Self-Assembled Polypeptide Nanogels with Enzymatically Transformable Surface as a Small Interfering RNA Delivery Platform.
Nanometer-size gel particles, or nanogels, have potential for delivering therapeutic macromolecules. A cationic surface promotes cellular internalization of nanogels, but undesired electrostatic interactions, such as with blood components, cause instability and toxicities. Poly(ethylene glycol) coating has been used to shield charges, but this decreases delivery efficiency. Technical difficulties in synthesis and controlling molecular weights make it unfeasible to, instead, coat with biodegradable polymers. Our proposed solution is cationized nanogels enzymatically functionalized with branched polysaccharide chains, forming a shell to shield charges and increase stability. Biodegradation of the polysaccharides by an endogenous enzyme would then expose the cationic charges, allowing cellular internalization and cargo delivery. We tested this concept, preparing maltopentaose functionalized cholesteryl poly(l-lysine) nanogel and using tandem enzymatic polymerization with glycogen phosphorylase and glycogen branching enzyme, to add branched amylose moieties, forming a CbAmyPL nanogel. We characterized CbAmyPL nanogels and investigated their suitability as small interfering RNA (siRNA) carriers in murine renal carcinoma (Renca) cells. The nanogels had neutral ζ potential values that became positive after degradation by α-amylase. Foster resonance energy transfer demonstrated that the nanogels formed stable complexes with siRNA, even in the presence of bovine serum albumin and after α-amylase exposure. The nanogels, with or without α-amylase, were not cytotoxic. Complexes of CbAmyPL with siRNA against vascular endothelial growth factor (VEGF), when incubated alone with Renca cells decreased VEGF mRNA levels by only 20%. With α-amylase added, however, VEGF mRNA knockdown by the siRNA/nanogels complexes was 50%. Our findings strongly supported the hypothesis that enzyme-responsive nanogels are promising as a therapeutic siRNA delivery platform.


Zoledronic Acid Add-on Therapy for Standard-Risk Ewing Sarcoma Patients in the Ewing 2008R1 Trial.
The phase III, open-label, prospective, multicenter, randomized Ewing 2008R1 trial (EudraCT2008-003658-13) was conducted in 12 countries to evaluate the effect of zoledronic acid (ZOL) maintenance therapy compared with no add-on regarding event-free survival (EFS, primary endpoint) and overall survival (OS) in standard-risk Ewing sarcoma (EWS). Eligible patients had localized EWS with either good histologic response to induction chemotherapy and/or small tumors (<200 mL). Patients received six cycles of VIDE induction and eight cycles of VAI (male) or eight cycles of VAC (female) consolidation. ZOL treatment started parallel to the sixth consolidation cycle. Randomization was stratified by tumor site (pelvis/other). The two-sided adaptive inverse-normal four-stage design (planned sample size 448 patients, significance level 5%, power 80%) was changed after the first interim analysis using the Müller-Schäfer method. Between April 2010 and November 2018, 284 patients were randomized (142 ZOL/142 no add-on). With a median follow-up of 3.9 years, EFS was not significantly different between ZOL and no add-on group in the adaptive design (HR, 0.74; 95% CI, 0.43-1.28, P = 0.27, intention-to-treat). Three-year EFS rates were 84.0% (95% CI, 77.7%-90.8%) for ZOL vs. 81.7% (95% CI, 75.2%-88.8%) for no add-on. Results were similar in the per-protocol collective. OS was not different between groups. The 3-year OS was 92.8% (95% CI, 88.4%-97.5%) for ZOL and 94.6% (95% CI, 90.9%-98.6%) for no add-on. Noticeable more renal, neurologic, and gastrointestinal toxicities were observed for ZOL (P < 0.05). Severe renal toxicities occurred more often in the ZOL arm (P = 0.003). In patients with standard-risk localized EWS, there is no additional benefit from maintenance treatment with ZOL.


Model for estimating the refractive-index structure constant in clear-air intermittent turbulence.
We explain discrepancies in comparing estimations of the refractive-index structure constant C(n)(2) in clear air by means of different techniques by taking into account atmospheric intermittency effects. We formulate a model of C(n)(2) in intermittent turbulence on the basis of the Tatarskii theory, and we calculate the mean value of C(n)(2) through a probabilistic approach. We deduce a factor, which gives a measure of the statistical reduction of turbulence that is due to intermittency, within the model framework. A procedure for estimating the mean value of C(n)(2) from data of a specific radiosonde observation is illustrated.


Vaccinia-virus-induced cellular contractility facilitates the subcellular localization of the viral replication sites.
Poxviruses, such as vaccinia virus (VV), replicate their DNA in endoplasmic-reticulum-enclosed cytoplasmic sites. Here, we compare the dynamics of the VV replication sites with those of the attenuated strain, modified VV Ankara (MVA). By live-cell imaging, small, early replication sites of both viruses undergo motility typical of microtubule (MT)-motor-mediated movement. Over time, growing replication sites of VV collect around the nucleus in a MT-dependent fashion, whereas those of MVA remain mostly scattered in the cytoplasm. Surprisingly, blocking the dynein function does not impair the perinuclear accumulation of large VV replication sites. Live-cell imaging demonstrates that in contrast to small replication sites, large sites do not display MT-motor-mediated motility. Instead, VV infection induces cellular contractility that facilitates the collection of growing replication sites around the nucleus. In a subset of cells (30-40%), this VV-induced contractility is alternated by phases of directed cell migration, suggesting that the two processes may be linked. The MVA-infected cells do not display contractility or cell migration, supporting the idea that these cellular activities facilitate the efficient accumulation of the VV replication sites around the nucleus. We propose that the recently described cytoskeletal rearrangements induced by VV are a prerequisite for the observed cell contractility and migration activities that apparently contribute to the organization of the complex cytoplasmic life cycle of VV.


A novel structure-specific endonuclease activity associated with polypyrimidine-tract binding (PTB) related protein from rat testis.
Nucleases are involved in the processing of various intermediates generated during crucial DNA metabolic processes such as replication, repair, and recombination and also during maturation of RNA precursors. An endonuclease, degrading specifically single-stranded circular DNA, was identified earlier in rat testis nuclear extract while purifying a strand-transfer activity. We are now reporting the purification of this endonuclease, which is a monomeric 42 kDa protein, from rat testis to near-homogeneity. In addition to degrading single-stranded circular DNA, it nicks supercoiled plasmid DNA to generate relaxed DNA and does not act on linear single-stranded or double-stranded DNA. It also makes specific incisions at the single-strand/duplex junction of pseudo-Y, 3'- and 5'-overhangs and 3'- and 5'-flap structures. Other structures such as mismatch, insertion loop, and Holliday junction are not substrates for the testis endonuclease. In contrast to FEN1, the testis endonuclease makes asymmetric incisions on both strands of the branched structures, and free single-stranded ends are not necessary for the structure-specific incisions. Neither 5'-3' nor 3'-5' exonuclease activity is associated with the testis endonuclease. The amino acid sequences of tryptic peptides of the 42 kDa endonuclease show near-identity to polypyrimidine-tract binding protein (PTB) that is involved in the regulation of splicing of eukaryotic mRNA. The significance of the results on the association of structure-specific endonucleae activities with PTB-related protein is discussed.


Expression and function of leptin receptor isoforms in myeloid leukemia and myelodysplastic syndromes: proliferative and anti-apoptotic activities.
The receptor for the gene product of the obesity gene, leptin, was recently reported to be expressed on murine and human hematopoietic progenitor cells. Therefore, we studied the expression of the leptin receptor, OB-R, in normal myeloid precursors, human leukemia cell lines, and primary leukemic cells using reverse-transcriptase polymerase chain reaction. In normal hematopoiesis, OB-R was expressed in CD34(+) cells. Normal promyelocytes (CD34(-)33(+) and CD34(-)13(+)) expressed only very low levels of the short, presumably nonsignaling isoform. Both the long and short isoforms of OB-R were expressed in 10 of 22 samples from patients with newly diagnosed primary or secondary acute myeloid leukemia (AML), with a higher incidence of the long isoform in primary AML (87.6% v 28.6%; P =.01). The incidence of OB-R expression was higher in recurrent than in newly diagnosed AML (P <.001), and samples from four patients with refractory AML showed strong expression of both isoforms. Both OB-R isoforms were also expressed in newly diagnosed and recurrent acute promyelocytic leukemia cells but were essentially absent in samples of chronic or acute lymphocytic leukemia. In vitro growth of myeloid leukemic cell lines and of blasts from 14 primary AMLs demonstrated that recombinant human leptin alone induced low level proliferation, significantly (P <.05) increased proliferation induced by recombinant human granulocyte colony-stimulating factor, interleukin 3, and stem cell factor in a subset of AML and increased colony formation (P <.005). Also, leptin reduced apoptosis induced by cytokine withdrawal in MO7E and TF-1 cells. Serum leptin levels correlated only with body mass index (P <. 001) and gender (P =.03). Results confirm the reported expression of leptin receptor in normal CD34(+) cells and demonstrate the frequent expression of leptin receptors in AML blasts. While normal promyelocytes lack receptor expression, leukemic promyelocytes express both isoforms. We also demonstrate proliferative effects of leptin alone and in combination with other physiologic cytokines, and anti-apoptotic properties of leptin. These findings could have implications for the pathophysiology of AML.


Surveillance activity for canine parvovirus in Italy.
Recent identification of unusual canine parvovirus (CPV) mutants in cats and dogs suggests that CPV type 2 (CPV-2), which emerged suddenly in the late 1970s, is undergoing continual genetic and antigenic variations. A peculiarity of parvoviruses is that single-nucleotide substitutions may determine drastic phenotypic changes. The effects of either natural or artificial mutations on CPV phenotypic properties have been largely investigated, and this sets up CPV as an interesting model to study virus evolution. By monitoring the evolution of CPV-2 in Italy, we observed the onset and quick spread of a Glu-426 mutant, antigenically different from the pre-existing variants that were partially displaced within a few years of the initial identification of the new mutant. The identification of CPV-2 variants raises several questions concerning their impact on the efficacy of the current CPV-2 vaccines, based on the original CPV-2 strain that no longer exists in the field.


Differing genetic trend estimates from traditional and genomic evaluations of genotyped animals as evidence of preselection bias in US Holsteins.
The objective of this study was to compare genetic trends from single-step genomic BLUP (ssGBLUP) and traditional BLUP models for milk production traits of US Holsteins. Phenotypes were 305-d milk, fat, and protein yields from 21,527,040 cows recorded between January 1990 and August 2015. The pedigree file included 29,651,623 animals and was limited to 3 generations back from recorded or genotyped animals. Genotypes for 764,029 animals were used, and analyses were by a 3-trait repeatability model as used in the US official genetic evaluation. Unknown-parent groups were incorporated into the inverse of a relationship matrix (H-1 in ssGBLUP and A-1 in BLUP) with the QP transformation. For ssGBLUP, 18,359 genotyped animals were randomly chosen as core animals to calculate the inverse of the genomic relationship matrix with the APY algorithm. Computations took 6.5 h and 1.4 GB of memory for BLUP, and 13 h and 115 GB of memory for ssGBLUP. For genotyped sires with at least 10 daughters, the average genetic levels for predicted transmitting ability (PTA) and genomic PTA were similar up to 2008, with a higher level for ssGBLUP later (approximately by 36 kg for milk, 2.1 kg for fat, and 1.1 kg for protein for bulls born in 2010). For genotyped cows, the average genetic levels were similar up to 2006, with a higher level for ssGBLUP (approximately by 91 kg for milk, 3.6 kg for fat, and 2.7 kg for protein for cows born in 2012). For all cows, the average levels were slightly higher for ssGBLUP, with much smaller differences than for genotyped cows. Trends for BLUP indicate bias due to genomic preselection for genotyped sires and cows. For official evaluations released in December 2016, traditional PTA had the same trend as multiple-step genomic PTA for both genotyped bulls and cows except for the youngest bulls, who had traditional PTA slightly lower than genomic PTA. For genotyped bulls born in recent years, genetic gain for official traditional and genomic evaluations was similar in contrast to ssGBLUP and BLUP differences. Official PTA for cows were adjusted so that the Mendelian sampling variance was comparable with that for bulls, and those adjustments likely removed bias due to genomic preselection from traditional PTA, especially for genotyped cows. The ssGBLUP method seems to account partially for that bias and is computationally suitable for national evaluations.


[Clinical manifestation of attic retraction pocket].
To analyze the clinical manifestation of attic retraction pocket. Ninety-two patients with 118 involved ears were studied. The clinical data including clinical manifestation, endoscopic findings and gradings,hearing tests, imaging findings and accompanying lesions. According to Tos's grades, 15 ears (14/118,12.7%) belonged to stage I, 48 (42/118, 40.7%) belonged to stage II, 23 (23/104,19.5%) belonged to stage III and 32 (32/118, 27.1%) belonged to stage IV. Patients always complained of ear blockage,hearing loss and tinnitus. Accompanying lesions including otitis media with effusion, middle ear atelectasis, adhesive otitis media and retraction pocket cholesteatoma could be found by endoscopy. The results of pure audiometry were related to the grades of lesions. Most patients had conductive hearing loss and a few patients had mixed hearing loss. The tympanometry in most cases was type B or C. Imaging examination (CT or X-ray) of 97 ears showed poor pneumatization of mastoids and there was a significant difference between involved group and control group (P < 0.01). The attic retraction pocket reflected the negative pressure of middle ear and its pathogenesis was related with the function of Eustachian tube, inflammation and pneumatization of mastoid. Since it had a risk to develop into retraction pocket cholesteatoma, following-up and interferon to early or middle stage lesion should be paid more attention.


Digital health in the management of allergic diseases.
In recent years there has been an important implementation in the medical field of both Mobile Health, such as the use of mobile communication devices, and of other telemedicine tools in general, with the aim of supporting the supervision of diseases from the moment of the first diagnosis to the therapeutic follow-up. In fact, Digital Health can also have a very positive impact on the management of allergic patients, who are known to have the greatest need for regular monitoring, simplifying contact between doctor and patient, but there is still a need to improve implementation regulations, define certification programs and adequate reimbursement systems, as well as to guarantee a high level of attention to the protection of sensitive data. The hope is that one positive outcome of the Covid-19 pandemic will be an acceleration, by all stakeholders involved, of the process of the modernization of health care.


Protein Stabilization and Delivery: A Case Study of Invasion Plasmid Antigen D Adsorbed on Porous Silica.
Approximately half of all vaccines produced annually are wasted because effectivity is dependent on protein structure and heat exposure disrupts the intermolecular interactions needed to maintain the structure. Thus, most vaccines require a temperature-controlled supply chain to minimize waste. A more sustainable technology was developed via the adsorption of invasion plasmid antigen D (IpaD) onto mesoporous silica, improving the thermal stability of this protein-based therapeutic. Seven silicas were characterized to determine the effects of pore diameter, pore volume, and surface area on protein adsorption. The silica-IpaD complex was then heated above the IpaD denaturing temperature and N,N-dimethyldodecylamine N-oxide was used to remove IpaD from the silica. Circular dichroism confirmed that the adsorbed IpaD after the heat treatment maintained a native secondary structure rich in α-helix content. In contrast, the unprotected IpaD after heat treatment lost its secondary structure. Isotherms using Langmuir, Freundlich, and Temkin models demonstrated that the adsorption of IpaD onto silicas is best fit by the Langmuir model. If pores are less than 15 nm, adsorption is negligible. If the pores are between 15 and 25 nm, then monolayer coverage is achieved and IpaD is protected from thermal denaturing. If pores are larger than 25 nm, the adsorption is a multilayer coverage and it is easier to remove the protein from the silica because of a less-developed hydrogen bond network. This case study provides strong evidence that IpaD is thermally stabilized via adsorption on mesoporous silica with the proper range of pore sizes.


A visual lamina in the medulla oblongata of the frog, Rana pipiens.
We have discovered a lamina of visually responsive units in the medulla oblongata of the frog. It spans the entire medial aspect of the rostrocaudal length of the medulla and extends dorsoventrally from the cell-dense dorsal zone into the cell-sparse ventral zone. Most visual units within this lamina have large receptive fields, with the majority extending bilaterally in the frontal visual field. Most of these neurons are binocular, have no apparent directional preference, respond equally well to stimuli of a variety of shapes and sizes, and exhibit strong habituation. More medial locations in the visual lamina represent ipsilateral visual space while more lateral locations within the lamina represent contralateral visual space. Many units in the caudal aspect of the visual lamina are bimodal, responding to both visual and somatosensory stimuli. HRP tracing reveals inputs to the lamina from many primary and secondary visual areas in the midbrain and diencephalon. There is no area-by-area segregation of the projections to the visual lamina. For example, most parts of the tectum project across the visual lamina. The only spatial order in the visual lamina is that at more medial sites there tends to be more input from contralateral tectum; and at more lateral sites there tends to be more input from ipsilateral tectum. There is bilateral input to the visual lamina from tectum, tegmentum, posterior nucleus of the thalamus, posterior tuberculum, and ventromedial thalamic nucleus. There is ipsilateral input to the visual lamina from torus semicircularis, pretectum, nucleus of Bellonci, and ventrolateral thalamic nucleus. There is contralateral input to the visual lamina from basal optic complex. Collectively, these results show the presence of visual influences in regions of the medulla that likely represent an important step in sensorimotor transformation.


Vertebrobasilar thrombosis in children: report of two cases and recommendations for treatment.
Two consecutive cases of children with vertebrobasilar thrombosis (VBT) were treated with high-dose intra-arterial urokinase within 4 h of presenting to the emergency room, after full evaluation by CT scan, MRI and MR angiography. Complete resolution of neurologic symptoms was achieved in both cases. Based on our limited pediatric experience, previous treatment of VBT at our institution and a review of the relevant literature, the authors suggest that VBT be specifically ruled out at initial diagnosis, and if present, full consideration be given to immediate treatment with intra-arterial thrombolytic therapy. This may lead to a significant reduction in the morbidity and mortality associated with VBT in children. A large prospective multi-institution study is needed to further evaluate the efficacy of this approach to childhood stroke.


A Reason to Rethink Fasting Guidelines? Marijuana-Induced Gastroparesis and the Implications for Aspiration Risk in the Nil Per Os (NPO) Patient: A Case Report.
BACKGROUND Tetrahydrocannabinol has been implicated in gastroparesis development, a disorder characterized by delayed gastric emptying in the absence of mechanical obstruction. While most patients with gastroparesis present with upper gastrointestinal symptoms, patients with significantly delayed emptying can be asymptomatic, creating a dangerous aspiration risk. CASE REPORT A 24-year-old man with a body mass index of 22 presented for right lower extremity open reduction and internal fixation. He endorsed daily marijuana use. His last meal was at 10 PM the prior night. After induction of general anesthesia and laryngeal mask airway (LMA) placement, 150 cc of gastric contents filled the LMA. The LMA was immediately removed and 25 to 50 cc of gastric contents was suctioned from the oropharynx. The airway was immediately secured with an endotracheal tube, after which 500 cc of clear gastric contents was suctioned from the stomach. Bronchoscopy revealed no obvious gastric contents in the tracheobronchial tree. The patient maintained adequate ventilation and oxygenation. He was safely extubated and brought to the post-anesthesia care unit, where recovery was uneventful. CONCLUSIONS Our case raises important questions and considerations for management of THC- dependent patients with potential for impacting NPO guidelines. It is unclear whether the quantity and chronicity of THC use effects gastroparesis development. The patient endorsed no signs of hyperemesis syndrome or gastric fullness; therefore, we cannot reliably deem patients risk-free based on their lack of symptomology alone. Until the true incidence of THC-induced gastroparesis is elicited in further studies, it is prudent to take aspiration precautions in all patients who endorse THC use.


[Overtreatment and undertreatment of depression].
The aim of this study was to examine what we know about the general practitioners' ability to diagnose and treat depression. Some studies suggest that clinicians might have considerable difficulty identifying depressions correctly, but that diagnostics become more accurate with increased severity of the disease. Several studies suggest that patients diagnosed with depression are undertreated, and some studies suggest that patients might receive antidepressants, even though they do not fulfil the International Classification of Diseases 10 criterion for depression.


The role of thermal tolerance in determining elevational distributions of four arthropod taxa in mountain ranges of southern Asia.
Understanding the role of thermal tolerances in determining species distributions is important for assessing species responses to climate change. Two hypotheses linking physiology with species distributions have been put forward-the climatic variability hypothesis and the climatic extreme hypothesis. The climatic variability hypothesis predicts the selection of individuals with broad thermal tolerance in more variable climatic conditions and the climatic extreme hypothesis predicts the selection of individuals with extreme thermal tolerance values under extreme climatic conditions. However, no study has tested the predictions of these hypotheses simultaneously for several taxonomic groups along elevational gradients. Here, we related experimentally measured critical thermal maxima, critical thermal minima and thermal tolerance breadths for 15,187 individuals belonging to 116 species of ants, beetles, grasshoppers, and spiders from mountain ranges in central and northern Pakistan to the limits and breadths of their geographic and temperature range. Across all species and taxonomic groups, we found strong relationships between thermal traits and elevational distributions both in terms of geography and temperature. The relationships were robust when repeating the analyses for ants, grasshoppers, and spiders but not for beetles. These results indicate a strong role of physiology in determining elevational distributions of arthropods in Southern Asia. Overall, we found strong support for the climatic variability hypothesis and the climatic extreme hypothesis. A close association between species' distributional limits and their thermal tolerances suggest that in case of a failure to adapt or acclimate to novel climatic conditions, species may be under pressure to track their preferred climatic conditions, potentially facing serious consequences under current and future climate change.


Renal cortical perfusion in rabbits: visualization with color amplitude imaging and an experimental microbubble-based US contrast agent.
To evaluate an experimental microbubble-based ultrasound (US) contrast agent (Imagent US, formulation AF0150) in the kidney to help identify focal perfusion abnormalities. Six incremental doses (0.007-0.2 mL/kg) of contrast material were injected intravenously in six New Zealand rabbits (3.1-3.5 kg), with experimental focal renal ischemia in five kidneys. Unenhanced and contrast material-enhanced color amplitude and gray-scale images of the kidney were analyzed for changes in mean pixel intensity. Kidneys were harvested and examined pathologically. On contrast-enhanced color amplitude images, mean pixel intensity increased significantly in cortex and medulla at every dose level (P < .0005) (with doses as low as 0.007 mL/kg), and differences were significant in cortical peak intensity and duration of enhancement with incremental increases in dose (P < .006). On contrast-enhanced gray-scale images, renal cortical enhancement reached statistical significance only at the highest dose (0.2 mL/kg). Duration of gray-scale tissue enhancement was not dose related. Color mean pixel intensity markedly increased in areas of normally perfused kidney (P = .002) but remained essentially unchanged in ischemic areas (P = .34). Color Doppler and gray-scale images enhanced with this microbubble-based US contrast agent depict renal cortical perfusion clearly in rabbits.


Synthesis, Structure, and Photophysical Properties of BN-Embedded Analogue of Coronene.
Two BN-embedded benzo[ghi]perylene (Bzp) and coronene derivatives (BN-Bzp and BN-Cor) have been successfully synthesized from binaphthyl precursors by new efficient one-pot-multibond routes, and their single crystal structures were analyzed. Both experimental spectra and DFT theoretical calculations indicated that the absorption and emission of these BN-embedded polycyclic aromatic hydrocarbons are significantly enhanced comparing with those of their all carbon analogues. Especially, the fluorescence quantum yield of BN-Cor is nearly 20 times higher than that of ordinary coronene.


Transforming growth factor-beta induced cell death in the developing chick retina is mediated via activation of c-jun N-terminal kinase and downregulation of the anti-apoptotic protein Bcl-X(L).
Cell death in general and especially in neuronal cells is regulated by a complex interplay between survival and death signals, generated by extracellular factors like neurotrophins and intracellular regulation mechanisms. The pleiotrophic transforming growth factor beta (TGF-beta) influences life and death decisions in cells depending on cell type and other growth factors present. It has been previously shown that TGF-beta is necessary to induce ontogenetic cell death during retinal development. In the present study, we analyzed the underlying intracellular signaling processes involved in TGF-beta mediated cell death. We established a cell culture system mimicking the situation of ontogenetic cell death in vivo with cultured retinal cells isolated from the retinae of embryonic day 7 white leghorn chick embryos. The neutralization of TGF-beta inhibits cell death of cultured retinal cells whereas exogenous application of TGF-beta is followed by enhanced apoptosis as observed by in situ cell death detection (terminal deoxynucleotidyl transferase-mediated nick end labeling) assay. TGF-beta induces the activation of c-jun N-terminal kinase in the mitogen-activated protein kinase (MAP kinase) pathway and provokes downregulation of the anti-apoptotic BCL-X(L) protein. Thus, TGF-beta influences cell death via activation of a pro-apoptotic MAP-kinase cascade accompanied by a downregulation of anti-apoptotic signals.


Intracellular signaling of transcription and secretion of type IV collagen after angiotensin II-induced cellular hypertrophy in cultured proximal tubular cells.
Physiologic concentrations of angiotensin II (AII) can induce cellular hypertrophy in murine proximal tubular epithelium (MCT cells). This response is characterized by an increase in cell size, new protein synthesis, and by the secretion of new basement membrane type IV collagen in the absence of cellular proliferation. The present study was undertaken to evaluate the second messengers of these AII-induced cellular events with special reference to the increase in type IV collagen secretion. In initial experiments we observed that pretreatment of MCT cells with agents that increase concentrations of intracellular cAMP, like forskolin, dibutyryl cAMP, and isobutyl-methyl-xanthine abolish AII-induced amino acid incorporation, but have no effect on control cells or on their proliferation. In addition, 10(-8) M AII significantly decreased the concentration of intracellular cAMP. Phorbolesters were without significant effect on the hypertrophy or proliferation of AII-stimulated MCT cells or their rested controls. The transfection of MCT cells with reporter genes containing regulatory elements for type IV collagen revealed that the stimulatory effects of AII on collagen type IV depend, at least to some extent, on an increase in gene transcription. Agents increasing intracellular cAMP concentrations inhibited the AII-induced increase in transcription and secretion of collagen type IV, but had no effect on MCT cells grown in media without AII. Our findings provide evidence that AII-induced changes in tubular epithelium leading to the secretion of type IV collagen are mediated by a decrease in intracellular cAMP.


Performance comparison of Zr-based and Bi-based erbium-doped fiber amplifiers.
In this Letter, we present a comprehensive comparison of the performance of a zirconia-based erbium-doped fiber amplifier (Zr-EDFA) and a bismuth-based erbium-doped fiber amplifier (Bi-EDFA). The experimental results reveal that a Zr-EDFA can achieve comparable performance to the conventional Bi-EDFA for C-band and L-band operations. With a combination of both Zr and Al, we could achieve a high erbium-doping concentration of about 2800 ppm (parts per million) in the glass host without any phase separations of rare earths. The Zr-based erbium-doped fiber (Zr-EDF) was fabricated using in a ternary glass host, zirconia-yttria-aluminum codoped silica fiber through a solution-doping technique along with modified chemical vapor deposition. At a high input signal of 0 dBm, a flat gain at average value of 13 dB is obtained with a gain variation of less than 2 dB within the wavelength region of 1530-1575 nm and using 2 m of Zr-EDF and 120 mW pump power. The noise figures are less than 9.2 at this wavelength region. It was found that a Zr-EDFA can achieve even better flat-gain value and bandwidth as well as lower noise figure than the conventional Bi-EDFA.


[The peri-operative cure of patients suffered allergic purpura before tonsillectomy].
To explore the peri-operative cure of patients suffered allergic purpura before tonsillectomy. To review the perioperative cure of 7 cases operated on tonsillectomy in retrospect. 7 cases didn't bleed during peri-operation and were discharged from hospital after recovery. The operation was arranged in allergic purpura catabasis and the acute inflammation controlled. The patients should avoid to contact all kinds of doubtful predisposition and were administered antihistamin, anti-inflammatory agent, drugs decreasing capillary permeability during peri-operation.


[Influence of individual characteristics and working conditions in the level of injury accident at work by registered in Andalusia, Spain, in 2003].
The study of the severity of occupational injuries is very important for the establishment of prevention plans. The aim of this paper is to analyze the distribution of occupational injuries by a) individual factors b) work place characteristics and c) working conditions and to analyze the severity of occupational injuries by this characteristics in men and women in Andalusia. Injury data came from the accident registry of the Ministry of Labor and social issues in 2003. Dependent variable: the severity of the injury: slight, serious, very serious and fatal; the independent variables: the characteristics of the worker, company data, and the accident itself. Bivariate and multivariate analysis were done to estimate the probability of serious, very serious and fatal injury, related to other variables, through odds ratio (OR), and using a 95% confidence interval (CI 95%). The 82.4% of the records were men and 17.6% were women, of whom the 78,1% are unskilled manual workers, compared to 44.9% of men. The men belonging to class I have a higher probability of more severe lesions (OR = 1.67, 95% CI = 1.17-2.38). The severity of the injury is associated with sex, age and type of injury. In men it is also related with the professional situation, the place where the accident happened, an unusual job, the size and the characteristics of the company and the social class, and in women with the sector.


Complications associated with use of a one-hole subpalpebral lavage system in horses: 150 cases (1977-1996).
To determine type and frequency of complications associated with use of a one-hole subpalpebral lavage (SPL) system in horses. Retrospective study. 150 horses with 156 SPL systems. Signalment, primary complaint, method used for placement, time SPL system was in place, and complications were retrieved from medical records. Complications were not associated with placement, maintenance, or removal of 66 of 156 (42%) SPL systems. A minor complication was reported in association with 53 (34%) SPL systems, and a serious complication was reported in association with 37 (24%). The 3 most common minor complications were mild swelling of the eyelid (31 horses), tearing of SPL system tubing (20), and loss of the injection cap (30). Serious complications included problems with the SPL system requiring its premature removal and possible replacement (26 horses), removal of the SPL system by the horse (6), infection of the eyelid (4), loss of the footplate in the eyelid (5), and cornea ulceration (1). Proper attention to the size of the footplate and placement of the SPL system in the eyelid should decrease the risk of serious complications. Minor eyelid swelling should be expected in the first 48 hours after SPL system placement.


The association between acute lower abdominal pain over a previous caesarean scar and uterine rupture.
Pregnant women with previous caesarean delivery might suffer from acute lower abdominal pain located at the site of previous caesarean scar (CS). The association between this complaint and uterine rupture (UR) is not fully understood. Therefore, we aimed to examine the risk of UR in women with acute persistent abdominal pain (APAP) over a previous CS and to investigate all the women with UR, with or without APAP and with or without previous CS, in order to determine risk factors, clinical presentation and management. We performed a retrospective analysis on two study groups: women who had APAP over previous CS and women who had UR. We found an incidence of UR in patients with APAP over the previous CS was 0.7%; which doubled the total UR rate among women with previous caesarean in our medical centre (0.35%). Forty percent of the women with APAP over a previous CS had preterm delivery. Twenty percent of the cases of UR occurred in preterm weeks. To conclude, APAP over a previous CS is associated with a doubled risk of UR. Considering this symptom as a preliminary sign of UR might lead to elevated rate of iatrogenic preterm deliveries.Impact statementWhat is already known on this subject? Women with UR may present with abdominal pain which may vary from non-specific mild discomfort to severe acute abdominal pain. Additionally, these women may suffer from acute persistent abdominal pain (APAP) located over the previous caesarean scar. The clinical significance of APAP in these women has not been fully investigated.What do the results of this study add? Lower abdominal pain located at the site of previous CS is associated with a doubled risk of UR. Considering this complaint as a major sign of UR might lead to an elevated rate of iatrogenic preterm deliveries.What are the implications of these findings for clinical practice and/or further research? Further studies are needed to explore whether women with a single complaint of APAP over CS could be managed expectantly and even offered a trial of labour after caesarean delivery (CD).


Molecular Ecological Basis of Grasshopper (Oedaleus asiaticus) Phenotypic Plasticity under Environmental Selection.
While ecological adaptation in insects can be reflected by plasticity of phenotype, determining the causes and molecular mechanisms for phenotypic plasticity (PP) remains a crucial and still difficult question in ecology, especially where control of insect pests is involved. Oedaleus asiaticus is one of the most dominant pests in the Inner Mongolia steppe and represents an excellent system to study phenotypic plasticity. To better understand ecological factors affecting grasshopper phenotypic plasticity and its molecular control, we conducted a full transcriptional screening of O. asiaticus grasshoppers reared in four different grassland patches in Inner Mongolia. Grasshoppers showed different degrees of PP associated with unique gene expressions and different habitat plant community compositions. Grasshopper performance variables were susceptible to habitat environment conditions and closely associated with plant architectures. Intriguingly, eco-transcriptome analysis revealed five potential candidate genes playing important roles in grasshopper performance, with gene expression closely relating to PP and plant community factors. By linking the grasshopper performances to gene profiles and ecological factors using canonical regression, we first demonstrated the eco-transcriptomic architecture (ETA) of grasshopper phenotypic traits (ETAGPTs). ETAGPTs revealed plant food type, plant density, coverage, and height were the main ecological factors influencing PP, while insect cuticle protein (ICP), negative elongation factor A (NELFA), and lactase-phlorizin hydrolase (LCT) were the key genes associated with PP. Our study gives a clear picture of gene-environment interaction in the formation and maintenance of PP and enriches our understanding of the transcriptional events underlying molecular control of rapid phenotypic plasticity associated with environmental variability. The findings of this study may also provide new targets for pest control and highlight the significance of ecological management practice on grassland conservation.


On Reliable and Efficient Data Gathering Based Routing in Underwater Wireless Sensor Networks.
This paper presents cooperative routing scheme to improve data reliability. The proposed protocol achieves its objective, however, at the cost of surplus energy consumption. Thus sink mobility is introduced to minimize the energy consumption cost of nodes as it directly collects data from the network nodes at minimized communication distance. We also present delay and energy optimized versions of our proposed RE-AEDG to further enhance its performance. Simulation results prove the effectiveness of our proposed RE-AEDG in terms of the selected performance matrics.


An analysis of naltrexone and naloxone's possible agonistic actions in the dog.
Naltrexone (0.01-2.0 mg/kg, i.v.) produced dose-dependent EEG slowing in the conscious dog as did morphine (0.5-8 mg/kg, i.v.) and fentanyl (5-20 micrograms/kg, i.v.). However, the dose-response curve for naltrexone was not parallel to the morphine or fentanyl dose-response curves. Morphine and fentanyl but not naltrexone also produced dose-dependent miosis and increased the skin twitch reflex latency. When administered into the fourth cerebral ventricle naltrexone (60 micrograms), morphine (80 micrograms) and ethylketazocine (30 micrograms) produced EEG slowing. Again, naltrexone did not alter the skin twitch latency whereas morphine lengthened it and ethylketazocine reduced it. The pharmacological profiles obtained from different routes of administration indicate that naltrexone is clearly different from morphine, fentanyl and ethylketazocine. However, naltrexone may act as a partial agonist in the production of EEG slowing at a previously unidentified opioid receptor.


Evidence for the Glutamine Synthetase/Glutamate Synthase Pathway during the Photorespiratory Nitrogen Cycle in Spinach Leaves.
Spinach leaf (Spinacia oleracea L.) discs infiltrated with [(15)N]glycine were incubated at 25 degrees C in the light and in darkness for 0, 30, 60 and 90 minutes. The kinetics of (15)N-incorporation into glutamine, glutamate, asparagine, aspartate, and serine from [(15)N]glycine was determined. At the beginning of the experiment, just after infiltration (0 min incubation) serine, and the amido-N of glutamine and asparagine were the only compounds significantly labeled in both light- and dark-treated leaf discs. Incorporation of (15)N-label into the other amino acids was observed at longer incubation time. The per cent (15)N-enrichment in all amino acids was found to increase with incubation. However, serine and the amido-N of glutamine remained the most highly labeled products in all treatments. The above pattern of (15)N-labeling suggests that glutamine synthetase was involved in the initial refixation of (15)NH(3) derived from [(15)N]glycine oxidation in spinach leaf discs.The (15)N-enrichment of the amino-N of glutamine was found to increase rapidly from 0 to 19% during incubation in the light. There was a comparatively smaller increase (4-9%) in the (15)N-label of the amino-N of glutamine in tissue incubated in darkness. Furthermore the total flux of (15)N-label into each of the amino acids examined was found to be greater in tissue incubated in the light than those in the dark. The above evidence indicates the involvement of the glutamine synthetase/glutamate synthase pathway in the recycling of photorespiratory NH(3) during glycine oxidation in spinach leaves.


A study of 10,296 pediatric and adolescent Papanicolaou smear diagnoses in northern New England.
This study analyzes pediatric and adolescent Papanicolaou (Pap) smear diagnoses to determine the prevalence rates of squamous intraepithelial lesion (SIL) as well as infectious and reactive processes in this age group. A total of 10 296 Pap smear diagnoses from patients 10 to 19 years of age collected over a 1-year period and classified according to the Bethesda system were reviewed. This population was almost exclusively white, the majority residing in rural or suburban areas of Maine, New Hampshire, and Vermont. The percentage of abnormal smear results was then compared with data generated for older age subsets. The following diagnoses were made on 10 296 Pap smears from patients 10 to 19 years of age: 7208 (70.01%) normal; 1689 (16.4%) benign cellular change; 1004 (9.75%) atypical squamous cells of undetermined significance; 388 (3.77%) squamous intraepithelial lesion (SIL); and 7 (0.06%) atypical glandular cells of undetermined significance. A total of 1503 (14.6%) of smears showed infectious processes. Compared with the results of adult Pap smears collected over the same time period, the age 20 to 29 subset with 27 067 Pap smears and the age 30+ subset with 42 617 Pap smears showed 11.79% and 8.43% infectious processes and 3.49% and 1.27% SIL, respectively. Therefore, the highest rate of infectious processes and SIL was found in the subset of patients age 10 to 19 years. Because the development of SIL and hence cervical cancer is causally related to sexually transmitted human papilloma virus (HPV) infection, this high rate of abnormal Pap smear results of both an infectious and precancerous nature in this population may reflect a high level of sexual activity among adolescent girls. These data reinforce the importance of implementing early cervical Pap smear screening in the sexually active pediatric and adolescent population.


[Comparative analysis of measles genome between vaccine strain and wild-type strain in Zhejiang province of China].
To explore the distinction between wild-type strain MVi/Zhejiang, CHN/7.05/4 and vaccine strain Shanghai-191 at genome level. After sequencing of measles wild-stain MVi/Zhejiang. CHN/7.05/4, the distinction between the wild-type strain and the vaccine strain was analysed by MEGA 3.1 software at genome level, and the antigen variation was studied by means of combining the epidemiological data. There were 822 nucleotide differences (5.17%) and 161 amino acid differences between these two strains, including three glycosylation sites variation found. Meanwhile, the antigen ratio between wild-type strain and vaccine strain was found to be 5.66. There should be certain differences between the contemporary wild-type strain MVi/Zhejiang, CHN/7.05/4 and vaccine strain Shanghai-191 at genome level, and the protective effects of measles vaccine should be studied further.


Role of protein kinase C in 1,25(OH)(2)-vitamin D(3) modulation of intracellular calcium during development of skeletal muscle cells in culture.
Regulation of muscle cell Ca(2+) metabolism by 1, 25-dihydroxy-vitamin D(3) [1,25(OH)(2)D(3)] is mediated by the classic nuclear mechanism and a fast, nongenomic mode of action that activates signal transduction pathways. The role of individual protein kinase C (PKC) isoforms in the regulation of intracellular Ca(2+) levels ([Ca(2+)](i)) by the hormone was investigated in cultured proliferating (myoblasts) and differentiated (myotubes) chick skeletal muscle cells. 1,25(OH)(2)D(3) (10(-9) M) induced a rapid (30- to 60-s) and sustained (>5-min) increase in [Ca(2+)](i) which was markedly higher in myotubes than in myoblasts. The effect was suppressed by the PKC inhibitor calphostin C. In differentiated cells, PKC activity increased in the particulate fraction and decreased in cytosol to a greater extent than in proliferating cells after 5-min treatment with 1,25(OH)(2)D(3). By Western blot analysis, these changes were correlated to translocation of the PKC alpha isoform from cytosol to the particulate fraction, which was more pronounced in myotubes than in myoblasts. Specific inhibition of PKC alpha activity using antibodies against this isoform decreased the 1, 25(OH)(2)D(3)-induced [Ca(2+)](i) sustained response associated with Ca(2+) influx through voltage-dependent calcium channels. Neomycin, a phospholipase C (PLC) inhibitor, blocked its effects on [Ca(2+)](i), PKC activity, and translocation of PKC alpha. Exposure of myotubes to 1,2-dioleyl-rac-glycerol (1,2-diolein), also increased [Ca(2+)](i), PKC activity, and the amount of PKC alpha associated with the particulate fraction. Changes in [Ca(2+)](i) induced by diolein were inhibited by calphostin C and nifedipine. The results indicate that PKC alpha activation via PLC-catalyzed phosphoinositide hydrolysis is part of the mechanism by which 1, 25(OH)(2)D(3) regulates muscle intracellular Ca(2+) through modulation of the Ca(2+) influx pathway of the Ca(2+) response to the sterol.


Expert Consensus on COVID-19 Vaccination in Korean Adolescents: A Modified Delphi Survey.
The effectiveness of coronavirus disease 2019 (COVID-19) vaccines had been shown in many studies in adult population, however, the real-world evidence in the childhood population was scarce. We aimed to organize the collective expert's opinions on adolescent vaccination against COVID-19 in Korea, therefore to guide the vaccination policy in the setting of available evidence. The Delphi panels responded that adolescents were greatly impacted by the quarantine measures, and COVID-19 is an important health problem for adolescents. Panels responded that in general, the benefits of the COVID-19 vaccine overweigh the potential risks in Korean adolescents. Continuing monitoring of available data is needed to provide the best vaccination practices in adolescents guided by the updated evidence.


Polycystic echinococcosis in the state of Acre, Brazil: contribution to patient diagnosis, treatment and prognosis.
The lack of knowledge regarding polycystic hydatid disease results in delayed or even incorrect diagnosis. The lack of systematic information regarding treatment also makes it difficult to assess the results and prognosis in patients with peritoneal and hepatic lesions caused by Echinococcus vogeli. Here we describe the clinical features of patients, propose a radiological classification protocol and describe a therapeutic option for the treatment of hydatid disease that previously had only been used for cases of cystic echinococcosis (Echinococcus granulosus). A prospective cohort study was initiated in 1999 and by 2009 the study included 60 patients. These patients were classified according to the PNM classification (parasite lesion, neighbouring organ invasion and metastases) and placed in one of three therapeutic modalities: (i) chemotherapy with albendazole at a dose of 10 mg/kg/day, (ii) surgical removal of cysts or (iii) percutaneous puncture of the cysts via puncture, aspiration, injection and re-aspiration (PAIR). The results were stratified according to therapeutic outcome: "cure", "clinical improvement", "no improvement", "death" or "no information". The PNM classification was useful in indicating the appropriate therapy in cases of polycystic hydatid disease. In conclusion, surgical therapy produced the best clinical results of all the therapies studied based on "cure" and "clinical improvement" outcomes. The use of PAIR for treatment requires additional study.


Two new alkaloids from Melodinus hemsleyanus Diels.
Two new monoterpenoid indole alkaloids, named 14,15-dihydro-14β,15β-epoxy-10-hydroxyscandine (1) and 15α-hydroxy-meloscandonine (2), together with 12 known compounds, were isolated from the aerial parts of Melodinus hemsleyanus Diels. The structures of 1 and 2 were elucidated on the bases of 1D and 2D NMR spectra and MS. Two new compounds were evaluated for their PTP1B and Drak2 inhibitory effects, and inactivity.


Antimicrobial Peptide Structures: From Model Membranes to Live Cells.
The rise in antibiotic resistance has led to a renewed interest in antimicrobial peptides (AMPs) that target membranes. The mode of action of AMPs involves the disruption of the lipid bilayer and leads to growth inhibition and death of the bacteria. However, details at the molecular level of how these peptides kill bacteria and the reasons for the observed differences in selectivity remain unclear. Structural information is crucial for defining the molecular mechanism by which these peptides recognize, self-assemble and interact with a particular lipid membrane. Solid-state NMR is a non-invasive technique that allows the study of the structural details of lipid-peptide and peptide-peptide interactions. Following on from studies of antibiotic and lytic peptides, gramicidin A and melittin, respectively, we investigated maculatin 1.1, an AMP from the skin of Australian tree frogs that acts against Gram-positive bacteria. By using perdeuterated phospholipids and specifically labelled peptides, 2 H, 31 P and {31 P}15 N REDOR solid-state NMR experiments have been used to localize, maculatin 1.1 in neutral and anionic model membranes. However, the structure, location and activity depend on the composition of the model membrane and current advances in solid-state NMR spectroscopy now allow structure determination of AMPs in live bacteria.


Effect of temperature on the plasma membrane and tonoplast ATPases of barley roots : comparison of results obtained with acridine orange and quinacrine.
The effect of temperature on the rate of proton transport and ATP hydrolysis by plasma membrane (PM) and tonoplast (TN) ATPases from barley (Hordeum vulgare L. cv CM 72) roots were compared. Rates of proton transport were estimated using the fluorescent amine dyes quinacrine and acridine orange. The ratio between rate of transport and ATP hydrolysis was found to depend on the dye, the temperature, and the type of membrane. The PM ATPase had an estimated Arrhenius energy of activation (Ea) of approximately 18 kilocalories per mole for ATP hydrolysis, and the Ea for proton transport was best estimated with acridine orange, which gave an Ea of 19 kilocalories per mole. The TN ATPase had an Ea for ATP hydrolysis of approximately 10 kilocalories per mole and the Ea for proton transport was best estimated with quinacrine, which gave an Ea of 10 kilocalories per mole. Acridine orange did not give an accurate estimate of Ea for the TN ATPase, nor did quinacrine for the PM ATPase. Reasons for the differences are discussed. Because it was suggested (AJ Pope, RA Leigh [1988] Plant Physiol 86: 1315-1322) that acridine orange interacts with anions to dissipate the pH gradient in TN vesicles, the complex effects of NO(3) (-) on the TN ATPase were also examined using acridine orange and quinacrine and membranes from oats and barley. Fluorescent amine dyes can be used to evaluate the effects of ions, substrates, inhibitors, and temperature on transport but caution is required in using rates of quench to make quantitative estimates of proton fluxes.


[The correction of the vaginal microflora in patients with a papillomavirus infection associated with cervical intraepithelial neoplasia by using the new bacterial preparation Zhlemik].
The microbiological study of vaginal microflora in 40 patients of the reproductive age (20-30 years) with papilloma virus infection in association with cervical intraepithelial neoplasia of the uterine neck revealed dysbiotic disturbances in vaginal microflora, manifested by a decrease in the isolation rate and amount of lacto- and bifidobacteria and by excessive growth of opportunistic microorganisms. The 10-day course of corrective therapy with the new bacterial preparation "Zhlemik" was carried out. Group 1 (81 women) received the preparation intravaginally on a tampon, group 2 (19 patients) received the preparation in the form of vaginal suppositories. The results of this treatment were indicative of a high sanative effect of the preparation irrespective of the method of its application. This was demonstrated by the results of the bacteriological study made after bacterial correction: the amount of the Lactobacillus was restored, and they could be isolated from all patients; the level of contamination of the cervicovaginal niche with opportunistic microbial strains considerably decreased. The positive clinical effect after bacterial therapy with "Zhlemik" was observed in 93-95% of cases, depending on the form of its application.


Community-building: a strategy to address inequality of health in Australia.
Given the improvements in the health of Australians in recent decades, not all sectors of populations have benefited equally. Inequality and poverty are major problems. Traditional hierarchical, top-down governance seems ineffective to address these complex problems. An innovative and more participatory community-based approach is called for to address these issues. Community-building approaches, characterized by wide-ranging community involvement, emphasizing strength and collaboration, can be a good vehicle and intervention strategy to address the poverty problem by empowering both the individual and the community. The relevance of the community-building approach to the poverty issue and Australian context is discussed.


Downregulation of lncRNA NEAT1_2 radiosensitizes hepatocellular carcinoma cells through regulation of miR-101-3p/WEE1 axis.
Radioresistance is a major obstacle in hepatocellular carcinoma (HCC) radiotherapy. Aberrant expression of long non-coding RNA (lncRNA) has been postulated to be implicated in the development of HCC radioresistance. We investigated the role of lncRNA nuclear enriched abundant transcript 1_2 (NEAT1_2) in radioresistance of HCC and its molecular mechanism in this study. We found that NEAT1_2 and WEE1 were upregulated, and miR-101-3p was downregulated in HCC tissues, as well as HCC cell lines. Downregulation of WEE1 sensitized the radiosensitivity of HCC cells, as evidenced by decreased survival fractions of Huh7 and PLC5 cells and increased percentage of apoptotic cells. Also, knockdown of NEAT1_2 exerted a reinforcing effect on the radiosensitivity of HCC cells. In addition, WEE1 was confirmed as a direct target of miR-101-3p. Upregulation of miR-101-3p obviously decreased the mRNA and protein levels of WEE1 compared with that in the miR-NC group, while transfection of anta-miR-101-3p presented the opposite effects. In parallel, NEAT1_2 was identified to interact with miR-101-3p, and NEAT1_2 upregulated the expression of WEE1 in Huh7 cells through sponging miR-101-3p. Besides, the reinforcing effect of NEAT1_2 silencing could be attenuated by downregulation of miR-101-3p. To conclude, our results support the concept that downregulation of lncRNA NEAT1_2 radiosensitizes hepatocellular carcinoma cells through regulation of miR-101-3p/WEE1 axis.


The role of the platelet pool of Plasminogen Activator Inhibitor-1 in well-controlled type 2 diabetes patients.
The main inhibitor of the fibrinolytic system, Plasminogen Activator Inhibitor -1 (PAI-1), irreversibly binds tissue-type Plasminogen Activator (t-PA) and thereby inhibits the protective action of tPA against thrombus formation. Elevated levels of plasma PAI-1 are associated with an increased risk of cardiovascular events and are observed in subjects with type 2 diabetes (T2D) and obesity. Platelets contain the majority of PAI-1 present in blood and exhibit the ability to synthesis active PAI-1. Diabetic platelets are known to be hyper-reactive and larger in size; however, whether these features affect their contribution to the elevated levels of plasma PAI-1 in T2D is not established. To characterize the PAI-1 antigen content and the mRNA expression in platelets from T2D subjects compared to obese and lean control subjects, in order to elucidate the role of platelet PAI-1 in T2D. Nine subjects with T2D and obesity were recruited from Primary Care Centers together with 15 healthy control subjects (8 lean subjects and 7 with obesity). PAI-1 antigen levels in plasma, serum and platelets were determined by ELISA, and PAI-1 mRNA expression was analyzed by qPCR. There was no significant difference in PAI-1 mRNA expression or PAI-1 antigen in platelets in T2D subject in comparison to obese and lean control subjects. An elevated level of plasma PAI-1 was seen in both T2D and obese subjects. PAI-1 gene expression was significantly higher in both obese groups compared to lean. Similar levels of protein and mRNA expression of PAI-1 in platelets from T2D, obese and lean subjects indicate a limited role of platelets for the elevated plasma PAI-1 levels. However, an increased synthesis rate of mRNA transcripts in platelets from T2D and an increased release of PAI-1 could also result in similar mRNA and protein levels. Hence, synthesis and release rates of PAI-1 from platelets in T2D and obesity need to be investigated to further elucidate the role of platelets in obesity and T2D.


Midodrine in Liver Cirrhosis With Ascites: A Systematic Review and Meta-Analysis.
Ascites is the most common complication of liver cirrhosis. Midodrine is a vasoconstrictor that improves splanchnic and systemic hemodynamics, reduces ascites, and improves clinical outcomes. Here, we aimed to examine the role of midodrine in cirrhosis-related ascites. Scopus, Embase, PubMed, and PubMed Central databases were searched for relevant randomized controlled trials comparing midodrine with other interventions in patients with cirrhotic ascites on November 25, 2020, using appropriate keywords like "midodrine", "ascitic cirrhosis", "peritoneal paracentesis" and suitable Boolean operators. Odds ratio (OR) and mean difference (MD) were used to analyze pool data as appropriate with a 95% confident interval (CI). A total of 14 studies were included in our analysis including 1199 patients. The addition of midodrine resulted in statistically significant improvement in mean arterial pressure (MAP) (MD, 3.95 mmHg; 95% CI, 1.53-6.36) and MELD (Model for End-Stage Liver Disease) score (MD, -1.27; 95% CI, -2.49 to -0.04) compared to standard medical treatment (SMT). There was also a significant improvement in plasma renin activity and plasma aldosterone concentration. However, there was no significant improvement in mortality or serum creatinine compared to SMT. In addition, there was no statistically significant improvement in MAP, plasma renin activity, plasma aldosterone concentration, MELD score, overall mortality, and paracentesis-induced circulatory dysfunction comparing midodrine with albumin. Midodrine alone leads to significant improvement in various clinical parameters in patients with cirrhotic ascites compared to standard medical care. At the same time, it was found to be non-inferior to albumin. Therefore, further well-designed studies need to be carried out on midodrine in addition to albumin for optimal clinical benefits among patients with ascites due to cirrhosis.


Affinity of calcium channel inhibitors, benzodiazepines, and other vasoactive compounds for the nucleoside transport system.
There is evidence to suggest that several different groups of drugs including the so-called coronary vasodilators, benzodiazepines, and calcium channel inhibitors may owe their vasoactivity, in part, to the potentiation of the vasorelaxant effects of endogenous adenosine. To measure the affinity of some of these agents for the membrane-located nucleoside transport system, competition binding assays have been performed using the high-affinity radioligand [3H]nitrobenzylthioinosine (NBMPR). Experiments were performed on human erythrocytes and cardiac membranes from guinea pigs and rats. Recognized nucleoside transport inhibitors had high affinity (less than 50 nM) for NBMPR recognition sites associated with the nucleoside transporter complex in human erythrocytes, whereas calcium channel inhibitors and benzodiazepines had predominantly low affinity (greater than 1 microM). Although some recognized transport inhibitors, such as dipyridamole, show marked differences in affinity for NBMPR sites in guinea pig and rat tissues, benzodiazepines and calcium channel blockers displayed no such species selectivity and had low affinity (greater than 1 microM) for NBMPR sites in both guinea pig and rat cardiac membranes. Consequently, it is unlikely that agents such as benzodiazepines and calcium channel inhibitors cause significant inhibition of adenosine transport, and hence potentiate adenosine actions, at the concentrations required to induce effects through occupation of their respective, specific high-affinity sites.


Inactivation of E. coli and L. innocua in milk by a thin film UV-C reactor modified with flow guiding elements (FGE).
In this study the suitability of a thin-film reactor (TFR) equipped with special flow guiding elements (FGE) was examined to analyse its capability to inactivate microorganisms in milk. Experiments were carried out with UHT-milk inoculated with Escherichia coli (E. coli), DH5α and Listeria innocua (L. innocua) WS 2258. Furthermore, the inactivation of microorganisms originally occurring in raw milk was investigated. E. coli, DH5α and L. innocua serving as biodosimeter were reduced by 4.58-log and 3.19-log, respectively. In milk, the original microorganisms showed a 4-log reduction. Without FGE the reduction was below 0.13-log. Thus, it can be derived that the efficacy of a UV-C thin-film reactor processing absorptive media like milk can be highly improved using FGE.


In vitro selection of highly darunavir-resistant and replication-competent HIV-1 variants by using a mixture of clinical HIV-1 isolates resistant to multiple conventional protease inhibitors.
We attempted to select HIV-1 variants resistant to darunavir (DRV), which potently inhibits the enzymatic activity and dimerization of protease and has a high genetic barrier to HIV-1 development of resistance to DRV. We conducted selection using a mixture of 8 highly multi-protease inhibitor (PI)-resistant, DRV-susceptible clinical HIV-1 variants (HIV-1(MIX)) containing 9 to 14 PI resistance-associated amino acid substitutions in protease. HIV-1(MIX) became highly resistant to DRV, with a 50% effective concentration (EC(50)) ∼333-fold greater than that against HIV-1(NL4-3). HIV-1(MIX) at passage 51 (HIV-1(MIX(P51))) replicated well in the presence of 5 μM DRV and contained 14 mutations. HIV-1(MIX(P51)) was highly resistant to amprenavir, indinavir, nelfinavir, ritonavir, lopinavir, and atazanavir and moderately resistant to saquinavir and tipranavir. HIV-1(MIX(P51)) had a resemblance with HIV-1(C) of the HIV-1(MIX) population, and selection using HIV-1(C) was also performed; however, its DRV resistance acquisition was substantially delayed. The H219Q and I223V substitutions in Gag, lacking in HIV-1(C(P51)), likely contributed to conferring a replication advantage on HIV-1(MIX(P51)) by reducing intravirion cyclophilin A content. HIV-1(MIX(P51)) apparently acquired the substitutions from another HIV-1 strain(s) of HIV-1(MIX) through possible homologous recombination. The present data suggest that the use of multiple drug-resistant HIV-1 isolates is of utility in selecting drug-resistant variants and that DRV would not easily permit HIV-1 to develop significant resistance; however, HIV-1 can develop high levels of DRV resistance when a variety of PI-resistant HIV-1 strains are generated, as seen in patients experiencing sequential PI failure, and ensuing homologous recombination takes place. HIV-1(MIX(P51)) should be useful in elucidating the mechanisms of HIV-1 resistance to DRV and related agents.


Higher Hospital Spending on Occupational Therapy Is Associated With Lower Readmission Rates.
Hospital executives are under continual pressure to control spending and improve quality. While prior studies have focused on the relationship between overall hospital spending and quality, the relationship between spending on specific services and quality has received minimal attention. The literature thus provides executives limited guidance regarding how they should allocate scarce resources. Using Medicare claims and cost report data, we examined the association between hospital spending for specific services and 30-day readmission rates for heart failure, pneumonia, and acute myocardial infarction. We found that occupational therapy is the only spending category where additional spending has a statistically significant association with lower readmission rates for all three medical conditions. One possible explanation is that occupational therapy places a unique and immediate focus on patients' functional and social needs, which can be important drivers of readmission if left unaddressed.


[Current status of Pasteurella multocida infection in Japan].
Recently, the case reports of Pasteurella multocida infection has been increasing in Japan. In 1989, the Japanese Government, Veterinary Sanitation Division, Ministry of Health and Welfare officially communicated this infection as a zoonosis to related institutions. The current status of Pasteurella multocida infection is not well known in Japan. Because of this, a nation wide questionnaire survey on Pasteurella multocida was conducted to clarify the status. A questionnaire was sent to 380 laboratories of the hospitals, and 258 (67.9%) replied. An infectious disease caused by Pasteurella multocida was found in Japan in 369 cases in 115 (44.6%) of 258 hospitals, or an average of 3.2 cases per hospital. The 369 cases were broken down into 123 males (from 1 month old to 87 years old), 118 females (from 7 months old to 88 years old), and 128 patients whose sex was unknown. The incidence of the infections tends to increase year by year. This incidence is higher than our expectation. It is considered that the contact with pets will in increase the infection with this agents. The organism was isolated in as many as nineteen different body specimens, including the appendix and urine, which in Japan has not been reported as organs harboring this organism. Some of the nineteen cases were severely infected. This organism was isolated most often from the sputum (48.5%). Pus was the next most common site (27.1%). This order was reversed in the U.K. and the U.S.. Possible explanations for the reversal are given below.(ABSTRACT TRUNCATED AT 250 WORDS)


Preferences for Patients with Type 2 Diabetes Mellitus for Medications in Shandong Province, China: A Discrete Choice Experiment.
To evaluate preferences for medications among patients with type 2 diabetes mellitus (T2DM) from urban community health stations or rural village clinics in Shandong province, China. We use a discrete choice experiment (DCE) to measure the medication preferences. Each patient completed six DCE choice sets. The attributes for the DCE questionnaire include mode of administration, out-of-pocket medication cost per month, fasting blood glucose control effect and frequency of hypoglycemia events. The conditional logit model (Clogit) and mixed logit model (MXL) were used to evaluate choice data. A total of 887 patients with T2DM completed the survey. The mean age of participants was 64 years, 36.42% experienced complications, and the mean duration of diabetes was about 8 years. Overall, patients' ideal medication would not have hypoglycemia events, provide normal fasting glucose levels, have oral medication three times a day and lower monthly medication cost. Patients prioritized the frequency of hypoglycemia events (β=15.055, P < 0.01) and were willing to spend CNY 393.10 per month to avoid hypoglycemia events. For patients with higher educational levels and with longer diagnosis time, the effect of fasting blood glucose was more relevant than all other outcomes. This study provides information on T2DM patients' preference for medications. Our results suggest that clinical doctors should present patients with a variety of pharmaceutical characteristics and include their preference into medication decision, which will improve patient adherence and health outcomes.


Sigma receptor scintigraphy with N-[2-(1'-piperidinyl)ethyl]-3-(123)I-iodo-4-methoxybenzamide of patients with suspected primary breast cancer: first clinical results.
The aim of this study was to investigate the potential of a new iodobenzamide, N-[2-(1'-piperidinyl)ethyl]-3-(123)I-iodo-4-methoxybenzamide (P-(123)I-MBA), to visualize primary breast tumor in humans in vivo. Tumor accumulation of benzamides is based on a preferential binding to sigma receptors that are overexpressed on breast cancer cells. P-(123)I-MBA (148-185 MBq) was administered to 12 patients with a mammographically suspicious breast mass. Two hours after administration, whole-body and spot images of the healthy and the diseased breast were obtained. A focal increased tracer accumulation was observed in 8 of 10 patients with histologically confirmed breast cancer (mean tumor-to-background ratio, 2.04). No uptake was seen in a case of lymphatic adenitis. This preliminary patient study shows that P-(123)I-MBA accumulates in most breast tumors in vivo. Future work should focus on the relationship between P-(123)I-MBA uptake and the proliferative activity of cells to anticipate use of this technique as a tool to noninvasively assess the degree of tumor proliferation.


Risk factors for mortality after cardiovascular surgery in patients with functional single ventricle and right isomerism.
Cardiac malformations are a major component of heterotaxy syndrome that results in significant mortality and morbidity, especially in patients with a single ventricle and right isomerism (RI). The goal of this study was to evaluate the mortality after cardiovascular surgery in patients with a functional single ventricle and RI over a long follow-up period (∼40 years) and to determine the predicted risk factors for mortality. We performed a retrospective review of the medical records of 129 consecutive patients with functional single ventricle and RI who underwent pulmonary flow control operations at Mt. Fuji Shizuoka Children's Hospital between 1979 and 2020. To evaluate mortality rates, the patients were divided into 2 groups (era 1: 1979-1999 and era 2: 2000-2020) based on the date of the first-stage palliation. The estimated survival rate at 10 years was 36.4% in era 1 and 57.8% in era 2. The estimated survival rate improved significantly (P = 0.0268) between the 2 eras. The rate of Fontan procedure completion was also significantly better in the current era (P = 0.0392; 22/59 in era 1 and 38/66 in era 2). In the multivariable analysis, the date of the first-stage palliation was the only predictor of mortality. The mortality rate after cardiovascular surgery in patients with a functional single ventricle and RI has improved over the past 20 years; however, it still remains high. This improvement may be attributed to our current surgical strategy and clinical management; however, further investigations are needed to prove this observation.


Mitochondrial adenylate kinase (AK2) from bovine heart. Homology with the cytosolic isoenzyme in the catalytic region.
The adenylate kinase isoenzyme located in the intermembrane space of mitochondria, AK2, is a monomeric protein of Mr 30000 which catalyzes the reaction ATP + AMP + AMP in equilibrium 2 ADP. The protein was reduced and S-carboxymethylated with iodo[14C2]acetate. Using a Laursen sequenator, the N-terminal sequence of S-carboxymethylated AK2 was determined as Ala-Pro-Asn-; in some batches of the isolated protein the N-terminal dipeptide portion was missing. The C-terminus of AK2 was found to be Met. Cleavage with CNBr yielded eight fragments which could be isolated in one step using high-performance size-exclusion chromatography. They ranged in size over 4-88 amino acid residues, the total being approximately 270 residues. All CNBr fragments were overlapped with Met-containing tryptic peptides of AK2. The N-terminal 111 residues of AK2 were sequenced. Except for an N-terminal extension of nine residues, this segment of AK2 could be aligned with the sequence 1-104 of cytosolic AK1. Allowing for two deletions in AK2, 43 of the 102 aligned residues are identical. Since this section contains the catalytic residues such as His-36 and Asp-93, we conclude that AK1 can serve as a three-dimensional model of AK2 in mechanistic and drug-designing studies. preliminary sequence results on AK2 beyond position 104 show that AK2 here contains a wing of approximately 50 residues which has no counterpart in AK1. The chain folds of the adenylate kinase isoenzymes are similar again from a position corresponding to residue 115 of AK1 onwards. The additional structural motifs of AK2 are probably related to the location of this isoenzyme in the mitochondrion.


The Short Health Scale: A Valid and Reliable Quality-of-Life Scale for Mainland Chinese Patients with Inflammatory Bowel Disease.
The aim of our study was to translate and validate the mainland Chinese version of the short health scale (SHS), a disease-specific quality-of-life (QoL) scale for patients with inflammatory bowel disease (IBD). The SHS was translated and validated according to the standard process: a translation and back-translation procedure and a reliability and validation study. Patients with IBD were enrolled, and their QoL was assessed using the SHS, the short inflammatory bowel disease questionnaire (SIBDQ), and the Bristol stool form scale. Reliability (internal consistency reliability, split-half reliability, and test-retest reliability) and validity analyses were performed to evaluate the psychometric characteristics of the SHS. The impacts of different severity of major symptoms on QoL were analyzed by comparing the scores of SHS. A total of 112 patients with IBD (69 with ulcerative colitis and 43 with Crohn's disease) completed the mainland Chinese version of the SHS, and 34 patients completed the SHS a second time within one to two weeks. Cronbach's alpha value of the SHS was 0.90, and its split-half coefficient was 0.83. Intraclass correlation coefficients of the four items ranged from 0.52 to 0.72. All four items of the SHS were significantly associated with the corresponding domains of the SIBDQ, with correlation coefficients ranging from -0.52 to -0.69 (p < 0.001). The results of confirmatory factor analysis indicated a good fit of the one-factor model, with comparative fit index (CFI) = 0.878, normed fit index (NFI) = 0.874, incremental fit index (IFI) = 0.880, and goodness of fit index (GFI) = 0.842. The patients with severe symptoms had higher scores in the SHS than those with no or mild symptoms. The SHS was simple and quick to be used. The SHS had good validity and reliability and was suitable for evaluating the QoL of patients with IBD in mainland China.


Prevention of nerve conduction deficit in diabetic rats by polyunsaturated fatty acids.
The influence of diets containing gamma-linolenic acid (GLA; 18:3n-6) on sciatic nerve conduction velocity (NCV) was determined in diabetic rats. NCV was lower in diabetic rats fed diets supplemented with olive oil or sunflower seed oil than in nondiabetic rats; rats supplemented with GLA during a 5-wk diabetic period, however, did not exhibit significantly lower NCV. The mean proportion of the phospholipid fatty acid linoleic acid (18:2n-6) was higher in the sciatic nerves of diabetic rats than in the nondiabetic groups irrespective of dietary lipid treatment. Additionally, the proportion of linoleic acid was higher in the diabetic rats fed sunflower oil than in all other groups. Dietary GLA supplementation did not significantly influence the fatty acid composition of nerve membrane phospholipids and there was no obvious correlation between the fatty acid composition of nerve membrane phospholipids and NCV. The content of fructose and glucose in sciatic nerves was higher, whereas that of myo-inositol was lower, in diabetic rats than in nondiabetic rats; however, this was not significantly influenced by dietary GLA. GLA administration did not significantly influence Na(+)-K(+)-exchanging ATPase or ouabain binding activity in sciatic nerve preparations, both of which remained nonsignificantly different in the diabetic and nondiabetic groups. The results suggest that dietary GLA can prevent the deficit in NCV induced by diabetes and that this effect is independent of the nerve phospholipid fatty acid profile, sugar and polyol content, Na(+)-K(+)-exchanging ATPase activity, and ouabain binding. GLA may prevent the deficit in NCV indirectly, possibly by its role as a precursor of vasodilatory prostaglandins. These results confirm that GLA is the active component of evening primrose oil.


Mental health training in primary care. Impact on physicians knowledge.
The objective of this study was to measure changes in the Primary Health Care (PHC) physicians` knowledge towards mental illnesses after a short-term training course. The interventive study consisted of a sample of 31 PHC physicians randomly selected from different PHC centers in Eastern Saudi Arabia. The physicians were exposed to a 4-day intensive training course in mental illnesses in June 1999. They were assessed using multiple choice questions testing before (pre-test) and after (post-test) to assess their knowledge regarding mental illnesses. The analysis of the accumulated data indicated that there was a significant improvement in the PHC physicians` knowledge of mental illnesses after the course (p<0.0001). The effect of recall bias was excluded in the post-test evaluation. In addition, multiple regression analysis indicated that the undergraduate psychiatric training courses had a positive contribution in both pre- and post-tests. The authors surmised that a mental training course could improve the knowledge base of PHC physicians. Furthermore, by giving substantial weight to the undergraduate and internship psychiatric training might improve the delivery of mental health services at the PHC level.


Excellence in role modelling: insight and perspectives from the pros.
Role modelling is an effective teaching method in medical education. We sought to better understand role modelling by examining the insights of respected physician role models. We conducted 30-minute in-depth interviews with 29 highly regarded role models at 2 large teaching hospitals. We coded the transcripts independently, and compared our coding for agreement. Content analysis identified several major categories of themes. The informants identified specific characteristics related to role modelling. Subcategories under the domain of personal qualities included interpersonal skills, a positive outlook, a commitment to excellence and growth, integrity and leadership. Under the domain of teaching, the subcategories were establishing rapport with learners, developing specific teaching philosophies and methods, and being committed to the growth of learners. Subjects thought there was some overlap between teaching and role modelling, but felt that the latter was more implicit and more encompassing. Being a strong clinician was regarded as necessary but not sufficient for being an exemplary physician role model. Perceived barriers to effective role modelling included being impatient and overly opinionated, being quiet, being overextended, and having difficulty remembering names and faces. Physician role models described role modeling consciousness, in that they specifically think about being role models when interacting with learners. Subjects believed that medical learners should emulate multiple role models. Highly regarded physician role models possess personal qualities, teaching abilities and exceptional clinical skills that outweigh their own barriers to serving as effective role models. Many of these positive attributes of role models represent behaviours that can be modified or skills that can be acquired.


Wrapping of a nanowire by a supported lipid membrane.
Internalization of particles by cells plays a crucial role for adsorbing nutrients and fighting infection. Endocytosis is one of the most important mechanisms of particle uptake, which encompasses multiple pathways. Although endocytosis is a complex mechanism involving biochemical signaling and active force generation, the energetic cost associated with the large deformations of the cell membrane wrapping around a foreign particle is an important factor controlling this process, which can be studied using quantitative physical models. Of particular interest is the competition between membrane-cytoskeleton and membrane-target adhesion. This competitive adhesion mechanism can be reproduced to some extent by studying particle wrapping by a membrane adhered to a substrate. We propose a theoretical analysis of this process. Here, we explore the wrapping of a lipid membrane around a long cylindrical object in the presence of a substrate mimicking the cytoskeleton. Using discretization of the Helfrich elastic energy, which accounts for the membrane bending rigidity and surface tension, we obtain a wrapping phase diagram as a function of the membrane-cytoskeleton and the membrane-target adhesion energy, which includes unwrapped, partially wrapped and fully wrapped states. We provide an analytical expression for the boundary between the different regimes. While the transition to partial wrapping is independent of the membrane tension, the transition to full wrapping is very much influenced by the membrane tension. We also show that target wrapping may proceed in an asymmetric fashion in the full wrapping regime.


Brain-computer interfaces in the completely locked-in state and chronic stroke.
Brain-computer interfaces (BCIs) use brain activity to control external devices, facilitating paralyzed patients to interact with the environment. In this chapter, we discuss the historical perspective of development of BCIs and the current advances of noninvasive BCIs for communication in patients with amyotrophic lateral sclerosis and for restoration of motor impairment after severe stroke. Distinct techniques have been explored to control a BCI in patient population especially electroencephalography (EEG) and more recently near-infrared spectroscopy (NIRS) because of their noninvasive nature and low cost. Previous studies demonstrated successful communication of patients with locked-in state (LIS) using EEG- and invasive electrocorticography-BCI and intracortical recordings when patients still showed residual eye control, but not with patients with complete LIS (ie, complete paralysis). Recently, a NIRS-BCI and classical conditioning procedure was introduced, allowing communication in patients in the complete locked-in state (CLIS). In severe chronic stroke without residual hand function first results indicate a possible superior motor rehabilitation to available treatment using BCI training. Here we present an overview of the available studies and recent results, which open new doors for communication, in the completely paralyzed and rehabilitation in severely affected stroke patients. We also reflect on and describe possible neuronal and learning mechanisms responsible for BCI control and perspective for future BMI research for communication in CLIS and stroke motor recovery.


FIB-5 versus FIB-4 index for assessment of hepatic fibrosis in chronic hepatitis B affected patients.
Chronic hepatitis B virus (HBV) infection is one of the major health problems worldwide. Use of non-invasive tests for assessment of hepatic fibrosis such as the FIB-4 index could be used to avoid liver biopsy. Another promising noninvasive test, FIB-5, could also be used to detect significant hepatic fibrosis. The aim of the study was to compare the use of FIB-5 and FIB-4 as noninvasive markers to assess chronic HBV-related hepatic fibrosis. This study was done on 176 chronic HBV patients who underwent liver biopsy. Grading and staging of liver fibrosis was done according to the METAVIR scoring system. FIB-5 and FIB-4 scores were calculated for all patients. As regards FIB-4 for differentiation between non-significant fibrosis (group I) and significant fibrosis (group II), at a cutoff level of 1.28 with positive predictive value (PPV) 41.4% and specificity 48% while at a cutoff level of 7.08 with PPV 98.8% and specificity 98% for FIB-5. As regards both scores, the FIB-5 score was more specific than FIB-4 for diagnosing significant from nonsignificant hepatic fibrosis in patients with chronic HBV infection.


Modulation of chloride, potassium and bicarbonate transport by muscarinic receptors in a human adenocarcinoma cell line.
1. Short-circuit current (I(SC)) responses to carbachol (CCh) were investigated in Colony 1 epithelia, a subpopulation of the HCA-7 adenocarcinoma cell line. In Krebs-Henseleit (KH) buffer, CCh responses consisted of three I(SC) components: an unusual rapid decrease (the 10 s spike) followed by an upward spike at 30 s and a slower transient increase (the 2 min peak). This response was not potentiated by forskolin; rather, CCh inhibited cyclic AMP-stimulated I(SC). 2. In HCO3- free buffer, the decrease in forskolin-elevated I(SC) after CCh was reduced, although the interactions between CCh and forskolin remained at best additive rather than synergistic. When Cl- anions were replaced by gluconate, both Ca2+- and cyclic AMP-mediated electrogenic responses were significantly inhibited. 3. Basolateral Ba2+ (1-10 mM) and 293B (10 microM) selectively inhibited forskolin stimulation of I(SC), without altering the effects of CCh. Under Ba2+- or 293B-treated conditions, CCh responses were potentiated by pretreatment with forskolin. 4. Basolateral charybdotoxin (50 nM) significantly increased the size of the 10 s spike of CCh responses in both KH and HCO3- free medium, without affecting the 2 min peak. The enhanced 10 s spike was inhibited by prior addition of 5 mM apical Ba2+. Charybdotoxin did not affect forskolin responses. 5. In epithelial layers prestimulated with forskolin, the muscarinic antagonists atropine and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP, both at 100 nM) abolished subsequent 10 microM CCh responses. Following addition of p-fluoro hexahydro-sila-difenidol (pF-HHSiD, 10 microM) or pirenzepine (1 microM), qualitative changes in the CCh response time-profile also indicated a rightward shift of the agonist concentration-response curve; however, 1 microM gallamine had no effect. These results suggest that a single M3-like receptor subtype mediates the secretory response to CCh. 6. It is concluded that CCh and forskolin activate discrete populations of basolateral K+ channels gated by either Ca2+ or cyclic AMP, but that the Cl- permeability of the apical membrane may limit their combined effects on electrogenic Cl- secretion. In addition, CCh activates a Ba2+-sensitive apical K+ conductance leading to electrogenic K+ transport. Both agents may also modulate HCO3- secretion through a mechanism at least partially dependent on carbonic anhydrase.


Parenting quality in drug-addicted mothers in a therapeutic mother-child community: the contribution of attachment and personality assessment.
Growing evidence shows that attachment is a key risk factor for the diagnosis and treatment of clinical diseases in Axis I, such as drug addiction. Recent literature regarding attachment, psychiatric pathology, and drug addiction demonstrates that there is a clear prevalence of insecure attachment patterns in clinical and drug addicted subjects. Specifically, some authors emphasize that the anxious-insecure attachment pattern is prevalent among drug-addicted women with double diagnosis (Fonagy et al., 1996). The construct of attachment as a risk factor in clinical samples of drug-addicted mothers needs to be studied more in depth though. The present explorative study focused on the evaluation of parenting quality in a therapeutic mother-child community using attachment and personality assessment tools able to outline drug-addicted mothers' profiles. This study involved 30 drug addicted mothers, inpatients of a therapeutic community (TC). Attachment representations were assessed via the Adult Attachment Interview; personality diagnosis and symptomatic profiles were performed using the Structured Clinical Interview of the DSM-IV (SCID-II) and the Symptom Check List-90-R (SCL-90-R), respectively. Both instruments were administered during the first six months of residence in a TC. Results confirmed the prevalence of insecure attachment representations (90%), with a high presence of U patterns, prevalently scored for dangerous and/or not protective experiences in infanthood. Very high values (>5) were found for some experience scales (i.e., neglect and rejection scales). Data also showed very low values (1-3) in metacognitive monitoring, coherence of transcript and coherence of mind scales. Patients' different profiles (U vs. E vs. Ds) were linked to SCID-II diagnosis, providing insightful indications both for treatment planning and intervention on parenting functions and for deciding if to start foster care or adoption proceedings for children.


Time-dependent mechanics and measures of glial activation and behavioral sensitivity in a rodent model of radiculopathy.
Nerve root compression induces persistent behavioral hypersensitivity and spinal glial reactivity. Viscoelastic properties of neural tissues suggest that physiologic outcomes may depend on the duration of an applied nerve root compression. This study evaluated the time-dependent properties of the root under compression in the context of pain-related behavioral and physiologic outcomes. The decrease in applied load measured by load relaxation under compression was quantified for rat cervical (C6-C8) roots in situ for durations of 30 sec, 3 min, or 15 min (n = 6). Immediately following compression, the change in the root width relative to its original width was quantified as a measure of its structural recovery. Both load relaxation and structural recovery were significantly (p < 0.05) correlated with duration of compression. After 30 sec of compression, load relaxed by 22 +/- 10%; increasing to 36 +/- 18% and 56 +/- 20% at 3 and 15 min, respectively. Following 30 sec, 3 min, and 15 min of compression, the root recovered to 91 +/- 5%, 88 +/- 5 and 72 +/- 13% of its original width, respectively. A companion in vivo study imposed these same compression durations and sham procedures to the C7 root to evaluate pain symptoms and spinal glial reactivity. Allodynia was assessed for 7 days to measure behavioral sensitivity. Immunohistochemistry and quantitative densitometry detected GFAP and OX-42 in the dorsal horn at day 7. Significant correlations were detected between compression duration and allodynia (p < 0.03), and astrocyte and microglial activation (p < 0.01). These biomechanical and glial results imply that a similar duration of compression may modulate both sustained pain and spinal glial reactivity.


Populations of selected microbial and fungal species growing on the surface of rape seeds following treatment with desiccants or plant growth regulators.
The aim of this study was to determine the effects of desiccants and plant growth regulators on selected microbial species affecting rape seeds, with special emphasis on the growth of fungi and identification of the genus and species composition. The experimental material in the study was seeds of winter rape cv. Californium that were collected from the field during combine harvest. The chemical agents applied, both desiccants and growth regulators, generally decreased the populations of bacteria occurring on the surface of rape seeds. The responses of fungi depended upon the type of agent applied and were manifested as either stimulation or inhibition of the growth of the fungal species. The fungi isolated from the surface of rape seeds were characteristic of those found in the field environment (Cladosporium and Penicillium) and typical for those present on the surface of rape seeds (Alternaria).


Potential for Dependence on Lisdexamfetamine - In vivo and In vitro Aspects.
Although lisdexamfetamine is used as a recreational drug, little research exists regarding its potential for dependence or its precise mechanisms of action. This study aims to evaluate the psychoactivity and dependence profile of lisdexamfetamine using conditioned place preference and self-administration paradigms in rodents. Additionally, biochemical techniques are used to assess alterations in the dopamine levels in striatal synaptosomes following administration of lisdexamfetamine. Lisdexamfetamine increased both conditioned place preference and self-administration. Moreover, after administration of the lisdexamfetamine, dopamine levels in the striatal synaptosomes were significantly increased. Although some modifications should be made to the analytical methods, performing high performance liquid chromatography studies on synaptosomes can aid in predicting dependence liability when studying new psychoactive substances in the future. Collectively, lisdexamfetamine has potential for dependence possible via dopaminergic pathway.


Gene and protein expressions of type I collagen are regulated tissue-specifically in rat hyaline cartilages in vivo.
The present study was designed to investigate how rat hyaline cartilages at various sites in vivo express the gene and protein of type I collagen using in situ hybridization and immunohistochemistry. The gene of pro alpha 1(I) collagen was expressed by chondrocytes in articular cartilage, and the protein of type I collagen was identified in the cartilage matrix. In contrast, growth plate cartilage expressed the gene of pro alpha 1(I) collagen, but no protein of type I collagen. Neither gene nor protein of type I collagen was expressed in cartilages of trachea and nasal septum. The present study suggested that expression of type I collagen in hyaline cartilages may be regulated tissue-specifically at gene and/or protein levels.


Comparative anatomy of stems and leaves of plectranthus L. (Lamiaceae) in Saudi Arabia and systematic implications.
A comparative anatomical study of the Plectranthus species growing in Saudi Arabia by using light microscopy method in order to evaluate their diagnostic value for systematic studies. Macro- and micromorphological characters, including stem, leaf, petiole anatomical, and trichome structures are presented. Descriptions of stem shape in cross-section, epidermal cell shape, cortex, pith, vascular bundles, leaf shape in cross-section, mesophyll, types and distribution of trichomes are presented. Taxonomic phylogenetic implications of stem, leaf, and trichomes anatomical micromorphology are also discussed in comparison with the available gross morphological and molecular data. Results of the present study offer useful data for evaluating the taxonomy of Plectranthus both at subgeneric and sectional levels. Moreover, our results indicate some degree of similarity among the species of subgenus Germanea and support the monophyly of this subgenus. A remarkable result from this study was identifying P. arabicus with distinctive characters and suggesting that it should be treated as a separate subgenus. Likewise, distinguishing differences between closely related endemic species P. asirensis and P. hijazensis are offered, and should be conserved as a different subgenus. A key for the identification of the investigated taxa based on studied characters is provided. Microsc. Res. Tech. 79:583-594, 2016. © 2016 Wiley Periodicals, Inc.