THE EFFECT OF NUCLEAR AND CYTOPLASMIC GENES ON FITNESS AND LOCAL ADAPTATION IN AN ANNUAL LEGUME, CHAMAECRISTA FASCICULATA.
The role of nuclear genes in local adaptation has been well documented. However, the role of maternally inherited cytoplasmic genes to the evolution of natural populations has been relatively unstudied. To evaluate the contribution of cytoplasmic and nuclear genomes and their interactions to local adaptation we created second-generation backcross hybrids between a Maryland and an Illinois population of the annual legume Chamaecrista fasciculata. Backcross progeny were planted in the sites native to each population for two years and we quantified germination, survivorship, fruit production, vegetative biomass, and cumulative fitness. We found limited evidence for the contribution of either cytoplasmic or nuclear genes to local adaptation. In Maryland plants had greater survivorship, biomass, fruit production, and cumulative fitness if their nuclear genome was composed predominately of native Maryland genes; cytoplasmic genes did not affect fitness. In Illinois local cytoplasm marginally enhanced fitness, whereas Maryland nuclear genes outperformed local nuclear genes. Interactions between cytoplasmic and nuclear genes influenced seed weight, vegetative biomass, and fitness and therefore may affect evolution of these characters. Genetic effects were stronger acting through seed size than directly on characters. However, seed size differences between the two populations were largely genetic and therefore selection on fitness components is likely to result in evolutionary change. The contribution of nuclear and cytoplasmic genes to fitness components varied across sites and years, suggesting that experiments should be replicated and conducted under natural conditions to understand the influence of these genomes and their interactions to population differentiation.


[Behavioral effects of amantadine on ambulatory activity and drinking in mice and on continuous and discrete avoidance responses in rats].
Effects of amantadine on ambulatory activity in mice and conditioned avoidance responses in rats were investigated. In addition, the change in diurnal drinking rhythm induced by free intake of amantadine solution was also examined in mice. Amantadine (10-40 mg/kg, i.p.) tended to increase the ambulatory activity in mice. Pretreatment with amantadine (10-40 mg/kg, i.p.) at 30 min or 4 hr before augmented the ambulation-increasing effect of apomorphine (0.5 mg/kg, s.c.), but attenuated that of methamphetamine (2 mg/kg, s.c.). When amantadine solution of 0.1 mg/ml or 0.4 mg/ml was freely given to mice for 7 or 21 days, the amount of fluid intake per day decreased slightly, and the daily doses of amantadine intake were estimated to be 15-80 mg/kg/day. However, no marked change in the diurnal pattern of drinking, showing a higher rate during the dark period and a lower rate during the light period, was observed. These mice also demonstrated an increased sensitivity to the ambulation-increasing effect of apomorphine and a decreased sensitivity to that of methamphetamine as compared with the control mice that drank water with no added drugs. Amantadine (10-20 mg/kg, i.p.) did not induce a marked change in the avoidance responses in rats. However, more than 40 mg/kg, i.p. of amantadine suppressed the avoidance responses with a general worsening of bodily condition. Amantadine (10-20 mg/kg, i.p.) did not modify the avoidance-facilitating effects of methamphetamine (0.13-1 mg/kg, s.c.), atropine (1.3-10 mg/kg, s.c.) and scopolamine (0.031-0.13 mg/kg, s.c.), but attenuated the avoidance-suppressing effects of chlorpromazine (0.25-2 mg/kg, s.c.), haloperidol (0.018-0.05 mg/kg, s.c.), tetrabenazine (0.25-1 mg/kg, s.c.), pilocarpine (2-8 mg/kg, s.c.) and physostigmine (0.1-0.4 mg/kg, s.c.) in a dose-dependent manner. The present results suggest that amantadine affects not only the central catecholaminergic neurons but also the other neurons.


Energy landscapes for diffusion: analysis of cage-breaking processes.
A wide spectrum of potential energy barriers exists for binary Lennard-Jones systems. Here we examine the barriers and cage-breaking rearrangements that are pertinent to long-term diffusion. Single-step cage-breaking processes, which follow high-barrier routes, are identified, and different methods and criteria for defining a cage-breaking process are considered. We examine the extent to which a description of cage-breaking within the energy landscape is a description of long-term diffusion. This description includes the identification of cage-breaks that are reversed, and those that are productive towards long-term diffusion. At low temperatures, diffusion is adequately described by productive cage-breaks, or by considering all cage-breaks and accounting for the effect of reversals. To estimate the diffusion constant we require only the mean square displacement of a cage-break, the average waiting time for a cage-break, and a measure of the number of reversed cage-breaks. Cage-breaks can be visualized within the potential energy landscape using disconnectivity graphs, and we compare the use of productive cage-breaks with previous definitions of "megabasins" or "metabasins."


Opportunistic case finding pivotal in diagnosing COPD.
The principle of COPD diagnosis is based on establishing the presence of obstructive post-bronchodilator spirometry in an at-risk individual with appropriate symptoms. Most individuals diagnosed with COPD have substantial cigarette smoke exposure, more than 10 and usually 20 pack years. Patients with COPD usually have symptoms of cough and phlegm, exertional breathlessness, wheezing, recurrent winter bronchitis or a combination of these symptoms. A current or ex-smoker, aged over 35, with respiratory symptoms should undergo spirometry testing. The hallmark of COPD is the presence of airflow obstruction after administration of a bronchodilator. However, while normal spirometry excludes COPD, obstructive spirometry is also seen in asthma and bronchiectasis emphasising the importance of clinical features. A significant change introduced in the 2010 NICE guideline is the new grading of COPD severity. A system based on FEV alone oversimplifies disease severity and full clinical assessment should include symptoms, frequency of exacerbations and impact of disease on functioning and health status. The most important single intervention is smoking cessation as this is proven to reduce decline in FEV1 and reduce mortality. All patients should also receive influenza and pneumococcal vaccination, weight management, pulmonary rehabilitation (if breathless) and short-acting bronchodilators. The evidence base for rehabilitation is very strong with positive effects shown on breathlessness, exercise capacity, activity level, exacerbation rate, leg muscle strength and quality of life.


Weather fluctuations: predictive factors in the prevalence of acute coronary syndrome.
Background: Meteorological parameters and seasonal changes can play an important role in the occurrence of acute coronary syndrome (ACS). However, there is almost no evidence on a national level to suggest the associations between these variables and ACS in Iran. We aim to identify the meteorological parameters and seasonal changes in relationship to ACS. Methods: This retrospective cross-sectional study was conducted between 03/19/2015 to 03/18/2016 and used documents and records of patients with ACS in Mazandaran ProvinceHeart Center, Iran. The following definitive diagnostic criteria for ACS were used: (1) existence of cardiac enzymes (CK or CK-MB) above the normal range; (2) Greater than 1 mm ST-segment elevation or depression; (3) abnormal Q waves; and (4) manifestation of troponin enzyme in the blood. Data were collected daily, such as temperature (Celsius) changes, wind speed and its direction, rainfall, daily evaporation rate; number of sunny days, and relative humidity were provided by the Meteorological Organization of Iran. Results: A sample of 2,054 patients with ACS were recruited. The results indicated the highest ACS events from March to May. Generally, wind speed (18 PM) [IRR = 1.051 (95% CI: 1.019 to1.083), P=0.001], daily evaporation [IRR = 1.039 (95% CI: 1.003 to 1.077), P=0.032], daily maximum (P<0.001) and minimum (P=0.003) relative humidity was positively correlated withACS events. Also, negatively correlated variables were daily relative humidity (18 PM) [IRR =0.985 (95% CI: 0.978 to 0.992), P<0.001], and daily minimum temperature [IRR = 0.942 (95%CI: 0.927 to 0.958), P<0.001]. Conclusion: Climate changes were found to be significantly associated with ACS; especially from cold weather to hot weather in March, April and May. Further research is needed to fully understand the specific conditions and cold exposures.


Transforming Growth Factor-Beta and Sonic Hedgehog Signaling in Palatal Epithelium Regulate Tenascin-C Expression in Palatal Mesenchyme During Soft Palate Development.
During palatogenesis, the palatal shelves first grow vertically on either side of the tongue before changing their direction of growth to horizontal. The extracellular matrix (ECM) plays an important role in these dynamic changes in palatal shelf morphology. Tenascin-C (TNC) is an ECM glycoprotein that shows unique expression in the posterior part of the palatal shelf, but little is known about the regulation of TNC expression. Since transforming growth factor-beta-3 (TGF-β3) and sonic hedgehog (SHH) signaling are known to play important roles in palatogenesis, we investigated whether TGF-β3 and SHH are involved in the regulation of TNC expression in the developing palate. TGF-β3 increased the expression of TNC mRNA and protein in primary mouse embryonic palatal mesenchymal cells (MEPM) obtained from palatal mesenchyme dissected at embryonic day 13.5-14.0. Interestingly, immunohistochemistry experiments revealed that TNC expression was diminished in K14-cre;Tgfbr2 fl/fl mice that lack the TGF-β type II receptor in palatal epithelial cells and exhibit cleft soft palate, whereas TNC expression was maintained in Wnt1-cre;Tgfbr2 fl/fl mice that lack the TGF-β type II receptor in palatal mesenchymal cells and exhibit a complete cleft palate. SHH also increased the expression of TNC mRNA and protein in MEPM cells. However, although TGF-β3 up-regulated TNC mRNA and protein expression in O9-1 cells (a cranial neural crest cell line), SHH did not. Furthermore, TGF-β inhibited the expression of osteoblastic differentiation markers (osterix and alkaline phosphatase) and induced the expression of fibroblastic markers (fibronectin and periostin) in O9-1 cells, whereas SHH did not affect the expression of osteoblastic and fibroblastic markers in O9-1 cells. However, immunohistochemistry experiments showed that TNC expression was diminished in the posterior palatal shelves of Shh-/+ ;MFCS4 +/- mice, which have deficient SHH signaling in the posterior palatal epithelium. Taken together, our findings support the proposal that TGF-β and SHH signaling in palatal epithelium co-ordinate the expression of TNC in the posterior palatal mesenchyme through a paracrine mechanism. This signal cascade may work in the later stage of palatogenesis when cranial neural crest cells have differentiated into fibroblast-like cells. The spatiotemporal regulation of ECM-related proteins by TGF-β and SHH signaling may contribute not only to tissue construction but also to cell differentiation or determination along the anterior-posterior axis of the palatal shelves.


Clinical Assessment of Prostate Displacement and Planning Target Volume Margins for Stereotactic Body Radiotherapy of Prostate Cancer.
Purpose: To assess the optimal planning target volume (PTV) margins for stereotactic body radiotherapy (SBRT) of prostate cancer based on inter- and intra-fractional prostate motion determined from daily image guidance. Methods and Materials: Two hundred and five patients who were enrolled on two prospective studies of SBRT (8 Gy × 5 fractions) for localized prostate cancer treated at a single institution between 2012 and 2017 had complete inter- and intra-fractional shift data available. All patients had scheduled kilovoltage planar imaging during SBRT with rigid registration to intraprostatic fiducials prior to each of four half-arcs delivered per fraction, as well as cone beam CT verification of anatomy prior to each fraction. Inter- and intra- fractional shift data were obtained to estimate the required PTV margins based on the classic van Herk formula. Inter- and intra-fractional motion were compared between patients with and without severe toxicities using the independent two-sample Wilcoxon test. Results: The margins required to account for inter-fractional motion were estimated to be 0.99, 1.52, and 1.45 cm in lateral (LR), longitudinal (SI), and vertical (AP) directions, respectively. The margins required to account for intra-fractional motion were estimated to be 0.19, 0.27, and 0.31 cm in LR, SI and AP directions, respectively. Large intra-fractional shifts were mostly observed in the SI and AP directions, with 2.0 and 5.4% of patients experiencing average intra-fractional motion >3 mm in the SI and AP directions, respectively, compared with none experiencing mean shifts >3 mm in the LR direction. Six patients experienced grade 3 gastrointestinal or genitourinary toxicity. There were no significant differences in mean inter- or intra-fractional motion in any of the cardinal directions compared to patients without severe toxicity (inter-fractional p = 0.46-0.99, intra-fractional p = 0.10-0.84). Conclusion: The inter- and intra-fractional margins estimated from this study are in line with prior reported values. Intra-fractional prostate motion was generally small with larger margins required for the SI and AP directions, notably just slightly exceeding the commonly used 3 mm posterior PTV margin even with realignment between half-arcs. Development of severe toxicity was not significantly associated with the degree of inter- or intra-fractional motion.


Recent Insights into the HIV/AIDS Pandemic.
Etiology, transmission and protection: Transmission of HIV, the causative agent of AIDS, occurs predominantly through bodily fluids. Factors that significantly alter the risk of HIV transmission include male circumcision, condom use, high viral load, and the presence of other sexually transmitted diseases. Pathology/Symptomatology: HIV infects preferentially CD4+ T lymphocytes, and Monocytes. Because of their central role in regulating the immune response, depletion of CD4+ T cells renders the infected individual incapable of adequately responding to microorganisms otherwise inconsequential. Epidemiology, incidence and prevalence: New HIV infections affect predominantly young heterosexual women and homosexual men. While the mortality rates of AIDS related causes have decreased globally in recent years due to the use of highly active antiretroviral therapy (HAART) treatment, a vaccine remains an elusive goal. Treatment and curability: For those afflicted HIV infection remains a serious illness. Nonetheless, the use of advanced therapeutics have transformed a dire scenario into a chronic condition with near average life spans. When to apply those remedies appears to be as important as the remedies themselves. The high rate of HIV replication and the ability to generate variants are central to the viral survival strategy and major barriers to be overcome. Molecular mechanisms of infection: In this review, we assemble new details on the molecular events from the attachment of the virus, to the assembly and release of the viral progeny. Yet, much remains to be learned as understanding of the molecular mechanisms used in viral replication and the measures engaged in the evasion of immune surveillance will be important to develop effective interventions to address the global HIV pandemic.


Relationship of fluid and mucin secretion to morphological changes in the perfused rat submandibular gland.
Transport of electrolytes/water and exocytosis are activated by elevation of the cytosolic Ca(2+) concentration and are potentiated by elevation of cytosolic cyclic AMP. To correlate mucin and fluid secretion with morphological changes, rat submandibular glands were vascularly perfused and the fluid secretion and N-acetylgalactosamine in the saliva were measured during stimulation with various concentrations of carbachol (CCh) and/or isoproterenol (ISP). Single stimulation with 1 microM CCh induced a transient increase of N-acetyl galactosamine followed by a decline to a low level during sustained stimulation. The overload of 1 microM ISP increased secretion of N-acetyl galactosamine to a higher sustained level of 40-50 microg/g-min. However, at 1 microM CCh, fluid secretion was maintained at the same level during stimulation and even overload of 1 microM ISP did not significantly affect its level, whereas addition of 0.5 microM ISP to the gland stimulated with 0.1 microM CCh increased fluid secretion. Morphological observation was carried out by HRSEM and TEM. Combination of CCh and ISP in different concentrations resulted in distinctive morphological changes which reflect fluid secretion and mucin secretion. The kinetics of ATP and creatine phosphate (PCr) were measured using P-31 NMR, which indicated that the potentiation of fluid secretion is limited under a higher level of CCh stimulation due to a limited energy supply.


Cancer incidence and cause-specific mortality in male and female physicians: a cohort study in Estonia.
To evaluate whether the presumed knowledge of physicians about healthier lifestyle decreases their risk of cancer and mortality, a retrospective cohort study of male and female physicians was conducted in Estonia. The cancer incidence and cause-specific mortality of 3,673 physicians (870 M, 2,803 F) in Estonia was compared with the rates of the general population. Information on cancer cases and deaths in the cohort between 1983 and 1998 was obtained from the Estonian Cancer Registry and the mortality database of Estonia. The standardized incidence ratio (SIR) for all cancers was 1.32 (95% confidence interval (CI) 1.15-1.48) in women and 0.92 (95% CI 0.73-1.13) in men. Female physicians had an elevated risk for breast cancer (SIR 2.03, 95% CI 1.62-2.51) and myeloid leukaemia (SIR 3.69, 95% CI 1.35-8.02). Male physicians had an excess of skin melanoma (SIR 4.88, 95% CI 1.58-11.38). A large deficit of lung cancer was observed (SIR 0.24, 95% CI 0.11-0.48). The very low all-cause mortality in the cohort (standardized mortality ratio 0.55, 95% CI 0.50-0.61) was mainly due to large deficits in deaths from lung cancer, cardiovascular diseases and external causes. The suicide rate in the cohort was lower than in the general population. No health risks were observed in the cohort that could be linked to the occupational exposures of physicians. The pattern of cancer incidence and mortality seen in physicians in Estonia is similar to the pattern seen among professional classes in other countries.


Drug interactions involving immunologic agents. Part II. Immunodiagnostic and other immunologic drug interactions.
This two-part review addresses pharmacists' need for information about immunologic drug interactions. Reports of interactions involving vaccines, immune globulins, and immunodiagnostic reagents are widely dispersed and, therefore, difficult to access. In the conclusion of this two-part article, immunodiagnostic drug interactions as well as interactions involving interferons and other biologic-response modifiers are discussed, as are epinephrine interactions that affect treatment of immunologic-induced anaphylaxis. Host factors of groups and individuals relating to the clinical significance of proposed interactions are covered, such as those involving influenza vaccine with theophylline, warfarin, or phenytoin, and other combinations. Vigilance by all pharmacists is needed to detect previously unreported immunologic drug interactions and further assess known interactions.


[A case of pathologically complete response of local recurrence in the mesorectum after multidisciplinary therapy].
The patient was a 68-year-old man who had undergone sigmoidectomy 1 year previously. Adjuvant therapy with tegafur- uraci(l UFT) and Leucovorin( UZEL) was administered. Seven months later, the carcinoembryonic antigen( CEA) level increased to 7.5 ng/mL. Enhanced computed tomography (CT) revealed a 4-cm mass in the mesorectum, and the patient was diagnosed as having local recurrence. Chemotherapy with oxaliplatin, 5-fluorouracil, and Leucovorin( mFOLFOX6) and radiation therapy( 60 Gy) were administered. As the tumor could not be detected after chemoradiotherapy, abdominoperineal resection was performed. Pathological examination showed no cancer and indicated a complete response. The patients CEA level has not increased and no recurrence has been detected on enhanced CT for 3.5 years. The tumor could be decreased in size by chemoradiotherapy and the operation could be safely performed without resecting other organs. Although surgery is generally performed for local recurrence, multidisciplinary therapy could be useful in performing such surgery safely.


Optical phantoms with adjustable subdiffusive scattering parameters.
A new epoxy-resin-based optical phantom system with adjustable subdiffusive scattering parameters is presented along with measurements of the intrinsic absorption, scattering, fluorescence, and refractive index of the matrix material. Both an aluminium oxide powder and a titanium dioxide dispersion were used as scattering agents and we present measurements of their scattering and reduced scattering coefficients. A method is theoretically described for a mixture of both scattering agents to obtain continuously adjustable anisotropy values g between 0.65 and 0.9 and values of the phase function parameter γ in the range of 1.4 to 2.2. Furthermore, we show absorption spectra for a set of pigments that can be added to achieve particular absorption characteristics. By additional analysis of the aging, a fully characterized phantom system is obtained with the novelty of g and γ parameter adjustment.


River bed carbon and nitrogen cycling: state of play and some new directions.
The significance of freshwaters as key players in the global budget of both carbon dioxide and methane has recently been highlighted. In particular, rivers clearly do not act simply as inert conduits merely piping carbon from catchment to coast, but, on the whole, their metabolic activity transforms a considerable fraction of the carbon that they convey. In addition, nitrogen is cycled, sometimes in tight unison with carbon, with appreciable amounts being 'denitrified' between catchment and coast. However, shortfalls in our knowledge about the significance of exchange and interaction between rivers and their catchments, particularly the significance of interactions mediated through hyporheic sediments, are still apparent. From humble beginnings of quantifying the consumption of oxygen by small samples of gravel, to an integrated measurement of reach scale transformations of carbon and nitrogen, our understanding of the cycling of these two macro elements in rivers has improved markedly in the past few decades. However, recent discoveries of novel metabolic pathways in both the nitrogen and carbon cycle across a spectrum of aquatic ecosystems, highlights the need for new directions and a truly multidisciplinary approach to quantifying the flux of carbon and nitrogen through rivers.


Extending the Compatibility of the SP3 Paramagnetic Bead Processing Approach for Proteomics.
The diversity in protein and peptide biochemistry necessitates robust protocols and reagents for efficiently handling and enriching these molecules prior to analysis with mass spectrometry (MS) or other techniques. Further exploration of the paramagnetic bead-based approach, single-pot solid-phase-enhanced sample preparation (SP3), is carried out toward updating and extending previously described conditions and experimental workflows. The SP3 approach was tested in a wide range of experimental scenarios, including (1) binding solvents (acetonitrile, ethanol, isopropanol, acetone), (2) binding pH (acidic vs neutral), (3) solvent/lysate ratios (50-200%, v/v), (4) mixing and rinsing conditions (on-rack vs off-rack rinsing), (5) Enrichment of nondenatured proteins, and (6) capture of individual proteins from noncomplex mixtures. These results highlight the robust handling of proteins in a broad set of scenarios while also enabling the development of a modified SP3 workflow that offers extended compatibility. The modified SP3 approach is used in quantitative in-depth proteome analyses to compare it with commercial paramagnetic bead-based HILIC methods (MagReSyn) and across multiple binding conditions (e.g., pH and solvent during binding). Together, these data reveal the extensive quantitative coverage of the proteome possible with SP3 independent of the binding approach utilized. The results further establish the utility of SP3 for the unbiased handling of peptides and proteins for proteomic applications.


Genetic diversity of donkey populations from the putative centers of domestication.
Donkey domestication drastically changed ancient transport systems in Africa and Asia, enabling overland circulation of people and goods and influencing the organization of early cities and pastoral societies. Genetic studies based on mtDNA have pointed to the African wild ass as the most probable ancestor of the domestic donkey, but questions regarding its center of origin remain unanswered. Endeavoring to pinpoint the geographical origin of domestic donkey, we assessed levels and patterns of genetic diversity at 15 microsatellite loci from eight populations, representing its three hypothesized centers of origin: northeast Africa, the Near East and the Arabian Peninsula. Additionally, we compared the donkey genotypes with those from their wild relative, the African wild ass (Equus africanus somaliensis) to visualize patterns of differentiation among wild and domestic individuals. Obtained results revealed limited variation in levels of unbiased expected heterozygosity across populations in studied geographic regions (ranging from 0.637 in northeast Africa to 0.679 in the Near East). Both allelic richness (Ar) and private allelic richness presented considerably higher values in northeast Africa and in the Arabian Peninsula. By looking at variation at the country level, for each region, we were able to identify Sudan and Yemen as the countries possessing higher allelic richness and, cumulatively, Yemen also presented higher values for private allelic richness. Our results support previously proposed northeast Africa as a putative center of origin, but the high levels of unique diversity in Yemen opens the possibility of considering this region as yet another center of origin for this species.


Lymphomatoid granulomatosis and other Epstein-Barr virus associated lymphoproliferative processes.
We now recognize that the Epstein-Barr virus (EBV), which is a member of the γ- herpesvirus family, plays a pivotal role in the development of several lymphomas and lymphoproliferative disorders that include B-cell, T-cell and NK-cell processes. While over recent years, EBV associated lymphomas that arise in patients with known defects in cellular immunity are relatively well characterized, these diseases are becoming increasingly recognized in patients without overt immunodeficiency. Improved understanding of the biology of these lymphomas including elucidating the role that EBV plays in their pathogenesis has paved the way for improved therapies targeted at critical signaling pathways as well as the development of novel cellular therapies. In this review, we focus on recent progress that has been made in the biology and treatment of the rare EBV-associated disorder lymphomatoid granulomatosis (LYG) and also discuss other EBV-associated processes that occur in both immunocompetent and immunocompromised hosts.


Interactions of adenosine and vecuronium in neuromuscular blockade in cats.
The effect of adenosine on the neuromuscular blockade induced by vecuronium and the capacity of neostigmine to reverse this combined blockade were studied in 30 cats on a standard sciatic nerve--tibialis anterior muscle preparation. Adenosine infused to 6 cats at a constant rate (3.9 +/- 1.1 mg/kg/min) to produce a stable 50% reduction of the mean arterial pressure did not affect neuromuscular transmission. At the same 50% reduction of the mean arterial pressure by adenosine or sodium nitroprusside infusion in another 15 cats, adenosine (n = 9) significantly potentiated vecuronium-induced neuromuscular blockade, but sodium nitroprusside (n = 6) did not. Neostigmine antagonized the neuromuscular blockade of similar degrees produced either by the combination of adenosine with vecuronium in the above 9 cats or by vecuronium alone in the remaining 9 cats. There was no significant difference in the doses of neostigmine given. Because no potentiation was found at the same level of hypotension induced by sodium nitroprusside, the potentiation effect of adenosine on neuromuscular blockade is not likely to be due to the hypotensive effect of adenosine, but may be due to impairment, by adenosine, of acetylcholine release from motor nerve endings. We conclude that adenosine potentiates neuromuscular blockade by vecuronium and that neostigmine can be expected to reverse this combined blockade.


Engineering Nucleotide Specificity of Succinyl-CoA Synthetase in Blastocystis: The Emerging Role of Gatekeeper Residues.
Charged, solvent-exposed residues at the entrance to the substrate binding site (gatekeeper residues) produce electrostatic dipole interactions with approaching substrates, and control their access by a novel mechanism called "electrostatic gatekeeper effect". This proof-of-concept study demonstrates that the nucleotide specificity can be engineered by altering the electrostatic properties of the gatekeeper residues outside the binding site. Using Blastocystis succinyl-CoA synthetase (SCS, EC 126.96.36.199), we demonstrated that the gatekeeper mutant (ED) resulted in ATP-specific SCS to show high GTP specificity. Moreover, nucleotide binding site mutant (LF) had no effect on GTP specificity and remained ATP-specific. However, via combination of the gatekeeper mutant with the nucleotide binding site mutant (ED+LF), a complete reversal of nucleotide specificity was obtained with GTP, but no detectable activity was obtained with ATP. This striking result of the combined mutant (ED+LF) was due to two changes; negatively charged gatekeeper residues (ED) favored GTP access, and nucleotide binding site residues (LF) altered ATP binding, which was consistent with the hypothesis of the "electrostatic gatekeeper effect". These results were further supported by molecular modeling and simulation studies. Hence, it is imperative to extend the strategy of the gatekeeper effect in a different range of crucial enzymes (synthetases, kinases, and transferases) to engineer substrate specificity for various industrial applications and substrate-based drug design.


Exaggerated natriuresis in primary aldosteronism.
Acute response in blood pressure (BP) and natriuresis to saline infusion was evaluated in 16 patients with primary aldosteronism caused by aldosteronoma (PA) and 12 patients with salt-sensitive essential hypertension (SSEH). Salt-sensitivity was defined by a decrease in mean BP exceeding 5% at the second hour after a 20 mg furosemide injection. Plasma renin activity (PRA), plasma aldosterone concentration (PAC) and urine electrolytes in response to saline infusion were determined. During a 2-liter isotonic saline infusion, a similar degree of natriuresis and change in BP were observed in PA and SSEH patients. A significantly inverse correlation between the increase in mean BP and the degree of natriuresis at the end of the infusion was found in patients with SSEH (r = -0.80, p less than 0.01). No correlation was observed between these parameters in patients with PA (r = 0.28, p greater than 0.05). These results suggest that hypernatriuresis in SSEH may play a protective mechanism against abrupt increases in BP and volume during acute saline loading. This protective mechanism was not evident in patients with PA.


Analysis of MUTYH alternative transcript expression, promoter function, and the effect of human genetic variants.
The human DNA repair gene MUTYH, whose mutational loss causes a colorectal polyposis and cancer predisposition, contains three alternative first exons. In order to analyze alternative transcription and the effect of genetic alterations found in humans, we established a cell-based minigene experimental model supporting transcription and splicing and thoroughly verified its functionality. We identified highly conserved promoter areas and inactivated them in the minigene, and also introduced six human variants. Moreover, the potential contribution of CpG island methylation and specific transcription factors on MUTYH transcription was addressed. The findings allowed to attribute regulatory roles to three conserved motifs in the promoter: an M4 motif, a transcription factor IIB recognition element, and a GC box. Moreover, the data showed that three patient variants compromised MUTYH expression and therefore have the potential to cause pathogenic effects. We did not find evidence for a biologically relevant contribution of CpG island methylation or a direct transcriptional activation by DNA damage. Besides insight into the regulation of MUTYH transcription, the work therefore provides a functional MUTYH minigene experimental system suitable as a diagnostic tool for analyzing patient variants, and a functional map of the promotor that also can facilitate pathogenicity classifications of human variants.


Self-produced biophotosensitizers enhance the degradation of organic pollutants in photo-bioelectrochemical systems.
Bioelectrochemical systems (BESs) with integrated photoactive components have been shown to be a promising strategy for enhancing the performance for bioenergy generation and pollutant removal. This study revealed an efficient photo-BES with an enhanced pollutant degradation rate by utilizing self-produced biomolecules as photosensitizers in situ. Results showed that the BES could increase the coulombic efficiency from 60.8% to 73.0% and the degradation rate of bisphenol A (BPA) from 0.030 to 0.189 h-1 when the suspension in the reactor was illuminated with simulated sunlight in the absence of any external photosensitizers. We identified that the regular BES released many organic substances into the reactor during operation. These substances, including dissolved biomolecules and solid cell residues, were photoactive for producing hydroxyl radicals during light illumination. Quenching experiments verified that the •OH generated from the self-produced biophotosensitizers contributed to the enhanced degradation of BPA. Additionally, the phototransformation of biophotosensitizers was also observed in photo-BES. The quantity of tyrosine protein-like components decreased, but that of the humic components remained relatively stable. Our findings imply that BESs with integrated self-produced biophotosensitizers may be promising for constructing advanced electrochemical and biological systems for synchronous bioelectricity production and degradation of organic pollutants in wastewater treatments.


Early prediction of high flow nasal cannula therapy outcomes using a modified ROX index incorporating heart rate.
The ROX index (ratio of pulse oximetry/FIO2 to respiratory rate) has been validated to predict high flow nasal cannula therapy (HFNC) outcomes in patients with pneumonia. We evaluated a modified ROX index incorporating heart rate (HR) in patients initiated on HFNC for acute hypoxemic respiratory failure and as a preventative treatment following planned extubation. We performed a prospective observational cohort study of 145 patients treated with HFNC. ROX-HR index was defined as the ratio of ROX index over HR (beats/min), multiplied by a factor of 100. Evaluation was performed using area under the receiving operating characteristic curve (AUROC) and cutoffs assessed for prediction of HFNC failure: defined as the need for mechanical ventilation. Ninety-nine (68.3%) and 46 (31.7%) patients were initiated on HFNC for acute hypoxemic respiratory failure and following a planned extubation, respectively. The majority (86.9%) of patients had pneumonia as a primary diagnosis, and 85 (56.6%) patients were immunocompromised. Sixty-one (42.1%) patients required intubation (HFNC failure). Amongst patients on HFNC for acute respiratory failure, HFNC failure was associated with a lower ROX and ROX-HR index recorded at time points between 1 and 48 h. Within the first 12 h, both indices performed with the highest AUROC at 10 h as follows: 0.723 (95% CI 0.605-0.840) and 0.739 (95% CI 0.626-0.853) for the ROX and ROX-HR index respectively. A ROX-HR index of > 6.80 was significantly associated with a lower risk of HFNC failure (hazard ratio 0.301 (95% CI 0.143-0.663)) at 10 h. This association was also observed at 2, 6, 18, and 24h, even with correction for potential confounding factors. For HFNC initiated post-extubation, only the ROX-HR index remained significantly associated with HFNC failure at all recorded time points between 1 and 24 h. A ROX-HR > 8.00 at 10 h was significantly associated with a lower risk of HFNC failure (hazard ratio 0.176 (95% CI 0.051-0.604)). While validation studies are required, the ROX-HR index appears to be a promising tool for early identification of treatment failure in patients initiated on HFNC for acute hypoxemic respiratory failure or as a preventative treatment after a planned extubation.


Correlation of Ventilation-Perfusion (V/Q) Scan Results as Compared with Clinical Probability of Pulmonary Embolism in African American Population.
Introduction Current guidelines suggest the use of the more specific Wells' score could safely reduce the number of unnecessary scans. There is a lack of research to support whether these guidelines apply to the African American population. This study aims to evaluate the correlation of clinical pretest probability of pulmonary embolism (PE) with ventilation-perfusion (V/Q) scan results in a predominantly African American population and to test whether current guidelines based on studies conducted in other populations hold true in this group. Material and Methods A retrospective descriptive study to determine the diagnostic utility of the V/Q scan was conducted among patients who were seen during January 2012 to January 2016. The study population included patients who underwent a V/Q scan for evaluation of PE. One hundred and seventy-five charts were reviewed and 49 were excluded due to poor quality data. A review of the initial history, as well as discharge summaries, was performed. Wells' probability of PE was compared with the results of the scan. Laboratory tests and imaging studies were reviewed and analyzed. Result The median age of the study population was 63.02 ± 16.12 years. The majority of the study population, 121 patients (92.4%), was African American. Sixty-four (48.9%) VQ scans were done for a low clinical probability for pulmonary embolism as defined by the Wells' clinical score. The most common clinical presentations were shortness of breath (SOB) - 74 (58%), leg pain or swelling - 39 (29.8%), chest pain - 36 (27.4%), and syncope - 4 (3.1%). Sixty-two (96.9 %) patients with low clinical probability had low probability VQ scans (P = 0.03). Among the patients who underwent CT angiography and V/Q scanning, a low probability scan was noted in 25 patients with no pulmonary embolism on CT (96.2 %) (P = 0.006). Conclusions This study showed a strong correlation between low clinical probability and low probability V/Q scans and its utility to safely rule out PE in a predominantly black population. Studies conducted in other populations have detected similar findings.


[Anti-angina effectiveness of the calcium antagonist nifedipine in relation to coronary involvement].
36 patients with chronic stable or the variant form of angina pectoris were subdivided according to their coronary angiogram into 4 groups: Group A with a single highgrade stenosis in one coronary artery, Groups B, C and D with different patterns of occluded, but collateralized coronary arteries supplying noninfarcted myocardium. All patients underwent multiple exercise step tests before (K) and after randomly assigned crossover treatment with 20 mg nifedipine (N), 20 mg isosorbiddinitrate (I), the combination of both (I + N) and Placebo (P). Peak and mean ischemic ST-segment depression, the occurrence of angina pectoris and heart rate were evaluated. The mean ischemic ST-segment depression decreased significantly after N in group A by -28% (p less than 0.01), but was not significantly altered in the groups B, C and D (B: -12%, C: +7%, D: +2%). After I, mean ST-segment depression decreased significantly in all groups (A: -36%, p less than 0.001; B: -27%, p less than 0.001; C: -22%, p less than 0.01; D: -29%, p less than 0.05). The combination of I + N was not better than I alone. Peak ST-depression and angina pectoris paralleled the results of mean ST-depression. The resting heart rate increased significantly after N only in group A (+9%, p less than 0.01) and increased after I in the groups A, B and C (A: +11%, p less than 0.05; B: +12%, p less than 0.05; C: +12%, p less than 0.01). During exercise, heart rate was not significantly different in any group or after any type of treatment.(ABSTRACT TRUNCATED AT 250 WORDS)


[Remission of nephrotic syndrome in patients treated with corticosteroids and other immunosuppressive agents].
Glomerular diseases are hybrid group, mostly immunologically related kidney diseases where pathologic changes start in glomerules, but afterwards within their further process they grasp the other kidney structures. The most common clinical manifestation of the primary glomerulonephritis in the adults is nephrotic syndrome. Therefore, this elaboration aims are to estimate the therapy effect onto the different pathohistologic forms of the primary nephrotic syndrome. Study has been conducted on 41 patients having the primary nephrotic syndrome, prior having normal kidney function. Different pathohistological forms of NS were verified by percutaneous kidney biopsy. Depending on the pathohistological finding the different therapeutic protocoles were applied. Each of the monitored patients were treated by giving steroids at least six weeks dosing them 2 mg/kg/bw/48 h, but not higher then 130 mg/48 h. In the cases of the patients having the given therapy, it did not lead them to the nephrotic syndrome remission, cyclophosphamid was induced in the dosage of 1.5-2.0 mg/kg/bw/48 h. According to the therapy reaction the patients were divided in three groups: the patients with the complete remission, with partial remission and the patients resistant to the treatment. 41 patients have been observed, 28 male and 13 female, aged between 16-69, average 32.3 years old. All patients had normal kidney function, and diseases duration from the moment of the pathohistomorphologic diagnosis had been lasting 0-2 years, from average 6.4 months. According to pathohistological diagnosis the patients were divided into four groups: a) "minimal changes" MCNS (n -11), b) focal--segmental glomerulosclerosis FSGS (n - 10) c) membraneous glomerulonephritis MGN (n-9) d) membranoproliferative MPGN (n-10). The best therapy effect had been resulted within the group with MCNS having 82% cases with the complete remission, while the group with MCNS had reacted the most adversely, 45% were resistant onto the therapy. 18 patients of the total 41 had the complete remission, 14 partial remission, and 9 of them were without remission. The patient having the minimal changes had the best response onto the steroids therapy, while the patients having the expressive proliferative changes did not respond with the complete remission.


Combined CT colonography and 18F-FDG PET of colon polyps: potential technique for selective detection of cancer and precancerous lesions.
The purpose of this study was to determine the feasibility of imaging the colon with fused CT colonography (CTC) and 18F-FDG PET and to correlate the findings with the histologic features of polyps. Eighteen patients with suspected colorectal polyps enrolled in this prospective study. Before colonoscopy, 17 of the patients underwent a combination of FDG PET and CTC. CTC consisted of 4-MDCT merged with PET. PET of the abdomen and pelvis was performed after each CTC scan. One radiologist and one nuclear medicine physician in consensus analyzed PET and CTC fusion data. PET standard uptake value was correlated with the findings at histologic examination of polyps. Patient feasibility was defined as the ability to tolerate prolonged scanning with good colonic distention. Technical feasibility was determined by how closely anatomically matched polyps overlapped on fusion images. Seventeen of 18 patients tolerated scanning. Eighty-five percent of colon segments were optimally distended. Twenty-three of 27 FDG-avid polyps measuring 10 mm or more had excellent overlap at fusion imaging. PET depicted 23 of 39 premalignant polyps and even showed increased tracer activity associated with four small tubular adenomas (4-6 mm). Sixteen benign polyps (10-25 mm) were not depicted on PET. All nine cases of cancer (tumors measuring 11-60 mm) were detected with both PET and CTC. The standard uptake value of malignant tumors ranged from 4 to 20 (mean, 9). However, six benign flat polyps did not exhibit FDG avidity. The novel combination of CTC and PET was feasible in 17 of 18 patients and allowed excellent image correlation in 23 of 27 proven polyps measuring 10 mm or more on PET-CTC fusion. This technique shows promise in accurate anatomic correlation of both malignant and premalignant lesions evaluated with FDG PET.


The Role of Hydrogen Sulfide in Regulation of Cell Death following Neurotrauma and Related Neurodegenerative and Psychiatric Diseases.
Injuries of the central (CNS) and peripheral nervous system (PNS) are a serious problem of the modern healthcare system. The situation is complicated by the lack of clinically effective neuroprotective drugs that can protect damaged neurons and glial cells from death. In addition, people who have undergone neurotrauma often develop mental disorders and neurodegenerative diseases that worsen the quality of life up to severe disability and death. Hydrogen sulfide (H2S) is a gaseous signaling molecule that performs various cellular functions in normal and pathological conditions. However, the role of H2S in neurotrauma and mental disorders remains unexplored and sometimes controversial. In this large-scale review study, we examined the various biological effects of H2S associated with survival and cell death in trauma to the brain, spinal cord, and PNS, and the signaling mechanisms underlying the pathogenesis of mental illnesses, such as cognitive impairment, encephalopathy, depression and anxiety disorders, epilepsy and chronic pain. We also studied the role of H2S in the pathogenesis of neurodegenerative diseases: Alzheimer's disease (AD) and Parkinson's disease (PD). In addition, we reviewed the current state of the art study of H2S donors as neuroprotectors and the possibility of their therapeutic uses in medicine. Our study showed that H2S has great neuroprotective potential. H2S reduces oxidative stress, lipid peroxidation, and neuroinflammation; inhibits processes associated with apoptosis, autophagy, ferroptosis and pyroptosis; prevents the destruction of the blood-brain barrier; increases the expression of neurotrophic factors; and models the activity of Ca2+ channels in neurotrauma. In addition, H2S activates neuroprotective signaling pathways in psychiatric and neurodegenerative diseases. However, high levels of H2S can cause cytotoxic effects. Thus, the development of H2S-associated neuroprotectors seems to be especially relevant. However, so far, all H2S modulators are at the stage of preclinical trials. Nevertheless, many of them show a high neuroprotective effect in various animal models of neurotrauma and related disorders. Despite the fact that our review is very extensive and detailed, it is well structured right down to the conclusions, which will allow researchers to quickly find the proper information they are interested in.


Henoch-Schönlein purpura in adulthood and childhood: two different expressions of the same syndrome.
To assess the possible differences between children (< or = 20 years) and adults (> 20 years) with Henoch-Schönlein purpura (HSP). A retrospective study of an unselected population of patients with HSP who presented to our teaching hospital between 1975 and 1994. Patients were classified as having HSP according to the criteria proposed by Michel et al. Following the above-mentioned criteria, 162 white patients (113 male and 49 female) were classified as having HSP; 46 of the patients were adults (mean +/- SD age 53.2 +/- 16.9 years) and 116 were children (6.9 +/- 3.1 years). We were unable to identify any precipitating event in 72% of the adults and 66% of the children. The frequency of previous drug treatment, primarily antibiotics or analgesics, was similar in both groups, whereas previous upper respiratory tract infection was more frequent among the children (P < 0.02). At symptom onset, cutaneous lesions were the main clinical manifestation in both groups. However, adults had a lower frequency of abdominal pain (P < 0.008) and fever (P < 0.01), and a higher frequency of joint symptoms (P < 0.001). During the clinical course, adults had more frequent (P < 0.001) and severe renal involvement. An increased erythrocyte sedimentation rate was also more frequent in the adults (P < 0.001). Adults required more aggressive therapy, consisting of steroids (P < 0.002) and/or cytotoxic agents (P < 0.001). The outcome was relatively good in both age groups, with complete recovery in 107 children (93.9%) and in 33 adults (89.2%) after a mean +/- SD followup of 19.4 +/- 27.7 (median 12) and 21.8 +/- 33.5 (median 15) months, respectively. In adulthood, HSP, as defined by the criteria proposed by Michel et al, represents a more severe clinical syndrome, with a higher frequency of renal involvement. However, the final outcome of HSP is equally good in patients of both age groups.


Protection of tomato seedlings against infection by Pseudomonas syringae pv. tomato by using the plant growth-promoting bacterium Azospirillum brasilense.
Pseudomonas syringae pv. tomato, the causal agent of bacterial speck of tomato, and the plant growth-promoting bacterium Azospirillum brasilense were inoculated onto tomato plants, either alone, as a mixed culture, or consecutively. The population dynamics in the rhizosphere and foliage, the development of bacterial speck disease, and their effects on plant growth were monitored. When inoculated onto separate plants, the A. brasilense population in the rhizosphere of tomato plants was 2 orders of magnitude greater than the population of P. syringae pv. tomato (10(7) versus 10(5) CFU/g [dry weight] of root). Under mist chamber conditions, the leaf population of P. syringae pv. tomato was 1 order of magnitude greater than that of A. brasilense (10(7) versus 10(6) CFU/g [dry weight] of leaf). Inoculation of seeds with a mixed culture of the two bacterial strains resulted in a reduction of the pathogen population in the rhizosphere, an increase in the A. brasilense population, the prevention of bacterial speck disease development, and improved plant growth. Inoculation of leaves with the mixed bacterial culture under mist conditions significantly reduced the P. syringae pv. tomato population and significantly decreased disease severity. Challenge with P. syringae pv. tomato after A. brasilense was established in the leaves further reduced both the population of P. syringae pv. tomato and disease severity and significantly enhanced plant development. Both bacteria maintained a large population in the rhizosphere for 45 days when each was inoculated separately onto tomato seeds (10(5) to 10(6) CFU/g [dry weight] of root). However, P. syringae pv. tomato did not survive in the rhizosphere in the presence of A. brasilense. Foliar inoculation of A. brasilense after P. syringae pv. tomato was established on the leaves did not alleviate bacterial speck disease, and A. brasilense did not survive well in the phyllosphere under these conditions, even in a mist chamber. Several applications of a low concentration of buffered malic acid significantly enhanced the leaf population of A. brasilense (>10(8) CFU/g [dry weight] of leaf), decreased the population of P. syringae pv. tomato to almost undetectable levels, almost eliminated disease development, and improved plant growth to the level of uninoculated healthy control plants. Based on our results, we propose that A. brasilense be used in prevention programs to combat the foliar bacterial speck disease caused by P. syringae pv. tomato.


Criticality in the brain: A synthesis of neurobiology, models and cognition.
Cognitive function requires the coordination of neural activity across many scales, from neurons and circuits to large-scale networks. As such, it is unlikely that an explanatory framework focused upon any single scale will yield a comprehensive theory of brain activity and cognitive function. Modelling and analysis methods for neuroscience should aim to accommodate multiscale phenomena. Emerging research now suggests that multi-scale processes in the brain arise from so-called critical phenomena that occur very broadly in the natural world. Criticality arises in complex systems perched between order and disorder, and is marked by fluctuations that do not have any privileged spatial or temporal scale. We review the core nature of criticality, the evidence supporting its role in neural systems and its explanatory potential in brain health and disease.


Endometrial stem cells repair injured endometrium and induce angiogenesis via AKT and ERK pathways.
Intrauterine adhesions are common acquired endometrial syndromes secondary to endometrial injury, with limited effective therapies. Recently, several studies have reported that bone marrow stem cells (BMSCs) could repair injured endometrium in animal experiments. However, the role of stem cells in endometrial injury repair and its therapeutic mechanisms remain unclear. Here, we established mouse endometrial injury model and examined the benefit of human endometrial mesenchymal stem cells derived from menstrual blood (MenSCs) in restoration of injured endometrium. Injured endometrium exhibited significantly accelerated restoration at Day 7 after MenSCs transplantation, with increased endometrial thickness and microvessel density. Moreover, the fertility of mice with injured endometrium was improved, with higher conception rate (53.57% vs 14.29%, P = 0.014) and larger embryo number (3.1 ± 0.6 vs 0.9 ± 0.7, P = 0.030) in MenSCs group than control group, while no difference was found in undamaged horns between two groups. Conditioned medium from MenSCs (MenSCs-CM) could decrease H2O2-induced apoptosis of human umbilical vein endothelial cells (HUVECs) and promote proliferation, migration and angiogenesis. Angiogenesis effect of MenSCs-CM was also confirmed in Matrigel plug assay in mice. Furthermore, we discovered that MenSCs-CM could activate AKT and ERK pathways and induce the overexpression of eNOS, VEGFA, VEGFR1, VEGFR2 and TIE2 in HUVECs, which are critical in MenSCs-CM-induced angiogenesis. Angiogenesis induced by MenSCs-CM could be reversed by inhibitors of AKT and/or ERK. Taken together, we concluded that MenSCs could restore injured endometrium and improve the fertility of the endometrial injury mice, which was partially attributed to angiogenesis induced by MenSCs.


The Antibacterial Mechanism of Silver Nanoparticles and Its Application in Dentistry.
Nanotechnology has recently emerged as a rapidly growing field with numerous biomedical science applications. At the same time, silver has been adopted as an antimicrobial material and disinfectant that is relatively free of adverse effects. Silver nanoparticles possess a broad spectrum of antibacterial, antifungal and antiviral properties. Silver nanoparticles have the ability to penetrate bacterial cell walls, changing the structure of cell membranes and even resulting in cell death. Their efficacy is due not only to their nanoscale size but also to their large ratio of surface area to volume. They can increase the permeability of cell membranes, produce reactive oxygen species, and interrupt replication of deoxyribonucleic acid by releasing silver ions. Researchers have studied silver nanoparticles as antimicrobial agents in dentistry. For instance, silver nanoparticles can be incorporated into acrylic resins for fabrication of removable dentures in prosthetic treatment, composite resin in restorative treatment, irrigating solution and obturation material in endodontic treatment, adhesive materials in orthodontic treatment, membrane for guided tissue regeneration in periodontal treatment, and titanium coating in dental implant treatment. Although not all authorities have acknowledged the safety of silver nanoparticles, no systemic toxicity of ingested silver nanoparticles has been reported. A broad concern is their potential hazard if they are released into the environment. However, the interaction of nanoparticles with toxic materials and organic compounds can either increase or reduce their toxicity. This paper provides an overview of the antibacterial use of silver nanoparticles in dentistry, highlighting their antibacterial mechanism, potential applications and safety in clinical treatment.


Accuracy of nurse aides' functional health assessments of nursing home residents.
Nurse aides provide assessments of nursing home residents' functional health for use in care planning and quality assurance. Nurse practitioner assessments can serve as a standard for analysis of nurse aides' accuracy. This study compared nurse aide to nurse practitioner assessments of nursing home residents' functional health with regard to possible bias and extent of correlation. Nurse aides' accuracy in assessing nursing home residents' activities of daily living was evaluated by comparisons to assessments performed by a master's-prepared nurse practitioner using four functional assessment instruments: the Barthel Index, the Katz Index of Activities of Daily Living, the Multidimensional Observation Scale for Elderly Subjects, and the Scaled Outcome Criteria. Data were collected in a 159-bed nonprofit nursing home licensed for skilled and intermediate care. Residents had a wide variety of functional and cognitive abilities and disabilities. Ninety-six nursing home residents provided data for the study. Functional health assessments by 24 nurse aides, each assessing 4 different nursing home residents, were compared to those of 1 nurse practitioner. Statistical analysis of accuracy used paired samples t-tests and Pearson product moment correlation coefficients. Nurse aide assessments and nurse practitioner assessments were highly correlated. Most functional health assessments evidenced no significant nurse aide bias. When bias was present it usually resulted from nurse aides electing more optimistic choices when using an assessment instrument that offered fewer response levels for rating residents. Nurse aides can accurately use well-calibrated instruments to assess nursing home residents' functional health. Demonstration of assessment accuracy in nurse aides, who provide the majority of direct care for nursing home residents, documented a valuable clinical resource for planning and evaluating resident care.


The cytologic diagnosis of low-grade transitional cell carcinoma.
Current theory suggests that transitional cell carcinoma (TCC) occurs as either of 2 disease processes, each of which has a distinct cytologic appearance and clinical course: low-grade and high-grade TCC. Urinary cytology has become a mainstay technique for monitoring disease recurrence in patients with TCC. Most cases of high-grade TCC can be diagnosed accurately in urinary cytology specimens. However, the cytologic diagnosis of low-grade TCC is difficult; these tumors exhibit subtle cytomorphologic alterations that are difficult to distinguish from benign or reactive processes. The cytologic criteria most useful for diagnosing low-grade TCC in urinary cytology specimens are reviewed. Additionally, the discussion includes some of the new ancillary tests that are emerging as possible diagnostic aids for the detection of low-grade urothelial neoplasms.


The usefulness of design of experimentation in defining the effect difficult airway factors and training have on simulator oral-tracheal intubation success rates in novice intubators.
This exploratory study examined novice intubators and the effect difficult airway factors have on pre- and posttraining oral-tracheal simulation intubation success rates. Using a two-level, full-factorial design of experimentation (DOE) involving a combination of six airway factors (curved vs. straight laryngoscope blade, trismus, tongue edema, laryngeal spasm, pharyngeal obstruction, or cervical immobilization), 64 airway scenarios were prospectively randomized to 12 critical care nurses to evaluate pre- and posttraining first-pass intubation success rates on a simulator. Scenario variables and intubation outcomes were analyzed using a generalized linear mixed-effects model to determine two-way main and interactive effects. Interactive effects between the six study factors were nonsignificant (p = 0.69). For both pre- and posttraining, main effects showed the straight blade (p = 0.006), tongue edema (p = 0.0001), and laryngeal spasm (p = 0.004) significantly reduced success rates, while trismus (p = 0.358), pharyngeal obstruction (p = 0.078), and cervical immobilization did not significantly change the success rate. First-pass intubation success rate on the simulator significantly improved (p = 0.005) from pre- (19%) to posttraining (36%). Design of experimentation is useful in analyzing the effect difficult airway factors and training have on simulator intubation success rates. Future quality improvement DOE simulator research studies should be performed to help clarify the relationship between simulator factors and patient intubation rates.


Differential modulation of spontaneous and evoked neurotransmitter release from hair cells: some novel hypotheses.
It has been generally accepted that even in the absence of mechanical stimulation of the transductional elements, a resting depolarizing current exists which is ultimately responsible for the spontaneous release of neurotransmitter. Movement of the transductional elements modulates this resting current and thereby the evoked release of neurotransmitter occurs. Recent data from our laboratory and others have led us to question whether the relationship between spontaneous and evoked neurotransmitter release is as simple as stated. Indeed, a variety of experimental manipulations appear to influence the two modes of release differently. Examination of our results and the results of others has led us to four hypotheses: 1. the two modes of neurotransmitter release are processed differently by the hair cells; 2. cyclic AMP is involved in spontaneous but not evoked neurotransmitter release; 3. there is a positive feedback step involving an excitatory amino acid and its receptor on the hair cell in evoked neurotransmitter release and; 4. different pools of calcium are involved according to the mode of release. Accordingly, there may be several biochemical steps between the transductional movement of the stereocilia at the apex of the hair cells and the ultimate release of the neurotransmitter at the base of these cells. Some of these biochemical steps are different depending on whether the mode of release is spontaneous or evoked. These biochemical steps may amplify or at least interact with the biophysical processes previously described in the hair cells.


Closing the Loop in Adults, Children and Adolescents With Suboptimally Controlled Type 1 Diabetes Under Free Living Conditions: A Psychosocial Substudy.
The objective was to explore psychosocial experiences of closed loop technology for adults, children, and adolescents with type 1 diabetes and their parents taking part in two multicenter, free-living, randomized crossover home studies. Participants using insulin pump therapy were randomized to either 12 weeks of automated closed-loop glucose control, then 12 weeks of sensor augmented insulin pump therapy (open loop), or vice versa. Closed loop was used for 24 hours by adults and overnight only by children and adolescents. Participants completed the Diabetes Technology Questionnaire (DTQ) periodically and shared their views in semistructured interviews. This analysis characterizes the impact of the technology, positive and negative aspects of living with the device, alongside participants' expectations, hopes, and anxieties. Participants were 32 adults, age 38.6 ± 9.6 years, 55% male, and 26 children, mean age 12 years (range 6-18 years), 54% male. DTQ results indicated moderately favorable impact of, and satisfaction with, both open and closed loop interventions, but little evidence of a comparative advantage of either. Key positive themes included perceived improved blood glucose control, improved general well-being, particularly on waking, improved sleep, reduced burden of diabetes, and visibility of data. Key negative themes included having to carry around the equipment and dislike of the pump and second cannula (ie, sensor) inserted. Overall, participants reported a positive experience of the closed loop technology. Results are consistent with previous research with size of equipment continuing to be a problem. Progress is being made in the usability of the closed-loop system.


[Circadian rhythm of melatonin in patients with hypertension].
Few reports, especially experimental ones indicate cause-effect connection between melatonin (Mel) and blood pressure. The aim of the study was to assess connection between a daily profile of Mel concentration in saliva and 24-hour cycle of blood pressure in 22 patients with moderate hypertension (12 men, 10 women), 20 patients with severe hypertension (9 men, 11 women) and 25 healthy subjects with normal blood pressure (13 men, 12 women). The age of studied subjects oscillated between 37 and 46 years. Concentrations of Mel were assessed in saliva collected during the day with intervals of 3 hours with the use of RIA method. Blood pressure assessment was done at the same time. A significant decrease of mesor's value and amplitude of Mel rhythm without essential changes in acrophase's value were observed in men and women with severe hypertension. Mean concentrations of Mel in saliva were significantly decreased from 8 pm to 5 am. Significant, negative correlation between nighttime Mel concentrations in saliva and both systolic and diastolic blood pressure was shown in patients with moderate and severe hypertension. Correlation coefficients values were in general higher in patients with severe hypertension. Results of our studies seem to confirm the conception that decreased Mel secretion can be one of the causes of hypertension.


Combined spinal epidural analgesia for labor with and without 3 ml of 1.5% epidural lidocaine.
Lidocaine is used as a 'test dose' to detect misplacement of epidural catheters. When administered immediately after the initiation of a combined spinal epidural for labor analgesia, it was found to interfere with motor function. The authors hypothesized that an epidural test dose of 3 ml of 1.5% lidocaine injected immediately after the initiation of combined spinal epidural analgesia and followed by a continuous epidural infusion would prolong the duration of analgesia and decrease the incidence of breakthrough pain. Sixty nulliparous parturients were recruited for this randomized controlled trial. Combined spinal epidural analgesia was performed with intrathecal levobupivacaine 2.5 mg and fentanyl 25 microg. The patients were randomized into three groups: group L receiving 3 ml of 1.5% epidural lidocaine, group S 3 ml of normal saline, and group C the control group receiving no test solution. An epidural infusion of 0.1% levobupivacaine and 2 microg of fentanyl/ml at 10 ml/hr was initiated immediately. Their need for supplementary analgesia, duration of analgesia, sensory block, motor block, side-effects, and overall satisfaction were recorded. The breakthrough pain rate in all groups were similar. Their mean duration of analgesia (mean survival times) were not significantly different. Group C had the lowest incidence of lower limb motor block. The satisfaction scores were significantly highest in group L. The administration of epidural lidocaine and saline immediately after intrathecal levobupivacaine 2.5 mg and fentanyl 25 microg did not reduce the incidence of breakthrough pain in combined spinal epidural labor.


[Hyponatremia induced by hypoglycemic sulfonamides: a study of 70 patients].
Sodium plasma level was followed every three months in 70 non insulin dependent diabetic patients; 112 prescriptions were performed. Five different oral hypoglycemic drugs were used but patients were taken no more than one drug in the same time. 21% of prescription were associated with a low sodium plasma level but it was beneath 129 meq in only 8% of them; a low sodium plasma level was observed with every oral hypoglycemic agent; when the drug was interrupted a normal sodium plasma level was restored; when patients were put under another agent, hyponatremia did not recidive. 67% of patients were free of any symptom. Interference with biological parameters, age of the patients, duration of diabetes and other therapy are studied.


SrFe0.5Ru0.5O2: square-planar Ru2+ in an extended oxide.
Low-temperature topochemical reduction of the cation disordered perovskite phase SrFe(0.5)Ru(0.5)O(3) with CaH(2) yields the infinite layer phase SrFe(0.5)Ru(0.5)O(2). Thermogravimetric and X-ray absorption data confirm the transition metal oxidation states as SrFe(0.5)(2+)Ru(0.5)(2+)O(2); thus, the title phase is the first reported observation of Ru(2+) centers in an extended oxide phase. DFT calculations reveal that, while the square-planar Fe(2+) centers adopt a high-spin S = 2 electronic configuration, the square-planar Ru(2+) cations have an intermediate S = 1 configuration. This combination of S = 2, Fe(2+) and S = 1, Ru(2+) is consistent with the observed spin-glass magnetic behavior of SrFe(0.5)Ru(0.5)O(2).


Disruption of neurogenesis in the subventricular zone of adult mice, and in human cortical neuronal precursor cells in culture, by amyloid beta-peptide: implications for the pathogenesis of Alzheimer's disease.
The adult mammalian brain contains populations of stem cells that can proliferate and then differentiate into neurons or glia. The highest concentration of such neural progenitor cells (NPC) is located in the subventricular zone (SVZ) and these cells can produce new olfactory bulb and cerebral cortical neurons. NPC may provide a cellular reservoir for replacement of cells lost during normal cell turnover and after brain injury. However, neurogenesis does not compensate for neuronal loss in age-related neurodegenerative disorders such as Alzheimer's disease (AD), suggesting the possibility that impaired neurogenesis contributes to the pathogenesis of such disorders. We now report that amyloid beta-peptide (Abeta), a self-aggregating neurotoxic protein thought to cause AD, can impair neurogenesis in the SVZ/cerebral cortex of adult mice and in human cortical NPC in culture. The proliferation and migration of NPC in the SVZ of amyloid precursor protein (APP) mutant mice, and in mice receiving an intraventricular infusion of Abeta, were greatly decreased compared to control mice. Studies of NPC neurosphere cultures derived from human embryonic cerebral cortex showed that Abeta can suppress NPC proliferation and differentiation, and can induce apoptosis. The adverse effects of Abeta on neurogenesis were associated with a disruption of calcium regulation in the NPC. Our data show that Abeta can impair cortical neurogenesis, and suggest that this adverse effect of Abeta contributes to the depletion of neurons and the resulting olfactory and cognitive deficits in AD.


Fingerprint Analysis and Identification of Strains ST309 as a Potential High Risk Clone in a Pseudomonas aeruginosa Population Isolated from Children with Bacteremia in Mexico City.
Pseudomonas aeruginosa is an opportunistic pathogen and is associated with nosocomial infections. Its ability to thrive in a broad range of environments is due to a large and diverse genome of which its accessory genome is part. The objective of this study was to characterize P. aeruginosa strains isolated from children who developed bacteremia, using pulse-field gel electrophoresis, and in terms of its genomic islands, virulence genes, multilocus sequence type, and antimicrobial susceptibility. Our results showed that P. aeruginosa strains presented the seven virulence genes: toxA, lasB, lecA, algR, plcH, phzA1, and toxR, a type IV pilin alleles (TFP) group I or II. Additionally, we detected a novel pilin and accessory gene, expanding the number of TFP alleles to group VI. All strains presented the PAPI-2 Island and the majority were exoU+ and exoS+ genotype. Ten percent of the strains were multi-drug resistant phenotype, 18% extensively drug-resistant, 68% moderately resistant and only 3% were susceptible to all the antimicrobial tested. The most prevalent acquired β-Lactamase was KPC. We identified a group of ST309 strains, as a potential high risk clone. Our finding also showed that the strains isolated from patients with bacteremia have important virulence factors involved in colonization and dissemination as: a TFP group I or II; the presence of the exoU gene within the PAPI-2 island and the presence of the exoS gene.


Controlling feeding behavior by chemical or gene-directed targeting in the brain: what's so spatial about our methods?
Intracranial chemical injection (ICI) methods have been used to identify the locations in the brain where feeding behavior can be controlled acutely. Scientists conducting ICI studies often document their injection site locations, thereby leaving kernels of valuable location data for others to use to further characterize feeding control circuits. Unfortunately, this rich dataset has not yet been formally contextualized with other published neuroanatomical data. In particular, axonal tracing studies have delineated several neural circuits originating in the same areas where ICI injection feeding-control sites have been documented, but it remains unclear whether these circuits participate in feeding control. Comparing injection sites with other types of location data would require careful anatomical registration between the datasets. Here, a conceptual framework is presented for how such anatomical registration efforts can be performed. For example, by using a simple atlas alignment tool, a hypothalamic locus sensitive to the orexigenic effects of neuropeptide Y (NPY) can be aligned accurately with the locations of neurons labeled by anterograde tracers or those known to express NPY receptors or feeding-related peptides. This approach can also be applied to those intracranial "gene-directed" injection (IGI) methods (e.g., site-specific recombinase methods, RNA expression or interference, optogenetics, and pharmacosynthetics) that involve viral injections to targeted neuronal populations. Spatial alignment efforts can be accelerated if location data from ICI/IGI methods are mapped to stereotaxic brain atlases to allow powerful neuroinformatics tools to overlay different types of data in the same reference space. Atlas-based mapping will be critical for community-based sharing of location data for feeding control circuits, and will accelerate our understanding of structure-function relationships in the brain for mammalian models of obesity and metabolic disorders.


A comparison of neural differentiation and retinal transplantation with bone marrow-derived cells and retinal progenitor cells.
Retinal progenitor cells (RPCs) are immature precursors that can differentiate into retinal neurons, including photoreceptors. Recently, it has been reported that bone marrow-derived cells may also be capable of differentiation into cells of central nervous system lineage, including retinal neurons. We compared these two cell types to evaluate their potential as a source of cells for retinal transplantation. Marrow stromal cells (MSCs) and macrophages were isolated from enhanced green fluorescence protein mice. MSCs were cultured with brain-derived neurotrophic factor, nerve growth factor, and basic fibroblast growth factor to induce neuronal differentiation. RPCs were cultured under the same conditions or with 10% fetal bovine serum. Neuronal marker expression was examined and compared between MSCs and RPCs. MSCs, macrophages, and RPCs were also cultured with explanted retinas from rhodopsin knockout mice to study their potential for retinal integration. MSCs expressed neuronal and retina-specific markers by reverse transcription-polymerase chain reaction and immunocytochemistry. Both types of cells migrated into retinal explants and expressed neurofilament 200, glial fibrillary acidic protein, protein kinase C-alpha, and recoverin. RPCs expressed rhodopsin, a photoreceptor marker we never detected in MSCs. A majority of bone marrow derived-macrophages differentiated into cells that resembled microglia, rather than neural cells, in the explanted retina. This study shows that RPCs are likely to be a preferred cell type for retinal transplantation studies, compared with MSCs. However, MSCs may remain an attractive candidate for autologous transplantation.


EUS-guided portal pressure gradient measurement in patients with acute or subacute portal hypertension.
EUS-guided portal pressure gradient (EUS-PPG) measurement is a novel method to evaluate portal hypertension severity. In this study, we determined the consistency between EUS-PPG and hepatic venous pressure gradient (HVPG) measurements in patients with acute or subacute portal hypertension. Twelve patients were prospectively enrolled. EUS-PPG measurements were performed using a 22-gauge FNA needle and a central venous pressure measurement monitor. The HVPG measurements were performed using the transjugular approach. If an HVPG measurement was not attainable and the patient underwent transjugular intrahepatic portosystemic shunt (TIPS) treatment, a PPG was recorded as a reference standard during the procedure. We assessed the feasibility and safety of EUS-PPG and calculated the correlation between the 2 measurements. EUS-PPG measurements were successful in 11 patients (91.7%). Subsequent HVPG measurements failed in 2 patients with Budd-Chiari syndrome (hepatic vein occlusion subtype), 1 of whom underwent TIPS treatment to obtain transjugular PPG data. A small shunt was found during 1 HVPG measurement that introduced inaccuracy. Nine patients were included in the statistical analysis. Mean EUS-PPG and HVPG/PPG (transjugular) were 18.07 ± 4.32 mm Hg and 18.82 ± 3.43 mm Hg, respectively. Pearson's correlation coefficient between the 2 methods was .923 (P < .001). EUS-PPG measurement using a 22-gauge FNA needle was a safe and accurate method to evaluate portal hypertension and has the potential to supplement the measurement of HVPG in liver diseases. (Clinical trial registration number: ChiCTR1800017317.).


[Up-date in spontaneous cerebral hemorrhage].
Non-aneurismatic spontaneous cerebral hemorrhage or intracranial hemorrhage accounts for 10-15% of total cerebral vascular accidents. Depending on its site it can may be intraparenchymal or intraventricular. The most frequent location is in the basal ganglia and its predominant etiology is poorly-controlled arterial hypertension. In Spain, the incidence of intracerebral hemorrhage is estimated to be 15 cases per 100,000 population/ year, this being more frequent in males over 55 years old. Intracranial hemorrhage is less frequent than ischemic stroke, but has higher mortality and morbidity, it being one of the first causes of severe disability. Cerebral hemorrhage is not a monophasic phenomenon which abates immediately, because the hematoma continues to increase in the first 24 hours. Due to this reason and because of their characteristics of the disease itself, these are critical patients who must be admitted in to Intensive Care Unit where hemodynamic and cardiorespiratory control should be made as well as strict monitoring of the awareness level and remaining neuromonitoring standard parameters. In this paper, we review some aspects of the epidemiology, physiopathology, clinical presentation, diagnosis and the different therapeutic options, performing an up-date on the treatment of intracranial hemorrhage from both the medical and surgical point of view.


Detection and characterisation of disseminated tumour cells in bone marrow of breast cancer patients by immunostaining of Her-2 and MUC-1 in combination with Thomsen-Friedenreich (CD176).
Disseminated tumour cells (DTCs) in the bone marrow derive from many primary tumours, such as breast cancer. Their mere existence hints to present or future metastasis and implicates a worse prognosis for the patient. DTCs may possess different characteristics in comparison to the primary tumour due to events like Epithelial-Mesenchymal-Transition. Therefore, these cells might be able to survive chemotherapy and cause relapses of the disease at a later point. We aimed to detect and further characterise DTCs by an immunostaining approach with three different antigen markers (Her-2, MUC-1 and TF, also known as CD 176). For that reason, bone marrow of 41 breast cancer patients was obtained during surgery; DTCs were enriched by density gradient centrifugation and cytospins were prepared. After fixation, immunofluorescent double-stainings were carried out with antibodies against CD176 in combination with HER-2 or MUC-1. Cells co-expressing two antigens were found in all staining combinations (Her-2 and CD176: 46.14%; MUC-1 and CD176: 18.15% of all cases). Cells that stained for a single antigen only were also found (Her-2: 36.86%; MUC-1: 34.45%; CD176: 29.65% of all cases). Significant correlations between the stainings of all markers could be shown (p<0,001). In conclusion, Thomsen-Friedenreich Antigen (TF, CD176) is a promising marker in combination with the established marker Her-2 and other markers like MUC-1. These results may serve as a basis for future DTC detection routines and help to individualize medical treatment, reducing side effects and increasing the efficiency of the therapy.


Current status of fetal age and growth assessment.
Reliable fetal age assessment is now possible using antenatal sonography. Crown-rump length, biparietal diameter, head perimeter and femur length are the most widely used parameters, while biocular distance and humerus length are of ancillary value. Techniques of sonographic measurement and clinical applications are reviewed and nomograms relating these parameters to gestational age are presented. Multiple sonographic parameters have been used to diagnose altered fetal growth. Clinical applications and pitfalls in the diagnosis of IUGR are discussed when biparietal diameter, abdominal perimeter, head to abdomen ratio, total intrauterine volume, qualitative amniotic fluid determination, fetal urine production rate and estimated fetal weight are used.


Intraoperative Radiotherapy of Breast Cancer and Its Biological Effects.
Conservative breast cancer surgery followed by radiation therapy is the standard treatment for this type of cancer. Numerous studies demonstrate that 90% of local recurrences after traditional surgery occur in the same quadrant as the primary cancer. The published data suggest that the wound healing process after surgery alters the area surrounding the original tumor and the modified microenvironment is more favorable for the tumor to recur. The majority of metastases within scar initiated much research, and the consequences of these studies led to clinical trials aimed at assessing whether localized radiotherapy, such as intraoperative radiotherapy (IORT), would be more effective in inhibiting formation of local recurrence than the standard postoperative whole breast radiotherapy. IORT involves irradiation of diseased tissue directly during surgery. The rationale for this approach is the fact that the increase in the radiation dose increases local tumor control, which is the primary goal of radiation therapy. The biological basis of this process are still not thoroughly understood. Gaining new knowledge about the recurrence formation at the molecular level could serve as a starting point for further analysis and to create an opportunity to identify new targets of therapy, and possibly new therapeutic agents.


A chimpanzee-passaged human immunodeficiency virus isolate is cytopathic for chimpanzee cells but does not induce disease.
The human immunodeficiency virus type 1 (HIV-1) readily infects both humans and chimpanzees, but the pathologic outcomes of infection in these two species differ greatly. In attempts to identify virus-cell interactions that might account for this differential pathogenicity, chimpanzee peripheral blood lymphocytes and bone marrow macrophages were assessed in vitro for their ability to support the replication of several HIV-1 isolates. Although the IIIb, RF, and MN isolates did not readily infect chimpanzee peripheral blood lymphocytes, an isolate of HIV-1 passaged in vivo in chimpanzees not only replicated well in both chimpanzee peripheral blood lymphocytes and bone marrow macrophages but also was cytopathic for chimpanzee CD4+ lymphocytes. Because no evidence of HIV-induced disease has been observed in chimpanzees infected with this isolate, in vitro replication to high titers with concomitant loss of CD4+ cells is not, in this instance, a correlate of pathogenicity. These observations, therefore, indicate that caution must be used when making extrapolations from in vitro data to in vivo pathogenesis.


Hospital information system-based pharmacovigilance for elderly patients: practices and results.
To evaluate the effect of a pharmacovigilance system on potentially inappropriate medication prescriptions for elderly patients. The retrospective study was conducted at Shaanxi Provincial People's Hospital, China, after approval from the ethics review committee, and comprised data from May 2020 to April 2021, and comprised prescriptions related to elderly patients aged at least 65 years. Number of medication risk assessment entries, number of intervened medical orders on outpatients and inpatients number of medical order prompts, and number of physician communication with prescription-checking pharmacists were noted. Potential drug interaction rate was compared between pre- implementation from May to October 2020 and post-implementation from November 2020 to April 2021. Besides, the usage of sedatives and hypnotics and potentially inappropriate medication was noted for the period from January to June 2021 to evaluate the sustained effect of pharmacovigilance system. Data was analysed using SPSS 19. A total of 118 drugs were involved in the 3911 entries of outpatient prescription warnings, of which 19 drugs accounted for 3156 (80%). Besides, a total of 113 drugs were involved in the 3999 entries of inpatient prescription warnings, of which 19 drugs accounted for 3199 (80%) The overall prevalence of potentially inappropriate medication related to sedatives and hypnotics decreased post-intervention as warning percentage was 16.1% in January and 6.7% in June among outpatients. On inpatients, the warning percentage was 30.6% in January and 6.1% in June. The pharmacovigilance system could reduce potentially inappropriate medication and provide deeper technical support for the safety of medical behaviour and individualised treatment of patients.


Effect of PTEN inactivating germline mutations on innate immune cell function and thyroid cancer-induced macrophages in patients with PTEN hamartoma tumor syndrome.
PTEN hamartoma tumor syndrome (PHTS) is caused by inactivating germline PTEN mutations with subsequent activation of Akt-mTOR signaling, leading to an increased risk of developing thyroid carcinoma (TC). Activation of Akt-mTOR signaling is essential for innate immune cell activation and reprogramming of TC-induced macrophages. Here, we aim to assess the effect of PTEN mutations on innate immune cell function in PHTS patients, especially in the context of TC, using a unique ex vivo model. Monocyte-derived cytokine responses were assessed in 29 PHTS patients and 29 controls. To assess the functional profile of TC-induced-macrophages, a co-culture model with two TC cell lines was performed. Rapamycin, a lactate transport blocker and metformin were used as modulators of the Akt-mTOR pathway and cell metabolism. Monocytes from PHTS patients showed increased production of IL-6, TNF-α, IL-8 and MCP-1, and higher lactate production. After co-culture with TC cell lines, TC-induced macrophages showed significantly increased production of cytokines in both patients and controls, especially after co-culture with a PTEN-deficient TC cell line; these effects were abolished after use of rapamycin or a lactate transport blocker. Metformin blocked the production of anti-inflammatory cytokines. In conclusion, innate immune cells from PHTS patients have increased lactate production and a more proinflammatory phenotype, especially after co-culture with PTEN-deficient TC. Metformin promotes a proinflammatory phenotype by blocking anti-inflammatory cytokine response, whereas rapamycin reduces production of proinflammatory cytokines. This indicates that PHTS patients may benefit from treatment with mTOR blocking agents to limit the inflammatory response in the tumor microenvironment.


Exogenously added BMP-6, BMP-7 and VEGF may not enhance the osteogenic differentiation of human adipose stem cells.
In the present study bone morphogenetic protein (BMP)-6 alone or in synergy with BMP-7 and vascular endothelial growth factor (VEGF) were tested with human adipose stem cells (hASCs) seeded on cell culture plastic or 3D bioactive glass. Osteogenic medium (OM) was used as a positive control for osteogenic differentiation. The same growth factor groups were also tested combined with OM. None of the growth factor treatments could enhance the osteogenic differentiation of hASCs in 3D- or 2D-culture compared to control or OM. In 3D-culture OM promoted significantly total collagen production, whereas in 2D-culture OM induced high total ALP activity and mineralization compared to control and growth factors groups, but also high cell proliferation. In this study, hASCs did not respond to exogenously added growth although various parameters of the study set-up may have affected these findings contradictory to the previous literature.


Colonization of cattle intestines by Campylobacter jejuni and Campylobacter lanienae.
The location and abundance of Campylobacter jejuni and Campylobacter lanienae in the intestines of beef cattle were investigated using real-time quantitative PCR in two studies. In an initial study, digesta and tissue samples were obtained along the digestive tract of two beef steers known to shed C. jejuni and C. lanienae (steers A and B). At the time of slaughter, steer B weighed 540 kg, compared to 600 kg for steer A, yet the intestine of steer B (40.5 m) was 36% longer than the intestine of steer A (26.1 m). In total, 323 digesta samples (20-cm intervals) and 998 tissue samples (3.3- to 6.7-cm intervals) were processed. Campylobacter DNA was detected in the digesta and in association with tissues throughout the small and large intestines of both animals. Although C. jejuni and C. lanienae DNA were detected in both animals, only steer A contained substantial quantities of C. jejuni DNA. In both digesta and tissues of steer A, C. jejuni was present in the duodenum and jejunum. Considerable quantities of C. jejuni DNA also were observed in the digesta obtained from the cecum and ascending colon, but minimal DNA was associated with tissues of these regions. In contrast, steer B contained substantial quantities of C. lanienae DNA, and DNA of this bacterium was limited to the large intestine (i.e., the cecum, proximal ascending colon, descending colon, and rectum); the majority of tissue-associated C. lanienae DNA was present in the cecum, descending colon, and rectum. In a second study, the location and abundance of C. jejuni and C. lanienae DNA were confirmed in the intestines of 20 arbitrarily selected beef cattle. DNA of C. jejuni and C. lanienae were detected in the digesta of 57% and 95% of the animals, respectively. C. jejuni associated with intestinal tissues was most abundant in the duodenum, ileum, and rectum. However, one animal contributed disproportionately to the abundance of C. jejuni DNA in the ileum and rectum. C. lanienae was most abundant in the large intestine, and the highest density of DNA of this bacterium was found in the cecum. Therefore, C. jejuni colonized the proximal small intestine of asymptomatic beef cattle, whereas C. lanienae primarily resided in the cecum, descending colon, and rectum. This information could be instrumental in developing efficacious strategies to manage the release of these bacteria from the gastrointestinal tracts of cattle.


Microscale ZnO with controllable crystal morphology as a platform to study antibacterial action on Staphylococcus aureus.
Nano- and microcrystalline ZnO is an inexpensive, easily synthesized material with a multitude of applications. Its usefulness in the present and future stems from its exceptional optoelectronic, structural, and chemical characteristics as well as a broad range of production techniques. One application comes from its ability to inhibit bacterial growth. Despite the well-documented, vigorously studied antimicrobial action of ZnO particles, the most fundamental physical and chemical mechanisms driving growth inhibition are still not well identified. Particularly, the nature of interactions between ZnO surfaces and extracellular material is not totally clear. This is important given the anisotropic lattice of ZnO leading to two characteristically different lattice terminations: polar and nonpolar, polar being electrically charged with many defect sites and nonpolar being electrically neutral while remaining relatively defect-free. In this work, we employ a hydrothermal growth protocol that allows us to produce ZnO microcrystals with dependable control of morphology and, particularly, the relative abundances of polar and nonpolar free surfaces. This functions as a platform for our investigations into surface-surface interactions behind the antibacterial action of ZnO microcrystals. In our studies, we produced ZnO crystals comparable in size or larger than Staphylococcus aureus bacteria. This was done intentionally to ensure that the ZnO particles would not internalize into the bacterial cells. Our experiments were performed in conjunction with surface photovoltage studies of ZnO crystals to characterize electronic structure and charge dynamics that might be contributing to the antibacterial properties of our samples. We report on the interactions between ZnO microcrystalline surfaces and extracellular material of Staphylococcus aureus bacteria.


Identification of Cell-Surface Proteins Endocytosed by Human Brain Microvascular Endothelial Cells In Vitro.
Cell-surface proteins that can endocytose into brain microvascular endothelial cells serve as promising candidates for receptor-mediated transcytosis across the blood-brain barrier (BBB). Here, we comprehensively screened endocytic cell-surface proteins in hCMEC/D3 cells, a model of human brain microvascular endothelial cells, using surface biotinylation methodology and sequential window acquisition of all theoretical fragment-ion spectra-mass spectrometry (SWATH-MS)-based quantitative proteomics. Using this method, we identified 125 endocytic cell-surface proteins from hCMEC/D3 cells. Of these, 34 cell-surface proteins were selectively internalized into human brain microvascular endothelial cells, but not into human umbilical vein endothelial cells (HUVECs), a model of human peripheral microvascular endothelial cells. Two cell-surface proteins, intercellular adhesion molecule-1 (ICAM1) and podocalyxin (PODXL), were identified as BBB-localized endocytic cell-surface proteins in humans, using open mRNA and protein databases. Immunohistochemical evaluation confirmed PODXL expression in the plasma membrane of hCMEC/D3 cells and revealed that anti-PODXL antibody-labeled cell-surface PODXL internalized into hCMEC/D3 cells. Immunohistochemistry further revealed that PODXL is localized at the luminal side of human brain microvessels, supporting its potential suitability for translational applications. In conclusion, our findings highlight novel endocytic cell-surface proteins capable of internalizing into human brain microvascular endothelial cells. ICAM1 or PODXL targeted antibody or ligand-labeled biopharmaceuticals and nanocarriers may provide effective targeted delivery to the brain across the BBB for the treatment of central nervous system (CNS) diseases.