Mild gestational hyperglycemia, the metabolic syndrome and adverse neonatal outcomes.
The aim of this study was to evaluate the prevalence of the metabolic syndrome and its effect on neonatal outcomes in pregnancies with different degrees of hyperglycemia. One hundred and fifty women with gestational diabetes, 100 with one abnormal value on the oral glucose tolerance test, 100 with a normal oral glucose challenge test and 350 with an abnormal challenge test and normal tolerance test were enrolled. The prevalence of the metabolic syndrome was: 0%, 4.9%, 20% and 18% in the normal challenge test, abnormal challenge and normal tolerance test, one abnormal value and gestational diabetes patients, respectively. Offspring birth weights, prevalence of large-for-gestational age babies and icterus were significantly higher in women with an abnormal challenge test (both with a normal tolerance test or one abnormal value or gestational diabetes). Metabolic syndrome was the best predictor of the presence of large-for-gestational age babies in patients with an abnormal challenge and normal tolerance test (OR = 3.15), one abnormal value (OR = 3.53) and gestational diabetes (OR = 4.15). Metabolic syndrome in mid-pregnancy was an independent predictor of macrosomia in women with any degree of gestational hyperglycemia; the oral glucose challenge test identifies pregnancies with metabolic abnormalities and adverse neonatal outcomes also in the presence of a normal oral glucose tolerance test.


Stereoselective analysis of metoprolol and its metabolites in rat plasma with application to oxidative metabolism.
We investigated the stereoselective kinetic disposition and metabolism of metoprolol (MET) in rats. The racemic MET (15 mg/kg) was given by oral gavage and blood samples were collected from 0 to 10h (n=6 at each time point). The enantiomeric concentrations of MET and its metabolites alpha-hydroxymetoprolol (alpha-OHM) and O-demethylmetoprolol (ODM) were determined by HPLC using a Chiralpak AD chiral column and fluorescence detection. The pharmacokinetic parameters of unchanged MET and the formation of ODM did not show to be stereoselective. In contrast, the AUC (ng h/mL) of alpha-hydroxymetoprolol isomers were higher to I'R [638.2(525.2-706.2) for 1'R2R and 659.6(580.4-698.1) for 1'R,2S, mean, (95%CI)] than to I'S products [58.3(47.4-66.1) for 1'S,2R and 57.1(44.7-67.9) for 1'S,2S, mean, (95%CI)]. We conclude that the kinetic disposition of unchanged MET and the formation of ODM are not enantioselective in rats but the metabolism of alpha-OHM yields predominantly the 1'R-product.


Intraocular lens power calculation with an improved anterior chamber depth prediction algorithm.
The accuracy of intraocular lens (IOL) power calculation was evaluated in a multicenter study of 822 IOL implantations using the Binkhorst II, Sanders/Retzlaff/Kraff (SRK I, SRK II, SRK/T), Holladay, and Olsen formulas. All but the first of these were optimized in retrospect with calculation of the SRK A-constant, the Holladay surgeon factor, and the Olsen pseudophakic anterior chamber depth (ACD) for each lens style. The ACD prediction of the Olsen formula was based on a previously described regression formula incorporating preoperative ACD, corneal height, axial length, and lens thickness. Among the optical IOL power calculation formulas, the highest IOL power prediction error was found with Binkhorst's and the lowest with Olsen's, which was more accurate than the SRK/T and the Holladay formulas (P < .05). The SRK/T formula was significantly more accurate than the original SRK regression formulas (P < .001). When analyzed for axial length dependence, all formulas showed the least error in the normal range. Error of the Olsen formula was lower than that of the others in the axial length interval 20 mm to 26 mm. No differences in accuracy were found between the optical IOL calculation formulas in eyes with an axial length above 26 mm (P < .05). The accuracy of IOL power calculation can be improved with optical formulas using newer-generation ACD-prediction algorithms.


Dentoalveolar injuries, bicycling accidents and helmet use in patients referred to a Norwegian Trauma Centre: A 12-year prospective study.
Despite its many benefits, bicycling carries the risk of accidents. Although numerous studies have reported the effect of helmet use on traumatic brain injury, it remains unclear if, and to what extent, helmet use reduces the risk of facial injuries. This is particularly true in regard to injuries of the lower face. In addition, there is limited evidence of the effect of helmet use on dentoalveolar injuries. Thus, the aim of this study was to determine the frequency and distribution of dentoalveolar injuries in bicycling accidents and to explore the influence of helmet use. A total of 1543 bicyclists were included from the trauma registry of a Norwegian tertiary trauma center over a 12-year period. Data were collected prospectively, including patient characteristics, type of injury, and helmet use. The prevalence of dentoalveolar injuries was assessed in conjunction with helmet use and facial fractures. Twenty-five percent of the patients had maxillofacial injuries, and 18% of those with facial fractures exhibited concomitant dentoalveolar injuries. The most common type of dentoalveolar injury was tooth fracture (39%). The most frequent location of facial fractures with combined dentoalveolar injuries was the maxilla, which had fractured in 32 patients. Women had a higher risk of sustaining dentoalveolar injuries compared to men (odds ratio 1.50, 95% confidence interval 1.02-2.22). There were 1257 patients (81%) who had reliable registration of helmet use; 54% of these wore a helmet, while 46% did not. Helmet users had an increased risk of dentoalveolar injuries compared to non-helmeted bicyclists (adjusted odds ratio 1.54, 95% confidence interval 1.02-2.31). Dentoalveolar injuries are fairly common in trauma patients admitted to a trauma center following bicycling accidents. Bicycling helmets are associated with an increased risk of dentoalveolar injuries.


Increased theophylline concentrations secondary to ciprofloxacin.
An 82-year-old man with a history of myasthenia gravis and heart failure was admitted to the hospital with respiratory failure. Aminophylline and eventually theophylline therapy were initiated to improve respiratory status. During the hospital stay, the patient developed a resistant pseudomonal pneumonia. After failure with conventional antibiotics, ciprofloxacin was initiated because of favorable sensitivity and the planned avoidance of aminoglycoside therapy. Seventy-two hours after initiation of ciprofloxacin, the patient's theophylline level rose from a steady-state baseline of 9.8 micrograms/ml to 34.7 micrograms/ml. After the theophylline dose was reduced by approximately 67 percent, the patient's theophylline serum concentration returned to baseline (10 micrograms/ml). Until more data concerning the interaction of theophylline and ciprofloxacin are available, we recommend close monitoring of theophylline serum concentrations in patients receiving concomitant ciprofloxacin.


Pollutant emission characteristics of rice husk combustion in a vortexing fluidized bed incinerator.
Rice husk with high volatile content was burned in a pilot scale vortexing fluidized bed incinerator. The fluidized bed incinerator was constructed of 6 mm stainless steel with 0.45 m in diameter and 5 m in height. The emission characteristics of CO, NO, and SO2 were studied. The effects of operating parameters, such as primary air flow rate, secondary air flow rate, and excess air ratio on the pollutant emissions were also investigated. The results show that a large proportion of combustion occurs at the bed surface and the freeboard zone. The SO2 concentration in the flue gas decreases with increasing excess air ratio, while the NOx concentration shows reverse trend. The flow rate of secondary air has a significant impact on the CO emission. For a fixed primary air flowrate, CO emission decreases with the secondary air flowrate. For a fixed excess air ratio, CO emission decreases with the ratio of secondary to primary air flow. The minimum CO emission of 72 ppm is attained at the operating condition of 40% excess air ratio and 0.6 partition air ratio. The NOx and SO2 concentrations in the flue gas at this condition are 159 and 36 ppm, which conform to the EPA regulation of Taiwan.


Antibacterial, antioxidant, antifungal and anti-inflammatory activities of crude extract from Nitraria schoberi fruits.
This study is the first comprehensive investigation of the antibacterial, antioxidant, antifungal and anti-inflammatory activities of a crude extract from Nitraria schoberi L. (Zygophyllaceae) fruits. The extract was tested against four Gram-negative (Pseudomonas aeruginosa, Enterobacter aerogenes, Klebsiella pneumoniae and Acinetobacter lwoffii) and one Gram-positive (Staphylococcus aureus) bacteria using the agar disc diffusion and microdilution methods. P. aeruginosa was inhibited the most (widest inhibition zone) while K. pneumonia showed the largest MIC value. The antioxidant activity of fruits (0.02 mg/mL vs. 0.04, 0.06 and 1.00 mg/mL for α-tocopherol, butylated hydroxyanisole and ascorbic acid, respectively) was determined by the paired diene method. The antifungal activity of N. schoberi fruits was tested against different fungi, including Aspergillus niger and Candida albicans, with 300 µg/mL of fruit extract being the most effective concentration. The percentage of anti-inflammatory activity assayed for N. schoberi fruit extract at 100, 200 and 500 µg/mL was 36.12, 59.89 and 88.33 %, respectively. N. schoberi fruits possess potent antibacterial, antioxidant, antifungal and anti-inflammatory properties, and may be used as an antibacterial and antifungal to treat diseases and/or as a protective agent against disorders associated with oxidative stress and inflammation.


Energy gap engineering of polymeric carbon nitride nanosheets for matching with NaYF4:Yb,Tm: enhanced visible-near infrared photocatalytic activity.
Molecularly grafted carbon nitride (CN) nanosheets, matching well with the emission energy of upconversion phosphors (UCPs), were acquired for the first time. As a result of energy gap engineering, the assembled composites successfully realized the full use of visible-NIR light and afforded much higher activity than any CN- or UCP-based photocatalyst ever reported.


Temperament, substance use and psychopathology in a prisoner population: implications for treatment.
The association between temperament and drug use or temperament and psychopathology has previously been restricted to community clinical or non-clinical samples. The aim of the present study was therefore to examine these associations in a large cohort of Australian offenders using Cloninger's Temperament and Character Inventory (TCI). A total of 1322 prisoners from New South Wales (NSW) who completed all dimensions of the TCI and were screened for mental illness using the Australian National Survey of Mental Health and Wellbeing were included in the study. A total of 15% of the sample fulfilled the criteria for a diagnosis for depression, 36% for anxiety disorders and 54% for a substance abuse disorder. Using logistic regression analysis, the TCI dimensions of harm-avoidance and low self-direction predicted depression. Being female, a poly-substance user and having high harm-avoidance, persistence, self-direction and self-transcendence predicted anxiety disorders. Significant stepped trends across age, gender, and type of drug use were found for all TCI dimensions. The TCI is useful in identifying prisoners with a history of psychopathology and substance misuse. This tool also provides clinically relevant information about at-risk individuals and has the potential to guide the development of intervention programmes for inmates.


Estrogen receptor inhibits interleukin-6 gene expression by disruption of nuclear factor kappaB transactivation.
The estrogen receptor (ER) suppresses interleukin-6 (IL-6) gene expression through interaction with nuclear factor kappaB (NF-kappaB) in a hormone-dependent manner. Classic ER binding to DNA is not required and the mechanism of repression is unclear. Previously reported studies suggest that the interference of NF-kappaB binding to DNA by ER may play an important role. An alternative model for repression would be the disruption of NF-kappaB transactivation. In the present study, gel shift assays were used to examine the binding of RelA and p50 dimers to the IL-6 promoter in the presence of ER. The effect of ER on NF-kappaB transactivation was studied independent of NF-kappaB binding to DNA using the mammalian one-hybrid system. ER had little effect on the binding of homodimers or heterodimers of RelA and p50 to the IL-6 promoter. In transfection experiments, both ERalpha and ERbeta inhibited NF-kappaB-mediated expression in a hormone dependent manner with repression also dependent upon dimerization of RelA with p50. Mutant ER that is unable to transactivate failed to repress NF-kappaB expression, but deletion of the N-terminal portion of the receptor had no effect. Taken together, these results suggest that the disruption of NF-kappaB-mediated transactivation plays a significant role in ER inhibition of IL-6 gene expression.


Sociocultural pressure, internalization, BMI, exercise, and body dissatisfaction in Korean female college students.
This research investigated the differential effects of sociocultural pressures from media, peers, and parents on the thin ideal internalization and body dissatisfaction of 472 Korean female college students using structural equation modeling. Results indicated that after controlling for body mass index and exercise, media pressure exerted the largest effects, followed by peer pressure and parental pressure, on thin ideal internalization, and in turn, body dissatisfaction. Furthermore, parent and media pressures were found to exert direct effects on body dissatisfaction as well as indirect effects through thin ideal internalization. The results and implications of the study are discussed.


Molecular imprinting-based separation methods for selective analysis of fluoroquinolones in soils.
Molecularly imprinted polymers (MIPs) for fluoroquinolone antibiotics (FQs) have been synthesised in one single preparative step by precipitation polymerisation using ciprofloxacin (CIP) as template. Combinations of methacrylic acid (MAA) or 4-vinylpyridine (VP) as functional monomers, ethylene glycol dimethacrylate as crosslinker and dichloromethane, methanol, acetonitrile or toluene as porogens were tested. The experiments carried out by molecularly imprinted solid-phase extraction (MISPE) in cartridges did not allow to detect any imprint effect in the VP-based polymers whereas it was clearly observed in the MAA-based polymers. Among them, the MIP prepared in methanol using MAA as monomer showed the best performance and was chosen for further experiments. The ability of the selected MIP for the selective recognition of other widely used FQs (enoxacin, norfloxacin, danofloxacin and enrofloxacin) and quinolones (Qs) (cinoxacin, flumequine, nalidixic acid and oxolinic acid) was evaluated. The obtained results revealed the high selectivity of the obtained polymer, which was able to distinguish between FQs, that were recognised and retained onto the MIP cartridge, and Qs, which were washed out during loading and washing steps. The MIP was then packed into a stainless steel column (50mmx4.6mm i.d.) and evaluated as chromatography column for screening of FQs in soil samples. The mobile phase composition, flow rate, and the elution profile were then optimised in order to improve peak shape without sacrifying imprinting factor. Finally, under optimised conditions, soil samples spiked with CIP or with a mixture of fluoroquinolones in concentration of 0.5microgg(-1) were successfully analysed by the developed MIP-based procedures.


Biofilm and cell adhesion strength on dental implant surfaces via the laser spallation technique.
The aims of this study are to quantify the adhesion strength differential between an oral bacterial biofilm and an osteoblast-like cell monolayer to a dental implant-simulant surface and develop a metric that quantifies the biocompatible effect of implant surfaces on bacterial and cell adhesion. High-amplitude short-duration stress waves generated by laser pulse absorption are used to spall bacteria and cells from titanium substrates. By carefully controlling laser fluence and calibration of laser fluence with applied stress, the adhesion difference between Streptococcus mutans biofilms and MG 63 osteoblast-like cell monolayers on smooth and rough titanium substrates is obtained. The ratio of cell adhesion strength to biofilm adhesion strength (i.e., Adhesion Index) is determined as a nondimensionalized parameter for biocompatibility assessment. Adhesion strength of 143 MPa, with a 95% C.I. (114, 176), is measured for MG 63 cells on smooth titanium and 292 MPa, with a 95% C.I. (267, 306), on roughened titanium. Adhesion strength for S. mutans on smooth titanium is 320 MPa, with a 95% C.I. (304, 333), and remained relatively constant at 332 MPa, with a 95% C.I. (324, 343), on roughened titanium. The calculated Adhesion Index for smooth titanium is 0.451, with a 95% C.I. (0.267, 0.622), which increased to 0.876, with a 95% C.I. (0.780, 0.932), on roughened titanium. The laser spallation technique provides a platform to examine the tradeoffs of adhesion modulators on both biofilm and cell adhesion. This tradeoff is characterized by the Adhesion Index, which is proposed to aid biocompatibility screening and could help improve implantation outcomes. The Adhesion Index is implemented to determine surface factors that promote favorable adhesion of cells greater than biofilms. Here, an Adhesion Index ≫ 1 suggests favorable biocompatibility.


Using a Novel Diagnostic Nomogram to Differentiate Malignant from Benign Parathyroid Neoplasms.
We sought to develop an immunohistochemical (IHC) tool to support the diagnosis of parathyroid carcinoma (PC) and help differentiate it from atypical parathyroid neoplasms (atypical) and benign adenomas. Distinguishing PC from benign parathyroid neoplasms can be challenging. Many cases of PC are histopathologically borderline for definitive malignancy. Recently, individual IHC biomarkers have been evaluated to aid in discrimination between parathyroid neoplasms. PC, atypical parathyroid neoplasms, and parathyroid adenomas treated at our institution from 1997 to 2014 were studied retrospectively. IHC analysis was performed to evaluate parafibromin, retinoblastoma (RB), protein gene product 9.5 (PGP9.5), Ki67, galectin-3, and E-cadherin expression. Receiver operating characteristic (ROC) analysis and multivariable logistic regression model for combinations of biomarkers were evaluated to classify patients as PC or atypical/adenoma. A diagnostic nomogram using 5 biomarkers was created for PC. Sixty-three patients were evaluated. The percent staining of parafibromin (p < 0.0001), RB (p = 0.04), Ki67 (p = 0.02), PGP9.5 (p = 0.04), and Galectin-3 (p = 0.01) differed significantly in the three diagnostic groups. ROC analysis demonstrated that parafibromin had the best performance in discriminating PC from atypical/adenoma; area under the curve (AUC) was 81% (cutoff, 92.5%; sensitivity rate, 64%; specificity rate, 87%). We created a diagnostic nomogram using a combination of biomarkers; AUC was 84.9% (95% confidence interval, 73.4-96.4%). The optimism-adjusted AUC for this model was 80.5% (mean absolute error, 0.043). A diagnostic nomogram utilizing an immunoexpression, a combination of immunohistochemical biomarkers, can be used to help differentiate PC from other parathyroid neoplasms, thus potentially improving diagnostic classification.


The Notch signalling pathway in hair growth.
The Notch signalling pathway is an important mediator of cell fate selection whose involvement in epidermal appendage formation is now becoming recognised. Hair follicle development and hair formation involve the co-ordinated differentiation of several different cell types in which Notch appears to have a role. We report intricate expression patterns for the Notch-1 receptor and three ligands, Delta-1, Jagged-1 and Jagged-2 in the hair follicle. Notch-1 is expressed in ectodermal-derived cells of the follicle, in the inner cells of the embryonic placode and the follicle bulb, and in the suprabasal cells of the mature outer root sheath. Delta-1 is only expressed during embryonic follicle development and is exclusive to the mesenchymal cells of the pre-papilla located beneath the follicle placode. Expression of Jagged-1 or Jagged-2 overlaps Notch-1 expression at all stages. In mature follicles, Jagged-1 and Jagged-2 are expressed in complementary patterns in the follicle bulb and outer root sheath, Jagged-1 in suprabasal cells and Jagged-2 predominantly in basal cells. In the follicle bulb, Jagged-2 is localised to the inner (basal) bulb cells next to the dermal papilla which do not express Notch-1, whereas Jagged-1 expression in the upper follicle bulb overlaps Notch-1 expression and correlates with bulb cell differentiation into hair shaft cortical and cuticle keratinocytes.


Synthesis and biological evaluation of new thiazolyl/benzothiazolyl-amides, derivatives of 4-phenyl-piperazine.
A series of thiazolyl-N-phenyl piperazines has been synthesised and tested for anti-inflammatory activity. Their R(M) values were determined as an expression of their lipophilicity. Theoretical calculation of their lipophilicity, as clog P and logPsk also performed. The effect of the synthesised compounds on inflammation, using the carrageenin induced mouse paw oedema model was studied. In general, the studied compounds were found to be potent anti-inflammatory agents (44-74.1%). Anti-inflammatory activity was influenced by some structural characteristics of the synthesised compounds. An attempt was made to correlate their biological activity with some physicochemical parameters using a quantitative structure-activity relationship approach (QSAR).


Congenital pulmonary airway malformation manifesting with severe respiratory distress and pneumothorax in a neonate.
Congenital pulmonary airway malformation (CPAM) is congenital pulmonary anomaly characterized by multicystic areas, over-distension, and proliferation of terminal bronchioles with lack of normal alveoli. Clinical presentation may vary from mild respiratory symptoms to severe respiratory distress and frequent pneumothoraxes. We report a rare case of neonatal CPAM type I manifested with neonatal respiratory distress and pneumothorax, which was managed successfully with left lower lobectomy. The online version contains supplementary material available at 10.1007/s12055-023-01510-x.


[The diagnostic value of RNA oncomarkers in evaluation of malignant breast tumors].
The levels of the RNA oncomarkers, telomerase (hTERT), cytokeratin-19 (CK-19) and mammaglobin (MAM) have been investigated in capillary blood of female patients with mammary ductal carcinoma. The study revealed overexpression of all three factors in patients with this pathology. This overexpression was not found in healthy donors and female patients with mammary fibroadenoma. Levels of the RNA oncomarkers return to the normal level within 10 days after successful tumor resection. These results have been used for the development of diagnostic kits, which may be applicable for differential diagnostics, screening and postoperation monitoring of patients with malignant breast tumors


Basic problems of serological laboratory diagnosis.
Serological laboratory diagnosis is inflicted with at least two kinds of basic problems. One type relates to the fact that the serological diagnosis of infectious diseases is double indirect: First, to diagnose an infectious disease, the identification of the microbial agent is sought that caused the disease. Second, to identify this infectious agent, the patient's immune response to potential agents is measured. So, the serological test is neither measuring directly disease nor the cause of the disease, but the patient's immune system. Another type of problem is based on the fact that each person's immune system is very individual. The exact physicochemical properties of antibodies are unique for each clone of antibodies. The way an individual's immune system sees an infectious agent depends not only on the genetic makeup of the person but also on the personal experience from former encounters with infectious agents. Both types of problems lead to complexities in selecting the appropriate test, in interpreting the results, and in standardizing serological tests. Therefore, a close collaboration of the laboratory with the clinic is mandatory to avoid erroneous conclusions from serological test results, which might lead to wrong decisions in patient care.


Intellectual disabilities, neuronal posttranscriptional RNA metabolism, and RNA-binding proteins: three actors for a complex scenario.
Intellectual disability (ID) is the most frequent cause of serious handicap in children and young adults and interests 2-3% of worldwide population, representing a serious problem from the medical, social, and economic points of view. The causes are very heterogeneous. Genes involved in ID have various functions altering different pathways important in neuronal function. Regulation of mRNA metabolism is particularly important in neurons for synaptic structure and function. Here, we review ID due to alteration of mRNA metabolism. Functional absence of some RNA-binding proteins--namely, FMRP, FMR2P, PQBP1, UFP3B, VCX-A--causes different forms of ID. These proteins are involved in different steps of RNA metabolism and, even if a detailed analysis of their RNA targets has been performed so far only for FMRP, it appears clear that they modulate some aspects (translation, stability, transport, and sublocalization) of a subset of RNAs coding for proteins, whose function must be relevant for neurons. Two other proteins, DYRK1A and CDKL5, involved in Down syndrome and Rett syndrome, respectively, have been shown to have an impact on splicing efficiency of specific mRNAs. Both proteins are kinases and their effect is indirect. Interestingly, both are localized in nuclear speckles, the nuclear domains where splicing factors are assembled, stocked, and recycled and influence their biogenesis and/or their organization.


Regulation of tetrapyrrole biosynthesis in higher plants.
Plant tetrapyrroles are the most abundant biomolecules on the earth and are cofactors of many apoproteins essential for plant function. The four end-products sirohaem, chlorophyll, haem and phytochromobilin are synthesized by a common branched pathway, which is tightly regulated to ensure a continuous supply to the cognate apoproteins. This may induce strong competition between different branches of the pathway for common substrates. In addition, the intermediates, which are phototoxic, must not be allowed to accumulate in the cell. The major control points are during the synthesis of the initial precursor, ALA (5-aminolaevulinic acid), and at the branch points for the insertion of metal ions into the porphyrin macrocycle. A recent study has also suggested that tetrapyrroles are involved in the communication between the chloroplast and the nucleus, strengthening the necessity for tight regulation. However, intermediates remain difficult to quantify mainly due to their low content and the different properties of the intermediates. In this paper, we summarize the regulation of this pathway and we detail why it is important to have an accurate method for the determination of tetrapyrroles in plants.


Some historiographical tools for the study of intellectual legacies.
Intellectual legacies are part of historians' concerns, when they study the evolution of ideas. There are, however, no guidelines to help characterize the reception of intellectual legacies. This article provides preliminary tools to fill this gap, with a typology (faithful, formal, substantial legacies), and with two criteria to assess the conformity between the heir's and her inspirer's proposals. The objective is not to judge the legitimacy of this or that reception, but to facilitate its characterization, for a better understanding of the transmission of ideas. One case study from the history of economic thought, Nicholas Georgescu-Roegen's bioeconomics and its legacies, is provided to illustrate the operability of the toolbox.


Reversible and non-competitive antagonist profile of CPCCOEt at the human type 1alpha metabotropic glutamate receptor.
In transfected CHO cells expressing the human metabotropic glutamate receptor mGlu1alpha, 7-(hydroxyimino)cyclopropan[b]-chromen-1a-carboxylic acid ethylester (CPCCOEt) was found to antagonize L-quisqualate-induced phosphoinositide hydrolysis in a non-competitive and reversible manner (apparent pKi value, 4.76+/-0.18; n=3). This suggests that CPCCOEt antagonizes type 1alpha metabotropic glutamate receptor activation by interacting with a site distinct from the agonist binding site.


Spectroscopic characterization of textilotoxin, a presynaptic neurotoxin from the venom of the Australian eastern brown snake (Pseudonaja t. textilis).
Spectroscopic behavior of textilotoxin, from the venom of Pseudonaja t. textilis, and its subunits were investigated using fluorescence, circular dichroism and Fourier transform infrared spectroscopy. Circular dichroism spectra of the B, C and D subunits indicate considerable similarity in their alpha-helix and beta-sheet contents. By contrast, the A subunit displays significantly more beta-sheet and 'remainder' structure. FTIR spectra confirm conclusions drawn from CD spectra. Fluorescence spectra indicate that, in general, tryptophan residues in the A, B and D subunits are relatively exposed to the solvent. The C subunit exhibits no fluorescence, suggesting a lack of tryptophan. Comparisons of individual subunit spectra with those of the intact toxin suggest that significant changes in secondary structure may occur when the toxin dissociates.


Norman K. Wessells: a life in science.
"In its triple role as locomotory organelle, as a site of deposition of new surface material for the elongating axon, and a source of microspikes (sensory probes), the growth cone becomes the key to axon elongation" Yamada et al. (1971).


Ranking the Leading Risk Factors for Sudden Unexpected Death in Epilepsy.
Sudden unexpected death in epilepsy (SUDEP) is rare in well-controlled epilepsy. However, SUDEP is a common cause of death in drug-resistant epilepsy. Over the last 30 years, multiple cohort and population studies have identified clinical risk factors associated with an increased risk for SUDEP. To identify and rank the leading SUDEP risk factors from major cohort and population-based studies. The incidence of SUDEP is also evaluated in special clinical situations, including antiepileptic drug treatment, epilepsy surgery, devices, and assignment to placebo in clinical trials. A PubMed search for English language human cohort studies for the terms Sudden, Death, and Epilepsy was performed for the years 1987-2017. Risk factors for SUDEP were identified and ranked by the weighted log adjusted odds ratio (OR)/relative risk ratio (RR). The top 10 leading risk factors ranked from highest to lowest log adjusted OR/RR are the following: ≥3 GTC seizures per year; ≥13 seizures in the last year; No Antiepileptic Drug (AED) treatment; ≥3 AEDs; ≥3 GTCs in the past year; 11-20 GTC seizures in the last 3 months; age of onset 0-15 years old; IQ < 70; 3-5 AED changes in the last year; ≥3 AEDs. Two risk factors from separate sources (≥3 GTC seizures and ≥3 AEDs) occur twice in the top 10 risk factors. The top 10 risk factors for SUDEP are identified and ranked. A ranking of the top risk factors could help clinicians identify patients at highest risk for SUDEP.


[Surgical management of talipes equinovarus as sequelae of a compartment and/or postischemic syndrome of the deep flexor compartment of the lower leg].
Cases of posttraumatic pes equinovarus after compartment syndrome have become more frequent in the last 3 decades because limb-saving procedures like compartment splitting, vascular repair, and microvascular free flaps have become well established in trauma surgery, thus reducing early below knee amputations. But if the deep flexor compartment is not split completely or if the muscles are crushed by direct trauma severe necrosis and subsequent muscle contractures result in a very severe clubfoot deformity. Metatarsalgia of fifth, fourth, and third metatarsal head even in well-fitted orthopaedic shoes occurs as well as painful bunions and fatigue fractures of the fifth metatarsal. Infected ulcers below the fifth/fourth metatarsal bone in a numb plantar sole often require head resection because of osteomyelitis.From 1994 to 2007 a total of 24 patients with severe pes equinovarus after compartment and/or postischemic syndrome were treated operatively. Only in 5 cases was a triple, Chopart, or Lisfranc arthrodesis necessary; 19 cases however could be treated only by soft tissue procedures like tenolysis, tendon lengthening, medial release of the scarred flexor retinacula and contracted capsules of the posterior ankle, subtalar and talonavicular joint to reorientate all axes of the foot. By temporary K-wire transfixation (6 weeks), initial external tibiotarsal transfixation of the foot (10 days), and additional tendon transfer for active foot elevation excellent and good long-term (5 years) results are achievable.The results according to the McKay Score are not significantly different regarding the triple arthrodesis group versus the pure soft tissue release group. Nevertheless, saving joints in the latter group seems to be very important.


Fructooligosaccharide intake promotes epigenetic changes in the intestinal mucosa in growing and ageing rats.
The aim of this study was to investigate the relationship between fructooligosaccharide (FOS) intake at different life stages of Wistar rats and its stimulatory effects on intestinal parameters. Recently weaned and ageing female rats were divided into growing and ageing treatments, which were fed diets that partially replaced sucrose with FOS for 12 weeks. Dietary FOS intake induced a significant increase in the numbers of Bifidobacterium and Lactobacillus in growing rats. FOS intake was associated with increased butyric acid levels and a reduced pH of the caecal contents at both ages. Differential gene expression patterns were observed by microarray analysis of growing and ageing animals fed the FOS diet. A total of 133 genes showed detectable changes in expression in the growing rats, while there were only 19 gene expression changes in ageing rats fed with FOS. These results suggest that dietary FOS intake may be beneficial for some parameters of intestinal health in growing rats.


Activity of 1-aryl-4-(naphthalimidoalkyl) piperazine derivatives against Leishmania major and Leishmania mexicana.
A series of 1-aryl-4-(phthalimidoalkyl) piperazines and 1-aryl-4-(naphthalimidoalkyl) piperazines were retrieved from a proprietary library based on their high structural similarity to haloperidol, an antipsychotic with antiparasitic activity, and assessed as potential antileishmanial scaffolds. Selected compounds were tested for antileishmanial activity against promastigotes of Leishmania major and Leishmania mexicana in dose-response assays. Two of the 1-aryl-4-(naphthalimidoalkyl) piperazines (compounds 10 and 11) were active against promastigotes of both Leishmania species without being toxic to human fibroblasts. Their activity was found to correlate with the length of their alkyl chains. Further analyses showed that compound 11 was also active against intracellular amastigotes of both Leishmania species. In promastigotes of both Leishmania species, compound 11 induced collapse of the mitochondrial electrochemical potential and increased the intracellular Ca2+ concentration. Therefore, it may serve as a promising lead compound for the development of novel antiparasitic drugs.


Design of a DT neutron source based PGNAA facility for element determination in aqueous solution.
A DT neutron source-based prompt gamma ray neutron activation (PGNAA) facility for bulk sample analysis was designed and developed in this research. The aqueous samples containing chlorine and boron with known concentration were determined for the calibration curve by the facility. The neutron self-shielding effect was corrected by internal standard method. The minimum detectable concentration of boron and chlorine are 1.37 ± 0.42 and 12.51 ± 3.80 mg/L, respectively. Finally, five mixture aqueous solution samples were measured for the facility performance test. The maximum relative deviation of boron and chlorine are 4.49% and 5.32% respectively.


In silico studies on the interaction of phage displayed biorecognition element (TFQAFDLSPFPS) with the structural protein VP28 of white spot syndrome virus.
White spot disease caused by the white spot syndrome virus (WSSV) incurs a huge loss to the shrimp farming industry. Since no effective therapeutic measures are available, early detection and prevention of the disease are indispensable. Towards this goal, we previously identified a 12-mer phage displayed peptide (designated as pep28) with high affinity for VP28, the structural protein of the white spot syndrome virus (WSSV). The peptide pep28 was successfully used as a biorecognition probe in the lateral flow assay developed for rapid, on-site detection of WSSV. To unravel the structural determinants for the selective binding between VP28 and pep28, we used bioinformatics, structural modeling, protein-protein docking, and binding-free energy studies. We performed atomistic molecular dynamics simulations of pep28-pIII model totaling 300 ns timescale. The most representative pep28-pIII structure from the simulation was used for docking with the crystal structure of VP28. Our results reveal that pep28 binds in a surface groove of the monomeric VP28 β-barrel and makes several hydrogen bonds and non-polar interactions. Ensemble-based binding-free energy studies reveal that the binding is dominated by non-polar interactions. Our studies provide molecular level insights into the binding mechanism of pep28 with VP28, which explain why the peptide is selective and can assist in modifying pep28 for its practical use, both as a biorecognition probe and a therapeutic.


Comparison of SARS-CoV-2 spike RNA sequences in feces and nasopharynx indicates intestinal replication.
Little is known of possible selection and replication of SARS-CoV-2 in the intestines and if viral load in feces is associated with severity of disease. Therefore, sequence variations of the spike region in strains collected from feces and nasopharynx (NPH) from the same patients were compared. It was also investigated whether viral load in feces related to severity of COVID-19 in hospitalized patients. SARS-CoV-2 RNA was found in 88 (79%) fecal samples from 112 patients. The complete spike region could be sequenced in 15 fecal and 14 NPH samples. Fourteen Alpha-variants and one Beta-variant of SARS-CoV-2 were identified. The majority of the viral genetic variants (viral populations) in two fecal samples, but none in NPH, had a reversion of the H69/V70 amino acid deletion normally seen in the Alpha variants. Nine fecal samples contained up to nine minority variants, each which may constitute a separate viral population. Five NPH samples had one genetic variant each, and one NPH sample contained nine minority populations of SARS-CoV-2 spike genes. The higher genomic diversity of SARS-CoV-2 in feces compared to NPH, and the reversion of the H69/V70 deletion in Alpha variants from feces indicate a selection of viral strains and replication of SARS-CoV-2 in the gastrointestinal tract.


Stage 2 Registered Report: The Bidirectional Relationship Between Brain Features and the Dysregulation Profile: A Longitudinal, Multimodal Approach.
Youth with symptoms of emotion dysregulation are at risk for a multitude of psychiatric diagnoses later in life. However, few studies have focused on the underlying neurobiology of emotion dysregulation. This study assessed the bidirectional relationship between emotion dysregulation symptoms and brain morphology throughout childhood and adolescence. A combined total of 8,235 children and adolescents drawn from 2 large population-based cohorts, the Generation R Study and Adolescent Brain Cognitive Development (ABCD) Study, were included. Data were acquired in 3 waves in Generation R (mean [SD] age = 7.8 [1.0] wave 1 [W1]; 10.1 [0.6] W2; 13.9 [0.5] W3) and in 2 waves in ABCD (mean [SD] age = 9.9 [0.6] W1; 11.9 [0.6] W2). Cross-lagged panel models were used to determine the bidirectional relationships between emotion dysregulation symptoms and brain morphology. The study was preregistered before performing analyses. In the Generation R sample, emotion dysregulation symptoms at W1 preceded lower hippocampal (β = -.07, SE = 0.03, p = .017) and temporal pole (β = -.19, SE = 0.07, p = .006) volumes at W2. Emotion dysregulation symptoms at W2 preceded lower fractional anisotropy in the uncinate fasciculus (β = -.11, SE = 0.05, p = .017) and corticospinal tract (β = -.12, SE = 0.05, p = .012). In the ABCD sample, emotion dysregulation symptoms preceded posterior cingulate (β = .01, SE = 0.003, p = .014) and nucleus accumbens volumes (left hemisphere: β = -.02, SE = 0.01, p = .014; right hemisphere: β = -.02, SE = 0.01, p = .003). In population-based samples, with relatively low psychopathology symptoms in the majority of children, symptoms of emotion dysregulation can precede differential development of brain morphology. This provides the foundation for future work to assess to what extent optimal brain development can be promoted through early intervention. The Bidirectional Relationship Between Brain Features and the Dysregulation Profile: A Longitudinal, Multimodal Approach; https://doi.org/10.1016/j.jaac.2022.03.008. We worked to ensure that the study questionnaires were prepared in an inclusive way. The author list of this paper includes contributors from the location and/or community where the research was conducted who participated in the data collection, design, analysis, and/or interpretation of the work.


[Care time as confrontations with the other].
Caregivers and patients are directly confronted with time, but it is the encounter in the framework of care which generates defensive expressions in relation to this unique head-to-head. Restoring some imagination can however support the care time.


Reaction rate studies of two enzymatic cholesterol reagent kits.
Reaction rate studies were carried out on various specimens using two commercial enzymatic cholesterol determination reagent kits (BMC and Worthington). The time required for complete colour formation differed between the two reagent systems and each kit required different incubation times for the standards and specimens to come to color completion. The choice of standards was found to be very important when using these enzymatic kits.


Identification and Functional Verification of MicroRNAs in the Obese Rat With Erectile Dysfunction.
Obesity is a potential risk factor for erectile dysfunction (ED). MicroRNAs (miRNAs) regulate the expression of genes involved in various pathophysiologic processes. To identify the miRNA profile in the corpus cavernosum (CC) of obese rats with ED and elucidate the potential function of miRNA in the pathogenesis of ED. Obesity was induced in rats by a high-fat diet. After the erectile function test, experimental animals were divided into two groups: obese rats with ED and obese rats with normal erectile function. The CCs from these rats were collected for miRNA microarray analysis. The results were verified by real-time polymerase chain reaction analysis. Subsequently, the targets of differentially expressed miRNAs were predicted. Bioinformatics analysis was applied to predict the functions of differentially expressed miRNAs in ED. Apomorphine-induced penile erection and intracavernous pressure measurements were used to evaluate the effects of miRNA on the erectile function of rats. MiRNA expression in the CC of obese rats with ED and those with normal erectile function was detected by miRNA microarray analysis. Candidate miRNAs were validated by real-time polymerase chain reaction. Bioinformatics analysis was used to predict the functions of miRNAs. Apomorphine-induced penile erection and intracavernous pressure measurements were used to reflect the erectile function of rats. Sixty-eight miRNAs were differentially expressed in the CC of obese rats with ED (≥1.5-fold change). The real-time polymerase chain reaction results were consistent with the miRNA microarray analysis results. Specifically, miR-328a was significantly upregulated in rats with ED compared with control rats and was chosen for functional evaluation in the pathogenesis of ED. Overexpression of miR-328a noticeably decreased the erectile response to apomorphine and the expression of heme oxygenase-1. MiRNAs are involved in the pathogenesis of obesity-related ED. MiR-328a might facilitate the induction of ED. Bai Y, Zhang L, Jiang Y, et al. Identification and Functional Verification of MicroRNAs in the Obese Rat With Erectile Dysfunction. Sex Med 2017;5:e261-e271.


Role of mycorrhizal infection in the growth and reproduction of wild vs. cultivated plants : II. Eight wild accessions and two cultivars ofLycopersicon esculentum Mill.
An experiment was conducted to determine whether wild accessions and cultivars ofLycopersicon esculentum Mill. differed in inherent morphological, physiological or phenological traits and whether such differences would result in variation in response to vesicular-arbuscular mycorrhizal infection. While distinctions between wild accessions and cultivars were apparent (the cultivars generally had higher phosphorus use efficiencies and shorter lifespans than the wild accessions) and the cultivars were, as a group, more responsive to mycorrhizal infection than the wild accessions, there was significant variation among wild accessions and among cultivars in response to infection. Regardless of cultivation status, non-mycorrhizal plant root density was significantly negatively correlated with response to infection. Phosphorus use efficiency was generally not significantly correlated with response to infection. Mycorrhizal infection decreased phosphorus use efficiency in all accessions, but had variable effects on root density, depending upon accession and time. Finally, the vegetative response was not necessarily of the same magnitude as the reproductive response.


Peer group reputation and smoking and alcohol consumption in early adolescence.
Research has shown that substance use might serve beneficial, developmental functions for adolescents, in particular concerning young people's functioning in friendships and intimate relationships. Nonetheless, a major problem in these studies is that information on social functioning is based on adolescent's self-reports, which undoubtedly might lead to an overestimation of the social functions of substance use. In the present study, we collected data of 3361 early and mid-adolescents at secondary schools in the Netherlands. Information on individual student's social behavior and psychological traits were gathered by using sociometric measures. All respondents were asked to evaluate all classmates on measures, such as sociability, self-confidence, achievement, withdrawal, and aggression. Person- and variable-centered analyses clearly showed that highest levels of smoking and drinking were found in adolescents who score high on sociability and self-confidence, and relatively low on aggression-inattentiveness, achievement-withdrawal, and emotionality-nervousness. This suggests that beneficial functions of substance use are not only in the eyes of the beholder, at least not in that of the individual drinker or smoker.


Urinary tract infection in the renal transplant patient.
Urinary tract infection (UTI) is the most common type of bacterial infection contracted by recipients of renal allografts in the post-transplantation period. Fungi and viruses can also cause UTIs, but infections caused by these organisms are less common than those caused by bacteria. Both the lower and upper urinary tract (encompassing grafted or native kidneys) can be affected. Factors that might contribute to the development of UTIs include excessive immunosuppression, and instrumentation of the urinary tract (e.g. urethral catheters and ureteric stents). Antimicrobials are the mainstays of treatment and should be accompanied by minimization of immunosuppression when possible. The use of long-term antimicrobial prophylaxis is controversial, however, as it might increase the likelihood of infective organisms becoming resistant to treatment. There are conflicting data on the associations of post-transplantation UTI with graft and patient survival.


When A is greater than B: Interactions between magnitude and serial order.
Processing ordinal information is an important aspect of cognitive ability, yet the nature of such ordinal representations remains largely unclear. Previously, it has been suggested that ordinal position is coded as magnitude, but this claim has not yet received direct empirical support. This study examined the nature of ordinal representations using a Stroop-like letter order judgment task. If ordinal position is coded as magnitude, then letter ordering and font size should interact. Experiments 1 and 2 identified a significant interaction between letter size and ordering. Specifically, a facilitation effect was observed for alphabetically ordered sequences with decreasing font size (e.g., ). This suggests an overlap in the mechanisms for order and magnitude processing. The finding also suggests that earlier ranks may be represented as "more" in such a magnitude-based code, and vice versa for later ranks.


Laryngeal sensory dysfunction in laryngeal hypersensitivity syndrome.
Diseases associated with laryngeal dysfunction include chronic refractory cough (CRC), paradoxical vocal fold movement (PVFM), muscle tension dysphonia (MTD) and globus pharyngeus. We hypothesized the presence of a common sensory laryngeal dysfunction, the 'laryngeal hypersensitivity' syndrome, in these conditions. The aim of the study was to compare symptoms and sensory function in patients with CRC, PVFM, MTD and globus. The 103 participants included healthy controls (n = 13) and four case groups: CRC (n = 33), PVFM (n = 28), globus pharyngeus (n = 11) and MTD (n = 18). Participants completed self-report questionnaires: Symptom Frequency and Severity Scale, Voice Handicap Index and the Laryngeal Paraesthesia Questionnaire; and quantitative sensory testing: capsaicin cough reflex sensitivity, hypertonic saline challenge, the timed swallow test, acoustic voice testing, cough frequency monitor and a voice stress test. All case groups reported a high-symptom burden in comparison to controls. The case groups showed a similar pattern of symptoms, with impairment in each of the cough, respiration, vocal and upper airway symptom domains. Objective testing revealed significant sensory impairment in the case groups compared to controls and also showed an overlap in sensory dysfunction between the four case groups. Furthermore, there was cross-sensory stimulation of symptoms whereby stimulation of a particular response resulted in symptoms in another domain. These discrete clinical laryngeal syndromes display considerable overlap in their clinical features and a common sensory dysfunction, supporting the 'laryngeal hypersensitivity' hypothesis. Reconceptualizing functional laryngeal disorders as a form of laryngeal hypersensitivity syndrome provides an alternative approach to management of these perplexing conditions.


3D-printed platform multi-loaded with bioactive, magnetic nanoparticles and an antibiotic for re-growing bone tissue.
Polymeric platforms obtained by three-dimensional (3D) printing are becoming increasingly important as multifunctional therapeutic systems for bone treatment applications. In particularly, researchers aim to control bacterial biofilm on these 3D-platforms and enhance re-growing bone tissue, at the same time. This study aimed to fabricate a 3D-printed polylactic acid platform loaded with hydroxyapatite (HA), iron oxide nanoparticles (IONPs) and an antibiotic (minocycline) with tuneable properties and multistimuli response. IONPs were produced by a facile chemical co-precipitation method showing an average diameter between 11 and 15 nm and a superparamagnetic behaviour which was preserved when loaded into the 3D-platforms. The presence of two types of nanoparticles (IONPs and HA) modify the nanomorphological/nanotopographical feature of the 3D-platforms justifying their adequate bioactivity profile and in vitro cellular effects on immortalized and primary osteoblasts, including cytocompatibility and increased osteogenesis-related gene expression (RUNX2, BGLAP and SPP1). Disk diffusion assays and SEM analysis confirmed the effect of the 3D-platforms loaded with minocycline against Staphylococcus aureus. Altogether results showed that fabricated 3D-platforms combined the exact therapeutic antibiofilm dose of the antibiotic against S. aureus, with the enhanced osteogenic stimulation of the HA and IONPs nanoparticles which is a disruptive approach for bone targeting applications.


Methicillin-resistant Staphylococcus aureus on hospital admission in Turkey.
A total of 900 patients were screened for methicillin-resistant Staphylococcus aureus (MRSA) on hospital admission, and 11 MRSA strains (1.2%) were detected. All 11 MRSA strains were positive for the mecA and PVL genes. Eight of the 11 MRSA-positive patients (72%) had a history of hospitalization within the previous 12 months.


How to prevent the postoperative pancreatic fistula with an ethylene vinyl alcohol copolymer (Onyx®): A proposal of a new technique.
Despite the advances in identifying risk factors, improving operative technique, and postoperative patient care, pancreatic leakage after pancreatic resection remains a highly debated topic. The aim of this study is to describe our technique and our initial experience with the intraoperative embolization of the main pancreatic duct with an Ethylene Vinyl Alcohol Copolymer (Onyx®). Two patients of 63 and 64 years underwent pancreaticoduodenectomy for a cholangiocarcinoma of the extrahepatic bile duct and a pancreatic adenocarcinoma, respectively. At the time of pancreatic parenchyma resection, a Wirsung duct was identified and catheterized. A wirsungography was done and then, embolization with Onyx® was carried out under fluoroscopic control. Neither of the patients developed a postoperative pancreatic fistula. They were discharged to home on the 17th and 18th postoperative day, respectively. At the last follow-up, no recurrence was found. The two patients became diabetics; both needed the support of supplementary pancreatic enzymes. To our knowledge, we are the first to describe this technique, which seems safe and reliable. Studies on this subject with more patients are needed to confirm the validity of this procedure.


Synthetic methyl hexagalacturonate hapten inhibitors of anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7.
A range of synthetic methyl hexagalacturonates were used as potential hapten inhibitors in competitive-inhibition enzyme-linked immunosorbent assays (ELISAs) with anti-homogalacturonan monoclonal antibodies LM7, JIM5 and JIM7. The selective inhibition of these antibodies by different haptens provides insight into the structures of the partially methyl-esterified pectin epitopes of these widely used monoclonal antibodies.


Nonunion of a midshaft clavicle fracture associated with subclavian vein compression. A case report.
Fractures of the clavicle are usually treated by closed means and heal uneventfully. In this report, we present an unusual case in which the subclavian vein was compressed by a fracture callus that had formed around a clavicular nonunion. Treatment included dissection of the subclavian vein from the fracture callus and compression plating with autogenous bone grafting of the fracture. Symptoms from the patient's venous obstruction slowly resolved without further treatment. The fracture united postoperatively.


Searching for differentially expressed gene combinations.
We propose 'CorScor', a novel approach for identifying gene pairs with joint differential expression. This is defined as a situation with good phenotype discrimination in the bivariate, but not in the two marginal distributions. CorScor can be used to detect phenotype-related dependencies and interactions among genes. Our easily interpretable approach is scalable to current microarray dimensions and yields promising results on several cancer-gene-expression datasets.


Novel approach using [18F]FTHA-PET and de novo synthesized VLDL for assessment of FFA metabolism in a rat model of diet induced NAFLD.
The worldwide prevalence of Non-alcoholic Fatty Liver Disease (NAFLD) raises concerns about associated risk factors, such as obesity and type 2 Diabetes Mellitus, for leading causes of disability and death. Besides Magnetic Resonance Imaging (MRI) and Spectroscopy (MRS), functional imaging with Positron Emission Tomography (PET) could contribute to a deeper understanding of the pathophysiology of NAFLD. Here we describe a novel approach using the PET tracer [18F]FTHA, which is an analog of long-chain free fatty acids (FFA) and is taken up by tissues to enter mitochondria or to be incorporated into complex lipids for further export as very-low-density lipoprotein (VLDL). Male Sprague Dawley rats, after 6 weeks on a high-fat diet (HFD), were used as a model of diet induced NAFLD, while a standard diet (SD) served as a control group. Liver fat was estimated by MR spectroscopy at a 9.4 T system for phenotyping. To measure hepatic FFA uptake, rats underwent 60 min dynamic [18F]FTHA-PET scans after unrestricted access to food (HFD: n = 6; SD: n = 6) or overnight (≤16h) fasting (HFD: n = 6; SD: n = 5). FFA removal was assessed from incorporated 18F-residual in de novo synthesized VLDL out of plasma. MRS of the liver confirmed the presence of NAFLD (>5.6% fat). Under non-fasting conditions, hepatic [18F]FTHA uptake was significantly increased in NAFLD: SUVmean (p = 0.03) within [0; 60] min interval, SUVmean (p = 0.01) and SUVmax (p = 0.03) within [30; 60] min interval. SUVs for hepatic uptake under fasting conditions were not significantly different between the groups. Analysis of FFA removal demonstrated elevated values of 18F-residue in the VLDL plasma fraction of the healthy group compared to the NAFLD (p = 0.0569). Our novel approach for assessing FFA metabolism using [18F]FTHA demonstrated differences in the hepatic FFA uptake and FFA incorporation into VLDL between healthy and NAFLD rats. [18F]FTHA-PET could be used to study metabolic disturbances involved in the progression of NAFLD.


Pharmacokinetics and relative bioavailability of levomepromazine after repeated administration of tablets and syrup.
Plasma levels of levomepromazine and its sulphoxide were measured in 8 psychiatric patients after repeated doses of levomepromazine tablets or syrup. The rate and extent of absorption of the drug were similar for the two dosage forms, although the extent of presystemic metabolism appeared to be slightly greater after administration of syrup than of tablets. The biological half-life of levomepromazine ranged from 16.5 h to 77.8 h, and a 13-fold variation was seen in the ratio of the total clearance to the absorbed fraction of the dose (Cl/Fpo). It is postulated that individual variation in the dose required for therapy was due in part to individual variation in the pharmacokinetics of the drug.


Mitochondrial F1FO-ATPase and permeability transition pore response to sulfide in the midgut gland of Mytilus galloprovincialis.
The molecular mechanisms which rule the formation and opening of the mitochondrial permeability transition pore (mPTP), the lethal mechanism which permeabilizes mitochondria to water and solutes and drives the cell to death, are still unclear and particularly little investigated in invertebrates. Since Ca2+ increase in mitochondria is accompanied by mPTP opening and the participation of the mitochondrial F1FO-ATPase in the mPTP is increasingly sustained, the substitution of the natural cofactor Mg2+ by Ca2+ in the F1FO-ATPase activation has been involved in the mPTP mechanism. In mussel midgut gland mitochondria the similar kinetic properties of the Mg2+- or Ca2+-dependent F1FO-ATPase activities, namely the same affinity for ATP and bi-site activation kinetics by the ATP substrate, in spite of the higher enzyme activity and coupling efficiency of the Mg2+-dependent F1FO-ATPase, suggest that both enzyme activities are involved in the bioenergetic machinery. Other than being a mitochondrial poison and environmental contaminant, sulfide at low concentrations acts as gaseous mediator and can induce post-translational modifications of proteins. The sulfide donor NaHS, at micromolar concentrations, does not alter the two F1FO-ATPase activities, but desensitizes the mPTP to Ca2+ input. Unexpectedly, NaHS, under the conditions tested, points out a chemical refractoriness of both F1FO-ATPase activities and a failed relationship between the Ca2+-dependent F1FO-ATPase and the mPTP in mussels. The findings suggest that mPTP role and regulation may be different in different taxa and that the F1FO-ATPase insensitivity to NaHS may allow mussels to cope with environmental sulfide.


Behavioral and neural network abnormalities in human APP transgenic mice resemble those of App knock-in mice and are modulated by familial Alzheimer's disease mutations but not by inhibition of BACE1.
Alzheimer's disease (AD) is the most frequent and costly neurodegenerative disorder. Although diverse lines of evidence suggest that the amyloid precursor protein (APP) is involved in its causation, the precise mechanisms remain unknown and no treatments are available to prevent or halt the disease. A favorite hypothesis has been that APP contributes to AD pathogenesis through the cerebral accumulation of the amyloid-β peptide (Aβ), which is derived from APP through sequential proteolytic cleavage by BACE1 and γ-secretase. However, inhibitors of these enzymes have failed in clinical trials despite clear evidence for target engagement. To further elucidate the roles of APP and its metabolites in AD pathogenesis, we analyzed transgenic mice overexpressing wildtype human APP (hAPP) or hAPP carrying mutations that cause autosomal dominant familial AD (FAD), as well as App knock-in mice that do not overexpress hAPP but have two mouse App alleles with FAD mutations and a humanized Aβ sequence. Although these lines of mice had marked differences in cortical and hippocampal levels of APP, APP C-terminal fragments, soluble Aβ, Aβ oligomers and age-dependent amyloid deposition, they all developed cognitive deficits as well as non-convulsive epileptiform activity, a type of network dysfunction that also occurs in a substantive proportion of humans with AD. Pharmacological inhibition of BACE1 effectively reduced levels of amyloidogenic APP C-terminal fragments (C99), soluble Aβ, Aβ oligomers, and amyloid deposits in transgenic mice expressing FAD-mutant hAPP, but did not improve their network dysfunction and behavioral abnormalities, even when initiated at early stages before amyloid deposits were detectable. hAPP transgenic and App knock-in mice develop similar pathophysiological alterations. APP and its metabolites contribute to AD-related functional alterations through complex combinatorial mechanisms that may be difficult to block with BACE inhibitors and, possibly, also with other anti-Aβ treatments.


Exercise and cardiac health: physiological and molecular insights.
The cardiac benefits of exercise have been recognized for centuries. Studies have undisputedly shown that regular exercise is beneficial for the cardiovascular system in young, old, healthy and diseased populations. For these reasons, physical activity has been recommended worldwide for cardiovascular disease prevention and treatment. Although the benefits of exercise are clear, understanding of the molecular triggers that orchestrate these effects remains incomplete and has been a topic of intense research in recent years. Here, we provide a comprehensive review of the cardiac effects of physical activity, beginning with a brief history of exercise in cardiovascular medicine and then discussing seminal work on the physiological effects of exercise in healthy, diseased and aged hearts. Later, we revisit pioneering work on the molecular mechanisms underlying the cardiac benefits of exercise, and we conclude with our view on the translational potential of this knowledge as a powerful platform for cardiovascular disease drug discovery.


Protection of the mouse from genetic radiation damage by an optimal-dose-ratio combination of ATP, AET, and serotonin.
The study concerned antiradiation effects in germ-cell genetic structures produced by a combination of ATP, AET, and serotonin at dose ratio optimal for lethality namely, 45:3:1, as arrived at in our previous work. Such a combination was found to reduce by a factor of 2 the translocation yields observed after 400 R X-rays to mouse spermatogonia. In terms of animal survival, ATP has been shown to contribute little to total protection achieved by the same combination; in terms of genetic damage; however, the role of ATP proved essential. Removal of ATP from the combination led to a significant reduction in protective effect.


[Retracted] Consecutive stimulation of HBsAg promotes the viability of the human B lymphoblastoid cell line IM‑9 through regulating the SIRT1‑NF‑κB pathway.
[This retracts the article DOI: 10.3892/ol.2017.6114.].


Downregulation of AHNAK2 inhibits cell cycle of lung adenocarcinoma cells by interacting with RUVBL1.
Lung adenocarcinoma (LUAD) is the leading cause of death among cancer diseases. The tumorigenic functions of AHNAK2 in LUAD have attracted more attention in recent years, while there are few studies which have reported its high molecular weight. The mRNA-seq data of AHNAK2 and corresponding clinical data from UCSC Xena and GEO was analyzed. LUAD cell lines were transfected with sh-NC and sh-AHNAK2, and cell proliferation, migration and invasion were then detected by in vitro experiments. We performed RNA sequencing and mass spectrometry analysis to explore the downstream mechanism and interacting proteins of AHNAK2. Finally, western blot, cell cycle analysis and CO-IP were used to confirm our assumptions regarding previous experiments. Our study revealed that AHNAK2 expression was significantly higher in tumors than in normal lung tissues and higher AHNAK2 expression led to a poor prognosis, especially in patients with advanced tumors. AHNAK2 suppression via shRNA reduced the LUAD cell lines proliferation, migration and invasion and induced significant changes in DNA replication, NF-kappa B signaling pathway and cell cycle. AHNAK2 knockdown also caused G1/S phase cell cycle arrest, which could be attributed to the interaction of AHNAK2 and RUVBL1. In addition, the results from gene set enrichment analysis (GSEA) and RNA sequencing suggested that AHNAK2 probably plays a part in the mitotic cell cycle. AHNAK2 promotes proliferation, migration and invasion in LUAD and regulates the cell cycle via the interaction with RUVBL1. More studies of AHNAK2 are still needed to reveal its upstream mechanism.


Imaging of Gastrointestinal Tract Perforation.
Gastrointestinal tract perforation involving the stomach, duodenum, small intestine, or large bowel occurs as a result of full-thickness gastrointestinal wall injury with release of intraluminal contents into the peritoneal or retroperitoneal cavity. Most cases are associated with high mortality and morbidity, requiring urgent surgical evaluation. Initial patient presentations can be nonspecific with a broad differential, which can delay timely management. This article provides brief overviews of different causes of perforation. Various imaging modalities and protocols are discussed, along with direct and indirect imaging findings of perforation. Specific findings associated with different causes are also described to aid in the diagnosis.


Influence of alcohol consumption on serum immunoglobulin E levels in atopic and nonatopic adults.
Total and specific serum immunoglobulin E (IgE) are routinely used as diagnostic tools in allergy clinics. Several studies have demonstrated an increase of total serum IgE concentrations in alcoholics, but the possible influence of lower quantities of ethanol intake on serum IgE values has not been fully evaluated. This study was aimed at analyzing the influence of alcohol intake on both total and specific serum IgE concentrations in patients studied in an allergy clinic. A total of 460 patients were included in the study. According to skin-prick tests to common aeroallergens, 325 were classified as atopics and 135 as nonatopics. Most atopic patients (253; 78%) were allergic to mites. Alcohol consumption was recorded as the number of standard (10-g) drinking units regularly consumed per week. Two hundred subjects (43%) were abstainers, and 260 (57%) were regular consumers of a median of 30 g of alcohol per week. Total serum IgE was measured in all patients by latex-enhanced nephelometry. Serum-specific IgE was assayed by fluoroenzymeimmunoassay. Total serum IgE increased along with ethanol consumption. On multivariate analysis, regular alcohol consumption greater than 70 g per week was associated with increased total serum IgE levels, even when adjusting for age, sex, atopy, and smoking. Among house-dust mite-allergic patients, specific serum IgE values against the house dust mite Dermatophagoides pteronyssinus were higher in regular alcohol consumers than in abstainers. This difference was not observed among patients allergic to grass pollen (Lolium perenne). Alcohol consumption, even in moderate quantities, is associated with increased total and specific IgE concentrations in subjects studied in an allergy clinic. Alcohol intake should be taken into account in epidemiological studies of total serum IgE levels.


Role of 11β-hydroxysteroid dehydrogenase type 1 in the development of atopic dermatitis.
Glucocorticoids (GCs) are potent anti-inflammatory drugs, the secretion of which is mediated and controlled by the hypothalamic-pituitary-adrenal axis. However, they are also secreted de novo by peripheral tissues for local use. Several tissues express 11β-hydroxysteroid dehydrogenase 1 (11β-HSD1), including the skin. The inactive GC cortisone is converted by 11β-HSD1 to active GC cortisol, which is responsible for delayed wound healing during a systemic excess of GC. However, the role of 11β-HSD1 in inflammation is unclear. We assessed whether 11β-HSD1 affects the development of atopic dermatitis (AD) in vitro and in vivo. The expression of 11β-HSD1 in the epidermis of AD lesions was higher than that in the epidermis of healthy controls. Knockdown of 11β-HSD1 in human epidermal keratinocytes increased the production of thymic stromal lymphopoietin. In an oxazolone-induced mouse model of AD, localized inhibition of 11β-HSD1 aggravated the development of AD and increased serum cytokine levels associated with AD. Mice with whole-body knockout (KO) of 11β-HSD1 developed significantly worse AD upon induction by oxazolone. We propose that 11β-HSD1 is a major factor affecting AD pathophysiology via suppression of atopic inflammation due to the modulation of active GC in the skin.


Etanercept reduces acute tissue injury and mortality associated to zymosan-induced multiple organ dysfunction syndrome.
It has been well demonstrated that TNF-alpha is integral to the pathogenesis of multiple organ dysfunction syndrome (MODS). In this study, we investigate the effects of etanercept (10 mg/kg, s.c.), a specific TNF-alpha-soluble inhibitor, on the acute phase and late mortality in a murine model of MODS of nonseptic origin induced by zymosan (500 mg/kg, suspended in saline solution, i.p.). Etanercept was administered 1 h after the injection of zymosan. Animals were killed after 18 h. In another set of experiments, mice were monitored for systemic toxicity, loss of body weight, and mortality for 12 days. Sham-treated and TNF receptor 1 (TNFR1)-deficient animals were used as control. Treatment of mice with Etanercept and TNFR1 gene deletion decreased the peritoneal exudation and the migration of neutrophils caused by zymosan. In addition, pharmacological and genetic neutralization of TNF-alpha attenuated pancreas and ileum injury (histology), the increase in myeloperoxidase activity in the ileum and in the lung, and the formation of TNF-alpha and IL-1beta. Immunohistochemical analysis for TNF-alpha, transforming growth factor beta, and vascular endothelial growth factor revealed a positive staining in pancreas and ileum sections. The degree of immunostaining was markedly reduced after etanercept treatment and in TNFR1 knockout mice. Furthermore, TNF-alpha neutralization decreased the potent apoptotic stimulus induced by zymosan. All of these findings ultimately led to an amelioration of organ functions at 18 h and to a better survival rate at 12 days. Therefore, we demonstrate that etanercept reduces acute tissue injury and mortality associated to MODS of nonseptic origin in mice.


The effects of bivalent inorganic salts on the mucoadhesive performance of a polymethylmethacrylate sodium salt.
The effects of bivalent inorganic salts, namely CaCl2, MgCl2, MgSO4, ZnCl2 and ZnSO4, on the mucoadhesive performance of a polymethylmethacrylate sodium salt (EuSNa) were investigated. The swelling properties and mucoadhesion of ten different blends made of EuSNa and 5 or 10% (m/m) inorganic salts were evaluated. Only the addition of Zn salts caused a significant reduction of mucoadhesive properties and an increase of swelling. Swelling and mucoadhesive properties of the linear polymethylmethacrylate salt were affected by a physical cross-linking due to bivalent cations. The extent of such interaction may be mainly ascribed to the different electronegativity of the bivalent cations. Attenuated total reflectance infrared spectroscopy (ATR-FTIR) and texture analysis supported this hypothesis. Adding magnesium and calcium salts, the intrinsic dissolution rate of EuSNa decreased of at least 25%. Mucoadhesive tablets were prepared with the most suitable blends and tested on human healthy volunteers. The addition of the magnesium and calcium increased the in vivo permanence time without affecting the mucoadhesive performances. The lack of swelling, mucosal irritation and unpleasant sensation allow the use of such blends in the preparation of mucoadhesive tablets.


Relationship of sebaceous cell stage to growth in culture.
The maturational stage at which sebaceous epithelial cells are irreversibly committed to autolysis has not been determined in a biologic system. The purpose of these experiments was to determine the relationship of sebocyte maturation to their ability to grow in culture. Single cell suspensions from adult rat preputial glands were prepared, and sebocyte stages were classified using lipid staining as one criterion of maturation. Cells were subjected to one-step, isokinetic discontinuous density gradient centrifugation in Percoll. Cells were cultured on 3T3 feeder layers in an epithelial cell medium containing 20% fetal calf serum, growth factors, and antibiotics. Growth in culture occurred from sebocytes in all gradient fractions. The least growth was found from cells in the lightest density fraction (1.020), which contained sebocytes with the most lipid, i.e., the most mature. Over fivefold more growth occurred from cells in the most dense fraction (1.080), which contained undifferentiated and immature sebocytes. Cell growth in culture was then correlated with the type of sebocyte in each fraction plated. Although cell growth in culture correlated significantly with the number of undifferentiated cells (r2 = 0.460), epithelial colonies were found in the absence of discernible undifferentiated cells in most fractions. Growth in culture correlated much better with the number of early plus mid-differentiated sebocytes (r2 = 0.702). These data suggest that, whereas mature sebocytes are not capable of attachment/proliferation, early differentiation of sebocytes is compatible with retention of the capacity to proliferate. If this property is shared by human sebocytes, it would contribute to the intransigence and diversity of acne-form lesions.


Risk factors for total ankle arthroplasty failure: A Dutch Arthroplasty Register study.
Studies concerning total ankle arthroplasty could be influenced by several forms of bias. Independent national arthroplasty registries represent objective data on survival and patient reported outcomes. The aim of this study was to determine survival and identify risk factors for early failure in a nationwide series of total ankle arthroplasties from the Dutch Arthroplasty Register (LROI). Data of 810 patients, who received 836 total ankle arthroplasties between 2014 and 2020 were obtained from the Dutch Arthroplasty Register (LROI) with a median follow-up of 38 months (range 1-84 months). Survival was expressed in Kaplan-Meier analysis and associated hazard ratios for implant failure were determined. Implant failure was defined as the need for revision surgery for any reason or (pan)arthrodesis. During follow-up, we recorded 39 failures (4.7%) resulting in a implant survival of 95.3% with a median follow-up of 38 months (range 1-84 months). Medial malleolus osteotomy (HR = 2.27), previous surgery (HR = 1.83), previous osteotomy (HR = 2.82) and previous ligament reconstruction (HR = 2.83) all showed potentially clinically meaningful associations with a higher incidence of implant failure, yet only previous OCD treatment (HR = 6.21), BMI (HR = 1.09) and age (HR = 0.71) were statistically significant. Excellent short-term survival (95.3%) with a median follow-up of 38 months was reported for TAA patients from the Dutch Arthroplasty Register. Patients with a lower age, a higher BMI or who had a prior surgical OCD treatment before TAA surgery appear to have a higher risk for revision after short-term clinical follow-up. Thorough patient selection with emphasis on risk factors associated with early implant failure might be essential to improve TAA survivorship.


Anisotropic and hydrogen bonding effects in phenylglyoxamides and mandelamides: theoretical and NMR conformational evaluation.
Interesting anisotropic effects were observed for phenylglyoxamides and their respective mandelamides. Such effects were observed in experimental (1)H and (13)C NMR (in CDCl(3), CD(3)OD, and DMSO-d(6) solvents) and in some cases with good correlation to theoretical (1)H and (13)C NMR DFT-GIAO (B3LYP/6-311++G**//B3LYP/6-31G*) calculations. A systematic conformational analysis of these compounds was performed in a two-step methodology, using PM3 and DFT (B3LYP/6-31G*) calculations; with good accomplishment and computational time economy. It was observed that intramolecular hydrogen bonding plays a significant role in the conformation of such compounds. Finally, a geminal nonequivalence of an N-CH(2) moiety, in one of the alkyl side chain (R1 = R2), was found for the tertiary mandelamides studied.


J or H mtDNA haplogroups in retinal pigment epithelial cells: Effects on cell physiology, cargo in extracellular vesicles, and differential uptake of such vesicles by naïve recipient cells.
Extracellular vesicles (EVs) are predicted to represent the internal state of cells. In polarized RPE monolayers, EVs can mediate long-distance communication, requiring endocytosis via protein-protein interactions. EV uptake from oxidatively stressed donor cells triggers loss in transepithelial resistance (TER) in recipient monolayers mediated by HDAC6. Here, we examine EVs released from RPE cells with identical nuclear genes but different mitochondrial (mt)DNA haplogroups (H, J). J-cybrids produce less ATP, and the J-haplogroup is associated with a higher risk for age-related macular degeneration. Cells were grown as mature monolayers to either collect EVs from apical surfaces or to serve as naïve recipient cells. Transfer assays, transferring EVs to a recipient monolayer were performed, monitoring TER and EV-uptake. The presence of known EV surface proteins was quantified by protein chemistry. H- and J-cybrids were confirmed to exhibit different levels of TER and energy metabolism. EVs from J-cybrids reduced TER in recipient ARPE-19 cells, whereas EVs from H-cybrids were ineffective. TER reduction was mediated by HDAC6 activity, and EV uptake required interaction between integrin and its ligands and surface proteoglycans. Protein quantifications confirmed elevated levels of fibronectin and annexin A2 on J-cybrid EVs. We speculate that RPE EVs have a finite set of ligands (membrane proteoglycans and integrins and/or annexin A2) that are elevated in EVs from stressed cells; and that if EVs released by the RPE could be captured from serum, that they might provide a disease biomarker of RPE-dependent diseases.


Immunocytochemical detection of T and B cell populations in routine blood smears.
Immunofluorescence labelling of peripheral blood mononuclear cells is usually done on cells in suspension. This paper describes a procedure based on immuno-alkaline phosphatase staining of routine blood smears. The advantages of this method are that a few drops of blood are sufficient for labelling multiple lymphocyte subpopulations; smears may be stored for long periods before labelling; it is unnecessary to isolate a mononuclear cell fraction before labelling; labelled preparations can be stored; and the morphological features of labelled cells are shown clearly. The technique was used to label T cells and their subsets, B cells, and HLA-DR antigen in blood smears from 15 normal donors, from 7 patients with infectious mononucleosis, from 1 patient with clinically proven AIDS, and from 1 symptom-free subject at risk of AIDS. The normal T helper/suppressor ratio of 1 X 95 was reversed in all of the last three groups of subjects, the mean being 0 X 34 for infectious mononucleosis; the value was 0 X 22 in the AIDS patients. Immuno-alkaline phosphatase labelling of routine blood smears seems to be a valuable method for studying abnormalities in circulating lymphocyte subpopulations and lends itself to mass screening for altered T helper and T suppressor subjects-for example, in blood donors.


The metamicrobiome: key determinant of the homeostasis of nutrient recycling.
The metamicrobiome is an integrated concept to study carbon and nutrient recycling in ecosystems. Decomposition of plant-derived matter by free-living microbes and fire - two key recycling pathways - are highly sensitive to global change. Mutualistic associations of microbes with plants and animals strongly reduce this sensitivity. By solving a fundamental allometric trade-off between metabolic and homeostatic capacity, these mutualisms enable continued recycling of plant matter where and when conditions are unfavourable for the free-living microbiome. A diverse metamicrobiome - where multiple plant- and animal-associated microbiomes complement the free-living microbiome - thus enhances homeostasis of ecosystem recycling rates in variable environments. Research into metamicrobiome structure and functioning in ecosystems is therefore important for progress towards understanding environmental change.


Treatment of Mandibular Class III/IV Furcation Defects with a Combination Plastic Regenerative Technique: A Case Series.
The aim of this present case series investigated the effect of a combination therapy utilizing connective tissue graft (CTG) in the treatment of periodontal regeneration of mandibular Class Ⅲ furcation involvement (FI). Six patients diagnosed with periodontitis stage Ⅲ or Ⅳ (grade A to C), presenting with Class Ⅲ or Ⅳ FI, were treated with fibroblast growth factor 2 and carbonate apatite in combination with CTG. The following clinical parameters were evaluated at baseline and after 6, 12 and 18 months: periodontal probing depth, clinical attachment level, furcation invasion, the radiographic vertical defect depth, and gingival phenotype. Significant improvements in clinical parameters were observed in all treated FI sites. Four Class Ⅲ Fls and one Class Ⅳ Fl obtained complete closure, and one Class Ⅳ furcation was improved to Class Ⅰ. This case series showed the potential of administering combination regenerative therapy for changing the prognosis of hopeless teeth with severe furcation defects.


Effects of platelet derived growth factor (PDGF) on fibronectin (FN) production by human skin and scar fibroblasts.
The fibroblast-type cell found in hypertrophic scars and keloids demonstrates an elevated fibronectin (FN) production, compared to the same type of cell in normal dermis. We wished to determine if the effects of platelet derived growth factor (PDGF) on FN production in these cell types would be equivalent or different. Cell lines were established from the dermis (reticularis) of hypertrophic scars, keloids, uninvolved normal skin adjacent to the lesions, including an assumed normal skin adjacent to a keloid (AS), and normal skin from a different uninjured patient (DS). Each parent tissue from which the cell lines originated was diagnosed histologically. Each hypertrophic scar, keloid and normal adjacent skin, with one exception, showed typical histologic findings confirming the clinical diagnosis. DS was also normal. AS, although assumed to be normal, in fact, demonstrated portions of nodules from the adjacent keloid. All cell lines were grown under standard conditions with subconfluent cells metabolically labeled for radioimmunoassays measuring FN at passage 3 (8 to 9 weeks in culture) in the absence and presence of PDGF. Significant differences in production of FN/cell and FN/PR/cell between two hypertrophic scars and their matched normal skins and for one keloid and its matched normal skin were observed. However, no significant difference was observed between the other keloid and AS, nor between the other hypertrophic scar and DS. PDGF significantly stimulated FN production in 2 of 4 NS cell lines, and in the AS cell line. By FN/cell values, 2 of 5 cell lines from the lesions were inhibited and one was increased. In terms of FN/PR/cell, 1 of 5 cell lines from the lesions was stimulated and the others showed no differences. The mixed results may be attributable to the likelihood that the cell lines represent mixed populations. This study demonstrates the importance of: 1) histological characterization of all parent tissues from which cell lines are derived, and 2) matching cell lines from lesions with cell lines from uninvolved normal dermis, in the same individual.


Cryogenic 3D printing of dual-delivery scaffolds for improved bone regeneration with enhanced vascularization.
Three-dimensional (3D) printing has been increasingly employed to produce advanced bone tissue engineering scaffolds with biomimetic structures and matched mechanical strengths, in order to induce improved bone regeneration in defects with a critical size. Given that the successful bone regeneration requires both excellent osteogenesis and vascularization, endowing scaffolds with both strong bone forming ability and favorable angiogenic potential would be highly desirable to induce improved bone regeneration with required vascularization. In this investigation, customized bone tissue engineering scaffolds with balanced osteoconductivity/osteoinductivity were produced via cryogenic 3D printing of β-tricalcium phosphate and osteogenic peptide (OP) containing water/poly(lactic-co-glycolic acid)/dichloromethane emulsion inks. The fabricated scaffolds had a hierarchically porous structure and were mechanically comparable to human cancellous bone. Angiogenic peptide (AP) containing collagen I hydrogel was then coated on scaffold surface to further provide scaffolds with angiogenic capability. A sequential release with a quick AP release and a slow but sustained OP release was obtained for the scaffolds. Both rat endothelial cells (ECs) and rat bone marrow derived mesenchymal stem cells (MSCs) showed high viability on scaffolds. Improved in vitro migration and angiogenesis of ECs were obtained for scaffolds delivered with AP while enhanced osteogenic differentiation was observed in scaffolds containing OP. The in vivo results showed that, toward scaffolds containing both AP and OP, the quick release of AP induced obvious angiogenesis in vivo, while the sustained OP release significantly improved the new bone formation. This study provides a facile method to produce dual-delivery scaffolds to achieve multiple functions.


[Otomycoses].
Otomycosis is a relatively frequent disorder (21.7% of the cases of external otitis) of fungi origin. Otomycoses can be discriminated from bacterial otitis by the following symptoms: itching, absence of otalgy, discharge and negative response to different local antibiotic therapies. Our study involves 148 cases of otomycosis and their therapy. Recovery was obtained in approximately 10 days.


Monitoring of DNA-protein interaction with single gold nanoparticles by localized scattering plasmon resonance spectroscopy.
We reported a sensitive detection system for measuring DNA-protein interaction at single plasmonic metal nanoparticles level by Localized Scattering Plasmon Resonance (LSPR) spectroscopy. As a proof of concept, DNA molecules were conjugated to gold nanoparticles (AuNPs) through gold-thiol chemistry and the resulted complex was served as single-particle probes of human topoisomerase I (TOPO). By recording the changes in Rayleigh light scattering signal of the individual nanoparticles upon protein binding, DNA-protein interaction was monitored and measured. The λmax shifts in LSPR spectrum of individual AuNP was found to be highly correlated with the amount of TOPO that bound onto. This technique provides a sensitive and high-throughput platform to screen and monitor accurately the specific biomolecular interactions. It is capable of revealing information such as particle-particle variations that might be buried in conventional bulk measurement.


Cathepsin D is released after severe tissue trauma in vivo and is capable of generating C5a in vitro.
In response to severe tissue trauma several danger sensing and signalling cascades are activated, including the complement and the apoptosis systems. In polytrauma patients, both the early activation of the complement cascade with an excessive generation of the potent anaphylatoxin C5a and the induction of apoptosis have been shown to modulate the post-traumatic immune response. However, little is known about a direct interaction between the complement and apoptosis systems after severe tissue trauma. Therefore the focus of the present study was to elucidate the interplay between the central complement component C5 and the pro-apoptotic aspartic protease cathepsin D. In vivo, the cathepsin D plasma concentration of multiple injured patients was markedly increased when compared to healthy volunteers. In vitro incubation of C5 with cathepsin D resulted in a concentration- and time-dependent generation of C5a, which was inhibited by the aspartate protease inhibitor pepstatin A. Immunoblotting and sequencing analysis indicated that the C5 cleavage product represents the native form of human C5a, also exhibiting chemotactic activity for human neutrophils. In conclusion, these data show for the first time that cathepsin D is increased in plasma early after severe tissue injury. Furthermore, the results provide in vitro evidence of cleavage of C5 by an aspartic protease with subsequent generation of functional C5a, which represents a new path of complement activation.


Dose to the developing dentition during therapeutic irradiation: organ at risk determination and clinical implications.
Irradiation of pediatric facial structures can cause severe impairment of permanent teeth later in life. We therefore focused on primary and permanent teeth as organs at risk, investigating the ability to identify individual teeth in children and infants and to correlate dose distributions with subsequent dental toxicity. We retrospectively reviewed 14 pediatric patients who received a maximum dose >20 Gy(relative biological effectiveness, RBE) to 1 or more primary or permanent teeth between 2003 and 2009. The patients (aged 1-16 years) received spot-scanning proton therapy with 46 to 66 Gy(RBE) in 23 to 33 daily fractions for a variety of tumors, including rhabdomyosarcoma (n=10), sarcoma (n=2), teratoma (n=1), and carcinoma (n=1). Individual teeth were contoured on axial slices from planning computed tomography (CT) scans. Dose-volume histogram data were retrospectively obtained from total calculated delivered treatments. Dental follow-up information was obtained from external care providers. All primary teeth and permanent incisors, canines, premolars, and first and second molars were identifiable on CT scans in all patients as early as 1 year of age. Dose-volume histogram analysis showed wide dose variability, with a median 37 Gy(RBE) per tooth dose range across all individuals, and a median 50 Gy(RBE) intraindividual dose range across all teeth. Dental follow-up revealed absence of significant toxicity in 7 of 10 patients but severe localized toxicity in teeth receiving >20 Gy(RBE) among 3 patients who were all treated at <4 years of age. CT-based assessment of dose distribution to individual teeth is feasible, although delayed calcification may complicate tooth identification in the youngest patients. Patterns of dental dose exposure vary markedly within and among patients, corresponding to rapid dose falloff with protons. Severe localized dental toxicity was observed in a few patients receiving the largest doses of radiation at the youngest ages; however, multiple factors including concurrent chemotherapy confounded the dose-effect relationship. Further studies with larger cohorts and appropriate controls will be required.


Cytokine therapy restores antitumor responses of NK cells rendered anergic in MHC I-deficient tumors.
Recent extraordinary advances in cancer immunotherapy rely primarily on marshaling T cell responses. Here we discuss how NK cell responses can be amplified. We find that MHC I-deficient tumors induce anergy of NK cells but that cytokine therapy restores NK cell activity and increases the survival of mice bearing MHC I-deficient tumors.


[Factor-free translation of poly(dT) by 80S ribosomes from Saccharomyces cerevisiae].
The conditions for preparation of 80S ribosomes from S. cerevisiae are suggested. The ribosomes can bind Phe-tRNAPhe in poly(U)-or poly(dT)-directed manner and are shown to be able to translate poly(dT) in the absence of elongation factor and GTP. Effects of different antibiotics on the factor-free translation have been studied.


Effect of marginal zinc deficiency on lipoprotein lipase activities in postheparin plasma, skeletal muscle and adipose tissues in the rat.
The activities of lipoprotein lipase in postheparin plasma, retroperitoneal adipose and gastrocnemius muscle tissues were determined in the rats fed 2.8 ppm of dietary zinc for eight weeks, as compared with pair-fed and ad libitum-fed rats given 30.8 ppm of zinc. The postheparin lipoprotein lipase activity, as determined by using a lipid emulsion labeled with [3H]triolein as the substrate, was significantly lower in the first group of rats, relative to that in the second and third groups. Tissue lipoprotein lipase activities were compared using the lipid emulsion and activator serum obtained from the zinc-deficient rats and the ad libitum-fed rats. The activator sera were devoid of very low density and low density lipoproteins, but enriched in high density lipoproteins. Muscle lipoprotein lipase activities were significantly lower when assayed with the activator serum from the zinc-deficient compared with the activities determined with the activator serum from the ad libitum-fed. Similarly, muscle lipoprotein lipase activities were lower in all groups when [3H]-triolein-labeled chylomicrons from the zinc-deficient were used as the substrate, compared with the activities determined using the chylomicrons from the ad libitum-fed. Lipoprotein lipase activities in the adipose tissues were not affected by the different sources of the activator sera and chylomicrons. The results strongly suggest that the decrease in postheparin lipoprotein lipase activity in zinc deficiency is not due to changes in tissue lipoprotein lipase enzyme per se, but to compositional alterations in chylomicrons and high density lipoprotein, particularly, with regard to C apolipoproteins, modulators of lipoprotein lipase activity.


A novel amplification-based approach to enable gene expression profiling from small clinical tumor specimens.
Glioblastoma is the most common and deadly type of brain cancer. Over the past decade, several divergent genetic pathways have been implicated in the initiation, progression and clinical outcome of this disease. As our understanding of GBM expands and identifies actionable targets specific to individual tumors, there will be a pressing need for the development of new tools that will maximize the use of limited clinical samples to enable the employment of personalized care paradigms. We used PrimePCR validated assays to generate a custom real-time PCR screening panel, containing 74 previously published mRNA targets showing gene expression changes in glioblastoma, and five house-keeping genes. A cohort of 19 frozen brain specimens were analyzed, including WHO Grade II oligodendroglioma (n = 3), WHO Grade II astrocytoma (n = 2), WHO Grade III astrocytoma (n = 1), and glioblastoma (n = 13). Four normal brain samples were also analyzed. We performed RNA extraction, followed by cDNA synthesis, multiplexed pre-amplification and SYBR-based qPCR, to generate expression profiles on all samples. We demonstrated that the workflow shows high tolerance to variation in RNA quality (RIN 8.5-4) and high sensitivity in detection. cDNA input that is equivalent to 3 ng of starting RNA was sufficient to conduct accurate semiquantitative analysis of the panel of 79 assays. Using principal component analysis, we were able to accurately separate glioblastoma from low-grade glioma. The two WHO Grade III tumors analyzed clustered with glioblastoma, but showed more similarity to Grade II gliomas. In this study, we have shown the feasibility of consolidating high-throughput data into a single functional panel capable of accurately classifying glioma specimens based solely on semiquantitative gene expression profiling.