QuEChERS-assisted ion pair chromatography/fluorescence detection method for determination of antimigraine combination therapy in rabbit plasma samples: Application to a pharmacokinetic study.
Antimigraine combination therapy has shown significant effectiveness in relieving pain, as well as reducing the frequency, duration, and severity of migraine attacks if compared to a single migraine medication. This work represents the first analytical investigation for emphasizing the synergistic effect of combining ophthalmic beta blockers with triptans in migraine treatment. The presented study was conducted to investigate the pharmacokinetic profile of almotriptan (ALM), a serotonin (5-HT1B/1D) receptor agonist used to treat migraine, when coadministered with timolol (TIM) or verapamil (VER) which are considered as an adjuvant therapy in migraine prevention. Ion pair chromatography (IPC) with online fluorescence detection was applied to simultaneously detect and quantify the binary mixtures of ALM/TIM and ALM/VER in rabbit plasma samples. The separation was achieved using a Platinum C18 analytical column with a mobile phase composed of methanol: 35 mmol L-1 phosphate buffer solution containing 10 mmol L-1 SDS at pH = 6.8 (60:40 v/v). Several parameters were evaluated during the optimization of separation conditions including mobile phase composition, buffer concentration, buffer pH and concentration of ion pair reagent. A thorough investigation of the retention mechanism was performed, and the results showed that Coulomb forces were the main contributors to the overall retention mechanism, which may be hydrophobically assisted. QuEChERS extraction technique was utilized to extract the investigated drugs from plasma samples and a detailed study was carried out to optimize partition/extraction solvents, pH, extraction salts, sample volume and clean-up step. The method had a limit of detection and quantitation of 5.6 and 16.9 ng mL-1 for ALM in ALM/TIM mixture and 2.5 and 7.6 ng mL-1 for ALM in ALM/VER mixture, with an overall recovery not less than 95.22%. This newly proposed method offers a faster alternative to existing chromatographic methods for extraction and determination of ALM in binary mixtures with TIM or VER in rabbit plasma and provides a platform for studying pharmacokinetic parameters. The coadministration of either TIM or VER with ALM resulted in a notable rise in Cmax (maximum plasma concentration) and AUC (area under the plasma concentration-time curve) of ALM, implying possible alterations in the absorption and overall exposure of ALM.


Nitric oxide: biosynthesis and biological significance.
The recent discovery that mammalian cells can synthesize nitric oxide coincided with the identification of this simple gas as a factor involved in cellular communication. Nitric oxide has now been shown to be derived from L-arginine in macrophages, endothelial cells and possibly other cell types. Its physiological role in macrophages may be as a cytotoxic agent. However, nitric oxide produced by endothelial cells is thought to trigger vascular smooth muscle relaxation through activation of the enzyme guanylate cyclase.


Radiotoxic fission products and radiological effects in the Mediterranean Sea biota from a hypothetical accident in Akkuyu Nuclear Power Plant.
A combined hydrodynamic/radiobiological model was used to calculate the distribution and dose rate of significant radionuclides in the Mediterranean Sea marine organisms in the event of an accident at the Akkuyu Nuclear Power Plant (ANPP). The Hybrid single-particle Lagrangian integrated trajectory (HYSPLIT) model was applied to simulate the dispersion and deposition of artificial radionuclide concentrations. Environmental Risk from Ionising Contaminants: Assessment and Management (ERICA) tools were utilized to assess issues related to sea health and potential hazards in case of an accident. The scenario source term profile was derived from the Fukushima nuclear power accident. Volumetric concentrations and deposition levels of pollutants increased from 1 MBqm-3 to 1 × 103 MBqm-3 and from 1 × 10-1 MBqm-2 to 1 × 10-7 MBqm-2 after 48 h. In terms of dose rates to the various marine biotas, polychaete worms had the greatest overall dose rate and the highest contribution to total dose rates attributable to 137Cs radionuclide.


PREDICT identifies precipitating events associated with the clinical course of acutely decompensated cirrhosis.
Acute decompensation (AD) of cirrhosis may present without acute-on-chronic liver failure (ACLF) (AD-No ACLF), or with ACLF (AD-ACLF), defined by organ failure(s). Herein, we aimed to analyze and characterize the precipitants leading to both of these AD phenotypes. The multicenter, prospective, observational PREDICT study (NCT03056612) included 1,273 non-electively hospitalized patients with AD (No ACLF = 1,071; ACLF = 202). Medical history, clinical data and laboratory data were collected at enrolment and during 90-day follow-up, with particular attention given to the following characteristics of precipitants: induction of organ dysfunction or failure, systemic inflammation, chronology, intensity, and relationship to outcome. Among various clinical events, 4 distinct events were precipitants consistently related to AD: proven bacterial infections, severe alcoholic hepatitis, gastrointestinal bleeding with shock and toxic encephalopathy. Among patients with precipitants in the AD-No ACLF cohort and the AD-ACLF cohort (38% and 71%, respectively), almost all (96% and 97%, respectively) showed proven bacterial infection and severe alcoholic hepatitis, either alone or in combination with other events. Survival was similar in patients with proven bacterial infections or severe alcoholic hepatitis in both AD phenotypes. The number of precipitants was associated with significantly increased 90-day mortality and was paralleled by increasing levels of surrogates for systemic inflammation. Importantly, adequate first-line antibiotic treatment of proven bacterial infections was associated with a lower ACLF development rate and lower 90-day mortality. This study identified precipitants that are significantly associated with a distinct clinical course and prognosis in patients with AD. Specific preventive and therapeutic strategies targeting these events may improve outcomes in patients with decompensated cirrhosis. Acute decompensation (AD) of cirrhosis is characterized by a rapid deterioration in patient health. Herein, we aimed to analyze the precipitating events that cause AD in patients with cirrhosis. Proven bacterial infections and severe alcoholic hepatitis, either alone or in combination, accounted for almost all (96-97%) cases of AD and acute-on-chronic liver failure. Whilst the type of precipitant was not associated with mortality, the number of precipitant(s) was. This study identified precipitants that are significantly associated with a distinct clinical course and prognosis of patients with AD. Specific preventive and therapeutic strategies targeting these events may improve patient outcomes.


Urinary excretion of estrone glucosiduronate, 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate. Significance of proportionate differences during the menstrual cycle.
The urinary excretion of estrone glucosiduronate, 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate in men and throughout the menstrual cycle in women was measured by specific radioimmunoassay. In 9 men the mean +/- SE excretion of these conjugates was 15.9 +/- 1.4, 2.7 +/- 0.3, and 3.2 +/- 0.2 microgram/24 h respectively. In 15 women studied in the midfollicular phase (day 8) of the menstrual cycle, the excretion was 19.4 +/- 1.7, 2.9 +/- 0.2, and 5.4 +/- 1.3 micrograms/24 h. Excretion of each conjugate was significantly (P less than 0.01) elevated in the midluteal phase (day 22) to 41.9 +/- 3.9, 6.3 +/- 0.8, and 12.2 +/- 1.5 micrograms/24 h respectively (n = 14). The mean excretion of estriol-16 alpha-glucosiduronate was greater than that of 17 beta-estradiol-17-glucosiduronate in the luteal phase (P less than 0.05) but not in the follicular phase or in men (P greater than 0.05). The excretion of each of these specific conjugates measured throughout the menstrual cycle in 7 women was characterized by a sharp midcycle peak and a lower, broader luteal phase peak. The ratios of estriol-16 alpha-glucosiduronate to 17 beta-estradiol-17-glucosiduronate, estrone glucosiduronate to 17 beta-estradiol-17-glucosiduronate, and estriol-16 alpha-glucosiduronate to estrone glucosiduronate throughout the menstrual cycle were analyzed. When the mean ratio during the follicular phase was set at 1, a significant increase (P less than 0.01) occurred in the mean luteal phase ratio in each case: 1.00 +/- 0.03 to 1.66 +/- 0.09, 1.00 +/- 0.04 to 1.30 +/- 0.04, and 1.00 +/- 0.03 to 1.24 +/- 0.04 (mean +/- SE) respectively. The marked alteration in the proportions of these urinary estrogen conjugates may be due to altered metabolism of 17 beta-estradiol, but it more likely reflects a change in the pattern of estrogen secretion or production between the two phases of the menstrual cycle.


Total thyroidectomy with ultrasonic dissector for cancer: multicentric experience.
We conducted an observational multicentric clinical study on a cohort of patients undergoing thyroidectomy for thyroid carcinoma. The aim of this study was to evaluate the benefits of the use of ultrasonic dissector (UAS) vs. the use of a conventional technique (vessel clamp and tie) in patients undergoing thyroid surgery for cancer. From June 2009 to May 2010 we evaluated 321 consecutive patients electively admitted to undergo total thyroidectomy for thyroid carcinoma. The first 201 patients (89 males, 112 females) presenting to our Department underwent thyroidectomy with the use of UAS while the following 120 patients (54 males, 66 females) underwent thyroidectomy performed with a conventional technique (CT): vessel clamp and tie. The operative time (mean: 75 min in UAS vs. 113 min in CT, range: 54 to 120 min in UAS vs. 68 to 173 min in CT) was much shorter in the group of thyroidectomies performed with UAS. The incidence of transient laryngeal nerve palsy (UAS 3/201 patients (1.49%); CT 1/120 patients (0.83%)) was higher in the group of UAS; the incidence of permanent laryngeal nerve palsy was similar in the two groups (UAS 2/201 patients (0.99%) vs. CT 2/120 patients (1.66%)). The incidence of transient hypocalcaemia (UAS 17/201 patients (8.4%) vs. CT 9/120 patients (7.5%)) was higher in the UAS group; no relevant differences were reported in the incidence of permanent hypocalcaemia in the two groups (UAS 5/201 patients (2.48%) vs. 2/120 patients (1.66%)). Also the average postoperative length of stay was similar in two groups (2 days). The only significant advantage proved by this study is represented by the cost-effectiveness (reduction of the usage of operating room) for patients treated with UAS, secondary to the significant reduction of the operative time. The analysis failed to show any advantages in terms of postoperative transient complications in the group of patients treated with ultrasonic dissector: transient laryngeal nerve palsy (1.49% in UAS vs. 0.83% in CT) and transient hypocalcaemia (8.4% in UAS vs. 7.5%in CT). No significant differences in the incidence of permanent laryngeal nerve palsy (0.8% in UAS vs. 1.04% in CT) and permanent hypocalcaemia (2.6% in UAS vs. 2.04% in CT) were demonstrated. The level of surgeons' expertise is a central factor, which can influence the complications rate; the use of UAS can only help surgical action but cannot replace the experience of the operator.


Physical map of DNA from a new cauliflower mosaic virus strain.
The restriction enzymes AluI, BamHI, BglII, EcoRI, HindIII, and SalI have been used to characterize and map a new cauliflower mosaic virus strain (Cabb-S). These fragments have been ordered by examining their overlapping regions after double enzymatic digestion. The single SalI cleavage site was chosen as the point of origin. We compare this strain with those already described.


Herbivory and intraspecific competition in a stream caddisfly population.
Field experiments were conducted to assess the effects of different population densities of the herbivorous caddisfly Helicopsyche borealis on periphyton biomass and on its own growth rate in Big Sulphur Creek, a third-order stream in northern California, USA. Stream enclosures were used to vary grazer density from one-eighth to twice natural density (1/8X-2X) in two experiments (35 d and 60 d), which spanned the period of most rapid larval growth. Periphyton biomass and chlorophyll a were inversely related to grazer density. Grazer densities of 1/8X-1/2X moderately reduced periphyton when compared to an ungrazed control, whereas densities of 1X-2X greatly depleted periphyton. The growth rate of H. borealis larvae declined with increasing larval density. Growth rate was highest at densities of 1/8X-1/2X; larvae grew more slowly at 1X and showed no growth at 2X. Weekly supplementation of periphyton to 1/4X, 1X, and 2X densities significantly increased the final body mass of larvae at 1X and 2X (by 26% and 50%, respectively, compared to unsupplemented larvae) but did not change the body mass of larvae at 1/4X. These results suggest that periphyton is a limiting resource in Big Sulphur Creek and that H. borealis larvae compete exploitatively for that resource. Intraspecific competition may be an important, but often overlooked, feature of many herbivore populations in streams.


Plant stress and insect performance: cottonwood, ozone and a leaf beetle.
Leaf area consumption rates, development rates, survivorship, and fecundity of the imported willow leaf beetle (Plagiodera versicolora Laich) were examined on two clones of eastern cottonwood which were previously exposed to ozone or charcoal-filtered air. P. versicolora consumed more ozone treated foliage, but were more fecund when reared on charcoal-filtered air treated plants. Beetle development rates and survivorship were not significantly different on treated and control cottonwoods. We concluded that: 1) Ozone fumigation of cottonwood reduced foliage quality, and the reproductive success and overall performance of P. versicolora. 2) increased foliage consumption by beetles was probably a mechanism compensating for decreases in foliage quality. 3) Reductions in beetle fecundity were due to an initial reduction in oviposition rates. 4) Beetle feeding preference did not correlate with the suitability of foliage for beetle performance. These results are discussed in relation to the impact of air pollution on plant-insect interactions.


Smart scan of medical device displays to integrate with a mHealth application.
The daily monitoring of the physiological parameters is essential for monitoring health condition and to prevent health problems. This is possible due to the democratization of numerous types of medical devices and promoted by the interconnection between these and smartphones. Nevertheless, medical devices that connect to smartphones are typically limited to manufacturers applications. This paper proposes an intelligent scanning system to simplify the collection of data displayed on different medical devices screens, recognizing the values, and optionally integrating them, through open protocols, with centralized databases. To develop this system, a dataset comprising 1614 images of medical devices was created, obtained from manufacturer catalogs, photographs and other public datasets. Then, three object detector algorithms (yolov3, Single-Shot Detector [SSD] 320 × 320 and SSD 640 × 640) were trained to detect digits and acronyms/units of measurements presented by medical devices. These models were tested under 3 different conditions to detect digits and acronyms/units as a single object (single label), digits and acronyms/units as independent objects (two labels), and digits and acronyms/units individually (fifteen labels). Models trained for single and two labels were completed with a convolutional neural network (CNN) to identify the detected objects. To group the recognized digits, a condition-tree based strategy on density spatial clustering was used. The most promising approach was the use of the SSD 640 × 640 for fifteen labels. Lastly, as future work, it is intended to convert this system to a mobile environment to accelerate and streamline the process of inserting data into mobile health (mhealth) applications.


Autologous rib cartilage as implanted stiffener of phalloplasty: comparing three different methods.
Maintaining the length, rigidity, and appearance of a reconstructed phallus in the long term has been challenging for phalloplasty and may depend on choosing the optimal implanted stiffener. The aim of this study was to determine the best type of stiffener for a reconstructed phallus regarding aesthetic and functional parameters. From 1987 to 2018, 376 individuals underwent phalloplasty at a single institution of which 81 met the following inclusion criteria: biological male gender, age between 18 and 60 years, radial forearm flap for tube-within-a-tube phalloplasty with rib cartilage and secondary glans sculpting and unimpaired urination preoperatively. Those were divided into three groups with regard to different types of autologous rib cartilage stiffener. A group with a cartilage strip, T-shaped cartilage, and mushroom-shaped cartilage group were evaluated regarding preserved length and aesthetic features of the phallus during a minimum period of 12 months. The rib cartilage length in the phallus was 10.5 cm in all groups. The length of the phallus was 10.8 ± 0.5 cm in the cartilage strip group, 11.3 ± 0.3 cm in the T-shaped cartilage group and 11.3 ± 0.3 cm in the mushroom-shaped cartilage group at 1-year postoperatively. The aesthetic appearance score of mushroom-shaped cartilage group was 1.67 ± 0.48, significantly higher than the cartilage strip group (1.38 ± 0.50, p = 0.03) at 1 year after surgery. The mushroom-shaped autologous rib cartilage group showed superior length preservation and aesthetic appearance and appeared as the best type of stiffener for one-phase phalloplasty.


Negative multiparametric magnetic resonance imaging for prostate cancer: further outcome and consequences.
EAU guidelines recommend multiparametric MRI of the prostate (mpMRI) prior to biopsy to increase accuracy and reduce biopsies. Whether biopsy can be avoided in case of negative mpMRI remains unclear. Aim of this study is to evaluate predictors of overall prostate cancer (PCa) in negative mpMRI. A total of 216 patients from 2018 to 2020 with suspicion of PCa and negative mpMRI (PI-RADS ≤ 2) were interviewed by telephone about outcome and further follow-up. Clinically significant PCa (csPCa) was defined as ISUP ≥ 2. Patients with vs. without biopsy and with vs. without PCa were compared. Univariate and multivariate analyses were performed to evaluate predictors of PCa occurrence in patients with negative mpMRI. 15.7% and 5.1% of patients with PI-RADS ≤ 2 on mpMRI showed PCa and csPCa, respectively. PCa patients had higher PSAD (0.14 vs. 0.09 ng/ml2; p = 0.001) and lower prostate volume (50.5 vs. 74.0 ml; p = 0.003). Patients without biopsy (25%) after MRI were older (69 vs. 65.5 years; p = 0.027), showed lower PSA (5.7 vs. 6.73 ng/ml; p = 0.033) and lower PSA density (0.09 vs. 0.1 ng/ml2; p = 0.027). Multivariate analysis revealed age (OR 1.09 [1.02-1.16]; p = 0.011), prostate volume (OR 0.982 [0.065; 0.997]; p = 0.027), total PSA level (OR 1.22 [1.01-1.47], p = 0.033), free PSA (OR 0.431 [0.177; 0.927]; p = 0.049) and no PI-RADS lesion vs PI-RADS 1-2 lesion (OR 0.38 [0.15-0.91], p = 0.032.) as predictive factors for the endpoint presence of PCa. Biopsy for selected patient groups (higher age, prostate volume and free PSA as well as lower PSA-Density) with negative mpMRI can be avoided, if sufficient follow-up care is guaranteed. Detailed counseling regarding residual risk for undetected prostate cancer should be mandatory.


[A case report of septicemia caused by methicillin cephem resistant Staphylococcus aureus (MRSA)].
The frequency of MRSA (Methicillin cephem resistant Staphylococcus aureus) infection has increased recently. A case of septicemia caused by MRSA is reported with discussion on its prophylaxis.


Successful treatment of lymphadenosis benigna cutis with topical photodynamic therapy with delta-aminolevulinic acid.
Photodynamic therapy (PDT) using topical delta-aminolevulinic acid (ALA) has been used for nonmelanoma skin cancers, including malignant cutaneous T-cell lymphomas. Moreover, PDT has been tried for benign inflammatory or infectious skin lesions. To evaluate the effects of ALA-PDT on skin lesions of lymphadenosis benigna cutis (LABC). Two 16-year-old females with solitary and infiltrated nodules were treated 5 times with topical ALA-PDT. Both patients responded well and showed dramatic clinical and histopathological improvement without visible scars. The results confirm that topical ALA-PDT is an effective and safe modality for the treatment of LABC, and that such treatment may be cosmetically beneficial.


Total chemical synthesis of fully functional Photoactive Yellow Protein.
The Photoactive Yellow Protein (PYP) is a structural prototype for the PAS superfamily of proteins, which includes hundreds of receptor and regulatory proteins from all three kingdoms of life. PYP itself is a small globular protein that undergoes a photocycle involving a series of conformational changes in response to light excitation of its p-coumaric acid chromophore, making it an excellent model system to study the molecular basis of signaling in the PAS super family. To enable novel chemical approaches to elucidating the structural changes that accompany signaling in PYP, we have chemically synthesized the 125 amino acid residue protein molecule using a combination of Boc chemistry solid phase peptide synthesis and native chemical ligation. Synthetic PYP exhibits the wildtype photocycle, as determined in photobleaching studies. Planned future studies include incorporation of site-specific isotopic labels into specific secondary structural elements to determine which structural elements are involved in signaling state formation using difference FTIR spectroscopy.


Fluence Rate Determines PDT Efficiency in Breast Cancer Cells Displaying Different GSH Levels.
Photodynamic therapy (PDT) appears as a promising alternative in the treatment of breast cancer since it can be highly effective in curing cancer while preserving normal tissue. However, predicting outcomes in PDT still constitutes a great challenge. One of the parameters that are usually empirically determined is the rate of photon flux delivered to the tissue (light fluence rate). In the present study, we intended to understand why monolayers of human cells derived from mammary adenocarcinomas (MDA-MB-231 and MCF-7) respond quite differently to fluence rates (cells were irradiated either for 6 or for 16 min) at a fixed light dose (4.5 J cm-2 ) delivered with an array of LEDs in a typical methylene blue PDT protocol. While death rates of MDA-MB-231 cells were insensitive to the fluence rate, MCF-7 cells showed a quite impressive (three times) decrease in cell death levels in the shorter irradiation protocol. Independent on cell type cell death was invariably correlated with the depletion of reduced glutathione intracellular levels and consequently with widespread redox misbalance. Our data show the potential to optimize fluence rates to provide exhaustion of the cell antioxidant responses in order to circumvent therapy resistance of breast tumors.


Laser lithotripsy. Patient care, staff education.
The high cost of laser equipment and the required specialization of staff members make extension of laser lithotripsy to every hospital impractical. At present, in Ontario, laser lithotripsy is available only in Toronto and Kingston, and ESWL is available only in Toronto and London. Laser lithotripsy is a viable alternative to invasive surgical treatment of urinary calculi that are resistant to ESWL. For the suitable candidate, laser lithotripsy is a welcome alternative to an extensive surgical procedure and recovery period. Hospitalization is reduced significantly, and patients can return to work after the calculus and residue have been passed. Patients can be managed safely and effectively, resulting in reduced morbidity and mortality and reduced cost in time and expense to the patient and hospital.


Amyloid-beta, tau alterations and mitochondrial dysfunction in Alzheimer disease: the chickens or the eggs?
Alzheimer disease (AD) is defined pathologically and diagnostically defined by amyloid-beta senile plaques and neurofibrillary tangles (NFT) composed of tau. From the time of their original description nearly a century ago, a major focus has been to understand the role that these lesions play in the pathogenesis of the disease. The majority favors the notion that these lesions cause the disease and therefore attempts at therapeutic intervention are focused on preventing lesions formation. However, this rationale may be misguided since new evidence from our laboratories and others suggest that the lesions not only occur as a by-product of the fundamental disease process but also that they may be protective.


Peripheral axon regrowth: new molecular approaches.
Peripheral nerves, essential connections between the brain, spinal cord and body, do not regenerate as well as generally reported. Identifying new strategies to facilitate regeneration is essential to reversing neurological deficits from nerve injuries or disease. This review will discuss several selected and novel molecular insights into peripheral nerve trunk repair and axon regrowth that have the potential to improve regenerative success. Of particular interest is the phosphatidylinositol 3-kinase (PI3K)-Akt pathway in peripheral neurons, inhibited by the constitutively expressed phosphatase tumor suppressor PTEN. Knockdown or inhibition of PTEN is associated with robust sprouting of adult sensory neurons in vitro and in vivo, additive to the accelerated outgrowth offered by the preconditioning effect. This sprouting response, if spatially and temporally constrained, may provide potent regrowth initiation, of interest in otherwise untreatable nerve damage.


Mycobacterial Trehalose 6,6'-Dimycolate-Induced M1-Type Inflammation.
Murine models of Mycobacterium tuberculosis (Mtb) infection demonstrate progression of M1-like (proinflammatory) and M2-like (anti-inflammatory) macrophage morphology following primary granuloma formation. The Mtb cell wall cording factor, trehalose 6,6'-dimycolate (TDM), is a physiologically relevant and useful molecule for modeling early macrophage-mediated events during establishment of the tuberculosis-induced granuloma pathogenesis. Here, it is shown that TDM is a major driver of the early M1-like macrophage response as seen during initiation of the granulomas of primary pathology. Proinflammatory cytokines tumor necrosis factor-α, IL-1β, IL-6, and IL-12p40 are produced in lung tissue after administration of TDM to mice. Furthermore, CD11b+CD45+ macrophages with a high surface expression of the M1-like markers CD38 and CD86 were found present in regions of pathology in lungs of mice at 7 days post-TDM introduction. Conversely, only low phenotypic marker expression of M2-like markers CD206 and EGR-2 were present on macrophages. These findings suggest that TDM plays a role in establishment of the M1-like shift in the microenvironment during primary tuberculosis.


C-kit expression in dedifferentiated and well-differentiated liposarcomas; immunohistochemistry and genetic analysis.
c-kit expression by immunohistochemistry has been utilized to identify cancer patients who can be treated with imatinib-mesylate. In gastrointestinal stromal tumors (GISTs), an activating mutation in c-kit predicts treatment response; its presence in other soft tissue tumors is unexplored. We evaluated seven cases of dedifferentiated liposarcomas (DDLS) and compared those with seven well-differentiated liposarcomas (WDLS). Immunohistochemical staining for c-kit was performed using a polyclonal antibody. Using PCR, exons 9, 10-11, 12-13 and 17 of c-kit were amplified and direct DNA sequencing performed. Two out of 7 (30%) DDLS showed focal weak immunoreactivity with c-kit; no (0%) WDLS stained with c-kit. Seven out of 7 (100%) DDLS showed an allelic variation in exon 10, with a single base pair substitution (A >C) at codon 541; 3/7 (43%) WDLS showed the same change. c-kit immunoreactivity did not correlate with the change in DNA sequence; DDLS showed a consistent allelic variation in c-kit that may have significant prognostic, diagnostic and therapeutic implications.


Continuous infusion aerosol delivery of prostacyclins during mechanical ventilation: challenges, limitations, and recent advances.
Critically ill mechanically ventilated patients routinely receive aerosol delivery of epoprostenol by continuous infusion of the nebulizer by syringe pump. This procedure is 'off-label' as no FDA approved drug presently exists. Without standardized protocols, therapy is based on prior experience with bronchodilators, limited studies of delivery systems and anecdotal clinical trials. Current protocols based upon patient body weight and drug concentration determines the infusion rate of drug dose delivered to the nebulizer , which is only distantly related to dose delivered to the lung and may be altered by many factors. This paper reviews the background of this technique as well as current methods of managing drug delivery, technical challenges, and limitations. A recent advance in aerosol laboratory bench testing, using radiolabeled aerosols, is presented to reveal important factors defining delivery. Off-label use of continuously nebulized prostacyclin in the ICU lacks the support of large clinical trials needed for FDA clearance. However, comprehensive bench studies afford the potential for clinicians to better understand and manage therapy at a level above simple dosing of the nebulizer by body weight. New research techniques are enhancing our basic comprehension of the interaction between aerosol devices and the mechanical ventilator.


Obstructive uropathy in a HIV+ infant under indinavir treatment.
The case is presented of a 10-year-old HIV+ male with renoureteral pain, who developed an obstructive uropathy with renal function impairment and required endoscopic placement of a ureteral stent. Certain aspects of the epidemiology, clinical presentation, diagnosis, treatment and prevention are discussed.


Expression of myoferlin in human and murine carcinoma tumors: role in membrane repair, cell proliferation, and tumorigenesis.
Cancer cells are often characterized by high proliferation rates, a consequence of increased mitotic signaling coupled with unchecked cellular growth. We recently demonstrated that vascular endothelial cells unexpectedly express ferlins, a family of muscle-specific proteins capable of regulating the fusion of lipid patches to the plasma membrane, and that these highly regulated membrane fusion events are essential to endothelial cell proliferation and homeostasis. Here, we show that human and mouse breast cancer cell lines also express myoferlin at various levels, and that the processes of transformation, epithelial-mesenchymal transition, and metastasis do not appear to have any effect on myoferlin expression in vitro. In vivo, we observed that solid mouse and human carcinoma tissues also express high levels of myoferlin protein. Loss-of-function studies performed in mice revealed that myoferlin gene knockdown can attenuate cancer cell proliferation in vitro and decrease tumor burden, and that accelerated tumor cell growth appears to rely on intact myoferlin-dependent membrane repair and signaling under exponential growth conditions. To our knowledge, these data provide the first evidence of myoferlin expression in solid human and mouse tumors. We have thus identified a novel membrane repair process that likely helps sustain the high growth rates characteristic of tumors, and we suggest that interfering with normal myoferlin expression and/or membrane repair and remodeling may provide therapeutically relevant antiproliferative effects.


Evaluation of monoclonality of cell lines from sequential dilution assays. Part II.
We extend the analysis in Koziol et al. (J. Immunol. Methods (1987) 105, 139) for the determination of the probability of achieving monoclonality in limiting dilution assays to three or more cloning stages. We also provide a computer program which will carry out this analysis for an arbitrary number of stages.


Genetic polymorphisms of IL17A and pri-microRNA-938, targeting IL17A 3'-UTR, influence susceptibility to gastric cancer.
We report an association between gastric cancer (GC) and polymorphisms in IL17A, rs2275913 (-197 G > A), rs3748067 (*1249 C > T), and pri-miR-938, rs2505901 (T > C). We employed the multiplex PCR-SSCP method to detect gene polymorphisms in 337 GC cases and 587 controls. The minor allele frequency of rs2275913 was significantly higher, and those of rs3748067 and rs2505901 significantly lower, in GC cases than controls. The rs2275913 AA homozygote was associated with an increased risk (OR, 2.38; 95%CI, 1.63-3.46; p < 0.0001) for the development of both intestinal and diffuse types of GC. The rs3748067 T polymorphism was associated with a decreased risk for intestinal GC (OR, 0.511; 95%CI, 0.272-0.962; p = 0.037), whereas rs2505901 C locus carried a decreased risk overall for GC (OR, 0.733; 95%CI, 0.545-0.985; p = 0.039). In addition, rs3748067 T allele was inversely correlated with lymph node metastasis. Our results suggest that polymorphisms in both IL17A and pri-miR-938 contribute to cancer risk susceptibility and therefore can affect the development of gastric cancer.


Investigations of Different Ion Intercalations on the Performance of FBG Hydrogen Sensors Based on Pt/MoO3.
α-MoO3 has been used as a hydrogen sensing material due to its excellent properties and unique crystalline layer structure. However, the low repeatability of α-MoO3 based hydrogen sensor restricts its practical application. In this paper, the effect of intercalated ion species and the amount in α-MoO3 is experimentally investigated and discussed. It is concluded that the repeatability of the sensor depends on the radius of intercalated ions and amount of ionic bonds. The optimal ion species is Na+ and the optimal amount of precursor is 1 mmol.


Roles of hydrogen sulfide and nitric oxide in the alleviation of cadmium-induced oxidative damage in alfalfa seedling roots.
Despite hydrogen sulfide (H(2)S) and nitric oxide (NO) are important endogenous signals or bioregulators involved in many vital aspects of plant growth and responses against abiotic stresses, little information was known about their interaction. In the present study, we evaluated the effects of H(2)S and NO on alfalfa (Medicago sativa L.) plants exposed to cadmium (Cd) stress. Pretreatment with an H(2)S donor sodium hydrosulfide (NaHS) and well-known NO donor sodium nitroprusside (SNP) decreased the Cd toxicity. This conclusion was supported by the decreases of lipid peroxidation as well as the amelioration of seedling growth inhibition and Cd accumulation, in comparison with the Cd-stressed alone plants. Total activities and corresponding transcripts of antioxidant enzymes, including superoxide dismutase, peroxidase and ascorbate peroxidase were modulated differentially, thus leading to the alleviation of oxidative damage. Effects of H(2)S above were reversed by 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO), the specific scavenger of NO. By using laser confocal scanning microscope combined with Greiss reagent method, further results showed that NO production increased significantly after the NaHS pretreatment regardless of whether Cd was applied or not, all of which were obviously inhibited by cPTIO. These decreases of NO production were consistent with the exaggerated syndromes associated with Cd toxicity. Together, above results suggested that NO was involved in the NaHS-induced alleviation of Cd toxicity in alfalfa seedlings, and also indicated that there exists a cross-talk between H(2)S and NO responsible for the increased abiotic stress tolerance.


Transoesophageal Doppler monitoring for fluid and hemodynamic treatment during lung surgery.
Patients undergoing lung resection are vulnerable to fluid overhydration. Recently, goal-directed fluid therapy using transoesophageal Doppler monitoring (TDM) has been shown to improve postoperative clinical outcome. The aim of this study was to assess the feasibility of TDM during open-chest procedures for guiding fluid and hemodynamic treatment. We performed an observational prospective study including 127 high-risk patients undergoing lung cancer resection. A restrictive fluid strategy was targeted to achieve a stroke volume index (SVI) > or = 30 ml/min/m(2). Besides standard hemodynamic measurements, stroke volume index (SVI), corrected flow time (FTc), maximal acceleration (MA) and velocity (PV) were recorded during two-lung ventilation (TLV) and one-lung ventilation (OLV). Doppler flow tracings could not be obtained in 4 patients during TLV (3.1%) and in 6 patients during OLV (4.9%). Preoperatively, 96 pts had SVI > or = 30 ml/min/m(2) (N-SVI group) whereas 21 patients had SVI < 30 ml/min/m(2) (L-SVI group) associated with lower FTc values. After OLV, SVI transiently decreased (-17 +/- 9%; P < 0.05) in the N-SVI group whereas in the L-SVI group, SVI increased steadily until the end of surgery (+40 +/- 12%). Other flow-related parameters as well as heart rate and mean arterial pressure remained unchanged. Surgical and medical characteristics did not differ between the two groups, except that larger volumes of colloids were administered intraoperatively in the L-SVI group (+2.2 +/- 0.6 ml/min/h compared with N-SVI group, P < 0.05). In thoracic surgical patients, TDM can be used to detect and correct low flow conditions and to guide hemodynamic support during the intraoperative period.


Prognostic value of sentinel lymph node biopsy in the pathologic staging of colorectal cancer patients.
Over the last decade, lymphatic mapping and sentinel lymph node (sN) biopsy have greatly increased the possibility of identifying nodal metastasis in clinically node-negative patients with melanoma and breast cancer, thus improving the accuracy of pathologic staging. Recently, sN biopsy has been applied also in colorectal cancer. This prospective study aimed to assess its feasibility and accuracy in predicting regional lymph nodes metastases in colorectal cancer patients as well as the impact on treatment decision-making. Lymphatic mapping was accomplished by means of blue dye, which was intraoperatively injected into the subserosa overlying the tumor site in 26 patients undergoing colorectal cancer surgery. Following bowel resection, the operative specimen was inspected to identify each blue-stained node, the sN, which was sent separately to the pathologist. One half of each sN was examined by multiple 200 microm sections, while the second half was examined by standard bi-valving technique with hematoxylin-eosin (H and E) staining; all the other regional non-sentinel nodes were routinely examined by standard bi-valving technique and H and E staining. At least one sN was detected in 24 of 26 patients (92.3%); two patients with rectal cancer had no sN identified. Overall, 70 sN were retrieved into the operative specimens, with a mean of 2.9 sNs/patient, and 19 sNs were tumor-positive. An agreement between sN and regional lymph-node status was observed in 20 of 24 patients (83.4%). The sN was histologically negative in two of nine patients with positive regional nodes (sensitivity = 77.8%; false-negative rate of 22.2%); in two of seven patients with tumor-positive sN (28.6%), the sN was the exclusive site of regional nodal metastasis. The negative predictive value was 88.2% (15 of 17 patients), and the overall accuracy was 91.7% (22 of 24 patients). As regards the contribution to the detection of nodal metastasis according to the pathologic technique, standard H and E bi-valving technique detected 16 of 19 tumor-positive sNs (84.2%) while, by means of serial sectioning, metastases were detected in the remaining 3 of 19 sNs (15.8%). The sN biopsy proved feasible, with a rather short learning curve. The focused analysis of the sN by means of serial sectioning improved the detection rate of nodal metastasis compared to standard bi-valving examination, so that a more accurate nodal staging should be expected; finally, an elective localization of metastasis within the sN was observed in almost one third of regional node-positive patients.


Solid-phase synthesis of a library of piperazinediones and diazepinediones via Kaiser oxime resin.
A combinatorial library of piperazinediones has been prepared by automated parallel solid-phase synthesis. The five-step reaction protocol makes use of Kaiser oxime resin, to enable cleavage from the polymeric support concomitant with an intramolecular displacement reaction, under very mild conditions. The methodology was also successfully extended to the preparation of the seven-membered ring homologs, diazepinediones.


Improvement of the reduction of condensable particulate matter in flue gas scrubbing process.
Condensable particulate matter (CPM) is characterized by complex composition, non-negligible emission concentration, and fine or ultrafine in size after conversion to particles, which is difficult to remove. Current methods to control CPM are not fully developed and mainly focus on synergistic removal of CPM in existing air pollution control devices, such as CPM reduction through scrubbing processes in wet flue gas desulfurization (WFGD) systems. In this work, an experimental system including a simulated WFGD scrubber, also referred to as the primary scrubber (PS), and a secondary scrubber (SS) was built to explore measures to improve the CPM reduction performance during scrubbing. The operating parameters of the liquid-to-gas (L/G) ratio and the spray temperature in the two scrubbers were tuned in the experiments. The results indicated that CPM could be reduced in the PS by conversion to filterable particulate matter (FPM), and captured by the spray droplets through the effects of dissolution and condensation, but the reduction was not very efficient. In the SS, the reduction performance of CPM could be further improved due to increased dissolution of CPM caused by increased opportunities for gas-liquid contact, and increased condensation of CPM due to lower spray temperature. The FPM transformed from the CPM in the PS could also be reduced in the SS by the effects of diffusiophoresis and thermophoresis contributed by water vapor condensation. An increase in the L/G ratio could improve the CPM reduction.


Utility of intravenous catheters for femoral arterial cannulation in infants having complicated sternal re-entry.
Many children undergoing surgery for congenital heart disease have had prior operations. Reoperative sternotomy carries with it the risk of cardiac injury and the need for emergency peripheral cannulation. Unfortunately, in smaller children, many of the standard sites are occluded, stenosed, or too small for the smallest manufactured cannulas. Using a model, we evaluated the feasibility of using peripheral intravenous catheters for establishing cardiopulmonary support. A model cardiopulmonary bypass circuit was created. Catheters of varying diameter and length were used to simulate arterial cannulation. With each catheter, flows were incrementally increased until a maximum arterial line pressure was achieved. Data were recorded. Catheter diameters increased from 24 to 14 gauge, and catheter lengths increased from 1 inch to 1.88 inches. Flow could not be established through the 24-gauge catheter without an unacceptably high arterial line pressure. From the next smallest catheter (22 gauge x 1 inch) to the largest catheter (14 gauge x 1.88 inches), maximum achievable flow rates ranged from 70 to 540 mL/min, respectively. Peripheral arterial cannulation may be lifesaving in cases of complicated sternal re-entry in children. Intravenous catheters can be used when vessels are too small for standard cannulas. Flow rates that provide adequate support while maintaining acceptable arterial line pressures are achievable with peripheral intravenous catheters until central cannulation can be accomplished safely.


Quantitative gas chromatographic/mass spectrometric analysis of morphine glucuronides in human plasma by negative ion chemical ionization mass spectrometry.
A sensitive and specific method for the determination of morphine glucuronides in human plasma is presented. Morphine glucuronides, namely morphine-6-glucuronide (M6G) and morphine-3-glucuronide (M3G), were extracted from plasma by solid-phase extraction on C(18) cartridges at pH 9.3 and derivatized to their pentafluorobenzyl ester trimethylsilyl ether derivatives. The compounds were measured by gas chromatography/negative ion chemical ionization mass spectrometry without any further purification. Using this detection mode, a diagnostic useful fragment ion at m/z 748 was obtained at high relative abundance for both target compounds. [(2)H(3)]-labeled morphine glucuronides were used as internal standards. Calibration graphs were calculated by polynomial fit within a range of 10-1280 and 15-1920 nmol l(-1) for the 6- and 3-glucuronide, respectively. At the limit of quantitation (LOQ), the inter-assay precision was 2.21% (M3G) and 2.23% (M6G) and the GC/MS assay variability was 1.8% (M3G) and 0.9% (M6G). The accuracy at the LOQ showed deviations of +4.92% (M3G) and +1.5% (M6G). The sample recovery after solid-phase extraction was 84.7% for both M3G and M6G. The method is rugged, rapid and robust and has been applied to the batch analysis of morphine glucuronides during pharmacokinetic profiling of the drugs.


Genotoxicity study of bojungchisup-tang, an oriental herbal decoction-in vitro chromosome aberration assay in Chinese hamster lung cells and in vivo supravital-staining micronucleus assay with mouse peripheral reticulocytes.
The toxicity evaluation of oriental herbal drugs is of great concern at present. Bojungchisuptang (BCST, in Korean), a decocted medicine of oriental herbal mixture, is now well used in clinic at oriental hospitals for the treatment of edema of several diseases in practice. However, the toxicity of the oriental herbal decocted medicines such as genetic toxicity is not well defined until now. In this respect, to clarify the genetic toxicity of BCST, in vitro chromosome aberration assay with Chinese hamster lung (CHL) fibroblasts and in vivo supravital micronucleus assay with mouse peripheral reticulocytes were performed in this study. In the chromosome aberration assay, we used 5,000 micrograms/ml BCST as maximum concentration because no remarkable cytotoxicity in CHL cells was observed both in the presence and absence of S-9 metabolic activation system. No statistical significant differences of chromosome aberrations were observed in CHL cells treated with 5,000, 2,500 and 1,250 micrograms/ml BCST for 6 hour both in the presence and absence of S-9 metabolic activation. However, very weak positive result (6.5-8.0% aberration) of BCST was obtained in the absence of S-9 metabolic activation system at 5,000 micrograms/ml BCST when treated for 24 hour, i.e. 1.5 normal cell cycle time. And also, in vivo clastogenicity of BCST was studied by acridine orange-supravital staining micronucleus assay using mouse peripheral reticulocytes. We used 2,000 mg/kg as the highest oral dose in this micronucleus assay because no acute oral toxicity of BCST was observed in mice. The optimum induction time of micronucleated reticulocytes (MNRETs) was determined as 36 hours after oral administration of 2,000 mg/kg BCST. No significant differences of MNRETs between control and BCST treatment groups were observed in vivo micronucleus assay. From these results, BCST revealed very weak positive result in chromosome aberration assay in vitro with CHL cells and no clastogenicity in micronucleus assay in vivo.


Simultaneous vs. sequential analysis for population PK/PD data II: robustness of methods.
A model can be fit to joint PK/PD data (concentration and effect) either simultaneously or sequentially. The results of a companion paper suggested that when the data-analytic and true models agree, a particular sequential approach is computationally faster than the simultaneous one, yet produces hardly less precise PD parameter estimates, and for suitable designs, about as accurate PD standard error estimates. In this paper, we compare the performance of various methods for the case that the data-analytic model is misspecified. We illustrate these methods by applying them to a set of real data. Using NONMEM, population PK/PD observations were simulated under various study designs according to a one- or two-compartment PK model and direct Emax or sigmoid Emax model. A one-compartment PK model and Emax PD model were fit to the simulated observations by simultaneous and sequential methods. Predictive performance (interpolation and extrapolation) of PD and the type-I error rate of a likelihood ratio test are compared. The real data set consists of PK and (more frequent) PD observations after administration of the muscle relaxant vecuronium. When only the PK data-analytic model is misspecified, the simultaneous method has greater precision than the sequential methods. However a sequential method that uses a non-parametric PK model performs better than both other methods when PK model misspecification is severe. When the PD data-analytic model is misspecified, sequential and simultaneous methods perform similarly. The analysis of the real data shows that the PK fitted with the simultaneous method can be quite sensitive to PD model misspecification, yielding a possible diagnostic for this type of misspecification.


Jigless knotless internal brace versus other open Achilles tendon repairs using a progressive rehabilitation protocol: a biomechanical study.
The jigless knotless internal brace surgery (JKIB), a modified minimal invasive surgery (MIS) for acute Achilles tendon injury, has advantages of preventing sural-nerve injury in MIS and superficial wound infection in open surgery, as demonstrated in previous clinical research. However, to date, biomechanical testing has not yet been validated. Sixty fresh porcine Achilles tendons were used to compare the JKIB with other open surgery techniques, the four-stranded Krackow suture (4sK) and the triple-bundle suture (TBS) in biomechanical testing with cyclic loading set at 1 Hz. This approach simulated a progressive rehabilitation protocol where 20-100 N was applied in the first 1000 cycles, followed by 20-190 N in the second 1000 cycles, and then 20-369 N in the third 1000 cycles. The cycles leading to repair gaps of 2 mm, 5 mm, and 10 mm were recorded. The survival cycles were defined as repair gap of 10 mm. With respect to survival cycles, a significant difference was found among the three groups, in which the TBS was the most robust, followed by the JKIB and the 4sK, where the mean survived cycles were 2639.3 +/- 263.55, 2073.6 +/- 319.92, and 1425.25 +/- 268.96, respectively. Significant differences were verified via a post hoc analysis with the Mann-Whitney U test after the Bonferroni correction (p < 0.017). The TBS was the strongest suture structure in acute Achilles tendon repair. However, the JKIB could be an option in acute Achilles tendon repair with the MIS technique due to it being more robust than the 4sK, which has been typically favored for use in open repair.


Factors predicting home medication management practices among chronically ill older population of selected districts of Nepal.
Older population often have multiple and complex needs that are consequently challenged by the presence of polypharmacy, adverse drug reactions and drug-drug interaction. We aimed to determine home medication management practices (MMP) and its associated factors among chronically ill older population of selected districts of Nepal. A community based cross-sectional survey was conducted among 386 chronically ill older individuals from selected areas of Nepal between April to September 2016. Appropriateness of MMP was assessed through scores of questions using interview method. Multivariate logistic regression analysis using potential variables from bivariate analysis were used to determine factors affecting MMP. The overall home MMP was mostly inappropriate (80.1%). Most participants had multiple prescribers for single disease (202, 52.3%) and inappropriate medication storage (188, 48.7%). Though the majority of them had drug administration schedule (378, 97.9%), expired medicines were also used (2, 0.5%). Regression analysis showed less than one year duration of disease (odds ratio [OR] = 3.901, 95% confidence interval [CI] = 1.528 to 9.959, P = 0.004), 1-2 years duration of disease (OR = 2.415, 95% CI = 1.210 to 4.821, P = 0.012) and smokers (OR = 2.025, 95% CI = 1.036 to 3.956, P = 0.039) as the major factors affecting appropriate home MMP. The home MMP was associated with duration of disease and smoking status among chronically ill older patients living in selected districts of Nepal. Proper counselling and monitoring of such patients might be necessary to improve the practice.


Effect of allograft amniotic membrane use on adhesion formation after Cesarean section in pregnant rat.
We aimed to investigate the effect of allograft amniotic membrane utilization to prevent the post-operative adhesion formation. In 24 pregnant inbreed Wistar-Albino rats, pregnancy was terminated by forming bilateral uterine horn defect via cesarean section at 20th gestational day. Rats were assigned in three groups randomly. In the first group, abdomen closure was achieved without administration of any intra-peritoneal material following standard surgical intervention. In the second group Seprafilm was used to cover the defect at anterior horn of uterus; whereas amniotic membrane of the rat itself was used in the third group. After 3 weeks, all rats were sacrificed and re-laparotomy was performed to determine adhesions scores. No significant difference was found in adhesion scores between the group 1 and group 3 and also between group 2 and group 3, supporting the previous findings in the literature. We observed that direct application of allograft amniotic membrane, which is an adhesion barrier used after cesarean section, to injured surface had no effect in the prevention of adhesions (Tab. 3, Fig. 5, Ref. 28).


Clinical, microbiological, and molecular epidemiological characteristics of Klebsiella pneumoniae-induced pyogenic liver abscess in southeastern China.
Klebsiella pneumoniae-induced pyogenic liver abscess (KP-PLA) has emerged as a life-threatening disease worldwide. However, to date, a limited number of scholars have attempted to systematically elucidate the characteristics of KP-PLA. The aim of the present study was to analyze clinical, microbiological, and molecular epidemiological characteristics of KP-PLA patients in Southeastern China. The KP-PLA cases from a tertiary teaching hospital in China from January 2016 to December 2017 were systemically studied and elucidated comprehensively. The virulence factors, resistant spectrum, and clones of K. pneumoniae isolates were identified with string test, polymerase chain reaction (PCR), antimicrobial susceptibility test, and multilocus sequence typing. Moreover, the characteristics in KP-PLA patients with and without other hepatobiliary diseases (OHD) were also been compared. A total of 163 KP-PLA cases were enrolled, in which the majority of those cases were senior males, and often associated with multiple underlying diseases, including diabetes (49.7%). The remaining cases belonged to healthy individuals (50.3%). The clinical symptoms were common but nonspecific, characterized by increased inflammatory parameters and abnormal liver function parameters. The abscess was often right-sided solitary presentation (58.3%). Cephalosporin or carbapenem plus metronidazole combined with percutaneous puncture or catheter drainage were favorable therapeutics. Although low resistance rates of commonly used antimicrobial drugs (< 10%) were observed, twelve strains were identified as multidrug resistant (MDR) strains, and were mainly isolated from the OHD patients. The hypermucoviscosity, as well as K1 and K2 serotypes accounted for 30.7, 40.5, and 19.0%, respectively. Except for iroN (24.5%) and magA (45.4%), the high prevalence of virulence genes (e.g. aerobactin, rmpA, mrkD, fimH, uge, ureA, entB, ybtA, kfuBC, and wcaG) was identified (68.7-100.0%). Additionally, ST23 was found as a predominant sequence type (ST; 38.7%), and three novel STs (ST3507, ST3508 and ST3509) were noted as well. The present study reported the abundant hvKp strains in KP-PLA, as well as convergence of hypervirulent and MDR K. pneumoniae isolates from the KP-PLA patients, particularly those cases with OHD. Given the various clinical manifestations and destructive pathogenicity, determination of the comprehensive characteristics of such isolates is highly essential to effectively carry out for optimal management and treatment of KP-PLA.


Single- Versus Multiple-Fraction Radiation Therapy for Painful Bone Metastases: A Systematic Review and Meta-analysis of Nonrandomized Studies.
Single-fraction radiation therapy (RT) is a convenient and cost-effective regimen for the palliation of painful bone metastases, but is still underused. Randomized controlled trials comparing single- versus multiple-fraction RT are limited by generalizability. We compared the pain response rates after single- versus multiple-fraction RT in nonrandomized studies. We searched PubMed and Scopus from the inception of each database through August 2018. We sought to identify nonrandomized studies in which data on pain response rates could be extracted for single- and multiple-fraction RT. Our primary outcomes of interest were the overall and complete pain response rates in evaluable patients. The analysis was performed using a random-effects model with the Mantel-Haenszel method. Of the 3933 articles identified through our search, 9 met our inclusion criteria. Five of 9 included studies did not exclude patients with features of complicated bone metastases. A 1 × 8 Gy radiation schedule was frequently used in single-fraction therapy, and schedules of 5 × 4 Gy and 10 × 3 Gy were frequently used in multiple-fraction therapy. In the 9 studies, the overall response rate was 67% (884 of 1321 patients) for patients in the single-fraction arm and 70% (953 of 1360 patients) for those in the multiple-fraction arm (pooled odds ratio [OR]: 0.85; 95% confidence interval [CI], 0.66-1.08). In 5 studies, the complete response rate was 26% (195 of 753 patients) for patients in the single-fraction arm and 35% (289 of 821 patients) for those in the multiple-fraction arm (pooled OR: 0.89; 95% CI, 0.70-1.13). There were no significant differences in the overall and complete response rates between single- and multiple-fraction RT. The effectiveness of single-fraction regimens was shown in nonrandomized settings, which better reflect daily practice than randomized studies. The CIs for the pooled ORs included clinically meaningful differences, and the study results are inconclusive.


Convergent pathways to biosynthesis of the versatile cofactor F420.
Cofactor F420 is historically known as the methanogenic redox cofactor, having a key role in the central metabolism of methanogens, and archaea in general. Over the past decade, however, it has become evident this cofactor is more widely distributed across archaeal and bacterial taxa, suggesting a broader role for F420 in various metabolic and ecological capacities. In this article, we focus on the recent findings that have led to a deeper understanding of F420 biosynthetic enzymes and metabolites across microorganisms.


Time course of hormonal independence for developmental events in neurons and other cell types during insect metamorphosis.
The ecdysteroid dependence of developmental events during the larval-pupal transformation of the tobacco hornworm, Manduca sexta, was studied using the technique of abdomen ligation to eliminate the ecdysteroid-secreting prothoracic glands (PTG) in the thorax. The time at which a particular developmental event no longer required the presence of the PTG for its successful completion, i.e., when the event became ecdysteroid independent, was determined for the dendritic regression and death of proleg motor neuron PPR, the degeneration of its target muscle PPRM, and the secretion of pupal cuticle by the abdominal epidermis. Groups of abdomens were ligated from synchronously developing larvae at 12-hr intervals over the 4-day period preceding entry into the pupal stage. PPR's dendritic regression became ecdysteroid independent over the first 24 hr of this period. When abdomens were ligated midway through this period PPR became arrested in a partially regressed state. Twelve hours after PPR's regression became independent, the degeneration of PPRM and the secretion of pupal cuticle by the epidermis acquired independence. Another 12 hr elapsed before PPR's programmed death became independent. Thus, different developmental events acquired ecdysteroid independence in a stereotyped temporal sequence, and even different events within a single cell, i.e., PPR's regression and death, became independent at different times.


Qualitative aspects on motility changes in mice induced by low doses of apomorphine and clonidine.
Single mice were treated with apomorphine (0.05, 0.2 or 0.8 mg/kg) or clonidine (0.025 or 0.05 mg/kg), and were placed in a new kind of motility meter, allowing classification of movements in terms of size, and also charting the motility with the help of an X/Y pen recorder. All treatments induced reduction of motility in this model. Apomorphine, 0.8 mg/kg, considerably changed the motility type, whereas the lower doses did not. This finding is consistent with earlier studies on gross lobomotor activity and suggests that the higher dose alone evokes a significant direct net stimulation of postsynaptic dopamine receptors, whereas the lower doses merely reduce the physiological stimulation of these receptors by inhibiting dopamine neurotransmission. Clonidine, 0.025 mg/kg, depressed motility at least as much as did 0.05 mg/kg, in agreement with previous behavioural studies.


The process of self-report of impairment in clinical research.
This paper examines how labels for impairment are negotiated by people with disabilities during clinical assessment. It builds on Robert Murphy's (1987) explanations of the disability experience as rooted in the individual's sense of having multiple past, present and intended future body-selves. Using transcripts of five consecutive daily clinical research assessments, it describes the conduct of clinical research assessments with an older man with stroke-related motor impairments and dysphoria. It also examines how the researcher as Other shapes a person's sense of identity, experience and quality of life by regulating the medical labels for personal experience, and by authoring socially authoritative scientific models of disabilities. Points of dissension (regarding identity, discourse and time perspectives) and collaboration are identified and then illustrated with excerpts from the transcripts. Analysis reveals how salient personal experiences are locally asserted in discourse, and selectively misrepresented in the clinical research record.


Receptor status of breast cancer diagnosed during pregnancy: A literature review.
The definition of PABC is inconsistently given as either breast cancer diagnosed exclusively during pregnancy, or combined with breast cancer diagnosed within six months to five years after delivery, and sometimes even longer. The longer away from the delivery date breast cancer is diagnosed, the less clear this association with pregnancy may become. Therefore, breast cancer diagnosed during pregnancy (BCdP) may not necessarily be the same disease entity as PABC. This review aims to provide an overview of BCdP receptor status, as this has not been assessed before. BCdP tumors were predominantly ER negative (56.6 %), PR negative (57.2 %) or both ER and PR negative (47.9 %). Moreover, HER2-overexpression was seen in 33.2 % of BCdP patients and 27.6 % had triple negative disease. This predominantly ER and PR negative profile with more often HER2 overexpression is aggressive and distinct from non-pregnant similar-aged patients, warranting future comparative research.


Application of cellulose nanofiber as a promising air filter for adsorbing particulate matter and carbon dioxide.
Pollution from particulate matter (PM) and toxic chemicals in the air cause some of the most critical health and environmental hazards in developed and developing countries. It can have a very destructive effect on human health and other living creatures. In particular, PM air pollution caused by rapid industrialization and population growth is a grave concern in developing countries. Oil and chemical-based synthetic polymers are non-environmentally friendly materials that lead to secondary environmental pollution. Thus, developing new and environmentally compatible renewable materials to construct air filters is essential. The goal of this review is to study the use of cellulose nanofibers (CNF) to adsorb PM in the air. Some of CNF's advantages include being the most abundant polymer in nature, biodegradable, and having a high specific surface area, low density, surface properties (broad possibility of chemical surface modification), high modulus and flexural stiffness, low energy consumption, which provide this new class of bio-based adsorbent with promising potential applications in environmental remediation. Such advantages have made CNF a competitive and highly in-demand material compared to other synthetic nanoparticles. Today, refining membranes and nanofiltration manufacturing are two important industries that could use CNF to provide a practical step in protecting the environment and saving energy. CNF nanofilters are capable of nearly eliminating most sources of air pollution, including carbon monoxide, sulfur oxides, nitrogen oxides, and PM2.5-10 μm. They also have a high porosity and low resistance air (pressure drop) ratio compared to ordinary filters made from cellulose fiber. If utilized correctly, humans do not need to inhale harmful chemicals.


Non-visual consciousness and visual images in blindsight.
In a recent response paper to Brogaard (2011a), Morten Overgaard and Thor Grünbaum argue that my case for the claim that blindsight subjects are not visually conscious of the stimuli they correctly identify rests on a mistaken necessary criterion for determining whether a conscious experience is visual or non-visual. Here I elaborate on the earlier argument while conceding that the question of whether blindsight subjects are visually conscious of the visual stimuli they correctly identify largely is an empirical question. I conclude by sketching a method for testing whether blindsight subjects have visual consciousness of stimuli presented to them in their blind field.


Association between retinal vein occlusion, axial length and vitreous chamber depth measured by optical low coherence reflectometry.
Results of ocular biometric measurements in retinal vein occlusion (RVO) eyes are still inconclusive and controversial. The aim of this study was to evaluate the association between ocular axial length (AL), vitreous chamber depth (VCD) and both central (CRVO) and branch retinal vein occlusions (BRVO) using optical low coherence reflectometry (OLCR). Both eyes of 37 patients with unilateral CRVO (mean age: 66 ± 14 years, male:female - 21:16) and 46 patients with unilateral BRVO (mean age: 63 ± 12 years, male:female - 24:22) were enrolled in this study. The control group consisted of randomly selected single eyes of 67 age and gender matched volunteers without the presence or history of RVO (mean age: 64 ± 14 years, male:female - 34:33). Optical biometry was performed by OLCR biometer (LenStar LS 900). Average keratometry readings, central corneal thickness (CCT), anterior chamber depth (ACD), lens thickness (LT), AL and VCD of eyes with RVO were compared with those of fellow eyes using paired t-tests and with those of control eyes using independent t-tests. Mean CCT, ACD and LT, average keratometry readings of affected RVO eyes, unaffected fellow eyes and control eyes was not statistically different in either groups. In eyes with CRVO mean AL and VCD of affected eyes were significantly shorter than those of control eyes (p < 0.001, p < 0.05), mean difference in AL and VCD between the affected and control eyes was 0.56 ± 0.15 mm and 0.45 ± 0.19 mm, respectively. In eyes with BRVO, mean AL of the affected eyes was significantly shorter with a mean difference of 0.57 ± 0.15 mm (p < 0.001) and the VCD was significantly shorter with a mean difference of 0.61 ± 0.15 mm (p < 0.001) comparing with the control eyes. Shorter AL and VCD might be a potential anatomical predisposing factor for development either of CRVO or BRVO.


Design, synthesis and biological evaluation of 2,4,6- trisubstituted triazine derivatives as new nonpeptide small-molecule SIRT5 inhibitors.
Human sirtuin 5 (SIRT5) participates in a variety of metabolic disorder-associated diseases, including cancer. Inhibition of SIRT5 has been confirmed to provide a new strategy for treatment of related diseases. Previously, we discovered a pyrimidine skeleton inhibitor XIV, which showed low micromolar inhibitory activity against SIRT5. Herein, we utilized the scaffold-hopping strategy to design and synthesize a series of 2,4,6- trisubstituted triazine derivatives. The SAR analysis led to the discovery of several new SIRT5 inhibitors with low micromolar inhibition levels. The most potent compounds 10 (IC50 = 5.38 µM), and 14 (IC50 = 4.07 µM) were further confirmed to be the substrate-competitive SIRT5 inhibitors through enzyme kinetic assays, which is consistent with the molecular docking analyses. Fluorescence-based thermal shift assays proved that these compounds may stabilize SIRT5 by binding withprotein.. In addition, compounds 10 and 14 were also revealed to have moderate selectivity to SIRT5 over SIRT1-3. This study will aid further efforts to develop highly potent and selective SIRT5 inhibitors for the treatment of cancer and other related diseases.


Research ethics committees: agents of research policy?
The purpose of this commentary is to describe the unintended effects ethics committees may have on research and to analyse the regulatory and administrative problems of clinical trials. The Finnish law makes an arbitrary distinction between medical research and other health research, and the European Union's directive for good clinical trials further differentiates drug trials. The starting point of current rules is that clinical trials are lesser in the interest of patients and society than routine health care. However, commercial interests are not considered unethical. The contrasting procedures in research and normal health care may tempt physicians to continue introducing innovations into practice by relying on unsystematic and uncontrolled observations. Tedious and bureaucratic rules may lead to the disappearance of trials initiated by researchers. Trying to accommodate the special legislative requirements for new drug trials into more complex interventions may result in poor designs with unreliable results and increased costs. Meanwhile, current legal requirements may undermine the morale of ethics committee members. The aims and the quality of the work of ethics committees should be evaluated, and a reformulation of the EU directive on good clinical trials is needed. Ethical judgement should consider the specific circumstance of each trial, and ethics committees should not foster poor research for legal reasons.


Activity and localization of NADH-dependant oxidoreductase (diaphorase) in boar spermatozoa.
Investigations were carried out into the activity and localization of NADH-dependant diaphorase in boar spermatozoa. Semen samples were collected from healthy boars, used in A.I. centers. The enzyme was extracted with distilled water and Triton X-100. Two forms of diaphorase were found-water-soluble and Triton X-100 soluble, showing low activity-0.36 U/ml and 0.26 U/ml. The enzyme was localized in the mitochondria, manifesting different intensities of reaction between sperm cells in the same ejaculate. It was found, that a part of the mitochondria and outer doublets showed positive reaction. It is suggested that the enzyme regulates the ratio between reduced and oxidized forms of NADH, takes part in the energy balance and possibly in the mechanism of sperm motility.


[Blood serum cortisol, insulin and somatotropin concentration in children with hypothalamo-hypophyseal insufficiency].
In 22 patients with restricted growth the consecutive stimulation with arginine and insuline was applied for the diagnosis of somatotropic insufficiency. 13 children showed GH insufficiency. Simultaneously cortisol level during hypoglycemia for ACTH reserve and insulin level after arginine application were estimated.


The optimal cut-off value of the preoperative prognostic nutritional index for the survival differs according to the TNM stage in hepatocellular carcinoma.
To establish the optimal cut-off value of the preoperative prognostic nutritional index (PNI) for prognosis according to the Tumor Node Metastasis (TNM) stage of hepatocellular carcinoma (HCC) after curative resection. This retrospective study reviewed the records of 375 patients. The optimal cut-off value of the PNI was established according to the TNM stage, and overall survival was compared between the low and high PNI groups. The optimal cut-off value of the PNI decreased with increasing TNM stage, with 52, 47, and 43 patients having stage I, II, and III HCC, respectively. A low preoperative PNI predicted a poorer overall survival than did a high PNI for stage I (P < 0.001) and II (P = 0.002), but not stage III disease (P = 0.052). Multivariate analysis revealed that the preoperative PNI was an independent predictor of overall survival for stage I and II HCC (hazard ratios: 6.96 and 3.57, P = 0.001 and P = 0.001, respectively). The findings of this study show that the optimal cut-off value for the PNI for prognosis differs among the TNM stages and that the preoperative PNI is a favorable prognostic factor for stage I HCC.


Effects of fitness and age on the response to vagotonic atropine.
Previous work indicates compromised cardiac vagal control plays a prominent role in reducing arterial baroreflex gain with age, however older fit individuals display cardiovagal baroreflex responses similar to young individuals. The purpose of this study was to test the hypothesis that chronic aerobic exercise mitigates against age-related declines in cardiac parasympathetic receptor function. In forty-four young and old (fit and unfit) individuals, we used the parasympathomimetic responses to low doses of atropine to probe cardiac cholinergic receptor responses. Data were collected before and after eight doses of atropine sulfate from 0.4 to 7.2 microg/kg. Chronotropic responses were assessed from average RR intervals and heart rate variabilities were derived in time and frequency domains. All subjects exhibited bradycardia with at least one dose of atropine and peak bradycardia occurred at a similar dose in each group. However, changes in heart rate variability did not consistently track the chronotropic responses within subjects (r-square from 0.90 down to 0). As expected, basal RR interval was longer in the fit groups and was unaffected by age. However, the degree of RR interval lengthening with parasympathomimetic atropine was unaffected by physical fitness and was significantly less in all older subjects. These data indicate there are certain prepotent age-related declines in the cardiac parasympathetic system that cannot be prevented by regular physical activity.


Prospective analysis of eosinophilia in spontaneously diabetic BB rats: correlation with islet inflammation but not with diabetes development.
A prospective analysis of eosinophil counts in diabetes prone BB rats showed that eosinophilia (greater than 5%) occurred between 70-100 days of age. The presence of eosinophilia did not predict diabetes onset because persistently normoglycaemic animals developed eosinophilia as well. Nevertheless a correlation with the disease process in BB rats was found. Eosinophilia decreases a few weeks after diabetes onset but persists in non diabetic rats. Histological analyzes showed that eosinophilia correlates with eosinophil infiltration of islets (p less than 0.005) and the latter correlates with severe insulitis (p less than 0.005). These findings indicate that eosinophilia is associated with a late stage of islet inflammation in diabetes prone BB rats independent of whether the animals develop diabetes or not.


TIMP-1 overexpression does not affect sensitivity to HER2-targeting drugs in the HER2-gene-amplified SK-BR-3 human breast cancer cell line.
Tissue inhibitor of metalloproteinases-1 (TIMP-1) has been suggested as a marker of prognosis and response to treatment in breast cancer. In vitro, TIMP-1 can regulate shedding of the extracellular domain of HER2 and signalling via the Akt pathway, and we hypothesize that TIMP-1 therefore can affect sensitivity to the HER2-targeting drugs trastuzumab and lapatinib. SK-BR-3 human breast cancer cells were stably transfected with TIMP-1, characterized with regard to TIMP-1 protein expression, proliferation, and functionality of the secreted TIMP-1, and the sensitivity to trastuzumab and lapatinib was studied in five selected single-cell subclones expressing TIMP-1 protein at various levels plus the parental SK-BR-3 cell line. Both trastuzumab and lapatinib reduced cell viability, as determined by MTT assay, but the sensitivity to the drugs was not associated with the expression level of TIMP-1 protein. Western blotting showed that the activation of Akt, PTEN, and HER2 as well as ADAM10 was similar in all clones. In conclusion, in this model, TIMP-1 overexpression does not affect HER2 cleavage by ADAM10 or signalling via the Akt pathway, and TIMP-1 does not influence sensitivity to trastuzumab and lapatinib.


Effect of isosporiasis prevention with toltrazuril on long-term pig performance.
The efficacy of toltrazuril treatment was assessed in two experiments in Polish swine herds. Experiment 1 included a toltrazuril treatment group, Group A (n = 410), and untreated control, Group B (n = 386). Time to sale in Group A was 108 days versus 120 days for Group B, with average body weights at sale of 114.2 kg and 108.8 kg, respectively (P < 0.05). In experiment 2, the health status and body weight gain of 238 piglets treated with toltrazuril (Group D) were compared to 235 untreated piglets (Group K). A similar difference was observed in average body weights of slaughtered animals, being on average 104 kg in Group D and 101 kg in Group K (P < 0.01). Animals from Group D were slaughtered 5 days earlier than animals from Group K (day 166 versus day 171). Data from clinical trials suggest treatment of coccidiosis with toltrazuril offering potential for improved animal welfare and yields, however this has remained unproven in field conditions in large swine production facilities. The present study confirms the efficacy of toltrazuril treatment when used in the field and the subsequent positive impact on time to weaning, time to market, and on weight gain at all time points.


Turning behaviour induced by injection of muscimol or picrotoxin into the substantia nigra demonstrates dual GABA components.
Injection of the GABA agonist muscimol into rat caudal substantia nigra caused contralateral turning, whereas injection into the rostral substantia nigra caused ipsilateral turning. The GABA antagonist picrotoxin had the opposite effect. These findings support the hypothesis that GABA has dual actions in the substantia nigra. Ipsilateral turning induced by injection of muscimol into rostral nigra was abolished by haloperidol pretreatment, indicating the involvement of dopaminergic mechanisms. Haloperidol pre-treatment did not prevent turning induced by muscimol injected into the caudal nigra, supporting the existence of a non-dopaminergic nigral output system.


Genetic diversity of penicillin-binding protein 2 genes of penicillin-resistant strains of Neisseria meningitidis revealed by fingerprinting of amplified DNA.
A 2-kilobase fragment containing the penicillin-binding protein 2 gene (penA) was amplified by using the polymerase chain reaction with DNA prepared from 35 penicillin-resistant strains of Neisseria meningitidis isolated in England, Ireland, and Spain (MICs, 0.16 to 1.28 micrograms of benzylpenicillin per ml) and from 10 penicillin-susceptible strains (MICs, less than or equal to 0.04 micrograms of benzylpenicillin per ml). The penA genes were digested with HpaII or TaqYI; and the resulting fragments were end-labeled, fractionated on a polyacrylamide gel, and autoradiographed, to produce gene fingerprints. The fingerprints of the penA genes of the 10 penicillin-susceptible strains were identical. In contrast, the fingerprints of the penA genes of all but two of the penicillin-resistant strains differed markedly from those of the penicillin-susceptible strains. The altered penA genes of group B penicillin-resistant strains were very diverse, and 14 different fingerprints were found among the 15 strains that we examined. The penA genes of the 19 resistant group C strains were less diverse, and two major fingerprint patterns predominated.


A Simple Spectrofluorimetric Method Based on Quenching of a Nickel(II)-Phthalocyanine Complex to Determine Iron (III).
A new nickel(II)-phthalocyanine complex (NiPc) was synthesized and used as a fluorescent ligand in determination of iron in real samples. The NiPc compound, when excited at 350 nm, decreases of emission with increases of the iron(III) concentration at 425 nm were used analytical response in a modified standard addition method. The method was validated by analyzing two certified reference materials (CRM-SA-C Sandy Soil C and Mixed Polish Herbs (INCT-MPH-2). Food and drug samples were digested in a closed microwave system using nitric acid and hydrogen peroxide. Therefore, all iron in the samples converted to iron(III) ion. These solutions were used directly in determination of iron(III) ion. No cleanup or enrichment of the solutions was required. The calibration graph was linear until 14.00 μg mL(-1). Detection limit and quantification limit were 1.29 μg mL(-1) and 3.88 μg mL(-1), respectively. The method provided accurate results for the majority of the food samples tested, including spanich, dill, mint, purslane, rocket, red lentils, dry beans and two iron medicinal tablets. Also, the high recovery (95.6 %) was obtained for a fortified stream water sample. The simple and cost-effective method is suitable for monitoring total iron concentration in foods and drug samples.


Fragile X syndrome.
The fragile X syndrome is the most common inherited form of mental retardation known. Its phenotype includes large or prominent ears, macroorchidism, and characteristic behavioral problems. It has attracted the interest of cytogeneticists and molecular biologists because of its characteristic fragile site on the X chromosome. It has puzzled geneticists because of its unusual inheritance pattern involving nonpenetrant males. This syndrome has also spearheaded an appreciation of cytogenetic abnormalities in the etiology of all degrees of developmental delay.


An improved multi-view spectral clustering based on tissue-like P systems.
Multi-view spectral clustering is one of the multi-view clustering methods widely studied by numerous scholars. The first step of multi-view spectral clustering is to construct the similarity matrix of each view. Consequently, the clustering performance will be greatly affected by the quality of the similarity matrix of each view. To solve this problem well, an improved multi-view spectral clustering based on tissue-like P systems is proposed in this paper. The optimal per-view similarity matrix is generated in an iterative manner. In addition, spectral clustering is combined with the symmetric nonnegative matrix factorization method to directly output the clustering results to avoid the secondary operation, such as k-means or spectral rotation. Furthermore, improved multi-view spectral clustering is integrated with the tissue-like P system to enhance the computational efficiency of the multi-view clustering algorithm. Extensive experiments verify the effectiveness of this algorithm over other state-of-the-art algorithms.


Long-term musculoskeletal morbidity after adult burn injury: a population-based cohort study.
To investigate if adults who are hospitalised for a burn injury have increased long-term hospital use for musculoskeletal diseases. A population-based retrospective cohort study using linked administrative health data from the Western Australian Data Linkage System. Records of 17,753 persons aged at least 20 years when hospitalised for a first burn injury in Western Australia during the period 1980-2012, and 70,758 persons who were age and gender-frequency matched with no injury admissions randomly selected from Western Australia's electoral roll. Admission rates and cumulative length of stay for musculoskeletal diseases. Negative binomial and Cox proportional hazards regression modelling were used to generate incidence rate ratios (IRR) and HRs with 95% CIs, respectively. After adjustment for pre-existing health status and demographic characteristics, the burn cohort had almost twice the hospitalisation rate for a musculoskeletal condition (IRR, 95% CI 1.98, 1.86 to 2.10), and spent 3.70 times as long in hospital with a musculoskeletal diagnosis (95% CI 3.10 to 4.42) over the 33-year period, than the uninjured comparison cohort. Adjusted survival analyses of incident post-burn musculoskeletal disease admissions found significant increases for the 15-year post burn discharge period (0-6 months: HR, 95% CI 2.51, 2.04 to 3.11; 6 months-2 years: HR, 95% CI 1.77, 1.53 to 2.05; 2-15 years: HR, 95% CI 1.32, 1.23 to 1.42). Incident admission rates were significantly elevated for 20 years post-burn for minor and severe burn injury for a range of musculoskeletal diseases that included arthropathies, dorsopathies, osteopathies and soft tissue disorders. Minor and severe burn injuries were associated with significantly increased post-burn incident admission rates, long-term hospital use and prolonged length of stay for a range of musculoskeletal diseases. Further research is required that facilitates identification of at-risk patients and appropriate treatment pathways, to reduce the long-term morbidity associated with burns.


Land use intensification alters the relative contributions of plant functional diversity and soil properties on grassland productivity.
Understanding the mechanisms of grassland productivity variation is critical for global carbon cycling and climate change mitigation. Heretofore, it is unknown how different environmental factors drive small-scale spatial variation in productivity, and whether land use intensification, one of the most important global changes, can regulate the processes that drive productivity change. Here we performed an 18-year exclosure experiment across six sites with high-intensity mowing/grazing history in northern China to examine the effects of land use intensification on plant functional diversity, soil properties, and their relative contributions to above-ground net primary productivity (ANPP). We found that plant functional diversity and soil properties contributed to the variation in ANPP both independently and equally in enclosed grasslands (plant diversity: 20.6%; soil properties: 19.5%). Intensive land use significantly decreased the Rao's quadratic entropy (RaoQ) and community-weighted mean value (CWM) of plant height, and further suppressed the contributions of plant functional diversity to ANPP. In contrast, intensive land use increased soil available N, P, pH, electrical conductivity, and homogeneity of soil available P, and strengthened their contributions to ANPP (31.5%). Our results indicate that high-intensity land use practices in grasslands decrease the role of plant functional diversity, but strengthen the effects of soil properties on productivity. We, therefore, suggest that plant functional diversity can be used effectively to boost productivity in undisturbed grasslands, while soil properties might be a more critical consideration for grassland management in an areas with increased land use.


Molecular cloning and expression analysis of three cadherin-8 isoforms in the embryonic chicken brain.
Three types of full-length cDNAs encoding chicken cadherin-8 (Cdh8) were identified and their expression in the embryonic chicken brain was investigated. The longest type of cDNA is closely similar to that of other classic cadherins, and the predicted protein shows a high similarity to rat and human Cdh8. The second type of cDNA is considerably shorter. The deduced protein lacks the cytoplasmic tail and the transmembrane domain, and contains a truncated fifth cadherin repeat in the extracellular domain (EC5; 68 amino acids), suggesting that it is a soluble isoform. The third type resembles the second one but is even shorter (only 30 amino acids in EC5). All types contain unique short sequences at their C terminus. Genomic analysis demonstrated that all of the three Cdh8 isoforms are located on chromosome 11, and consist of 12, 10, and 9 exons, respectively. Semi-quantitative RT-PCR with type-specific primers showed that the transcription of the three Cdh8 isoforms was temporally and spatially regulated in different parts of the embryonic chicken brain. The distinct regulation of gene expression suggested that the three isoforms likely play different roles during brain development. Northern blot analysis revealed that the transcription of the long isoform was much stronger than that of the two shorter ones. In situ hybridization showed that the long isoform of Cdh8 is expressed by a specific subset of brain nuclei, regions and layers in all major parts of the brain.


Laser-ablative aqueous synthesis and characterization of elemental boron nanoparticles for biomedical applications.
Boron-based nano-formulations look very promising for biomedical applications, including photo- and boron neutron capture therapies, but the fabrication of non-toxic water-dispersible boron nanoparticles (NPs), which contain the highest boron atom concentration, is difficult using currently available chemical and plasma synthesis methods. Here, we demonstrate purely aqueous synthesis of clean boron NPs by methods of femtosecond laser ablation from a solid boron target in water, thus free of any toxic organic solvents, and characterize their properties. We show that despite highly oxidizing water ambience, the laser-ablative synthesis process follows an unusual scenario leading to the formation of boron NPs together with boric acid (H3BO3) as an oxidation by-product coating the nanoparticles, which acts to stabilize the elemental boron NPs dispersion. We then demonstrate the purification of boron NPs from residual boric acid in deionized water, followed by their coating with polyethylene glycol to improve colloidal stability and biocompatibility. It was found that the formed NPs have a spherical shape with averaged size of about 37 nm, and are composed of elemental boron in mostly amorphous phase with the presence of certain crystalline fraction. The synthesized NPs demonstrate low toxicity and exhibit strong absorption in the NIR window of relative tissue transparency, promising their use in photoacoustic imaging and phototherapy, in addition to their promise for neutron capture therapy. This combined potential ability of generating imaging and therapy functionalities makes laser-synthesized B NPs a very promising multifunctional agent for biomedical applications.


Paroxysmal cold haemoglobinuria in children: 3 cases encountered within a period of 7 months.
3 children who fulfilled the criteria for the diagnosis of paroxysmal cold haemoglobinuria of the classical Donath-Landsteiner type are described. The benign clinical course of this illness is confirmed. A serological study of two of the Donath-Landsteiner antibodies revealed the anti-P specificity in one, but not in another serum.


Involvement of NPY-immunoreactive neurons in the cerebral cortex of amyotrophic lateral sclerosis patients.
We performed a quantitative histochemical study of neuropeptide Y (NPY) immunoreactivity in postmortem human brain to further define patterns of neuronal degeneration in amyotrophic lateral sclerosis (ALS). The density of NPY fibers was decreased in ALS motor cortex but not in other cortical regions or striatum. Although, the density of cortical NPY neurons was unchanged in high grade cases, neurons were shrunken and atrophic with pruned dendrites. NPY neurons are less severely affected than parvalbumin neurons which are severely depleted in ALS cortex. This differential involvement of local circuit neurons suggests that a simple excitotoxic model may not adequately explain patterns of neuronal loss in ALS.


Glutamate dehydrogenase from Escherichia coli: induction, purification and properties of the enzyme.
When Escherichia coli was grown in a minimum medium with glucose as sole carbon source and a proper level of ammonia, NADP+ specific glutamate dehydrogenase (L-glutamate: NADP+ oxidoreductase (deaminating), ED 220.127.116.11) was induced. The enzyme was solubilized by French press treatment and purified to homogeneity by (NH4)2SO4 fractionation, heat treatment followed by DEAE-cellulose, hydroxylapatite and Bio-Gel chromatography with an overall yield of 30%. The enzyme proved to be heat stable and relatively resistant to protein denaturants. The optimum of enzymic activity for the reductive amination is at pH 8 and at pH 9 for the oxidative deamination. The activity is affected by adenine nucleotides. The molecular weight (about 250 000 for the native form and 46 000 for the inactive subunit) and amino acid composition, suggest strict similarities with the NADP+ enzyme from fungal origin.


Selective intraarterial chemotherapy of tumors in the lingual artery territory by a new approach.
Selective intraarterial neoadjuvant chemotherapy of carcinoma in the region of the lingual artery was performed in 23 patients (bilaterally in 4 patients). The superficial temporal artery (STA) was surgically exposed and a preshaped infusion catheter was inserted. The catheter tip was maneuvered through the STA into the lingual artery under fluoroscopic guidance. Two combinations of cytostatic agents were then continually infused through the catheter for 5-9 days. Treatment response was observed in 16 patients, 9 of whom showed a complete response. The combination of cisplatin and 5-fluorouracil was found most effective. The catheter became dislocated six times among 27 catheterizations and in each instance the position could be corrected. No major complications were encountered. We conclude that selective infusion technique is safer than previously reported semiselective infusions techniques.


Evaluation of the therapeutic effect of percutaneous nephroureterolithotomy by Tc-99m diethylenetiaminepentaacetic acid (DTPA) renal scintigraphy--alteration of the renal fraction of blood flow, split-GFR, and renal mean transit time.
To evaluate the therapeutic effects of percutaneous nephroureterolithotomy, the renal function of eleven patients with renal calculi was studied, pre- and post-intervention. Renal function was determined, by renal scintigraphy with the renal agent, Tc-99m diethylenetriaminepentaacetic acid (DTPA). In each renal scintigram the renogram curve was analyzed and the following were determined by deconvolution analysis; the renal fraction of blood flow (RFBF), DTPA-glomerular filtration ratio (GFR), and the renal mean transit time (MTT). The successful results in percutaneous nephroureterolithotomy (PNL) was proven using the radionuclide technique in most cases. From these results it can be concluded that renal scintigraphy is an effective procedure to evaluate the effect of PNL for treating renal calculi and secondary hydronephrosis.


A novel production process for optically pure L-lactic acid from kitchen refuse using a bacterial consortium at high temperatures.
We investigated L-lactic acid production in static batch fermentation of kitchen refuse using a bacterial consortium from marine-animal-resource (MAR) composts at temperatures ranging from 30 to 65 °C. At relatively low temperatures butyric acid accumulated, whereas at higher temperatures L-lactic acid was produced. In particular, fermentation at 50 °C produced 34.5 g L(-1) L-lactic acid with 90% lactic acid selectivity and 100% optical purity. Denaturing gradient gel electrophoresis indicated that dominant bacteria present in the original MAR composts diminished rapidly and Bacillus coagulans strains became the dominant contributors to L-lactic acid production at 45, 50 and 55 °C. This is the first report of the achievement of 100% optical purity of L-lactic acid using a bacterial consortium.


Bioactivity assay of porcine relaxin based on cAMP accumulation in THP-1 cells quantified by LC-MS/MS.
This study describes a reliable bioactivity assay of porcine relaxin (pRLX) based on cyclic adenosine 3',5'-monophosphate (cAMP) accumulation in the human monocyte cell-line quantified by liquid chromatography-tandem mass spectrometry (LC-MS/MS). As a result, the LC-MS/MS was based on a positive selected reaction monitoring of cAMP with a stable internal standard, 8-Br-cAMP and a protein precipitation procedure by HClO4. The standard curve of cAMP was linear from 5.0 ng mL(-1) to 992.0 ng mL(-1), with lower limits of detection and quantification of 0.5 ng mL(-1) and 5.0 ng mL(-1), respectively. The satisfactory validation data including stability assay were obtained. When measured by the LC-MS/MS, the pRLX sample showed a time- and dose-dependent stimulation of cAMP with the concentration for 50% of the maximal effect (EC50) of 40.6 ng mL(-1). The developed method indicated higher precision and selectivity than the commercial enzyme linked immunosorbent assay kits, which showed EC50 of 66.6 ng mL(-1).