatment benefit was performed through serial serum HER2-shed antigen assessments. Ninety patients (CMF: 19; CMF+T: 71) were enrolled between 2002 and 2006. Median age was 54 years. 42 patients had prior chemotherapy (33 with anthracyclines) and 41/71 patients who received CMF+T continued trastuzumab monotherapy for a median duration of 40 weeks. Overall response rate was 50% for CMF+T (35/70) and 32% for CMF (6/19). Median duration of response was 10.3 months and 5.4 months, respectively. Median progression-free survival was 9.4 months (95% CI 8.1-11.6) and 4.8 months (95% CI 2.8-7.9), respectively. In the CMF+T arm, 13(18%) patients had an absolute LVEF decline, including 3 patients developing any grade of New York Heart Association cardiac dysfunction. Patients with an increase of 30% over baseline shed antigen had a higher progression risk (95% CI 7.6, 3.9-14.8). CMF+T is effective, with an acceptable cardiotoxicity profile. LVEF declines were mostly asymptomatic and occurred irrespective of previous anthracycline exposure. CMF+T can be considered for these patients, if other cytotoxics are contraindicated.


[Prostate cancer treated with androgen deprivation therapy: Care and monitoring in daily practice].
Our purpose was to identify measures implemented by urologists and radiation oncologists at the initiation of a 6-month formulation of luteinizing hormone releasing hormone (LHRH) agonist in patients with advanced PCa. This cross-sectional cohort survey was conducted during 6 months. Participants completed a questionnaire of 15 items on the first prescription of an androgen deprivation therapy (ADT), the parameters prescribed for monitoring and information provided to patients. The median age of the 1100 enrolled patients was 75 years (range: 51-98 years); 245 patients (29.0%) were metastatic and 411 (39.4%) had a Gleason score ≥ 8. Prior to the treatment initiation, the dosage of the total testosterone was not very often performed (4.8%). Associated comorbidities such as arterial hypertension (53.6%) and hypercholesterolemia (31.8%) did not constitute a barrier to the initiation of ADT, alone (60.5%) or combined with anti-androgens (61%). According to the recommendations of the French Association of Urology (AFU), fasting glycemia was required in 427 patients (41.1%), lipid profile in 380 (36.1%), a blood count in 219 (21.1%) and bone densitometry in 111 (10.8%). The advice given to patients involved diet and lifestyle rules (61%). The potential risks of adverse events (AEs) mentioned were mainly hot flashes (95.5%). Some recommendations seem insufficiently followed by the French specialists on information and monitoring procedures of ADT, especially in the cardiovascular field.


Immunohistochemical localization of substance P, vasoactive intestinal polypeptide and gastrin-releasing peptide in vas deferens and seminal vesicle, and the effect of these and eight other neuropeptides on resting tension and neurally evoked contractile activity.
Immunohistochemical studies of the vas deferens and seminal vesicle of mouse, guinea-pig, and rabbit showed the presence of nerve fibres containing vasoactive intestinal polypeptide (VIP), substance P (SP), and gastrin-releasing peptide (GRP) supplying the smooth muscle layers as well as blood vessels. The nerve supply was better developed in the seminal vesicle than in the vas deferens. The motor activity of the vas deferens and seminal vesicle of the guinea-pig was studied in vitro. The vas deferens responded to transmural electrical stimulation with a twitch followed by a slow contraction. The twitch was blocked by guanethidine and tetrodotoxin, but not by atropine, propranolol, phenoxybenzamine, or fluphenazine. The slow contraction exhibited features of an alpha-receptor-mediated response. SP, physalaemin and eledoisin contracted the smooth muscle and also potentiated the twitch response to electrical nerve stimulation in a concentration-dependent manner. The SP blocking agent, (D-Pro2,D-Trp7,9)-SP, affected neither the resting tension nor the response to electrical stimulation. It is therefore suggested that the SP fibres act mainly prejunctionally. VIP, Leu-enkephalin, cholecystokinin octapeptide (CCK-8), angiotensin II, vasopressin, neurotensin, bombesin, and GRP had no effect on either the resting tension or the response to electrical nerve stimulation. The seminal vesicle responded to electrical stimulation with a contraction which was unimpaired by atropine, propranolol, phenoxybenzamine, and guanethidine, but abolished by tetrodotoxin. Hence, this contraction is mediated by a non-adrenergic, non-cholinergic neurotransmitter. Bombesin, GRP, SP, physalaemin and eledoisin contracted the smooth muscle and potentiated the response to electrical stimulation. VIP, Leu-enkephalin, CCK-8, angiotensin II, vasopressin, and neurotensin had no effect on the resting tension or on the response to transmural electrical stimulation. The SP antagonist abolished the contraction elicited by SP but did not influence the response to nerve stimulation. The results suggest that the SP and GRP nerves may have prejunctional and facilitating postjunctional effects in the seminal vesicle.


Epigenetics and Control of RNAs.
Histone modifications are epigenetic marks that fundamentally impact the regulation of gene expression. Integrating histone modification information in the analysis of gene expression traits (eQTL mapping) has been shown to significantly enhance the prediction of eQTLs. In this chapter, we describe (1) how to perform quantitative trait locus (QTL) analysis using histone modification levels as traits and (2) how to integrate these data with information on RNA expression for the elucidation of the epigenetic control of transcript levels. We will provide a comprehensive introduction into the topic, describe in detail how ChIP-seq data are analyzed and elaborate on how to integrate ChIP-seq and RNA-seq data from a segregating disease animal model for the identification of the epigenetic control of RNA expression.


Dibutyl phthalate: maternal effects versus fetotoxicity.
Dibutyl phthalate (DBP), a plasticizer, is a teratogen in mice and rabbits but produces fetal loss in the rat. Long-term dosing studies indicating reduced fertility in the rat suggested a maternal effect of the compound. The decidual cell response (DCR) and pregnant rats were used to examine whether DBP affects maternal physiological parameters independent of the compound's fetotoxic effect. DBP has no effect on the DCR, pregnant uterine weight, number of implantation sites, ovarian weight, or serum progesterone concentration during early pregnancy or pseudopregnancy. These data show that short-term dosing with DBP has no direct maternal effect in the rat and suggest that the viability of preimplantation embryos is not compromised.


In vivo characterization of [18F]AVT-011 as a radiotracer for PET imaging of multidrug resistance.
Multidrug resistance (MDR) impedes cancer treatment. Two efflux transporters from the ATP-binding cassette (ABC) family, ABCB1 and ABCG2, may contribute to MDR by restricting the entry of therapeutic drugs into tumor cells. Although a higher expression of these transporters has been correlated with an unfavorable response to chemotherapy, transporter expression does not necessarily correlate with function. In this study, we characterized the pharmacological properties of [18F]AVT-011, a new PET radiotracer for imaging transporter-mediated MDR in tumors. AVT-011 was radiolabeled with 18F and evaluated with PET imaging in preclinical models. Transport of [18F]AVT-011 by ABCB1 and/or ABCG2 was assessed by measuring its uptake in the brains of wild-type, Abcb1a/b-/-, and Abcg2-/- mice at baseline and after administration of the ABCB1 inhibitor tariquidar (n = 5/group). Metabolism and biodistribution of [18F]AVT-011 were also measured. To measure ABCB1 function in tumors, we performed PET experiments using both [18F]AVT-011 and [18F]FDG in mice bearing orthotopic breast tumors (n = 7-10/group) expressing clinically relevant levels of ABCB1. At baseline, brain uptake was highest in Abcb1a/b-/- mice. After tariquidar administration, brain uptake increased 3-fold and 8-fold in wild-type and Abcg2-/- mice, respectively, but did not increase further in Abcb1a/b-/- mice. At 30 min after injection, the radiotracer was > 90% in its parent form and had highest uptake in organs of the hepatobiliary system. Compared with that in drug-sensitive tumors, uptake of [18F]AVT-011 was 32% lower in doxorubicin-resistant tumors with highest ABCB1 expression and increased by 40% with tariquidar administration. Tumor uptake of [18F]FDG did not significantly differ among groups. [18F]AVT-011 is a dual ABCB1/ABCG2 substrate radiotracer that can quantify transporter function at the blood-brain barrier and in ABCB1-expressing tumors, making it potentially suitable for clinical imaging of ABCB1-mediated MDR in tumors.


Effect of plasma membranes on solute transport in 3T3 cells.
Sparse cultures of Swiss 3T3 cells are arrested early in the G1 phase of growth by the addition of a plasma membrane fraction obtained from confluent 3T3 cells. We have examined whether the changes in solute transport which are usually associated with cessation of growth at confluency also take place when cell growth is arrested by the addition of plasma membranes. We find that the rate of uptake of alpha-aminoisobutyric acid and uridine is decreased after the addition of plasma membranes to 3T3 cells, but the rate of uptake of 2-deoxyglucose and phosphate is not. We conclude from these observations that uptake of uridine and alpha-aminoisobutyric acid are related to contact inhibition of growth, while the decline in the rate of uptake of 2-deoxyglucose and phosphate observed at high cell density must be due to changes other than cell to cell contact.


Differential Diagnosis of Periapical Radiopacities and Radiolucencies.
Periapical pathoses represent changes noted at the apices of teeth within the alveolar process that are suspected on examination, visualized via imaging, and confirmed via histopathology. They can be bone forming or destructive. There are pathologic entities that show both types of changes at the apical regions. These lesions must be identified if they are odontogenic in origin because treatment modalities differ. This article discusses identification of radiopaque and radiolucent lesions noted on radiographs. The common opacities and lucencies are described. When required, advanced imaging is used to depict changes within the bone near the periapical regions of mandibular and maxillary teeth.


CSF protein biomarkers predicting longitudinal reduction of CSF β-amyloid42 in cognitively healthy elders.
β-amyloid (Aβ) plaque accumulation is a hallmark of Alzheimer's disease (AD). It is believed to start many years prior to symptoms and is reflected by reduced cerebrospinal fluid (CSF) levels of the peptide Aβ1-42 (Aβ42). Here we tested the hypothesis that baseline levels of CSF proteins involved in microglia activity, synaptic function and Aβ metabolism predict the development of Aβ plaques, assessed by longitudinal CSF Aβ42 decrease in cognitively healthy people. Forty-six healthy people with three to four serial CSF samples were included (mean follow-up 3 years, range 2-4 years). There was an overall reduction in Aβ42 from a mean concentration of 211-195 pg ml(-1) after 4 years. Linear mixed-effects models using longitudinal Aβ42 as the response variable, and baseline proteins as explanatory variables (n=69 proteins potentially relevant for Aβ metabolism, microglia or synaptic/neuronal function), identified 10 proteins with significant effects on longitudinal Aβ42. The most significant proteins were angiotensin-converting enzyme (ACE, P=0.009), Chromogranin A (CgA, P=0.009) and Axl receptor tyrosine kinase (AXL, P=0.009). Receiver-operating characteristic analysis identified 11 proteins with significant effects on longitudinal Aβ42 (largely overlapping with the proteins identified by linear mixed-effects models). Several proteins (including ACE, CgA and AXL) were associated with Aβ42 reduction only in subjects with normal baseline Aβ42, and not in subjects with reduced baseline Aβ42. We conclude that baseline CSF proteins related to Aβ metabolism, microglia activity or synapses predict longitudinal Aβ42 reduction in cognitively healthy elders. The finding that some proteins only predict Aβ42 reduction in subjects with normal baseline Aβ42 suggest that they predict future development of the brain Aβ pathology at the earliest stages of AD, prior to widespread development of Aβ plaques.


Chemo-embolization of experimental liver metastases. Part I: distribution of biodegradable microspheres of different sizes in an animal model for the locoregional therapy.
An experimental in vivo rat model was established to simulate the embolization therapy of non-resectable liver metastases with microparticulate systems. The effects of biodegradable microspheres on a transplanted liver-tumor cell line in rat livers were investigated with respect to particle size. The distribution of fluorescence-labelled microspheres was investigated to characterize the effects of long-term embolization with biodegradable microspheres, and the suitability and relevance of this animal model. Novikoff hepatoma cells were implanted into the central liver lobe of Sprague-Dawley rats. After seven days, four batches of fluorescence-marked microspheres (17 microm, 25 microm, 30 microm, and 40 microm) were administered into the proper hepatic artery. Liver(including the tumor), lung and spleen were isolated and frozen sections and tissue cubes of liver-, border-, and tumor tissue were prepared. The sections were examined by fluorescence microscopy. The cubes were extracted and the fluorescence of the marker quantified. During operation, microspheres smaller than 40 microm did not cause complete embolization. The slides showed the spreading of the smaller particles to the spleen and lung. Only 40 microm particles accumulated in the liver and were rarely detectable in other organs. The extraction showed a high concentration of the 40 microm particles in the border tissue. Apparently they were trapped proximal to the tumor capillary system and therefore showed a high ratio of border/tumor concentration. Microparticles smaller than 40 microm had a border/tumor concentration ratio of less than one and were distributed to the spleen and lung. In conclusion, a mean particle diameter of at least 40 microm is required for embolization. Thus, the model shows similarity to clinical situations in the treatment of human liver tumor and metastases.


Integrated analysis of industrial limitations and image quality: an end-to-end design approach.
There is a trend in optical system design toward explicitly considering real-world industrial demands in the metrics to be optimized, from which emerges a cost-performance trade-off. Another relevant recent tendency is the so-called end-to-end design, where the design metric is an expected quality index of the final image, after digital restoration. We propose an integrated approach for analyzing the cost-performance trade-off in end-to-end designs. We exemplify it with a simple optical model where the cost is determined by the inclusion of an aspherical surface. We show that the resulting optimal trade-off configurations when applying an end-to-end design are substantially different from a conventional design. Such differences, as well as the increase in performance, are especially significant for lower-cost configurations.


Evidence against HLA and immunological dependence of disease outbreak in SLE. Immunological characterisation of identical twins clinically discordant for SLE.
Identical female twins clinically discordant for 20 years for SLE were studied. Their HLA-haplotype was A1,28; B8w6,w35; Cw3,w7; Dr3,4. Both twins had a raised erythrocyte sedimentation rate, autoantibodies, and circulating immune complexes. The diseases sibling had a reversed OKT4/OKT8 ratio (0.43), decreased helper T cell number, defective pokeweed mitogen (PWM) induced plasma cell differentiation, and overactive hydrocortisone sensitive suppressor cells. Immunological abnormalities may be only partly HLA related (B8; Dr3), but are most probably secondary to the disease process in the sibling with SLE. Exogenous and/or endogenous factor(s) other than genetic or immunological are suggested as being operative in the predisposition to and expression of SLE.


Otorhinolaryngologic Problems in Liver and Kidney Transplant Recipients.
The aim of this study was to define the otorhinolaryngologic manifestations and clinical characteristics of patients who received kidney or liver transplants at a university hospital. Medical records of patients who received a kidney or liver transplant between 2000 and 2013 and who were referred or applied to the ear, nose, and throat clinic were retrospectively reviewed. Otorhinolaryngologic complaints, signs, examination findings, and diagnoses of patients were noted. Our analyses included 540 visits to the ear, nose, and throat clinic by 101 liver and 191 kidney transplant recipients. Mean duration between date of transplant and otorhinolaryngologic examination was 747.9 ± 37.1 days. The most common complaint was rhinorrhea (n = 112), whereas the most common diagnosis was acute rhinosinusitis (n = 85). Acute upper respiratory tract infections, including rhinosinusitis, were diagnosed more frequently during the late postoperative period (ie, > 180 days after transplant). Epistaxis was more frequent during the first 30 days after transplant. A diverse variety of otorhinolaryngologic conditions, including emergencies and potentially life-threatening infections, were seen in our kidney and liver transplant recipients, both during the early and the late follow-up period. All transplant team members should be familiar with the clinical presentation of frequently seen otorhinolaryngologic diseases.


The unexpected high practical value of medical ontologies.
Ontology is no longer a mere research topic, but its relevance has been recognized in several practical fields. Current applications areas include natural language translation, e-commerce, geographic information systems, legal information systems and biology and medicine. It is the backbone of solid and effective applications in health care and can help to build more powerful and more interoperable medical information systems. The design and implementation of ontologies in medicine is mainly focused on the re-organization of medical terminologies. This is obviously a difficult task and requires a deep analysis of the structure and the concepts of such terminologies, in order to define domain ontologies able to provide both flexibility and consistency to medical information systems. The aim of this special issue of Computers in Biology and Medicine is to report the current evolution of research in biomedical ontologies, presenting both papers devoted to methodological issues and works with a more applicative emphasis.


Relationship between white blood cell count and incident hypertension.
Elevated white blood cell (WBC) count is considered to be prospectively associated with cardiovascular disease. However, its relationship to hypertension, independent of smoking and other established cardiovascular risk factors, is not clear, especially among women. We used data from a large population-based study in Wisconsin (Beaver Dam Eye study) to examine the prospective association between elevated WBC count and incident hypertension among 2459 hypertension-free women (48.6%) and men (51.4%) after adjusting for, and stratifying by smoking and several other potential confounding factors. In multivariable proportional hazards models, increasing tertiles of WBC count was associated with increased risk ratios (RR) of hypertension in the whole cohort (WBC count tertiles 1-3; RR 1, 1.2, 1.7; P <.01), and separately among women (WBC count tertiles 1-3; RR 1, 1.1, 1.4; P <.05) and men (WBC count tertiles 1-3; RR 1, 1.3, 1.9; P <.01). Results from subsequent analyses stratified by smoking and several other related factors were consistent with this finding. Elevated WBC count is associated with incident hypertension among women and men independent of smoking and most traditional cardiovascular risk factors in this predominantly white cohort. Further research is required to determine whether this association is true among racial minorities (eg, African Americans), and independent of C-reactive protein, a more specific marker of inflammation.


Nitric oxide suppresses growth and development in the unicellular green alga Micrasterias denticulata.
Nitric oxide (NO), a key molecule in inter- and intracellular signalling, is implicated in developmental processes, host defense, and apoptosis in higher plants. We investigated the effect of NO on development in the unicellular green alga, Micrasterias denticulata, using two different NO donors, S-nitroso-N-acetyl-dl-penicillamine (SNAP) and sodium nitroprusside (SNP). Investigations at the light microsopic level revealed that both NO donors suppressed cell growth. Ultrastructural analyses were performed with SNAP- as well as SNP-treated cells and, additionally, with the control compound N-acetyl-d-penicillamine (NAP). Cells incubated with NO donors lacked a secondary wall and dictyosomal function was impaired, whereas NAP-treated cells showed no difference in development and organelle structure compared to control cells. Moreover, cisternae of the Golgi stacks were slightly involute and no vesicles were pinched off after SNAP and SNP incubation. The NO scavenger cPTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, potassium salt) abrogated the effect of SNP, thus confirming that inhibition of cell growth is due to nitric oxide. Addition of iodoacetic acid, an inhibitor of cysteine-containing enzymes, like glyceraldehyde-3-phosphate dehydrogenase (GAPDH), evoked similar effects on cell growth and secondary wall formation as obtained by treatment with NO donors. Therefore, we hypothesize that NO inhibits activity of enzymes involved in the secretory pathway, such as GAPDH, via S-nitrosylation of the cysteine residue and, consequently, modulates cell growth in M. denticulata.


Single-Dose Pharmacokinetic Study of Diphenhydramine HCl in Children and Adolescents.
Diphenhydramine pharmacokinetics were characterized following a single oral dose in children aged 2 to 17 years using a weight- and age-based dosing schedule with more tiers than the current age-based dosing schedule recommended by the nonprescription drug monograph. This study was conducted in 42 subjects, aged 2 to 17 years. Doses were based on a weight-age dosing schedule, ranging from 6.25 to 50 mg. An oral dose was administered with water about 2 hours after a light breakfast. Plasma samples were obtained up to 48 hours after dosing and analyzed for diphenhydramine. Pharmacokinetic parameters were estimated using noncompartmental methods, and the relationship of oral clearance with age was assessed using linear regression. Over an 8-fold range of doses, Cmax and AUC increased ∼90 % to ∼140% across age groups, with a similar Tmax (1.5 hours). Oral CL/F increased with age, but after allometric scaling, no maturation-related change in CL/F was apparent. Mild somnolence was the most commonly reported adverse event (95% of the subjects). A weight-age dosing schedule using an 8-fold range of doses achieved Cmax and AUC that increased about 2-fold across age groups. No effect of maturation on CL/F was observed after allometric scaling.


Comprehensive Transcriptome Profiling of Balding and Non-Balding Scalps in Trichorhinophalangeal Syndrome Type I Patient.
Trichorhinophalangeal syndrome (TRPS) patients tend to have alopecia that appears to be androgenetic, and this genetic model might give clues to the pathogenesis of hair loss or hair morphogenesis. This study was conducted to identify additional genetic evidence of TRPS and hair morphogenesis from a TRPS patient. From one TRPS type I patient, we extracted RNA and profiled whole transcriptome in non-balding and balding scalp areas using high-throughput RNA sequencing. We found a total of 26,320 genes, which comprised 14,892 known genes with new isoforms and 4,883 novel genes from the non-balding and balding areas. Among these, a total of 1,242 genes showed different expression in the two scalp areas (p<0.05 and log2 fold-change >0). Several genes related to the skin and hair, alopecia, and the TRPS1 gene were validated by qRT-PCR. Twelve of 15 genes (KRT6C, KRTAP3-1, MKI67, GPRC5D, TYRP1, DSC1, PMEL, WIF1, SOX21, TINAG, PTGDS, and TRPS1) were down-regulated (10 genes: p<0.01; SOX21 and PTGDS: p>0.05), and the three other genes (HBA2, GAL, and DES) were up-regulated (p<0.01) in the balding scalp. Many genes related to keratin and hair development were down-regulated in the balding scalp of the TRPS type I patient. In particular, the TRPS1 gene might be related to androgen metabolism and hair morphogenesis. Our result could suggest a novel perspective and evidence to support further study of TRPS and hair morphogenesis.


Vitamin D autocrine system and prostate cancer.
25-Hydroxyvitamin D-1alpha-hydroxylase (lalpha-OHase) is expressed in prostate cells. The expression suggests that local production of 1,25-dihydroxyvitamin D could provide an important cell growth regulatory mechanism. However, there is differential expression of 1alpha-OHase activity among the primary cultures of prostate cells derived from cancerous, benign prostatic hypertrophy and normal tissue, and among noncancerous (PZHPV-7) and various cancer cell lines (PC-3, DU145). No activity was found in cancer cell line LNCaP. The observed marked decrease in 1alpha-OHase activity in prostate cancer cells suggests some defect of the 1alpha-OHase in these cells. Using luciferase reporter gene assay, we observed a step-wise decrease in the basal promoter activity in two truncated promoter fragments, AN2 (-1,100 bp) and AN5 (-394 bp), with the highest basal activities found in PZHPV-7 and with loss of promoter activity in LNCaP. In order to understand the mechanism underlying the differential promoter activities among different prostate cells, we investigated the possible role of phosphorylation of cyclic AMP response element binding protein (CREB) on the regulation of 1alpha-OHase promoter activity in the four prostate cell lines. First we compared the levels of CREB phosphorylation among PZHPV-7, DU145, PC-3 and LNCaP cells by Western blot analysis using antibody against phosphorylated CREB. We observed that CREB was phosphorylated to a greater extent in PZHPV-7 than in DU145 cells. No significant phosphorylation of CREB was found in PC-3 and LNCaP cells. Next, we utilized activators and inhibitors of protein kinase A (PKA), protein kinase C (PKC), mitogen-activated protein kinase kinase (MAPKK) and calcium/calmodulin-dependent protein kinase II (CaMKII) to determine which kinases might be involved in phosphorylating the CREB in PZHPV-7 cells. We demonstrated that forskolin (an activator of PKA) increased the AN2 basal promoter activity 50%, whereas H-89 (an inhibitor of PKA) inhibited the basal and forskolin-stimulated AN2 promoter activity 40% and 70%, respectively. We also showed that PD98059 (an inhibitor of MAPKK) decreased the AN2 promoter activity 70%. Phorbol 12-myristate 13-acetate (an activator of PKC), GF109203 (an inhibitor of PKC) and KN-93 (an inhibitor of CaMKII) had no effect on AN2 promoter activity in PZHPV-7 cells. Thus, our results suggest that differential phosphorylation of CREB through PKA and MAPK pathways may be involved in the regulation of 1alpha-OHase promoter activity.


The pain ward follow-up analyses.
Two-year and 3-year follow-up analyses show that those who did not receive surgery for pain relief did as well as those who did receive such surgery during their stay, both groups showing consistently lower pain levels and analgesic intake than on admission, while activity levels progressively increased. The surgical patients were more likely to be readmitted for their pain problem than were the others, whereas the others were more likely to be readmitted for other medical problems.