Mutual paracrine effects of colorectal tumour cells and stromal cells: modulation of tumour and stromal cell differentiation and extracellular matrix component production in culture.
Interactions of tumour and stromal cells influence tumour cell proliferation and differentiation, stromal cell phenotypic transdifferentiation and secretion of extracellular matrix (ECM) components. In this study, we established a monolayer and a three-dimensional cell-to-cell interaction model between canine mammary stromal cells and human colonic carcinoma cell lines (Caco-2 and HT-29) to investigate mutual paracrine effects of tumour cells and stromal cells on (i) tumour cell differentiation, (ii) production of ECM components and (iii) phenotypic transdifferentiation of stromal cells. We showed that when Caco-2 or HT-29 cells are cultured in collagen gels, they form a few small solid cell clusters with no lumina, but when cocultured with stromal cells, the tumour cells formed glandular structures with central lumina. This fibroblast-induced organization and differentiation of Caco-2 cells (not HT-29 cells) appeared to be mediated by transforming growth factor-beta (TGF-beta). Culturing of stromal cells, Caco-2 cells or HT-29 cells alone in both monolayers and gels resulted in weak tenascin-C expression in stromal cells and HT-29 cells and no expression in the Caco-2 cells. Coculturing of stromal cells with tumour cells resulted in increased tenascin-C expression in the stromal cells and HT-29 cells and induced expression of tenascin-C in the Caco-2 cells. This induction and increased expression of tenascin-C appeared to be mediated by TGF-beta. Culturing of stromal cells, Caco-2 cells or HT-29 cells alone on monolayers and in gels resulted in a weak expression of chondroitin sulfate (CS), chondroitin-6-sulfate (C-6-S) and versican in stromal cells and no expression in Caco-2 and HT-29 cells. Coculturing of stromal cells with tumour cells on monolayers and in gels resulted in increased CS, C-6-S and versican expression in stromal cells. This tumour cell-induced expression of CS, C-6-S and versican appeared to be mediated by TGF-beta and platelet-derived growth factor (PDGF). Coculturing of Caco-2 and HT-29 and stromal cells promoted the transdifferentiation of stromal cells into myofibroblasts, and this appeared to be mediated by TGF-beta. These results suggest that TGF-beta and PDGF are part of a paracrine system involved in stromal-epithelial cell interaction important in stromal cell differentiation and ECM component production.


Three large scale surveys highlight the complexity of cervical cancer under-screening among women 45-65years of age in the United States.
Large scale United States (US) surveys guide efforts to maximize the health of its population. Cervical cancer screening is an effective preventive measure with a consistent question format among surveys. The aim of this study is to describe the predictors of cervical cancer screening in older women as reported by three national surveys. The Behavioral Risk Factor Surveillance System (BRFSS 2016), the Health Information National Trends Survey (HINTS 2017), and the Health Center Patient Survey (HCPS 2014) were analyzed with univariate and multivariate analyses. We defined the cohort as women, without hysterectomy, who were 45-65years old. The primary outcome was cytology within the last 3years. Overall, Pap screening rates were 71% (BRFSS), 79% (HINTS) and 66% (HCPS), among 41,657, 740 and 1571 women, respectively. BRFSS showed that women 60-64years old (aPR=0.88, 95% CI: 0.85, 0.91), and in rural locations (aPR=0.95, 95% CI: 0.92, 0.98) were significantly less likely to report cervical cancer screening than women 45-49-years old or in urban locations. Compared to less than high school, women with more education reported more screening (aPR=1.20, 95% CI: 1.13, 1.28), and those with insurance had higher screening rates than the uninsured (aPR=1.47, 95% CI: 1.33, 1.62). HINTS and HCPS also showed these trends. All three surveys show that cervical cancer screening rates in women 45-65years are insufficient to reduce cervical cancer incidence. Insurance is the major positive predictor of screening, followed by younger age and more education. Race/ethnicity are variable predictors depending on the survey.


Ontogeny of metabolism, thermoregulation and torpor in the house martin Delichon u. urbica (L.) and its ecological significance.
Special energetic adaptations are of great evolutionary significance for birds that encounter transient problems in finding food during the breeding season. House martins, as aerial insectivores, encounter such problems during spells of bad weather, when they must survive on body reserves. This species employs the following behavioural and physiological adaptations to save energy: Low basal metabolic rate (only 43% of the values predicted by allometric equations); low thermal conductance 51% (day) and 67% (night) of the predicted values; clustering behaviour; high tolerance of the young to periods of low food supply; and the ability to become torpid, found in adults and young from the age of 11 days on. House martins are the first passerine birds in which torpor has been found. These adaptations might have played a role in the great success of the house martin, one of the 10-15 most abundant bird species in Europe.


Effect of pregnancy on gastric motility in vivo and in vitro in the guinea pig.
Experiments were designed to examine the effects of pregnancy on gastric emptying and on the in vitro contractile response of gastric smooth muscle to cholinergic stimulation. Gastric emptying of a radio-labeled liquid marker (51Cr in saline) was determined at regular intervals (5, 15, 30, and 60 min) in nonpregnant and pregnant (third trimester) guinea pigs. Gastric emptying after 30 min also was determined for animals 1 and 4 days postpartum. The in vitro contractile responses of muscle strips from the fundus, corpus, and antrum of nonpregnant and pregnant animals after stimulation with acetylcholine were analyzed with respect to the maximal force developed and the pD2 value (negative logarithm of the dose that gives a one-half maximal response). The results can be summarized as follows. Pregnancy was characterized by an increase in the amount of marker retained within the stomach at each time point. For nonpregnant animals the time to one-half emptying was approximately 10 min; in pregnant animals this was prolonged to 30 min. Gastric emptying was reduced in animals 1-day postpartum but returned to normal by 4 days after delivery. In contrast, pregnancy had no effect on the maximal contractile response to acetylcholine of muscle strips from the orad and caudad stomach or on their respective pD2 values. The results suggest that pregnancy is associated with decreased gastric emptying of liquids in the guinea pig, but that the delay cannot be attributed to alterations in the maximal force-generating capacity of gastric smooth muscle.


Anophthalmic socket and orbital implants. Role of CT and MR imaging.
The role of CT and MR imaging examination of orbital implants and the anophthalmic socket is expanding constantly. As the imaging techniques gain refinement and resolution, the list of potential clinical application grows. Frequent clinical-radiologic queries regarding the anophthalmic socket include neuro-radiologic findings with congenital anophthalmia or bilateral microphthalmia, the vascularization or position of an orbital implant following enucleation, or orbital recurrence of tumor following enucleation. Common clinical-radiologic questions regarding orbital implants relate to the associated findings of trauma, infection, bleeding, or recurrence of tumor in relationship to the existing orbital implant.


Investigation of novel alginate-magnesium aluminum silicate microcomposite films for modified-release tablets.
Physicochemical properties of sodium alginate-magnesium aluminum silicate (SA-MAS) composite films were investigated and a potential as a film former of SA-MAS dispersion for modifying drug release from tablets was evaluated as well. Interaction between SA and MAS in the composite films was revealed using FTIR spectroscopy. Thermal behavior of the composite films was changed due to the complexation of SA and MAS. Powder X-ray diffractometry data suggested that a higher crystallinity of the composite film and a phase-separated microcomposite were formed. The composite films in the ratios of 1:0.5 and 1:1 showed the increases of tensile strength and percentage of elongation when compared with SA films. Water vapor permeability of the composite films tended to increase with increasing ratio of MAS. The decreases in water uptake and drug permeability in 0.1 M HCl were found in the composite films. A positive charge drug, propranolol HCl, provided a higher affinity on the composite films than a weakly acidic nonelectrolyte, acetaminophen, resulting in a longer lag time and a higher partition coefficient depending on the content of MAS in the composite films. This was due to the complex formation of propranolol HCl and MAS. Using SEM, the tablets coated with SA-MAS dispersion had a smooth surface, while those with SA dispersion showed a pinholing on the surface, resulting in a faster drug release. The drug release profiles of the tablets could be modified by coating with the composite film at different coating levels. This finding suggests that MAS could improve physicochemical properties of the SA films, leading to a novel coating material of the SA-MAS dispersion for modifying drug release from tablets.


Therapeutic concentrations of anti-epileptic drugs do not inhibit the activity of the oncolytic adenovirus Delta24-RGD in malignant glioma.
The oncolytic adenovirus Delta24-RGD is currently being tested in phase I trials for the treatment of glioblastoma (GBM). Literature suggests that frequently prescribed anticonvulsants for these patients, phenytoin (PHE), valproic acid (VPA) and levetiracetam (LEV), may interfere with cellular mechanisms of cancer or oncolytic virus activity. We therefore investigated the direct effects of these drugs on Delta24-RGD infection and oncolytic activity. The anticonvulsants PHE, VPA, and LEV were combined with Delta24-RGD treatment in established glioma cell lines as well as on a panel of patient-derived GBM cultures. Effects on infection efficiency were assessed using luciferase-encoding adenoviral vectors. Oncolytic activity was determined by WST-1 assay and viral progeny production was quantified by dilution titration. IC50 values of the anti-epileptic drugs on the four glioma cell lines were far above clinically-relevant concentrations. At therapeutic concentrations, the anti-epileptics generally did not alter the infection efficiency of RGD-modified adenovirus, nor affect progeny production or oncolytic activity of Delta24-RGD. The only exception was found in U373 cells, where VPA slightly antagonised the oncolytic effect of Delta24-RGD (from 29% to 55% viability, p<0.01) as well as viral progeny production (60% decrease, p<0.01). Oncolysis by Delta24-RGD was not inhibited by the anti-epileptics in any of the patient-derived glioma cultures (n=6). In fact, in one culture a slight enhancement of viral oncolysis by PHE and LEV was found, from 89.7% viability to 76% and 62.4%, respectively (p<0.01) Therapeutic levels of valproic acid, phenytoin and levetiracetam do not negatively interfere with the infection efficiency or oncolytic activity of Delta24-RGD in patient-derived GBM cells. Therefore, there is no indication that the choice of anticonvulsant for seizure control in glioma patients should take treatment with Delta24-RGD into account.


The VLDL receptor: an LDL receptor relative with eight ligand binding repeats, LR8.
The discovery in 1992 of a member of the low density lipoprotein receptor (LDLR) family with eight ligand binding repeats (LR8) has raised more questions than have been answered to date. Here, we summarize the current status of knowledge about this intriguing molecule, generally termed VLDL receptor, at the molecular biological, cell biological, and physiological levels. On one hand, the wealth of reports concerning the role(s) of this receptor in lipoprotein metabolism in mammalian systems has revealed partially conflicting details, particularly in regards to its natural ligand(s) and site of action. On the other hand, molecular genetic and biochemical studies in the chicken have clearly demonstrated the multiple roles of LR8 in the physiology and reproduction of egg-laying species, and have generated insights into the evolutionary aspects of the LDLR gene family.


The effects of arsenic compounds on human and bovine lymphocyte mitogenesis in vitro.
The response of human and bovine peripheral blood lymphocytes to PHA stimulation was measured in the presence of low concentrations of sodium arsenite and sodium arsenate. In bovine lymphocytes, 41% augmentation of the response occurred at 10(-6) M arsenite with a return to the normal response at 2.5 X 10(-6) M. Complete inhibition of mitogenesis occurred at 6 X 10(-6) M. In the presence of sodium arsenate, similar results were obtained but at the higher concentrations of 2 X 10(-5) (for 57% augmentation), 5.2 X 10(-5), and 1.9 X 10(-4) M, respectively. The possible significance of these findings in view of the known relationship between chronic arsenicalism and human skin cancer is discussed. It is suggested that arsenic compounds may, by potentiating mitogenesis, increase the possibility of errors in DNA replication, some of which could be potentially carcinogenic. Additionally, interference with the immune response could enable potentially cancerous cells to escape immune surveillance.


The Role of Allergy in Chronic Rhinosinusitis.
The role of allergy in chronic rhinosinusitis (CRS) has long been debated and remains controversial. The 2 diseases frequently co-occur; however, direct causality has never been proved. The literature is largely mixed as to the manner and degree by which allergy contributes to CRS and this is in large part due to heterogeneity in the definitions of allergy and of CRS. In this review, the potential role of allergy in the disease processes of CRS without polyps, CRS with polyps, and allergic fungal rhinosinusitis is discussed.


Association between methylenetetrahydrofolate reductase tagging polymorphisms and susceptibility of hepatocellular carcinoma: a case-control study.
Polymorphisms in one-carbon metabolism genes may influence the susceptibility to hepatocellular carcinoma (HCC). In the present study, we studied methylenetetrahydrofolate reductase (MTHFR) tagging polymorphisms in 584 HCC cases and 923 controls. Polymerase chain reaction was harnessed to detect MTHFR genotype. Overall, our results showed that genotype distribution of MTHFR rs4846048 and rs4845882 polymorphisms was not different between HCC patients and controls. MTHFR rs9651118 and rs1801133 loci were protective factors for HCC (rs9651118: CT vs. TT: adjusted odds ratio (OR) = 0.67, 95% confidence interval (CI): 0.49-0.90, P=0.008 and TC/CC vs. TT: adjusted OR = 0.70, 95% CI: 0.53-0.93, P=0.015; rs1801133: GA vs. GG: adjusted OR = 0.72, 95% CI: 0.54-0.97, P=0.031, AA/GA vs. GG: adjusted OR = 0.76, 95% CI: 0.57-0.99, P=0.045). However, MTHFR rs3753584 locus was a candidate for susceptibility to HCC (CT vs. TT: adjusted OR = 1.67, 95% CI: 1.20-2.32, P=0.003 and TC/CC vs. TT: adjusted OR = 1.59, 95% CI: 1.15-2.20, P=0.005). Results of haplotype analysis suggested that MTHFR Grs1801133Trs3753584Grs4845882Ars4846048Trs9651118 was associated with the risk of HCC (OR = 1.55, 95% CI: 1.16-2.07, P=0.003). The power of our study also confirmed these associations (the value of power >0.80). In summary, our findings suggested that MTHFR rs3753584, rs9651118 and rs1801133 polymorphisms may affect the risk of HCC in Chinese Han population. In future, our findings should be further validated in additional case-control studies.


Usefulness of three-dimensional thoracoscope for prone position thoracoscopic esophagectomy improves mediastinal lymph node dissection and prognosis for esophageal cancer.
This study aimed to assess the superiority of 3D flexible thoracoscope against 2D thoracoscope for lymph node dissection (LND) and prognosis for prone-position thoracoscopic esophagectomy (TE) in esophageal cancer. Three hundred and sixty-seven esophageal cancer patients who underwent prone-position TE with 3-field LND between 2009 and 2018 were evaluated. 2D and 3D thoracoscope was used in 182 (2D group) and 185 cases (3D group), respectively. Short-term surgical outcomes, numbers of retrieved mediastinal lymph node (LN), and rates of LN recurrence were compared. Risk factors for mediastinal LN recurrence and long-time prognosis were also evaluated. No differences in postoperative complications were observed between the groups. The numbers of retrieved mediastinal LN were significantly higher, and the rates of LN recurrence were significantly lower in the 3D group compared to 2D group. Use of 2D thoracoscope was a significant independent factor of middle mediastinal LN recurrence by multivariable analysis. Survival was compared by cox regression analysis, and the 3D group had a significantly better prognosis than the 2D group. Prone position TE using 3D thoracoscope may improve the accuracy of mediastinal LND and prognosis without increasing postoperative complications for esophageal cancer.


[RESEARCH PROGRESS OF TISSUE ENGINEERING TECHNIQUE IN ESOPHAGEAL DEFECT REPAIR AND RECONSTRUCTION].
To review the research progress of the tissue engineering technique in the esophageal defect repair and reconstruction. The recently published clinical and experimental literature at home and abroad on the scaffold materials and the seeding cells used in the tissue engineered esophageal reconstruction was consulted and summarized. A large number of basic researches and clinical applications show that the effect of the tissue engineered esophagus is close to the autologous structure and function of the esophagus and it could be used for the repair of the esophageal defect. However, those techniques have a long distance from the clinical application and need an acknowledged rule of technology. Tissue engineering technique could provide an innovative theory for the esophageal defect reconstruction, but its clinical application need further research.


Sixteen failures in a single model of bipolar polyurethane-insulated ventricular pacing lead: a 44-month experience.
During a 44-month period, 105 Medtronic model 6972-58 polyurethane-insulated, bipolar ventricular pacing leads were implanted, of which 16 failed clinically at a mean of 20.5 months post-implant. Pacing dysfunction presented as either intermittent or complete loss of sensing and/or capture and, rarely, oversensing . Thirteen of the 16 patients were asymptomatic, and problems were detected in 11 instances during routine telephonic surveillance. At the time of lead replacement, low or widely fluctuating lead resistance values and high current drain were uniformly observed. The single lead capable of total extraction revealed rupture of both internal and external insulation. A brief summary of the currently much-discussed "polyurethane controversy" is presented, and the issue of patient management is discussed.


Response phase of the flicker electroretinogram (ERG) is influenced by cone excitation strength.
We measured electroretinogram (ERG) response phases at different cone contrasts in trichromats and dichromats to investigate the dynamics of the long-wavelength-sensitive (L-) and middle-wavelength-sensitive (M-) cone pathways. ERG responses to stimuli, temporally modulated at 30 Hz, were recorded. The stimuli were generated on a computer controlled colour monitor. Thirty-two different combinations of L- and M-cone excitation strength, expressed as cone contrasts, were presented. The short-wavelength-sensitive (S-) cones were not stimulated (S-cone contrast = 0%). The response phase of the fundamental stimulus component was obtained from Fourier analysis. The ERG response phase lags decreased with increasing cone contrast. This was observed in all subjects with a normal appearing fundus. In dichromats and trichromats at low and intermediate contrasts, the phase lags to M-cone isolating conditions were smaller than those to L-cone isolating stimuli. In one dichromat with extreme myopia and cupping of the optic disc, the ERG phase lags increased with increasing cone contrast. The ERG response phase may be potentially useful for detecting retinal abnormalities.


Exploring the perception of key stakeholders toward khat policy approaches in Ethiopia: a qualitative study.
Khat is an amphetamine-like plant, produced and commonly chewed in Ethiopia by a large group of the population. Although significant multidimensional harms of khat use have been reported, currently, there are no policies or organized activities against khat use in Ethiopia. Therefore, the current study aimed to explore the perception of key stakeholders toward khat policy approaches for Ethiopia. A qualitative study was conducted using focus group discussion (FGD) and in-depth interview (IDI). Ten stakeholders participated in the IDIs, and another 15 individuals participated in the FGDs. Key stakeholders were appropriately mapped and purposively selected based on their experience related to khat use and khat harm reduction. Interviews and FGDs were led by the study authors and were audio-recorded. The audio-recorded data were transcribed verbatim and then translated into English language. The data were analyzed using thematic analysis approach. Participants preferred prevention and law regulation measures for khat policy approaches for Ethiopia. Proposed prevention and law regulation measures included regulating the transportation of khat, limiting the minimum age to buy and sell khat, prohibiting khat use at some public places, banning khat advertising and promotion, and imposing excise tax. Individual level khat harm reduction strategies were also proposed to be useful. However, the participants asserted that total khat ban in Ethiopia is not likely to be feasible and acceptable. Prevention, treatment or care for individuals with problematic khat use, law regulation, and harm reduction were preferred approaches by stakeholders for khat policy in Ethiopia instead of total banning.


Urethral Stone Disease Leading to Retention After Hair-bearing Neophalloplasty.
A 35-year-old male patient with a past history of traumatic penile amputation and subsequent penile reconstruction with a radial artery free flap phalloplasty presented to the urology clinic for urinary retention and complaint of a firm penile mass. The patient had been lost to follow-up for 2 years before this presentation. Patient had a suprapubic tube in place from initial surgery, with imaging showing 2 large uroliths encrusted around the end. Urethral stricture was suspected in the patient. On cystoscopy, an additional obstructing urolith was found in penile urethra, appearing to have formed in situ.


Association of Time since Migration from Rural to Urban Slums and Maternal and Child Outcomes: Dhaka (North and South) and Gazipur City Corporations.
This study analyzes data from a new Urban Health and Demographic Surveillance (UHDSS) in five slums in Dhaka (North and South) and Gazipur City Corporations to examine the relationship between migration status and maternal and child health service utilization. Migration status was determined by duration in urban slums (<= 9.99 years, 10-19.99 years, 20+ years, and urban-born). Compared to those born in the city, migrants were characterized by significant disadvantages in every maternal, neonatal, and child health (MNCH) indicator under study, including antenatal care, facility-based delivery, doctor-assisted delivery, child immunization, caesarean-section delivery, and use of modern contraceptives. We found that the level of service coverage among migrants gradually converged-but did not fully converge-to that of the urban-born with increasing duration in the city. We observed a strong positive association between wealth and total MNCH coverage, with a more modest association with higher levels of schooling attainment. Women who were engaged in market employment were less likely to receive adequate coverage, suggesting a tradeoff between livelihood attainment and mother-and-child health. After controlling for these socioeconomic and neighborhood variations in coverage, the duration gradient was diminished but still significant. In line with existing studies of healthcare access, this study highlights the persistent and widespread burden of unequal access to maternal and child health care facing migrants to slum areas, even relative to the overall disadvantages experienced in informal settlements.


How to Tell When a Digital Technology Is Not Ready for You.
The stages of digital technology readiness are viewed through the lens of three contemporary and widely discussed examples, namely distributed ledger technology, machine learning, and the internet of things. I use these examples to clarify when there is really just an old technology being re-branded, when there is something genuinely new and useful, and whether there may be over-claiming.


NLRP3 inflammasome-dependent pyroptosis is proposed to be involved in the mechanism of age-dependent isoflurane-induced cognitive impairment.
Wang Z et al. recently published a paper, titled "Critical role of NLRP3-caspase-1 pathway in age-dependent isoflurane-induced microglial inflammatory response and cognitive impairment". The finding in this paper is consistent with our previous study on NLRP3-caspase-1 pathway. Here, we propose that NLRP3 inflammasome-dependent pyroptosis may be involved in the mechanism of age-dependent isoflurane-induced cognitive impairment and discuss that inhibiting NLRP3 inflammasome activation with a novel inhibitor MCC950 may ameliorate age-dependent isoflurane-induced neuro-inflammation.


Maintenance of viability and transport function after preservation of isolated rat hepatocytes in various simplified University of Wisconsin solutions.
Rat hepatocytes were preserved for 24 hr with high recovery and good maintenance of viability and transport function both in University of Wisconsin (UW) solution and in various simplified UW solutions. Cell quality is somewhat affected after 48 hr of preservation in both the original UW solution and the simplified solutions. ATP content and uptake rate of taurocholic acid are more sensitive markers of cell viability than Trypan blue exclusion or the MTT test. A much less expensive solution than UW, containing only K(+)-lactobionate, KH2PO4, MgSO4 and raffinose, can be used successfully for preservation of rat hepatocytes for 24 hr for drug transport studies.


Residual and recurrent acoustic neuromas.
A retrospective study was conducted on all 23 patients who underwent operation for residual or recurrent acoustic neuromas during the 10-year period January 1976 through December 1985. The most common symptoms at the time of reoperation were ataxia (16 patients), facial paresthesias (13 patients), and headaches (9 patients). Primary procedures had consisted of suboccipital posterior fossa approaches in 22 patients and a middle fossa approach in one. Reoperation for recurrent or residual tumor consisted of a retrosigmoid approach in 17 patients and a translabyrinthine or translabyrinthine/retrosigmoid approach in six patients. This study confirms that residual or recurrent acoustic tumors are not common. It also suggests that long-term follow-up, for at least 7 to 8 years, is indicated.


In Situ Fabricated Non-Flammable Quasi-Solid Electrolytes for Li-Metal Batteries.
Lithium metal batteries (LMBs) are viewed as one of the most promising high energy density battery systems, but their practical application is hindered by significant fire hazards and fast performance degradation due to the lack of a safe and compatible configuration. Herein, nonflammable quasi-solid electrolytes (NQSEs) are designed and fabricated by using the in situ polymerization method, in which 1,3,2-dioxathiolan-2,2-oxide is used as both initiator to trigger the in situ polymerization of solvents and interphase formation agent to construct robust interface layers to protect the electrodes, and triethyl phosphate as a fire-retardant agent. The NQSEs show a high ionic conductivity of 0.38 mS cm-1 at room temperature and enable intimate solid-electrolyte interphases, and demonstrate excellent performance with stable plating/striping of Li metal anode, and high voltage (4.5 V) and high temperature (>60 °C) survivability. The findings provide an effective strategy to build high-temperature, high-energy density, and safe quasi-solid LMBs.


Zymogen/enzyme discrimination using peptide chloromethyl ketones.
Glutamylglycinylarginyl chloromethyl ketone, tyrosylglycinylarginyl chloromethyl ketone, and phenylalanylprolylarginyl chloromethyl ketone have been labeled at their amino termini using fluorescein, rhodamine-X, lissamine-rhodamine, pyrene, and the 1,5-, 2,5-, and 2,6-dimethylaminonaphthalene-1-sulfonyl moieties. These peptidyl chloromethyl ketones have also been modified by incorporation of biotin and epsilon-amino caproyl biotin. The ability of these various chloromethyl ketones to be incorporated into a collection of zymogen-enzyme pairs has been evaluated using a variety of coagulation and fibrinolytic proteins. All labeled chloromethyl ketones were efficiently incorporated into the proteases tested, with the exception of urokinase which was refractory to inhibition by phenylalanylprolylarginyl chloromethyl ketone derivatives. No modification of any zymogen species was observed even under conditions designed to detect minimal reactivity. When enzymes were modified using chloromethyl ketones labeled with epsilon-amino caproyl biotin, the modified proteins readily reacted with avidin under a variety of different conditions. The observed reactivity with avidin was used in enzyme "blotting" following electrophoretic resolution of polypeptide chains and to remove active enzyme present in enzyme-zymogen mixtures. These reagents have been used to evaluate the potential for active site expression by the single-chain human factor VII molecule. Studies conducted with tissue factor, phospholipids, and calcium using factor X as substrate demonstrate that no activity can be obtained without initial activation of either factor X to factor Xa or factor VII to factor VIIa by an external source. We thus conclude that factor VII is a true zymogen, inert in the blood clotting process prior to its cleavage to factor VIIa.


A boron-doped helicene as a highly soluble, benchtop-stable green emitter.
The high-yield synthesis of a boron-doped [4]helicene was achieved through a Ni-mediated Yamamoto C-C-coupling reaction; the moderate distortion of the molecular scaffold confers excellent solubility to the air- and water-stable green luminophore.


Transient tritanopia of a second kind.
Using a psychophysical method that allows the tracking of very rapid changes in sensitivity, we demonstrate an anomaly in the time-course of light adaptation for the short-wave mechanism: after the onset of a yellow (581 nm) field of approximately 10(5.3) td the threshold for short-wave targets does not recover monotonically but continues to rise for several seconds before falling to its equilibrium value. The phenomenon is absent when the adapting field has a wavelength of 511 nm and has been adjusted to give a similar equilibrium value for the short-wave threshold.


In vitro and ex vivo models of multiple sclerosis.
Multiple sclerosis (MS) is an autoimmune disorder of the central nervous system (CNS). Current therapies suppress a misdirected myelin-destructive immune response. To combat the progressive, neurodestructive phase of MS, the therapeutic research focus is currently on compounds that might boost the endogenous potential of the brain to remyelinate axons, thereby achieving lesion repair. Here, we describe the testing of fingolimod on cultures of oligodendrocytes (OLs) and organotypic brain slices. We detail the protocols, pros, and cons of these in vitro and ex vivo approaches, along with the potential benefit of exploiting skin-punch biopsies from patients with MS, before concluding with a summary of future developments.


Pancreatic morphology and function in relationship to pain in chronic pancreatitis.
The correlation of pancreatic pain and the degree of morphological and functional changes was prospectively investigated in 64 patients (45 males, 19 females) with chronic pancreatitis. All patients underwent clinical examination and diagnostic investigations by means of endoscopic retrograde pancreatography (ERP), computed tomography (CT) and exocrine pancreatic function test (secretin-ceruletide; SC). The individual tests were performed within 4 weeks in all patients. No significant correlation was found between the degree of pain and the severity of ERP and CT findings. Severe pain was found in 62% of patients with advanced ductal changes, and in 41% of patients with advanced parenchymal lesions. 58% of patients with severe pain had marked exocrine insufficiency. Ductal changes characterized by obstruction of the main pancreatic duct were most frequently associated with severe pain (58%). Less often, severe pain was associated with duct dilatation (34%) or with slight duct changes (26%). Among parenchymal lesions as revealed in CT, large cysts were most frequently related to severe pain (62%). A notable finding was that 89% of patients presenting with calcifications in CT still experienced pain and 39% of them had severe pain. The combined diagnostic approach is useful for the individual therapeutic management, but does not provide insight into the mechanisms of pain in chronic pancreatitis.


Chlorophyll a variations and responses to environmental stressors along hydrological connectivity gradients: Insights from a large floodplain lake.
Understanding the key drivers of eutrophication in floodplain lakes has long been a challenge. In this study, the Chlorophyll a (Chla) variations and associated relationships with environmental stressors along the temporal hydrological connectivity gradient were investigated using a 11-year dataset in a large floodplain lake (Poyang Lake). A geostatistical method was firstly used to calculate the hydrological connectivity curves for each sampling campaign that was further classified by K-means technique. Linear mixed effect (LME) models were developed through the inclusion of the site as a random effect to identify the limiting factors of Chla variations. The results identified three clear hydrological connectivity variation patterns with remarkable connecting water area changes in Poyang Lake. Furthermore, hydrological connectivity changes exerted a great influence on environmental variables in Poyang Lake, with a decrease in nutrient concentrations as the hydrological connectivity enhanced. The Chla exhibited contrast variations with nutrient variables along the temporal hydrological connectivity gradient and generally depended on WT, DO, EC and TP, for the entire study period. Nevertheless, the relative roles of nutrient and non-nutrient variables in phytoplankton growth varied with different degrees of hydrological connectivity as confirmed by the LME models. In the low hydrological connectivity phase, the Chla dynamics were controlled only by water temperature with sufficient nutrients available. In the high hydrological connectivity phase, the synergistic influences of both nutrient and physical variables jointly limited the Chla dynamics. In addition, a significant increasing trend was observed for Chla variations from 2008 to 2018 in the HHC phase, which could largely be attributed to the elevated nutrient concentrations. This study confirmed the strong influences of hydrological connectivity on the nutrient and non-nutrient limitation of phytoplankton growth in floodplain lakes. The present study could provide new insights on the driving mechanisms underlying phytoplankton growth in floodplain lakes.


The Sense of Coherence, Self-Perception of Aging and the Occurrence of Depression Among the Participants of the University of the Third Age Depending on Socio-Demographic Factors.
The aim of the study was to assess the relationship between the sense of coherence and self-perception of aging and the risk of depression among attendees of the University of the Third Age (U3A). The influence of socio-demographic factors was studied. The study involved 315 attendees of the U3A aged on average 68.4 years, age range 60-82 years old. The overwhelming majority were women (91.7%). Cross-sectional research was conducted using standardized questionnaires - the SOC-29 scale, the Geriatric Depression Scale (GDS - brief version) and the author's own questionnaire regarding self-perception of aging. The study participants showed an average level of sense of coherence (130.7± 23.2). The highest number of people obtained average results of sense of coherence, both in terms of global SOC (75.2%) and its components. The highest scores on the SOC-29 scale were found for the sense of comprehensibility (41.3%). The mean value of the self-perception of aging was 69.0±16.0 and indicates the lower range of results for positive attitude. Nearly ¾ (72.1%) of the respondents had a positive self-perception of aging. The values of the scale of self-perception of aging showed a statistically significant relation to the global level of sense of coherence and its components (p<0.001). In people without depression, positive self-perception of aging was significantly more frequent than among participants with depression of increasing intensity. Education was significantly correlated with the results of the SOC-29 scale, the self-perception of aging scale and the GDS scale. From the U3A research group, better educated elderly people have a higher sense of coherence and show a more positive self-perception of aging. Moreover, these people are less likely to show signs of senile depression.


Lipid peroxidation-derived modification and its effect on the activity of glutathione peroxidase 1.
Oxidative stress and the resulting lipid peroxidation are associated with various pathological states, including neurodegenerative diseases and cancer. The end products of lipid peroxidation, such as 4-oxo-2(E)-nonenal (ONE), 4-hydroxy-2(E)-nonenal (HNE), and methylglyoxal (MG), exert several biological effects through modification of various cellular components, including DNA and proteins. Glutathione peroxidase 1 (GPx1) is an intracellular antioxidant enzyme that uses glutathione (GSH) to reduce a variety of peroxides, thereby modulating cellular oxidative stress and redox-mediated responses. GPx1 contains nucleophilic amino acids at its active (one Sec) and GSH-binding (four Arg and one Lys) sites. We found that lipid peroxidation-derived reactive aldehydes (ONE, HNE, and MG) modified the GSH-binding site, resulting in the inhibition of GPx1 activity. Mass spectrometry-based proteomic analysis identified the sites modified by each aldehyde (ONE, 14 sites; HNE, 7 sites; MG, 9 sites). The GSH-binding sites modified were as follows: ONE, Arg57, 103, 184, and 185; HNE, Lys91; MG, Arg103. Upon incubation of GPx1 with each aldehyde, ONE reduced GPx1 activity more significantly than did HNE or MG in a dose- and time-dependent manner. The addition of GSH to GPx1 3 h after incubation with ONE prevented further inhibition by trapping ONE as a ONE-GSH adduct. However, the activity of GPx1 was not restored to the initial level, indicating that ONE modified GPx1 irreversibly. This study suggests that oxidative damage to lipids, resulting in the formation of reactive aldehydes, can amplify cellular oxidative stress via direct inactivation of GPx1, which increases the production of intracellular peroxides.


On the morphological variability of Limotettix (Limotettix) striola (Falln, 1806) (Homoptera: Auchenorrhyncha: Cicadellidae: Deltocephalinae: Limotettigini) with notes on the taxonomic status and distribution of L. striola ferganensis Dubovskiy, 1966.
Investigation of the male calling signals of Limotettix (Limotettix) striola (Falln, 1806) in populations from central and southern European Russia, Northern Caucasus, and West Tien Shan, Kyrgyzstan demonstrated that they belong to the same species. Males of L. striola from West Tien Shan have a somewhat different penis shape and belong to a separate subspecies, L. striola ferganensis Dubovskiy, 1966. L. striola ferganensis from West Tien Shan differs from L. ferganensis sensu Hamilton (1983, 1994) from the Nearctic in the shape of penis in dorsal view. Apparently, L. ferganensis sensu Hamilton (1983, 1994) is a separate, possibly, undescribed Nearctic taxon.


Histamine stimulates normal human melanocytes in vitro: one of the possible inducers of hyperpigmentation in urticaria pigmentosa.
Isolated normal human melanocytes became enlarged and more dendritic in association with an increase in the activity of tyrosinase and the amount of b-locus protein when they were cultured with 0.1-10 microM histamine in vitro. However, histamine did not exert a proliferative effect on them. The stimulatory effect of histamine was observable even 6 h after starting the treatment. This stimulation seems not to be pharmacologically mediated through histamine receptors, because it was inhibited neither by pyrilamine, a histamine H-1 antagonist, nor by cimetidine, a H-2 antagonist. Imidazole derivatives that are rapidly metabolized from histamine in vivo and in vitro also stimulated the melanocytes. We propose that high concentrations of histamine and its imidazole metabolites continuously produced in the lesions of urticaria pigmentosa are probable causative factors of its characteristic skin pigmentation.


Mini-laparoscopic cholecystectomy: evolution of a new technique.
Since the first laparoscopic cholecystectomy (LC) in 1985, there has been much advancement in laparoscopic surgery in terms of reduction in number and size of ports. We report a new technique of performing mini laparoscopic cholecystectomy using only three ports, 5 mm each. The indications of this procedure include GB polyps, GB dyskinesia, microlithiasis, and idiopathic pancreatitis. In this case report, we present a new technique that has been performed safely in a 49-year-old male patient with pancreatitis caused by microlithiasis. This was performed using a novel three port procedure consisting of only 5 mm ports, and he was discharged as a day case without complications. Informed patient consent was obtained. The fundamentals of this mini-LC technique remain the same as that of a standard laparoscopic cholecystectomy throughout the procedure. It is a feasible option in selected cases, and it has the potential to further augment the inherent benefits of minimal access surgery namely less analgesia, improved cosmesis and faster recovery. Further trials will help ascertain its potential advantages.


Exploring autoantibody signatures in brain tissue from patients with severe mental illness.
In recent years, studies have shown higher prevalence of autoantibodies in patients with schizophrenia compared to healthy individuals. This study applies an untargeted and a targeted affinity proteomics approach to explore and characterize the autoantibody repertoire in brain tissues from 73 subjects diagnosed with schizophrenia and 52 control subjects with no psychiatric or neurological disorders. Selected brain tissue lysates were first explored for IgG reactivity on planar microarrays composed of 11,520 protein fragments representing 10,820 unique proteins. Based on these results of ours and other previous studies of autoantibodies related to psychosis, we selected 226 fragments with an average length of 80 amino acids, representing 127 unique proteins. Tissue-based analysis of IgG reactivities using antigen suspension bead arrays was performed in a multiplex and parallel fashion for all 125 subjects. Among the detected autoantigens, higher IgG reactivity in subjects with schizophrenia, as compared to psychiatrically healthy subjects, was found against the glutamate ionotropic receptor NMDA type subunit 2D (anti-GluN2D). In a separate cohort with serum samples from 395 young adults with a wider spectrum of psychiatric disorders, higher levels of serum autoantibodies targeting GluN2D were found when compared to 102 control individuals. By further validating GluN2D and additional potential autoantigens, we will seek insights into how these are associated with severe mental illnesses.


Benazepril hydrochloride improves diabetic nephropathy and decreases proteinuria by decreasing ANGPTL-4 expression.
This study aimed to investigate the effects of benazepril hydrochloride (BH) on proteinuria and ANGPTL-4 expression in a diabetic nephropathy (DN) rat model. A total of 72 Wistar male rats were randomly divided into three groups: normal control (NC), DN group and BH treatment (BH) groups. The DN model was induced by streptozotocin (STZ). Weight, glucose, proteinuria, biochemical indicators and the kidney weight index were examined at 8, 12 and 16 weeks. In addition, ANGPTL-4 protein and mRNA expressions were assessed by immunohistochemistry and qRT-PCR, respectively. Relationships between ANGPTL-4 and biochemical indicators were investigated using Spearman analysis. Weight was significantly lower but glucose levels were significantly higher in both the DN and BH groups than in the NC group (P < 0.05). Compared with the DN group, proteinuria, urea, creatinine, triglycerides and total cholesterol levels were decreased, whereas the albumin level was increased after BH treatment (all P < 0.05). Furthermore, BH diminished kidney volume and ameliorated the pathological changes associated with DN. ANGPTL-4 expression was significantly decreased after BH treatment, and ANGPTL-4 expression was highly correlated with biochemical indicators of DN (P < 0.05). Benazepril hydrochloride improves DN and decreases proteinuria by decreasing ANGPTL-4 expression.


Orbital "blow-in" fracture: MRI.
A case of traumatic orbital encephalocele following a "blow-in" fracture is presented. Computed tomography (CT) and magnetic resonance imaging (MRI) findings are shown. Although CT was useful for identifying the orbital roof fracture, bone fragments and soft tissue abnormalities, MRI was more sensitive to brain herniation and an intraorbital haematoma.


Application of synthetic oligonucleotides to the diagnosis of human genetic diseases.
Under appropriate conditions synthetic oligonucleotide hybridization probes display essentially absolute hybridization specificity. That is, every nucleotide must form a Watson-Crick base pair in order that the probe forms a stable duplex. All of the non-Watson-Crick base pairs, including G-T, have a destabilizing effect. Thus, it is possible to choose stringent conditions of hybridization such that, while a perfectly matched duplex between an oligonucleotide and complementary DNA will form, duplexes mismatched at one or more position will not. Mutations in a single base in the DNA sequence of a gene can and do result in genetic diseases. The hybridization of oligonucleotides to the region of DNA containing these base changes would be affected by the mutations and thus, oligonucleotide hybridization provides a means of detecting single base changes. In an attempt to develop a non-radioactive method for the detection of human genetic diseases, we have prepared biotinylated-oligonucleotides by an enzymatic method. An oligonucleotide probe (23-mer) containing a single biotinylated deoxyuridine residue at the 3'-terminus was prepared by a primer extention reaction using E. coli DNA polymerase I (Klenow fragment). The probe could be specifically and tightly bound with Avidin D in 1 M NaCl. It could be hybridized to a plasmid DNA containing a perfectly matched complementary sequence, but not to a DNA containing 5 non-consecutive non-complementary bases. The hybridized biotinylated probe could be visualized by Avidin D and biotinylated alkaline phosphatase, even when 1.8 ng of the plasmid DNA (0.5 fmol) was used. A general approach to the enzymatic synthesis of oligonucleotides containing a single biotinylated deoxyuridine at the 3' end is described.


Effects of Ginkgo biloba on alertness and chemosensory function in healthy adults.
Ginkgo biloba is reported to enhance cognitive function in patients with selected neural disorders. Its effects in healthy, young adults are less well characterized. This work explored whether Ginkgo biloba could ameliorate decrements in alertness post-prandially and/or enhance chemosensory function. Both are functions that could be influenced by enhanced cerebral blood flow and neuronal metabolism, reported properties of the compound. A double-blind placebo-controlled study was conducted with 19 males and 20 females with a mean age of 23.6 +/- 5.4 years and mean weight of 70.0 +/- 1.9 kg. Participants were supplemented for 13 weeks with either Ginkgo biloba (mean dose 184.5 mg/d (range 130-234 mg/d)) or placebo and administered various alertness, performance, affective state and chemosensory tests at weeks 1, 5, 9 and 13. Participants did experience the post-prandial affective state decrement (i.e. post-lunch dip), but not the performance decrement. Performance on the chemosensory tests improved over the 13-week study. However, Ginkgo biloba was ineffective at alleviating the symptoms of the post-lunch dip or at enhancing taste and smell function.


Patterns of coordination and clinical outcomes: a study of surgical services.
To test the hypothesis that surgical services combining relatively high levels of feedback and programming approaches to the coordination of surgical staff would have better quality of care than surgical services using low levels of both coordination approaches as well as those surgical service using low levels of either coordination approach. A study sample of 44 academically affiliated surgical services that are part of the Department of Veterans Affairs. In a cross-sectional analysis, surgical services were assigned to one of three groups based on their scores on feedback and programming coordination measures: high on both measures; high on one measure, low on the other; and low on both. Univariate and multivariate analyses were used to assess differences among these groups with respect to three quality indicators: risk-adjusted mortality, risk-adjusted morbidity, and staff perceptions of quality. Risk-adjusted mortality and morbidity came from an outcomes reporting program within the Department of Veterans Affairs that entails the prospective collection of clinical data from patient charts. Data on coordination practices and perceived quality came from a survey of surgical staff at each of the 44 participating surgical services. The group of surgical services using high feedback and high programming had the best perceived quality. This group also had the lowest morbidity, but the difference was statistically significant with respect to only one of the two other groups: the group with low feedback and low programming. No significant group differences were found for mortality. Study results provide partial support for the hypothesis that high levels of feedback and programming should be combined for optimal quality of care. Study results also suggest that staff coordination is more important for improving morbidity than mortality in surgical services.


Effects of selegiline alone or with donepezil on memory impairment in rats.
Selegiline, a monoamine oxidase-B inhibitor, is reported to improve memory and learning in dementia of Alzheimer's type. However, only a few studies have reported its use in animal models. Here, we evaluated the effects of selegiline only or its combined use with donepezil, a selective acetylcholinesterase inhibitor on memory impairment, using a Morris water maze. Selegiline dose-dependently attenuated ethylcholine aziridinium ion-induced memory impairment. Co-administration of selegiline and donepezil, at doses that do not exert efficacy individually, significantly ameliorated scopolamine+p-chlorophenylalanine-induced memory deficits. These results suggest that selegiline improves memory impairment mediated by the cholinergic system, and provide evidence of the usefulness of co-treatment with selegiline and donepezil for treating spatial deficits in dementia.


Plasticity of aggregation pheromones in insects.
Pheromone plasticity is widely observed in insects and enhances their survival, adaptation, and reproductive success. Aggregation pheromones, which cause notable individual aggregation and consequently impact agriculture and human life, are renowned for their special function. Here, we present a review of research progress regarding pheromone plasticity in three typical aggregative insects: locusts, bark beetles, and cockroaches. These insects are major pest species with considerable impacts on the social economy and public health. Numerous studies have demonstrated the plasticity of aggregation pheromones in different populations of these insect species. Although pheromone chemicals and compositions vary across the three groups, the plasticity of aggregation pheromones is significantly impacted by population density, location, food resources, and gut symbiotic microorganisms, indicating the complexity of pheromone plasticity regulated by multiple factors. Finally, we discuss the potential application of pheromone plasticity in basic research and pest management.


A comparative study between freeze-dried and spray-dried goat milk on lipid profiling and digestibility.
Different drying techniques impart distinguishing characteristics to goat milk, particularly to its fat globules. Here, we investigated the difference between freeze-dried and spray-dried goat milk (FGM and SGM) fat globules on lipid profiling and in vitro infant gastrointestinal digestibility. The former presented higher levels of MUFA (31.76%) and lower cholesterol content (1.20 ± 0.02 mg/g). Some important long-chain polyunsaturated triacylglycerols such as POL (16:0/18:1/18:2), PSL (16:0/18:0/18:2), and POO (16:0/18:1/18:1), also had better preservation in FGM. Moreover, we detected more species of lysophospholipid in FGM than SGM, accounting for 2.51% and 0.71% of total phospholipids, respectively. More intriguingly, FGM, which has better membrane integrity and larger particle size, showed longer lag during gastric digestion and lower level of final lipolysis throughout gastrointestinal digestion. Therefore, our results showed the effects of different drying techniques on lipid profiling and digestibility of goat milk, providing significant insight for appropriate utilization of goat milk in infant nutrition.


Acute metabolic acid-base disorders.
Metabolic acid-base disturbances commonly and predictably complicate the course of many intensive care unit patients and plague the intensivists, surgeons, and anesthesiologists beset with the task of caring for them. In this article, we offer a systematic approach to the patient with the metabolic acid-base disorders that are likely to be encountered in the setting of the intensive care unit. The discussions are in no way presented as through treatments of all metabolic disorders, but rather focus on the more practical aspects, namely, clinical scenarios, diagnostic clues, and therapeutic goals. Additionally, we interject a discussion of some of the more controversial topics with regard to the therapy of metabolic disorders.


Familial amyloid polyneuropathy: alanine-for-threonine substitution in the transthyretin (prealbumin) molecule.
A previously reported family with amyloid polyneuropathy (FAP) was reinvestigated to determine the type of mutation in the transthyretin (prealbumin) molecule. Transthyretin was isolated from amyloid-laden myocardium and serum, and tryptic peptides were resolved by high-performance liquid chromatography. Amino acid sequencing of an anomalous peptide revealed an alanine-for-threonine substitution corresponding to position No. 60 of the transthyretin monomer. Detection of the FAP gene in asymptomatic carriers was accomplished by hybrid isoelectric focusing of transthyretin in the presence of dithiothreitol and high concentrations of urea, and by Southern blotting of Pvull-digested leukocyte deoxyribonucleic acid. This type of FAP was found to be identical to the previously described Appalachian amyloid. Patients with FAP and their asymptomatic gene-carrying offspring had significantly reduced levels of serum transthyretin and retinol-binding protein.


Hypoxic preconditioning potentiates the trophic effects of mesenchymal stem cells on co-cultured human primary hepatocytes.
Mesenchymal stem/stromal cells (MSCs) improve the metabolic function of co-cultured hepatocytes. The present study aimed to further enhance the trophic effects of co-culture with hepatocytes using hypoxic preconditioning (HPc) of the MSCs and also to investigate the underlying molecular mechanisms involved. Human adipose tissue-derived MSCs were subjected to hypoxia (2 % O2; HPc) or normoxia (20 % O2) for 24 h and then co-cultured with isolated human hepatocytes. Assays of metabolic function and apoptosis were performed to investigate the hepatotrophic and anti-apoptotic effects of co-culture. Indirect co-cultures and co-culture with MSC-conditioned medium investigated the role of paracrine factors in the hepatotrophic effects of co-culture. Reactive oxygen species (ROS) activity was antagonised with N-acetylcysteine to investigate whether HPc potentiated the effects of MSCs by intracellular ROS-dependent mechanisms. Tumour necrosis factor (TNF)-α, transforming growth factor (TGF)-β1, and extracellular collagen production was determined and CASP9 and BAX/BCL-2 signalling pathways analysed to investigate the role of soluble factors, extracellular matrix deposition, and apoptosis-associated gene signalling in the effects of co-culture. HPc potentiated the hepatotrophic and anti-apoptotic effects of co-culture by ROS-dependent mechanisms. There was increased MSC TGF-β1 production, and enhanced MSC deposition of extracellular collagen, with reduced synthesis of TNF-α, as well as a downregulation of the expression of pro-apoptotic CASP9, BAX, BID and BLK genes and upregulated expression of anti-apoptotic BCL-2 in hepatocytes. HPc potentiated the trophic and anti-apoptotic effects of MSCs on hepatocytes via mechanisms including intracellular ROS, autocrine TGF-β, extracellular collagen and caspase and BAX/BCL-2 signalling pathways.


A naturalistic study of orally administered vs. inhaled legal market cannabis: cannabinoids exposure, intoxication, and impairment.
Published studies examining the effects of cannabis have largely utilized forms of cannabis that are not representative of the legal market products currently available. The present study aimed to characterize naturalistic use of legal market flower and edible products by examining associations among blood cannabinoids and amount of THC consumed as well as physiological, cognitive, and subjective effects in users of edible and flower forms. Eighty-four participants who used cannabis at least 1 × /week (55 flower cannabis using participants; 29 edible cannabis using participants mean age = 31.95 years, 44% female) participated. At the experimental appointment in our mobile laboratory, participants completed a blood draw to assess plasma cannabinoids, measures of heart rate, subjective drug effects, and cognition both before and after ad libitum use of legal market flower or edible cannabis. Average self-reported THC consumed was 15.97 mg (SD = 22.40) in edible users and 51.25 mg (SD = 45.23) in flower users. In the edible group, but not the flower group, strong correlations emerged between self-reported ad libitum THC consumed and plasma THC. Plasma THC was significantly higher after use of inhaled cannabis, but similar levels of plasma THC metabolites and similar levels of subjective intoxication and verbal memory impairment were observed in both flower and edible users. Findings support strong correlations among ad libitum THC consumed and THC plasma levels after edible cannabis use and suggest few differences in intoxication and impairment between edible and flower cannabis users after ad libitum use. This novel study provides important preliminary data on the pharmacology and effects of legal market edible cannabis.


Templated Reactive Crystallization of Active Pharmaceutical Ingredient in Hydrogel Microparticles Enabling Robust Drug Product Processing.
Commercialization of most promising active pharmaceutical ingredients (APIs) is impeded either by poor bioavailability or challenging physical properties leading to costly manufacture. Bioavailability of ionizable hydrophobic APIs can be enhanced by its conversion to salt form. While salt form of the API presents higher solution concentration than the non-ionized form, poor physical properties resulting from particle anisotropy or non-ideal morphology (needles) and particle size distribution not meeting dissolution rate targets can still inhibit its commercial translation. In this regard, API physical properties can be improved through addition of non-active components (excipients or carriers) during API manufacture. In this work, a facile method to perform reactive crystallization of an API salt in presence of the microporous environment of a hydrogel microparticle is presented. Specifically, the reaction between acidic antiretroviral API, raltegravir and base potassium hydroxide is performed in the presence of polyethylene glycol diacrylamide hydrogel microparticles. In this bottom-up approach, the spherical template hydrogel microparticles for the reaction lead to monodisperse composites loaded with inherently micronized raltegravir-potassium crystals, thus improving API physical properties without hampering bioavailability. Overall, this technique provides a novel approach to reactive crystallization while maintaining the API polymorph and crystallinity.


Cystocele repair by vaginal route: comparison of three different surgical techniques of mesh placement.
Different techniques of mesh placement for cystocele repair are known. Our goal was to compare anatomical and functional outcomes of three different techniques of mesh placement over a 3-year follow-up. Between March 2003 and June 2004, 230 patients (stage 2-4 pelvic organ prolapse (POP)) were included in a prospective study. For cystocele repair, mesh was implanted either with two arms into the retropubic space (RP) or with two to four arms into the obturator foramen (TO), or fixed to the arcus tendineous fascia pelvis (FG). Patients' distribution is as follows: 142 TO, 32 RP, and 31 FG. Anatomical success (cystocele < stage 2 in the POP staging system) was clearly poorer after the retropubic free technique, with success rates of 69% (RP), 90.1% (TO), and 96.6% (FG) (p = 0.004). POP distress inventory (p < 0.005) and POP impact questionnaire scores were both significantly poorer after RP. RP technique is less effective than TO and FG techniques.


Use of the C-terminal octapeptide of cholecystokinin in clinical radiology. I. The gallbladder.
Two 30-min. infusion doses of the octapeptide of chloecystokinin (C8-CCK) (1 and 4 ng/kg/min) produced equal and consistent gallbladder contractions (75 +/- 8% mean and SEM decrease in size) in 24 human volunteers. The long 30-min infusion method of C8-CCK administration appears better than bolus injection (30 to 180 s) for cholecystokinetic cholecystography.


SUMO E3 ligase CBX4 regulates hTERT-mediated transcription of CDH1 and promotes breast cancer cell migration and invasion.
hTERT, the catalytic component of the human telomerase enzyme, is regulated by post-translational modifications, like phosphorylation and ubiquitination by multiple proteins which remarkably affects the overall activity of the enzyme. Here we report that hTERT gets SUMOylated by SUMO1 and polycomb protein CBX4 acts as the SUMO E3 ligase of hTERT. hTERT SUMOylation positively regulates its telomerase activity which can be inhibited by SENP3-mediated deSUMOylation. Interestingly, we have established a new role of hTERT SUMOylation in the repression of E-cadherin gene expression and consequent triggering on the epithelial-mesenchymal-transition (EMT) program in breast cancer cells. We also observed that catalytically active CBX4, leads to retention of hTERT/ZEB1 complex onto E-cadherin promoter leading to its repression through hTERT-SUMOylation. Further through wound healing and invasion assays in breast cancer cells, we showed the tumor promoting ability of hTERT was significantly compromised upon overexpression of SUMO-defective mutant of hTERT. Thus our findings establish a new post-translational modification of hTERT which on one hand is involved in telomerase activity maintenance and on the other hand plays a crucial role in the regulation of gene expression thereby promoting migration and invasion of breast cancer cells.


Identification of anaplastic lymphoma kinase fusions in renal cancer: large-scale immunohistochemical screening by the intercalated antibody-enhanced polymer method.
Several promising molecular-targeted drugs are used for advanced renal cancers. However, complete remission is rarely achieved, because none of the drugs targets a key molecule that is specific to the cancer, or is associated with "oncogene addiction" (dependence on one or a few oncogenes for cell survival) of renal cancer. Recently, an anaplastic lymphoma kinase (ALK) fusion, vinculin-ALK, has been reported in pediatric renal cell carcinoma (RCC) cases who have a history of sickle cell trait. In this context, ALK inhibitor therapy would constitute a therapeutic advance, as has previously been demonstrated with lung cancer, inflammatory myofibroblastic tumors, and anaplastic large cell lymphomas. Anti-ALK immunohistochemistry was used to screen 355 tumor tissues, using the intercalated antibody-enhanced polymer (iAEP) method. The cohort consisted of 255 clear cell RCCs, 32 papillary RCCs, 34 chromophobe RCCs, 6 collecting duct carcinomas, 10 unclassified RCCs, 6 sarcomatoid RCCs, and 12 other tumors. Two patients (36- and 53-year-old females) were positive for ALK as determined by iAEP immunohistochemistry. Using 5'- rapid amplification of complementary DNA ends, we detected TPM3-ALK and EML4-ALK in these tumors. The results of this study were confirmed by fluorescence in situ hybridization assays. The 2 ALK-positive RCCs were unclassified (mixed features of papillary, mucinous cribriform, and solid patterns with rhabdoid cells) and papillary subtype. They comprised 2.3% of non-clear cell RCCs (2 of 88) and 3.7% of non-clear cell and nonchromophobe RCCs (2 of 54). The results of this study indicate that ALK fusions also exist in adult RCC cases without uncommon backgrounds. These findings confirm the potential of ALK inhibitor therapy for selected cases of RCC.


Tetra-substituted phthalocyanines bearing thiazolidine derivatives: synthesis, anticancer activity on different cancer cell lines, and molecular docking studies.
In the first step, (4R)-2-(2-hydroxyphenyl)thiazolidine-4-carboxylic acid (c) and 2-(2-(3,4-dicyanophenoxy)phenyl)thiazolidine-4-carboxylic acid (1) were prepared. Then, the peripherally tetra-substituted metallophthalocyanines [ZnPc (2), CuPc (3), and CoPc (4)] were synthesized by using 1. The structures of the obtained compounds were characterized by common spectroscopic methods. Aggregation behaviors of the tetra-substituted metallophthalocyanines (2-4) were investigated by UV-Vis and fluorescence spectroscopy in the presence/absence of soft metal ions. The electronic spectra of the newly synthesized metallophthalocyanines [ZnPc (2), CuPc (3), and CoPc (4)] were analyzed by the Bayliss method. The fluorescence quantum yield of diamagnetic ZnPc (2) was obtained in DMSO at room temperature. Also, the anticancer activity of the newly synthesized metallophthalocyanine derivatives was studied on C6, DU-145, and WI-38 cell lines and investigated using six concentrations (3.125; 6.25; 12.5; 50; 75; 100 μg L-1). The cell cycle and apoptosis analyses of CuPc (3) were performed. In addition, the chemical and biological activities of 2-(2-(3,4-dicyanophenoxy)phenyl)thiazolidine-4-carboxylic acid (1) and its novel type metallophthalocyanines [ZnPc (2), CuPc (3), and CoPc (4)] were compared with many parameters obtained from the Gaussian software and molecular docking methods.


Gender differences of amplitude of low-frequency fluctuations in bipolar disorder: A resting state fMRI study.
The clinical and epidemiological features of bipolar disorder (BD) between females and males have many differences. The association between brain function and gender in BD is unknown. This research aimed to investigate the association between brain function and gender in BD by using amplitude of low-frequency fluctuations (ALFFs). Ninety-eight patients (49 females and 49 males) with BD and 171 matched healthy controls (HCs, 89 females and 82 males) were recruited for resting-state functional magnetic resonance imaging. ALFF was used to estimate brain function. A main effect of diagnosis in ALFF was observed in the dorsal lateral prefrontal cortex (DLPFC), ventral prefrontal cortex (VPFC), caudate and occipital lobe. A main effect of gender in ALFF was found in the right VPFC, DLPFC, thalamus, and occipital lobe. A main effect of diagnosis gender interaction in ALFF was observed in the left DLPFC. Analyses of two-sample t-test indicated that male patients with BD had increased ALFF in the right hippocampus, right amygdala, left caudate, and left DLPFC, and decreased ALFF in the occipital lobe compared with male HC. Female patients with BD demonstrated increased ALFF in the right VPFC and right DLPFC compared with female HC. Male patients with BD exhibited increased ALFF in the right VPFC and left DLPFC and decreased ALFF in the occipital lobe compared with female patients with BD. This study did not consider the effect of medications and emotional states on brain activity. Results suggested gender differences in the dysfunctions of the cortico-limbic neural system in BD.


Combination treatment of cis- and carboplatin in cancers restricted to the peritoneal cavity in the rat.
In the present study, cisplatin (cDDP) and carboplatin (CBDCA) were combined in different in vitro and in vivo assays to determine whether combined cDDP and CBDCA treatment would eventually lead to a better antitumor response. Co-incubation of CC531 cells with cDDP and CBDCA led to higher intracellular Pt concentrations (30.5 +/- 3.4 ng Pt/10(6) cells) than did cDDP (16.9 +/- 9.4 ng Pt/10(6) cells) or CBDCA (1.28 +/- 0.72 ng Pt/10(6) cells) incubation alone. In survival assays an additive cell kill was seen after combined treatment with cDDP and CBDCA. DNA binding experiments using isolated salmon-sperm DNA exposed to the drugs separately or in combination were in agreement with the survival studies (for cDDP a binding of 12.42 micrograms Pt/mg DNA; for CBDCA, 0.49 microgram Pt/mg DNA; and for combined CBDCA and cDDP, 12.9 micrograms Pt/mg DNA at 76 h). Toxicity studies in rats treated with cDDP plus CBDCA required a dose reduction for cDDP amounting to 20% of the MTD, whereas the CBDCA dose could be maintained. Pharmacokinetics studies showed higher AUCs and t1/2 beta in plasma as well as the peritoneal cavity after combined treatment with cDDP and CBDCA (both given i.p.) or following cDDP given i.p. and CBDCA given i.v. Pt concentrations in peritoneal tumors corresponded with these observations, with higher Pt concentrations following combined treatment than after single-agent injection. In addition, combined administration of cDDP i.p. and CBDCA i.v. led to higher Pt concentrations in peritoneal tumors than did administration of both drugs i.p. (3.93 +/- 0.9 vs 2.76 +/- 0.2 mg Pt/g tissue). The higher Pt concentrations in the peritoneal tumors after combined treatment was associated with a significantly better antitumor response in comparison with that observed after single-agent treatment (a growth delay of 30.2 +/- 5.6 days for cDDP i.p. plus CBDCA i.v. vs 16.1 +/- 5.4 days for cDDP alone and 10.8 +/- 4.2 days for CBDCA alone).


Autumn leaf colouration: a new hypothesis involving plant-ant mutualism via aphids.
Several recent hypotheses on the adaptive significance of autumn leaf colours have focused on specialist aphids. However, these hypotheses have overlooked several factors: the preferential investment by healthy vigorous trees in growth rather than defence against herbivores, variation among aphid species in their responses to bright autumn leaves and plant defences and the occurrence of tritrophic interactions in tree crowns. I incorporate these factors into a hypothesis that autumn leaf colours signal tree quality to myrmecophilous specialist aphids, with the aphids, in turn, attracting aphid-tending ants during the following spring, and the ants defending the trees from other aphids and herbivores. Therefore, bright autumn leaves may have adaptive significance, attracting myrmecophilous specialist aphids and their attending ants and, thus, reducing herbivory and competition among aphids.


Delayed perforation after endoscopic resection of a colonic laterally spreading tumor: A case report and literature review.
Endoscopic mucosal resection (EMR) and endoscopic submucosal dissection (ESD) have been widely used for the treatment of early gastrointestinal cancer. Endoscopic piecemeal mucosal resection (EPMR) is derived from the combination of EMR and ESD. Delayed perforation with peritonitis after colonic EPMR is a rare but severe complication, sometimes requiring surgery. There are some associated risk factors, including patient- (location, diameter, and presence of fibrosis) and procedure-related factors. Early recognition and timely treatment are crucial for its management. We report a case in which delayed perforation with peritonitis was treated using endoscopic closure. A 54-year-old man was diagnosed with a 30-mm-diameter laterally spreading tumor in the colonic hepatic curvature. Fifteen hours after endoscopic resection, peritonitis caused by delayed perforation occurred and gradually aggravated. Conservative treatment was ineffective and no obvious perforation was observed. After timely endoscopic closure, the patient was discharged on postoperative day 4. In occasion of localized peritonitis aggravating without macroscopic perforation, endoscopic closure is an effective treatment for delayed perforation with stable vital signs in the early stage.


Atomic force microscopy analysis of the effect of plasma treatment on gas permeable contact lens surface topography.
Using atomic force microscopy (AFM) to investigate anterior surface topography (AST) in worn and unworn, plasma surface-treated (PST) and untreated (UT) gas permeable (GP) lenses, and influence of surface topography on in vivo comfort. GP lens AST evaluated with AFM in tapping mode, using an uncoated, 40 nm symmetric tip (sampling frequency: 300 kHz), at five randomised locations, over a 100μm2 area, to produce mean average roughness (Ra) and root mean square (RMS) values for each sample. Four unworn lenses (two PST, two UT) were examined (Quasar/Boston EO material). Twenty worn lenses (ten PST, ten UT) of same design and material as unworn lenses collected after 3 months lens wear. General wearing comfort reported by visual analogue scale (VAS) at 3 months visit. For sample preparation, two worn UT GP lenses were divided into four segments; each segment underwent a different lens rinse and drying method. Unworn: UT lenses had significantly higher mean RMS and Ra values compared to PST (Mann-Whitney, p < 0.05). Worn: UT Median RMS values were significantly higher than PST (Mann-Whitney, p < 0.05). Comfort: no correlation found between general comfort and RMS or Ra scores. Sample preparation: Method 4 (purified, distilled water rinse/nitrogen gas dry) produced optimum median RMS and Ra values. Unworn PST GP lenses had lower Ra and RMS values compared with unworn UT GP lenses. After 3 months wear, PST lenses had smoother surface topographies than UT lenses. No relationship was found between surface topography and lens wear comfort. Sample preparation protocol directly impacts AFM results.


Effects of reserpine and chlorpromazine on the serotonin, norepinephrine, and epinephrine levels in the pineal-paraphyseal complex of soft-shelled turtles (Lissemys punctata punctata).
Turtles were injected intramuscularly with reserpine (0.5 mg/100 g body wt) and chlorpromazine (10 mg/100 g body wt) at 2400, 1800, and 0600 hr and were sacrificed at 6, 12, and 24 hr, respectively after the treatments, i.e., at 0600 hr. The spectrofluorometric findings revealed that the serotonin, norepinephrine, and epinephrine contents of the pineal-paraphyseal complex decreased at 6 hr, increased at 12 hr, and declined again at 24 hr after the treatments. It is suggested that reserpine and chlorpromazine act largely by favoring release with transient resynthesis of these amines in the pineal-paraphyseal complex of soft-shelled turtles.


Osteochondral regeneration by a bilayered construct in a cell-free or cell-based approach.
A bilayered construct with or without adipose-derived stem cells (ASCs) was applied to repair full-thickness defects in the patellar groove of 18 rabbits. Non-treated and treated defects were divided into three groups: a control group (n = 12), a cell-free group (n = 12) and a cell-based group (n = 12). Histological appearance and grading were evaluated at 8 and 12 weeks. At 12 weeks, osteochondral-like tissues completely filled in the defects and integrated with host tissues in the cell-based group. The semi-quantitative score of the cell-based group (4.2 ± 1.2), which is a total score ranging from 0 (best) to 20 (worst), was significantly better than that of the other two groups (cell-free: 13.8 ± 2.5; control: 10.3 ± 2.4). This finding indicated that the bilayered constructs combined with ASCs could be an effective way to enhance osteochondral regeneration.


Ganoderma lucidum Polysaccharides Reduce Lipopolysaccharide-Induced Interleukin-1 β Expression in Cultured Smooth Muscle Cells and in Thoracic Aortas in Mice.
The expression of inflammatory cytokines on vascular walls is a critical event in vascular diseases and inflammation. The aim of the present study was to examine the effects of an extract of Ganoderma lucidum (Reishi) polysaccharides (EORPs), which is effective against immunological disorders, on interleukin- (IL-) 1 β expression by human aortic smooth muscle cells (HASMCs) and the underlying mechanism. The lipopolysaccharide- (LPS-) induced IL-1 β expression was significantly reduced when HASMCs were pretreated with EORP by Western blot and immunofluorescent staining. Pretreatment with 10 μ g/mL EORP decreased LPS-induced ERK, p38, JNK, and Akt phosphorylation. But the increase in IL-1 β expression with LPS treatment was only inhibited by pretreatment with the ERK1/2 inhibitor, while the JNK and p38 inhibitors had no effect. In addition, EORP reduced the phosphorylation and nuclear translocation of nuclear factor- (NF-) κ B p65 in LPS-treated HASMCs. Furthermore, in vivo, IL-1 β expression was strongly expressed in thoracic aortas in LPS-treated mice. Oral administration of EORP decreased IL-1 β expression. The level of IL-1 β expression in LPS-treated or in LPS/EORP-treated group was very low and was similar to that of the saline-treated group in toll-like receptor 4-deficient (TLR4(-/-)) mice. These findings suggest that EORP has the anti-inflammatory property and could prove useful in the prevention of vascular diseases and inflammatory responses.


The mTOR-inhibitor Sirolimus decreases the cyclosporine-induced expression of the oncogene ATF3 in human keratinocytes.
Due to their immunosuppressive therapy, organtransplant recipients (OTRs) exhibit a high incidence for the development of cutaneous squamous cell carcinoma (cSCC). Randomized studies of kidney-transplanted patients indicate a significant lower susceptibility for cSCC among patients receiving the mTOR-inhibitor Sirolimus, compared to patients without mTOR-regimen. The exact mechanism, how mTOR inhibition affects keratinocyte carcinogenesis remains unclear. Our aim was to investigate the impact of Sirolimus on the expression level of the oncogene ATF3, which is involved in the development and progression of cSCC. We incubated human keratinocytes, cSSC cell lines and 3D skin equivalents with Sirolimus, exposed the cells to calcineurin inhibitors (CNI) and UVA-radiation and measured the expression level of ATF3 by real-time PCR and western blot. We show that Sirolimus downregulates the expression of ATF3 induced by cyclosporine or cyclosporine plus UV-radiation in keratinocytes. In line with this we demonstrate a decrease in ATF3 expression, by incubating 3D skin equivalents with Sirolimus prior to cyclosporine and UV-light. However, Sirolimus has no significant impact on the ATF3 expression levels of cyclosporine stimulated cSCC cell lines. Taken together, our study demonstrates that Sirolimus downregulates the CNI or UV-induced ATF3 expression in human keratinocytes, which could be a potential molecular mechanism how Sirolimus reduces cSCC in OTRs. The lack of ATF3 suppression by Sirolimus in cSCC cell lines fits to observations from clinical studies which demonstrated a clinical benefit from the switch to a mTOR-regimen in patients with low tumor burden in early stage of disease.


[Results of large clinical studies presented at the European Society of Cardiology Congress 2012].
The article contains short characteristics of 6 novel European guidelines on the treatment of cardiovascular diseases and accounts of main scientific sessions (Hot Line, Clinical Trial and Registry Update) of the European Society of Cardiology Congress 2012.


Revealing the modulation of boundary conditions and governing processes on ozone formation over northern China in June 2017.
In this study, ozonesonde data were used to evaluate the impact of different boundary conditions on the vertical distribution of ozone over urban Beijing. The comparison shows that the clean and static boundary conditions, referred to as PROFILE, apparently underestimate the ozone concentration over the upper troposphere and stratosphere, whereas the global chemical transport model (CTM) provides much more reasonable performance. Further investigation reveals that the boundary conditions exert larger impacts over areas with high altitudes and close distances to boundaries, such as the Tibetan Plateau, while they yield weak impacts on regions relatively far from the boundary, such as the North China Plain (NCP). Process analysis was conducted to investigate the modulation of physical and chemical processes on ozone formation in June 2017, illustrating that during the daytime of the high-O3 period, the photochemical reactions within the planetary boundary layer (PBL) almost become the only source favorable to ozone accumulation. Motivated by this phenomenon, we constructed a linear regression and found that the maximum daily 8-hr ozone (MDA8) ozone concentration was highly correlated with the surface ozone change rate and chemical reactions in the PBL during the pollution period, with MDA8 ozone exceeding 70 ppbv over NCP. Based on this relationship as well as the design of numerical experiments, we propose a strategy of dynamic emission control. Firstly, the emission reduction during the peak ozone formation period may weaken the fast chemical reactions in the PBL and subsequent surface ozone concentration. Secondly, emission reduction one or two days prior to an episode might achieve larger ozone reduction through the accumulation effect. Lastly, emission control outside of the NCP may surpass the local impact under favorable meteorological conditions. Therefore, the efficacy of dynamic emission control was striking when both the accumulation and transport effect were taken into consideration.


White matter hypointensities and hyperintensities have equivalent correlations with age and CSF β-amyloid in the nondemented elderly.
T1- and T2-weighted sequences from MRI often provide useful complementary information about tissue properties. Leukoaraiosis results in signal abnormalities on T1-weighted images, which are automatically quantified by FreeSurfer, but this marker is poorly characterized and is rarely used. We evaluated associations between white matter hyperintensity (WM-hyper) volume from FLAIR and white matter hypointensity (WM-hypo) volume from T1-weighted images and compared their associations with age and cerebrospinal fluid (CSF) β-amyloid and tau. A total of 56 nondemented participants (68-94 years) were recruited and gave informed consent. All participants went through MR imaging on a GE 1.5T scanner and of these 47 underwent lumbar puncture for CSF analysis. WM-hypo was calculated using FreeSurfer analysis of T1 FSPGR 3D, and WM-hyper was calculated with the Lesion Segmentation Toolbox in the SPM software package using T2-FLAIR. WM-hyper and WM-hypo were strongly correlated (r = .81; parameter estimate (p.e.): 1.53 ± 0.15; p < .0001). Age was significantly associated with both WM-hyper (r = .31, p.e. 0.078 ± 0.030, p = .013) and WM-hypo (r = .42, p.e. 0.055 ± 0.015, p < .001). CSF β-amyloid levels were predicted by WM-hyper (r = .33, p.e. -0.11 ± 0.044, p = .013) and WM-hypo (r = .42, p.e. -0.24 ± 0.073, p = .002). CSF tau levels were not correlated with either WM-hyper (p = .9) or WM-hypo (p = .99). Strong correlations between WM-hyper and WM-hypo, and similar associations with age, abnormal β-amyloid, and tau suggest a general equivalence between these two imaging markers. Our work supports the equivalence of white matter hypointensity volumes derived from FreeSurfer for evaluating leukoaraiosis. This may have particular utility when T2-FLAIR is low in quality or absent, enabling analysis of older imaging data sets.


Sequential variation in the components of reproductive success in the distylousJasminum fruticans (Oleaceae).
A 2-year study of three natural populations of the distylousJasminum fruticans showed that mean fruit and seed production were significantly greater in shortstyled plants (thrums) than in long-styled plants (pins). In this study, we investigated the role of four sequential factors which may differentially influence fruit and seed set in the two floral morphs: (1) differences in flowering phenology, (2) a limitation of pollen transfer towards pins, (3) a differential capacity of the two morphs to act through famale and male function and (4) differential fruit abortion in the two morphs. Fruit set was significantly influenced by differences in flowering phenology although there were no differences in flowering time between the two morphs. supplementary pollinations in a natural population significantly increased fruit set and reduced the difference in fruit set between the two morphs in relation to controls, indicating a limitation on pollen transfer which was most severe towards pin stigmas. In reciprocal crosses, seed set was significantly dependent on the paternal and maternal identity of the pin parent. There was no significant variation among thrums for their performance as male or female parent. Furthermore, individual pin plants with relatively high percentage seed set as female parents gave poor seed set as male parents and vice-versa. Whereas fruit removal had no effect on seed number in thrum plants, a greater proportion of viable seeds were produced on pin plants which were left to naturally mature their fruits than on pins which had fruits artificially removed, suggesting the occurrence of selective fruit abortion in pins but not in thrums. The initially greater maternal fitness of thrums due to their greater success as pollen recipients may thus be opposed by increased viable seed set in the pins due to factors acting after the pollination stage. The relative reproductive success of floral morphs in the distylousJ. fruticans is thus differentially influenced by ecological factors occurring at different stages of the reproductive process.


Androgen-receptor blockade enhances pulsatile luteinizing hormone production in late pubertal males: evidence for a hypothalamic site of physiologic androgen feedback action.
To determine potential mechanisms by which androgens alter gonadotropin secretion and elimination in the pubertal male, we administered the potent nonsteroidal androgen receptor blocking agent, flutamide, to eight males with Tanner IV or V genital development. Venous blood samples were obtained every 10 min for 24 h and assayed for LH by a sensitive and high-precision fluorimmunoassay. Subjects were studied before and after the administration of flutamide. Deconvolution analysis was used to assess specific pulsatile LH secretory characteristics and estimate LH production and metabolic clearance rates quantitatively. After antagonism of endogenous androgen action, mean 24-h serum LH concentrations increased significantly. An increased mean 24-h LH production rate, without evident changes in serum LH half-life, accounted for the increase in average serum LH levels. The increased daily secretion rate of LH was in turn due to both an augmented mass of LH released per secretory episode and increased frequency of secretory events. There was no demonstrable change in the maximal rate of LH secretion attained within each secretory event. Serum concentrations of total testosterone, free-testosterone, and 17 beta-estradiol all increased during blockade of androgen action. Administration of the antiandrogen had no measurable effect on the pituitary response to a single maximally effective dose of exogenous gonadotropin releasing hormone (GnRH). These results indicate that, in the late pubertal male, endogenous androgen exerts negative feedback control of gonadotropin secretion primarily at a hypothalamic site reflected by regulation of the frequency of pulsatile LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)


Increased blood-brain barrier permeability on perfusion CT might predict malignant middle cerebral artery infarction.
Perfusion CT has been used to assess the extent of blood-brain barrier breakdown. The purpose of this study was to determine the predictive value of blood-brain barrier permeability measured using perfusion CT for development of malignant middle cerebral artery infarction requiring hemicraniectomy (HC). We retrospectively identified patients from our stroke registry who had middle cerebral artery infarction and were evaluated with admission perfusion CT. Blood-brain barrier permeability and cerebral blood volume maps were generated and infarct volumes calculated. Clinical and radiographic characteristics were compared between those who underwent HC versus those who did not undergo HC. One hundred twenty-two patients (12 HC, 110 no HC) were identified. Twelve patients who underwent HC had developed edema, midline shift, or infarct expansion. Infarct permeability area, infarct cerebral blood volume area, and infarct volumes were significantly different (P < 0.018, P < 0.0211, P < 0.0001, P < 0.0014) between HC and no HC groups. Age (P = 0.03) and admission National Institutes of Health Stroke Scale (P = 0.0029) were found to be independent predictors for HC. Using logistic regression modeling, there was an association between increased infarct permeability area and HC. The OR for HC based on a 5-, 10-, 15-, or 20-cm² increase in infarct permeability area were 1.179, 1.390, 1.638, or 1.932, respectively (95% CI, 1.035 to 1.343, 1.071 to 1.804, 1.108 to 2.423, 1.146 to 3.255, respectively). Increased infarct permeability area is associated with an increased likelihood for undergoing HC. Because early HC for malignant middle cerebral artery infarction has been associated with better outcomes, the infarct permeability area on admission perfusion CT might be a useful tool to predict malignant middle cerebral artery infarction and need for HC.


Novel application of rhein and its prodrug diacerein for reversing cancer-related multidrug resistance through the dual inhibition of P-glycoprotein efflux and STAT3-mediated P-glycoprotein expression.
Multidrug resistance (MDR) is a multifactorial issue in cancer treatment. Drug efflux transporters, particularly P-glycoprotein (P-gp), are major contributors to such resistance. In the present study, we evaluated the P-gp-inhibiting and MDR-reversing effects of two compounds, namely rhein, an anthraquinone, and diacerein, the acetylated prodrug of rhein. ABCB1/Flp-In-293 was used as a model for investigating the related molecular mechanisms, and the multi-drug-resistant cancer cell line KB/VIN was used as a platform for evaluating the reversal of MDR0. The results indicated that at a concentration of 2.5 μM, both diacerein and rhein significantly inhibited P-gp efflux function. They also downregulated P-gp expression by interacting with the signal transducer and activator of transcription 3. Further investigation of the inhibitory mechanism of these compounds revealed that both stimulated P-gp ATPase activity dose dependently and engaged in the noncompetitive inhibition of rhodamine 123 efflux. Furthermore, rhein was revealed to be a potent reverser of MDR in cancer, and the combination of 30 μM rhein and 1000 nM vincristine exerted a strong synergistic effect, achieving a high combination index (CI) of 0.092. Diacerein demonstrated potential applications as a selective cytotoxic agent against multi-drug-resistant cancer cells at a concentration of > 18.92 μM and as a mild MDR reverser at doses of < 10 μM. In conclusion, diacerein and rhein are potential candidates for P-gp inhibition and MDR reversal in cancer cells.


MiR-93 blocks cell cycle progression and promotes apoptosis in uterine leiomyoma cells by targeting CCND1.
Uterine leiomyoma (UL) is the most common type of benign tumor in the women's reproductive system. A number of genes has been found to play an important role in the initiation and progression of UL, including miRNAs. In this study, our results exhibited that miR-93, a member of mir-106b-25 cluster, significantly reduced the cell viability, promoted cell cycle arrest, caused apoptosis, and inhibited migration in UL cells (p < .01). Moreover, our results have provided experimental evidence that miR-93 regulated the biological functions of UL cells by targeting CCND1.


Modification of the Lowry assay to measure proteins and phenols in covalently bound complexes.
It is well established that phenols interfere with many routine protein assays and a number of protocols have been developed to overcome this. One such method is based on the differences in response obtained with the Lowry assay in the presence and absence of copper. This assumes that the phenol response with the Lowry assay is not affected by copper. However ortho-diphenols such as catechol, methylcatechol, caffeic acid, chlorogenic acid, and phaselic acid show decreased responses in the presence of copper. Three methods of estimating protein were compared for their accuracy in measuring proteins in the presence of covalently bound ortho-diphenols; the Lowry assay, the modified Lowry assay, and a new method including a calculation to take into account differences in ortho-diphenol response in the presence and absence of copper. The ortho-diphenols were caffeic acid and phaselic acid, which were bound to bovine serum albumin and red clover protein either chemically or enzymatically. For all assays, the new method gave values within 4 to 8% of control values for protein (without bound phenols) as determined by the modified Lowry method. Values for the Lowry and modified Lowry methods varied by 20-50% from control protein values. The new method also gave a good approximation of protein-bound phenol content.


Targeting of tissue plasminogen activator to the regulated pathway of secretion.
Tissue plasminogen activator is a serine protease that plays the dominant role in removal of fibrin from the vascular tree by activating plasminogen to the primary fibrinolytic enzyme, plasmin. Tissue plasminogen activator has a widespread neuroendocrine distribution in addition to its expression by endothelial cells. Within neuroendocrine cells, secretory proteins are sorted into one of two pathways: regulated or constitutive. Proteins entering the regulated pathway are concentrated and stored in vesicles, and subsequently released upon stimulation by a secretagogue. In contrast, in the constitutive pathway, newly synthesized protein is not stored but is transported directly to the cell surface and secreted even in the absence of an extracellular signal. The focus of this article is to review recent studies demonstrating that tissue plasminogen activator is targeted to the regulated secretory pathway in neuroendocrine cells and to discuss the physiological implications of the trafficking of tissue plasminogen activator to regulated secretory vesicles.


Vision, Aphasia, Neglect Assessment to Predict Neurosurgical Intervention in Patients with Nontraumatic Intracerebral Hemorrhage.
The Vision, Aphasia, and Neglect (VAN) screening tool is a simple bedside test developed to identify patients with large vessel occlusion stroke. In the setting of intracerebral hemorrhage (ICH), there are very few bedside predictors of need for neurosurgical interventions other than age and Glasgow Coma Scale (GCS). We aimed to assess the utility of the VAN screening tool in predicting the need for neurosurgical intervention in patients with ICH. We accessed sensitivity, specificity, positive predictive value, negative predictive value (NPV), and area under receiver operating characteristics curve of VAN for identifying ICH patients who require neurosurgical intervention. Among 228 ICH patients, 176 were VAN positive and 52 were VAN negative. On unadjusted analyses, VAN positive patients had a significantly higher ICH volume, GCS score, and National Institutes of Health Stroke Scale score (P < .001 for all). As compared to VAN negative patients, significantly higher proportion of VAN positive ICH patients (15.4% versus 32.4%) underwent a neurosurgical procedure such as external ventricular drain (EVD) and/or hematoma evacuation with craniotomy or craniectomy. The VAN screening tool had high sensitivity and NPV (100%) in predicting the need for craniectomy or hematoma evacuation, but had lower sensitivity (87.7%) for any neurosurgical procedure, as 15.4% of VAN negative patients received EVD. Our study suggests that VAN screening tool can identify high-risk ICH patients who are more likely to undergo craniotomy or craniectomy but is less sensitive to rule out need for EVD.