Ginsenoside Rh2 inhibits CBP/p300-mediated FOXO3a acetylation and epilepsy-induced oxidative damage via the FOXO3a-KEAP1-NRF2 pathway.
Epilepsy is a chronic disease that affects a wide range of people. Furthermore, a third of patients suffering from epileptic seizures do not respond to antiepileptic drugs. In recent years, increasing attention has focused on the role of oxidative stress in acquired epilepsy, and adjuvant antiepileptic drugs to reduce oxidative stress may be a new therapeutic strategy. In this study ginsenoside Rh2 was resistant to oxidative stress induced by epileptic activity in vivo and in vitro. Using online databases, we identified forkhead box O3a (FOXO3a) overexpression in epilepsy tissue and validated this in vitro, in vivo, and in clinical tissues of patients with epilepsy. An in vitro epilepsy model revealed that the overexpression of FOXO3a led to more severe oxidative stress, while the knockdown of FOXO3a had a protective effect on SH-SY5Y cells. Moreover, our results showed that the positive effect of FOXO3a on oxidative stress was caused by the transcriptional activation of Kelch-like ECH-associated protein 1 (KEAP1), a negative regulator of nuclear factor erythroid 2-related factor 2 (NRF2). We also found that ginsenoside Rh2 can directly inhibit the activation of FOXO3a by selectively blocking CREB-binding protein (CBP)/p300-mediated FOXO3a acetylation and play a role in regulating the KEAP1-NRF2 pathway to resist oxidative stress.


Induction of neutralising antibodies and cellular immune responses against SARS coronavirus by recombinant measles viruses.
Live attenuated recombinant measles viruses (rMV) expressing a codon-optimised spike glycoprotein (S) or nucleocapsid protein (N) of severe acute respiratory syndrome-associated coronavirus (SARS-CoV) were generated (rMV-S and rMV-N). Both recombinant viruses stably expressed the corresponding SARS-CoV proteins, grew to similar end titres as the parental strain and induced high antibody titres against MV and the vectored SARS-CoV antigens (S and N) in transgenic mice susceptible to measles infection. The antibodies induced by rMV-S had a high neutralising effect on SARS-CoV as well as on MV. Moreover, significant N-specific cellular immune responses were measured by IFN-gamma ELISPOT assays. The pre-existence of anti-MV antibodies induced by the initial immunisation dose did not inhibit boost of anti-S and anti-N antibodies. Immunisations comprising a mixture of rMV-S and rMV-N induced immune responses similar in magnitude to that of vaccine components administered separately. These data support the suitability of MV as a bivalent candidate vaccine vector against MV and emerging viruses such as SARS-CoV.


Could a "body fragmentation index" be useful in reconstructing events prior to burial: Case studies of selected primary and secondary mass graves from eastern Bosnia.
This study analyses variable levels of body fragmentation among secondary mass grave sites with similar formation process history. The study is based on data from 10 commingled secondary mass grave sites and two primary sites related to the war in Bosnia in 1995. The aim was to investigate differences in level of body fragmentation between mass graves of similar origin and taphonomy. In order to quantify the degree of fragmentation (and level of commingling) within a grave, we introduced a fragmentation index (FI). FI represents the ratio between the number of complete bodies and number of body parts from the same context. Results show high discrepancies in body fragmentations between different sites. FI for secondary sites of similar formation history varied from 0.01 to 0.59 (max = 1), while two primary sites have values 0.92 and 0.90 respectively. Variable levels of fragmentation among similar secondary sites suggest a possibility of different peri mortem circumstances of buried, so we tested whether the "body fragmentation index" could assist in elucidating the manner of death. Unusually high levels of body fragmentation (FI value below 0.1) in some secondary sites may indicate that body disarticulation was most likely caused peri-mortem by explosives, land mines, mortars or tank fire, all suggesting a combat situation.


Lignified and nonlignified fiber cables in the lacunae of Typha angustifolia.
The leaves of Typha are noteworthy in terms of their mechanical properties. We determined the mechanical properties of the fiber cables within the leaf. We found that in vegetative plants, the lignified fiber cables isolated from the leaf sheath and nonlignified fiber cables isolated from the leaf blade of Typha angustifolia differ in their diameter, swelling capacity, Young's modulus, tensile strength, and break load. These differing properties are related to their contributions to stability in the two regions of the leaf.


Is the high intensity symptoms experienced by patients admitted with chronic obstructive pulmonary disease documented by health professionals? - a prospective survey with comparison of patient reported outcomes and medical records.
Context: Patients with chronic obstructive pulmonary disease (COPD) have a high symptom burden and reduced quality of life. There is an increasing attention on palliation for patients with COPD. Recognition of symptoms is a prerequisite for palliation. Objectives: We aim to investigate the extent to which symptoms in patients with COPD are recognized in the documentation of the health professionals, indicated in 'Doctors Symptom Recognition Rate' (DSR), 'Nurses Symptom Recognition Rate' (NSR) or 'Doctors and/or Nurses Symptom Recognition rates '(DNSR) as a team, respectively. Methods: Patients with COPD (n = 40) admitted in two respiratory units, responded within 48 h on two symptom-screening-tools that access quality of life; COPD assessment test (CAT) used for the treatment of COPD and EORTC-QLQ-C15-PAL used for palliation in patients with cancer. Patient-described symptomatology was compared to the symptoms as recognized in the documentation of doctors and/or nurses. Results: There was a significant discrepancy between the symptomatology indicated by patients with COPD on CAT and EORTC-QLQ-C15-PAL, and the degree by which it was recognized in the medical records indicated in DSR or NSR. In 30 out of 44 items DSR or NSR were < 70%. There was a significant difference between DNSR versus DSR or NSR, respectively, in 19 out of 22 items.Conclusion: A team-based symptom recognition DNSR is superior when compared to DSR or NSR. Team-based systematic screening is suggested as a pathway to increase symptom recognition in patients with COPD. Increased rates of symptom recognition may improve symptom alleviation and thus palliation.


Breakdown of the regulatory control of pyrimidine biosynthesis in human breast cancer cells.
The activity of the de novo pyrimidine biosynthetic pathway in the MCF7 breast cancer cells was 4.4-fold higher than that in normal MCF10A breast cells. Moreover, while pyrimidine biosynthesis in MCF10A was tightly regulated, increasing as the culture matured and subsequently down-regulated in confluency, the biosynthetic rate in MCF7 cells remained elevated and invariant in all growth phases. The flux through the pathway is regulated by carbamoyl phosphate synthetase, a component of the multifunctional protein, CAD. The intracellular CAD concentration was 3.5- to 4-fold higher in MCF7 cells, an observation that explains the high rate of pyrimidine biosynthesis but cannot account for the lack of growth-dependent regulation. In MCF10A cells, up-regulation of the pathway in the exponential growth phase resulted from MAP kinase phosphorylation of CAD Thr456. The pathway was subsequently down-regulated by dephosphorylation of P approximately Thr456 and the phosphorylation of CAD by PKA. In contrast, the CAD P approximately Thr456 was persistently phosphorylated in MCF7 cells, while the PKA site remained unphosphorylated and consequently the activity of the pathway was elevated in all growth phases. In support of this interpretation, inhibition of MAP kinase in MCF7 cells decreased CAD P approximately Thr456, increased PKA phosphorylation and decreased pyrimidine biosynthesis. Conversely, transfection of MCF10A with constructs that elevated MAP kinase activity increased CAD P approximately Thr456 and the pyrimidine biosynthetic rate. The differences in the CAD phosphorylation state responsible for unregulated pyrimidine biosynthesis in MCF7 cells are likely to be a consequence of the elevated MAP kinase activity and the antagonism between MAP kinase- and PKA-mediated phosphorylations.


Fate of aflatoxins B1 and G 1 residues during biscuit processing.
Effect of biscuit processing on the destruction of aflatoxins B1 and G1 with and/or without some commonly leavening agents used namely sodium bicarbonate, ammonium bicarbonate and sodium bisulfite and sodium chloride.It was found that mixing step reduced the concentration of aflatoxins B1 and G1 by 80.7% and 82.7%, while the effect of baking step being 28.9% and 21.5%. The effect of mixing was found to be more pronounced than that baking step.The highest destruction effect on aflatoxin B1 was observed by adding a mixture composed of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium chloride, sodium bisulfite, ammonium bicarbonate and/or sodium bicarbonate alone, where the reduction values of toxin after mixing were 93.4,91.9,91.7, 88.8 and 86.6% respectively, while the baking effect ranged 17.2 to 34.5% in the presence of different leaving agents added.Concerning aflatoxin G1; the highest destructive effect of toxin was adsorbed by adding a mixture of sodium and ammonium bicarbonate and sodium bisulfite followed by sodium bisulfite, sodium chloride, ammonium bicarbonate and/or sodium bicarbonate alone since the destruction values of such toxin after mixing were 96.2%, 92.8%, 92.6%, 89.0% and 87.7% respectively, while the baking effect ranged 20.9 to 34.5% in all leavening agents added.


[On Cheyletiella infestation in domestic cats (author's transl)].
Two Persian cats kept in common, 3 months and 2 years old, presented a heavy infestation with Cheyletiella mites. The cats showed itching and loss of hair. The animals' owners suffered from severe itching and redish papulae, particularly on those parts of the body which frequently got in contact with the cats. After three treatments with Alugan powder (Hoechst) the cats were freed from the mite infestation. The owners' skin affection disappeared soon without any treatment. This is the first recovery of Cheyletiella mites on cats in Austria. The mites were identified as C. blakei, although obvious differences between the first description and our material were encountered. C. blakei could be primarily separated from C. parasitivorax according to the female tectum and the peritremata. Typically the sense organs on the genua of the first pair of legs are ovoid in shape. This feature, although morphologically variable, could be used as a diagnostic aid, if enough adult specimens are available.


A comparison of the incidence and understanding of altitude illness between porters and trekkers in the Solu Khumbu Region of Nepal.
Altitude illness can occur in anyone who ascends to high altitude. Better understanding of altitude illness is associated with a lower incidence of acute mountain sickness (AMS). The purpose of this study is to compare, for the first time, the incidence and understanding of altitude illness between foreign trekkers and indigenous porters in Nepal. Interviews and questionnaires were completed at the International Porter Protection Group Rescue Post at Machermo (4470 m). Participants completed the Lake Louise acute mountain sickness self-assessment questionnaire. They were also asked about their actions in response to high altitude illness scenarios as well as their perception of the vulnerability of porters vs trekkers to altitude illness. Ascent profile, age, gender, ethnic origin, and altitude of home residence were also obtained. Trekkers (n=131) had a significantly higher incidence of AMS (21% vs 8%) than porters (n=92; P < .02). Lowland porters (whose home villages were below 3050 m, n=61) had a numerically higher, though not significantly different, incidence of AMS (10% vs 3%) compared to highland porters (n=31). The majority of trekkers and porters recognized the symptoms of altitude illness and the most appropriate action to be taken. Despite the lower incidence of AMS in porters, around half felt that they were at greater risk than trekkers. Porters had a lower incidence of AMS, which may be attributable to repeated ascents through the trekking season, or differences in reporting symptoms. Both trekkers and porters demonstrated appropriate knowledge of actions to be taken in response to altitude illness.


Synthesis and antitumor activity of 1-beta-D-arabinofuranosylcytosine conjugates of optical isomers of ether and thioether lipids.
Four 1-beta-D-arabinofuranosylcytosine conjugates (ara-C) (1a, b and 2a, b) of sn-1 and sn-3 isomers of 1-O-octadecyl-2-O-palmitoylglycerol and its 1-S-alkyl analogue have been synthesized, and their antitumor activity against L1210 lymphoid leukemia in mice were compared with those of the previous conjugates (3a, b) of racemates in order to determine the significance of chirality of the glycerol moieties for activity. Administration (i.p.) of a single dose (300 mg/kg) of conjugates of sn-1 (1a), sn-3 (2a) and rac (3a) isomers of the ether lipid increased lifespan of i.p. implanted L1210 lymphoid leukemic DBA/2J mice by 169, 175 and 236%, respectively. The sn-1 (1b), sn-3 (2b), and rac (3b) isomers of the thioether lipid with a single dose of 300 mg/kg produced an increase in lifespan values of 238, 263 and 250%, respectively. The results indicate that chirality of the glycerol moieties appears not to be critical for the activity, and racemates 3a and 3b are promising prodrugs of ara-C for further clinical investigations.


Comparison of the production of a human monoclonal antibody against HIV-1 by heterohybridoma cells and recombinant CHO cells: A flow cytometric study.
The production of human monoclonal antibodies for therapeutic use is of increasing importance for treatment of viral infections such as AIDS. As human x mouse heterohybridomas rarely reach the growth rates and cell specific production rates of mouse hybridomas the transfection of standard cell lines, such as CHO or BHK, is a promising alternative. This has the additional advantage that the IgG subtype can be changed to suit the desired application. However, the use of a cell line that has not originally developed to produce antibodies, as lymphocytes and myeloma cells have, might have unrecognised drawbacks. This will be especially significant in the case of antibodies as each molecule consists of 4 chains linked by disulphide bonds which require specific intracellular factors to be properly folded and processed (Heavy chain binding protein, Protein Disulfide Isomerase a.o.). In this study we have therefore compared two cell lines: a human x mouse heterohybridoma producing IAM-2F5, a human IgG(3) antibody specific for HIV-1 with neutralising properties and a Chinese Hamster Ovary cell transfected with dihydrofolate reductase and with the heavy and light chain genes of IAM-2F5 modified to IgG(1). From each cell line three subclones were selected with low, medium and high specific production rates. Batch cultures were performed and the following cellular parameters analysed by flow cytometry; 1) total RNA content (translational activity); 2) total protein content; 3) cell cycle phase distribution; 4) concentration of light and heavy chains; 5) concentration of helper proteins such as BiP and PDI. The production rate of heterohybridoma cells was best reflected in the intracellular concentration of kappa chain, while the gamma chain concentration was comparable for all three subclones. In the CHO cells the gamma chain expression and thus gene copy number appeared to be the limiting factor. The GRP78/BiP concentration in CHO remained unchanged in spite of a 5-fold higher concentration of gamma chain in the high producing subclone. The PDI concentration in CHO cells was much lower compared to the heterohybridoma cells, irrespective of production rates.


Royal College of Ophthalmologists' National Ophthalmology Database study of cataract surgery: report 8, cohort analysis of the relationship between intraoperative complications of cataract surgery and axial length.
To describe the relationships between axial length and intraoperative complications in patients undergoing cataract surgery. Cohort analysis of the Royal College of Ophthalmologists' National Ophthalmology Database (RCOphth NOD). 110 National Health Service Trusts in England, Health Boards in Wales, Independent Sector Treatment Centres and Guernsey. 820 354 patients, aged 18 years or older, undergoing cataract surgery. Eligible operations were those from centres with at least 50 operations with a recorded axial length measurement and age at surgery between 1 April 2010 and 31 August 2019. Phacoemulsification where the primary intention was cataract surgery alone. Posterior capsule rupture (PCR) and other recorded intraoperative complications. 1 211 520 eligible operations were performed by 3210 surgeons. The baseline axial length was <21 mm (short eyes) for 17 170 (1.4%) eyes, 21-28 mm (medium eyes) for 1 182 513 (97.6%) eyes and >28 mm (long eyes) for 11 837 (1.0%) eyes. The median age at surgery was younger for patients with long eyes than those with short or medium eyes. The rate of any intraoperative complication was higher for short eyes than medium or long with complication rates of 4.5%, 2.9% and 3.3%, respectively (p<0.001). PCR occurred in 1.40% surgeries overall, and in 1.53%, 1.40% and 1.61% of short, medium and long eyes, respectively (p=0.043, not significant at the 1% level). Overall PCR rates for cataract surgery in RCOphth NOD contributing centres are lower than previously reported and there is little change in PCR rates by axial length. Short eyes were more likely to have an intraoperative complication than medium or long eyes.


Somatosensory graviception inhibits the soleus H-reflex in standing man - a parabolic flight experiment-.
The purpose of this study was to investigate how gravity level affects the excitability of the soleus muscle (SOL) motoneuron pool to la afferent input while erect posture is maintained in humans. Three healthy male subjects participated in an experiment whereby three different gravity conditions (micro gravity (MG), normal gravity (NG), and hyper gravity) were imposed using a parabolic flight procedure. The SOL H-reflex was evoked every 2 seconds while the subjects kept an erect posture. The background electromyographic activity (BGA) of the SOL was almost absent during MG. The SOL H-reflex amplitude was significantly larger during MG than during NG. These results suggest that the somatosensory systems detecting a load at the lower limbs and/or vertebral column play a role in reducing the excitability of the SOL motoneuron pool to la afferent inputs by presynaptic inhibition.


Role of regulated upon activation normal T-cell expressed and secreted in a model of retransplantation acute rejection mediated by alloreactive memory CD4+ T cells.
It is unknown what role Regulated Upon Activation Normal T-Cell Exposed and Secreted may play in retransplantation or T-cell memory-transfer models. The experiment observed the influence of the chemokine RANTES in a mouse model of acute cardiac allograft rejection induced by adoptive transfer of alloreactive CD4(+) memory T (Tm) cells. Alloreactive CD4(+) Tm cells from spleens of skin-grafted C57BL/6 were adoptively transferred to naïve C57BL/6 recipients prior to heterotopic heart transplantation. We measured the median survival time of cardiac grafts and performed some tests. Spleens from skin-grafted C57BL/6 contained 26.83% CD4(+) Tm cells. The median graft survival time of heterotopic heart transplantations (n = 6) was 5.17 ± 0.17 days for hosts receiving CD4(+) Tm cells compared with 7.76 ± 0.21 days among controls (n = 6; P < .001). The mean rejection activity in histological sections of cardiac allografts at day 5 postgrafting was 3.92 ± 0.08 in the CD4(+) Tm cell recipient group (n = 6) compared with 2.67 ± 0.14 in the controls (n = 6; P < .001). Gene expression of Ccl5, interferon (IFN)-γ and interleukin2 was significantly higher among CD4(+) Tm recipients compared with controls. Serum concentrations of RANTES and IFN-γ were higher in the heterotopic heart transplantation group receiving CD4(+) Tm compared with controls. Alloreactive CD4(+) Tm cells contribute to increased expression and secretion of RANTES, and to the Tm and other inflammatory cells migration into the graft.


Transitory macrophages in the white matter of the developing visual cortex. I. Light and electron microscopic characteristics and distribution.
Injections of horseradish peroxidase (HRP) into the occipital cortex of the kitten and diffusing to the white matter label a widely distributed microglial population and in addition, cells with light and electron microscopic features of 'gitter cells'. The latter are concentrated in a complex and highly consistent system of interconnected clusters in the white matter of the lateral, postlateral, middle suprasylvian and posterior ectosylvian gyri, as well as on the roof of the lateral ventricle. The 'gitter cells' have the ultrastructural and (as described by others) chemical characteristics of macrophages, and may be involved in the elimination of transitory axons.


Synaptic pruning through glial synapse engulfment upon motor learning.
Synaptic pruning is a fundamental process of neuronal circuit refinement in learning and memory. Accumulating evidence suggests that glia participates in sculpting the neuronal circuits through synapse engulfment. However, whether glial involvement in synaptic pruning has a role in memory formation remains elusive. Using newly developed phagocytosis reporter mice and three-dimensional ultrastructural characterization, we found that synaptic engulfment by cerebellar Bergmann glia (BG) frequently occurred upon cerebellum-dependent motor learning in mice. We observed increases in pre- and postsynaptic nibbling by BG along with a reduction in spine volume after learning. Pharmacological blockade of engulfment with Annexin V inhibited both the spine volume reduction and overnight improvement of motor adaptation. These results indicate that BG contribute to the refinement of the mature cerebellar cortical circuit through synaptic engulfment during motor learning.


Determination of rhodium by resonance light-scattering technique coupled with solid phase extraction using Rh(III) ion-imprinted polymers as sorbent.
A resonance light-scattering method (RLS) for the determination of Rh(III) was initially developed, based on the reaction among Rh(III), WO4(2-) and ethylrhodamine B. The method possesses high sensitivity, but lacks selectivity. Therefore, a Rh(III) ion-imprinted polymer (IIP), prepared by precipitation polymerization using 2-(allylthio)nicotinic acid (ANA) as functional monomer, was used as sorbent to construct a ion-imprint based solid-phase extraction (IIP-SPE) method for separation of rhodium from complicated matrices prior to its determination by RLS. The experimental parameters affecting the extraction efficiency and selectivity of IIP-SPE were studied carefully. Under the optimal conditions, the IIP-SPE column with the enrichment factor (EF) of 10 could be used at least 20 times without decreasing its extraction recovery (above 90%) significantly. The calibration graph for the determination of rhodium by RLS coupled with IIP-SPE procedure was linear in the range of 0.06-1.5 ng mL(-1) with the detection limit of 0.024 ng mL(-1). There is no metal ions tested at the concentration below 10 ng mL(-1) interfered in the determination of 0.8 ng mL(-1) Rh(III). The proposed IIP-SPE-RLS method was successfully applied to the extraction and measurement of trace rhodium in catalyst, water and geochemical samples with the relative standard deviation (RSD) of less than 4.0% (n=4).


Single-step purification of equine chorionic gonadotrophin directly from plasma using affinity chromatography.
Equine chorionic gonadotrophin (eCG) is a hormone widely used in superovulation protocols because of its follicle-stimulating action, which increases reproductive efficiency in animals of productive interest. It contains 45% carbohydrate, 10% of which is N-acetylneuraminic acid (sialic acid). The eCG purification procedures from equine serum or plasma are mainly based on chromatographic methods. However, before these procedures, it is necessary to follow sample pre-conditioning steps, such as several precipitation stages and/or ultrafiltration/diafiltration processes. In this work, an efficient affinity chromatographic matrix for eCG purification directly from plasma was developed. The matrix consisted of chitosan mini-spheres with immobilized wheat germ agglutinin (WGA). The matrix allowed 98% adsorption of eCG directly from plasma without any pre-treatment with an overall yield of around 60%. The matrix chosen was able to maintain the efficient performance of the purification process for three consecutive cycles. Also, the process was scaled-up 500 times in volume and tested over seven consecutive cycles maintaining its chromatographic performance. The results presented here suggest the potential application of this matrix to one-step purification of eCG from plasma.


Polymers for biodegradable medical devices. II. Hydroxybutyrate-hydroxyvalerate copolymers: hydrolytic degradation studies.
The hydrolytic degradation of poly(hydroxybutyrate) together with a series of hydroxybutyrate-hydroxyvalerate copolymers has been studied. The effects of copolymer composition and molecular weight are presented together with the results of varying pH and temperature on the degradation rate. Degradation has been monitored by weight loss and water uptake measurements together with goniophotometric, surface energy and scanning electron microscopic studies. Some comparisons with the more widely used so-called 'biodegradable' polymers, poly(glycolic acid), poly(dioxanone) and the glycolic-lactic acid (90:10) copolymers are presented together with the effect of blood plasma on the degradation process.


Delayed glucose peak and elevated 1-hour glucose on the oral glucose tolerance test identify youth with cystic fibrosis with lower oral disposition index.
Alternate methods for characterizing oral glucose tolerance tests (OGTT) have emerged as superior to the 2-hour glucose in identifying individuals at risk for type 2 diabetes. The significance of these methods in cystic fibrosis (CF) is unclear. We compared 3 OGTT classifications in youth with CF: 1. curve shape (biphasic vs. monophasic), 2. time to glucose peak (≤30minutes vs. >30minutes), 3. 1-hour glucose (1hG) <155 mg/dL vs. ≥155 mg/dL to traditional OGTT criteria to determine which best identifies lower oral disposition index (oDI), pulmonary function, and body mass index (BMI). Youth 10-18 years with CF, not on insulin, underwent 2-hour OGTT. Glucoses were classified by traditional criteria and 3 alternate methods as normal (biphasic curve, glucose peak ≤30minutes, and/or 1hG <155 mg/dL) or abnormal (monophasic curve, glucose peak >30minutes, and/or 1hG ≥155 mg/dL). oDI was calculated [1/fasting insulin*(ΔInsulin0-30 min/ΔGlucose0-30 min)]. Mean oDI, BMI, forced expiratory volume in 1 second (FEV1), and forced vital capacity (FVC) were compared by OGTT classification. Fifty-two youth with CF participated (mean±SD age 13±4years; 37% male; BMI z-score 0.0±0.8; FEV1 88±16.3%; FVC 97±14.8%). Late time to peak glucose and 1hG ≥155 mg/dL identified individuals with lower oDI (p=0.01); traditional OGTT criteria for prediabetes did not. No OGTT classification identified individuals with worse BMI nor pulmonary function. oDI was not associated with BMI, FEV1, or FVC. Alternate OGTT measures including time to peak glucose and 1hG better identify oDI abnormalities than traditional criteria. Further studies are required to determine whether these alternate methods identify individuals with CF at risk for future clinical decline.


Laparoscopic adrenalectomy for nonmalignant disease: improved safety, morbidity, and cost-effectiveness.
Laparoscopic adrenalectomy has rapidly gained widespread acceptance for treatment of benign adrenal neoplasms. A number of authors have compared various anatomic approaches to laparoscopic adrenalectomy, comparing length of inpatient stay, transfusion requirements, and perioperative complications. Separate studies have found inpatient stay reduced 40-60% with the use of laparoscopic adrenalectomy vs. an open procedure. There have been no studies designed specifically to examine and compare perioperative morbidity, length of stay, and patient charges in patients undergoing laparoscopic adrenalectomy. This report examines the Johns Hopkins Hospital experience with laparoscopic adrenalectomy in 22 patients, comparing length of stay, perioperative morbidity, and patient charges. These data are compared with those seen in 17 patients undergoing open adrenalectomy within our institution and 70 patients at all other nonfederal hospitals in the state of Maryland. Outcomes after laparoscopic versus open adrenalectomy were compared. Resumption of diet (1.6 vs. 6.1 days), independent activity (1.6 vs. 7.9 days), inpatient length of stay (1.7 vs. 7.8 days), and total hospital patient charges ($8,698 vs. $12,610) were all significantly reduced in patients undergoing laparoscopic adrenalectomy at our institution. Similar findings were obtained when our data were compared against adrenalectomy performed at other hospitals within the state of Maryland. Length of stay (1.7 vs. 8.9 days) and total hospital patient charges ($8,698 vs. $13,867) were both significantly reduced compared to state-wide data in patients treated with laparoscopic adrenalectomy. Although a technically challenging procedure, laparoscopic adrenalectomy provides clear advantages over open procedures for the vast majority of adrenal neoplasms. Our data support the conclusion that laparoscopic adrenalectomy should be considered for all patients with benign adrenal neoplasms.


Cl- interference with the epithelial Na+ channel ENaC.
The cystic fibrosis transmembrane conductance regulator (CFTR) is a protein kinase A and ATP-regulated Cl- channel that also controls the activity of other membrane transport proteins, such as the epithelial Na+ channel ENaC. Previous studies demonstrated that cytosolic domains of ENaC are critical for down-regulation of ENaC by CFTR, whereas others suggested a role of cytosolic Cl- ions. We therefore examined in detail the anion dependence of ENaC and the role of its cytosolic domains for the inhibition by CFTR and the Cl- channel CLC-0. Coexpression of rat ENaC with human CFTR or the human Cl- channel CLC-0 caused inhibition of amiloride-sensitive Na+ currents after cAMP-dependent stimulation and in the presence of a 100 mM bath Cl- concentration. After activation of CFTR by 3-isobutyl-1-methylxanthine and forskolin or expression of CLC-0, the intracellular Cl- concentration was increased in Xenopus oocytes in the presence of a high bath Cl- concentration, which inhibited ENaC without changing surface expression of alpha beta gammaENaC. In contrast, a 5 mM bath Cl- concentration reduced the cytosolic Cl- concentration and enhanced ENaC activity. ENaC was also inhibited by injection of Cl- into oocytes and in inside/out macropatches by exposure to high cytosolic Cl- concentrations. The effect of Cl- was mimicked by Br-, Br-, NO3(-), and I-. Inhibition by Cl- was reduced in trimeric channels with a truncated COOH terminus of betaENaC and gammaENaC, and it was no longer detected in dimeric alpha deltaCbeta ENaC channels. Deletion of the NH2 terminus of alpha-, beta-, or gammaENaC, mutations in the NH2-terminal phosphatidylinositol bisphosphate-binding domain of betaENaC and gammaEnaC, and activation of phospholipase C, all reduced ENaC activity but allowed for Cl(-)-dependent inhibition of the remaining ENaC current. The results confirm a role of the carboxyl terminus of betaENaC for Cl(-)-dependent inhibition of the Na+ channel, which, however, may only be part of a complex regulation of ENaC by CFTR.


Peripartum changes in the activity and expression of neutrophils may predispose to the postpartum occurrence of metritis in dairy cows.
Metritis is a postpartum uterine pathology that causes a huge economic loss due to increased culling risk and impaired milk yield and reproduction in cows. The present study was carried out to study the changes in the activity and expression of blood neutrophils in crossbred dairy cows with and without metritis. Collection of blood samples was done at -3, -2 and - 1 weeks before calving, at calving and during the first day of metritis diagnosis in metritis group (n = 8) or at day 8-10 post calving in healthy group (n = 8). Neutrophils were studied for its percentage (microscopically), respiratory burst (nitro blue tetrazolium assay), myeloperoxidase (MPO) concentrations (sandwich ELISA) and expression of CXCR1, CXCR2, TLR2, TLR4, GRα, CD11b, CD14, CD25, CD44, CD47 and CD62L (RT-PCR). Immunocytochemistry was used to investigate MPO concentration and CD14 activity, and western blotting was used for estimating MPO. Although most of these parameters changed in the cows that developed metritis one week before calving, MPO and CD14 got altered much earlier. Myeloperoxidase concentrations and expression of CD14 were considerably lower starting from -2 weeks before calving in cows that developed metritis compared to healthy cows. Further studies are warranted to study the possible use of MPO and CD14 to identify transition cows more vulnerable to develop metritis several weeks before disease occurrence.


A mimotope of Aβ oligomers may also behave as a β-sheet inhibitor.
Beta-amyloid (Aβ) oligomers are strongly associated with the cascade of harmful events leading to neurodegeneration in Alzheimer's disease (AD). Elimination of Aβ oligomers or inhibition of Aβ assembly is a valuable therapeutic approach for the treatment of AD. Here, we obtained a mimotope of Aβ oligomers, AOEP2, by screening a peptide library using oligomer-specific antibodies. The antibodies induced by AOEP2 specifically recognize Aβ oligomers rather than monomers and fibrils. Interestingly, the AOEP2 peptide binds to Aβ monomers and inhibits the formation of Aβ oligomers and β-sheet structure, reduces Aβ42-induced neurotoxicity, and decreases the release of proinflammatory cytokines. Taken together, AOEP2, a novel multifunctional peptide directly or indirectly targeting Aβ, has promising therapeutic potential for AD.


Nasopharyngeal Lymphoma: Rare Case Report.
A case of Nasopharyngeal Non Hogkins lypmhomas, diagnosed on Histopathology (HPE) and immunohistochemistry. In this case report we present the clinical presentation, diagnosis and treatment protocol of nasopharyngeal NHL.


Production of transgenic models in hypertension.
This chapter describes the generation of transgenic mice and rats by microinjection of DNA constructs into the pronucleus of a zygote. The transgene DNA is randomly integrated as several tandem copies at one site into the genome and is transmitted to the offspring of the founder animal derived from the injected embryo, thereby creating a stable transgenic line. The technology includes the following steps: design and generation of the transgene construct, superovulation of donor animals, isolation of fertilized oocytes, microinjection of the transgene construct into one pronucleus of the zygotes, transfer of injected embryos into the oviduct of a foster mother, and identification of transgenic animals in the offspring.


Differential scanning calorimetry of chromaffin granule membranes.
Differential scanning calorimetry thermograms of native chromaffin granule membranes exhibit several peaks in the 15-35 degrees C region. Extraction of cholesterol increases the size of the melting peaks. Addition of Ca2+ ions does not seem to influence the lipid transitions.


Impaired glucagon secretion in patients with fulminant type 1 diabetes mellitus.
Fulminant type 1 diabetes mellitus (FT1DM), characterized by rapid and almost complete destruction of pancreatic β-cells, is a newly identified subtype of type 1 diabetes mellitus. Although, the pathophysiology of this condition remains still unclear, histological evidence suggests that not only β-cells but also α-cells of pancreatic islets are reduced in number in FT1DM. However, the ability of glucagon secretion in patients with this condition has remained largely uncharacterized. We therefore examined glucagon secretion in patients with FT1DM and compared that with patients with other types of diabetes mellitus. Fasting glucagon levels as well as glucagon secretion induced by intravenous administration of arginine were measured in hospitalized 83 patients with diabetes mellitus, including 4 with FT1DM, 18 with type 1 diabetes mellitus (T1DM), 40 with type 2 diabetes mellitus (T2DM), 5 with slowly progressive insulin-dependent diabetes mellitus (SPIDDM), and 16 with pancreatic diabetes mellitus (PDM). The area under the curve for serum glucagon levels after arginine infusion in FT1DM patients was significantly smaller than that in T1DM, T2DM, or SPIDDM patients but was similar to that in PDM patients. The fasting serum glucagon level of FT1DM patients was lower than that of T1DM or T2DM patients but did not significantly differ from that of SPIDDM or PDM patients. These results suggest that glucagon secretion is impaired in patients with FT1DM.


Diagnostic and Treatment Considerations in Restless Legs Syndrome Complicated by Diabetic Neuropathy.
Diabetic neuropathy from type I diabetes (DMI), type II diabetes (DMII), or gestational diabetes commonly occurs concurrently with restless legs syndrome (RLS) with an estimated prevalence in DMII specifically of 17.7 to 45%. The diagnosis of RLS can be obscured by symptoms of peripheral neuropathy, which may be similar to those of RLS, but also by other RLS mimics, including nocturnal leg cramps or akathisia. The purpose of this review is to outline a framework for considering comorbid diabetic neuropathy and RLS by discussing RLS and its association with peripheral nerve disease, diabetes, and sleep disruption. Treatment for patients with both diabetic neuropathy and RLS may differ from treatment of persons with solely RLS or diabetic neuropathy; key points in this difference are discussed. Finally, a treatment algorithm is provided to guide the approach to treating patients with comorbid RLS and diabetic neuropathy. There have been several revisions of the RLS diagnostic criteria which in addition to the main four essential criteria of having (1) an urge to move the legs which is (2) exacerbated by rest, (3) relieved temporarily by movement, and (4) occurs in the evening or night time, now includes a consideration for RLS mimics, requiring that (5) symptoms not be explained by any other symptoms. Additionally, the treatment of both diabetic neuropathy and RLS has evolved to focus first on controlling the blood glucose, avoiding exacerbating medications, supplementing with iron if serum ferritin is low, and starting pharmacotherapy with alpha-2-delta ligands such as gabapentin or pregabalin. Knowledge of the prevalence of peripheral neuropathy with RLS, revised RLS diagnostic criteria, and a general treatment algorithm for the combination of peripheral neuropathy and RLS is critical to providing appropriate care to patients suffering from these two diseases.


Cardiovascular effects of anatoxin-A in the conscious rat.
The effects of anatoxin-A on mean arterial pressure (MAP), heart rate, cardiac index (CI), and blood flow (BF) in hindquarter (HQ), renal (R), and mesenteric (M) vascular beds were studied after intravenous (iv) and intracerebroventricular (icv) administration in the conscious rat. The pharmacological profile of anatoxin-A was further compared to nicotine administered iv and icv. MAP and heart rate were measured from femoral artery, CI by thermodilution method, and blood flow by Doppler velocimetry. Anatoxin-A and nicotine (30, 100 and 300 micrograms/kg iv) produced an increase in MAP with concomitant bradycardia. The highest doses increased CI. MBF and RBF decreased due to a vasoconstriction in M and R vasculature. These effects were attenuated by the ganglion blocker chlorisondamine (5 mg/kg, iv). Anatoxin-A (100 micrograms/kg, iv) increased plasma epinephrine levels by 2-fold with virtually no effect on norepinephrine whereas nicotine (100 micrograms/kg, iv) increased plasma epinephrine and norepinephrine by 20- to 30-fold. Central administration of anatoxin-A and nicotine (30-100 micrograms/kg icv) increased MAP with no effect on heart rate and produced M and R vasoconstriction. In summary, the present study demonstrates that anatoxin-A acts as a nicotinic cholinergic agonist in the conscious rat after both systemic and central administration. Anatoxin-A and nicotine produced pressor and reno-splanchnic vasoconstrictor responses and at high doses increased cardiac output. These effects were mediated by activation of the nicotinic receptors in the adrenal medulla and sympathetic ganglia. However, marked differences were found in the potency of anatoxin-A versus nicotine to stimulate the sympathoadrenomedullary axis.


Roles for central leptin receptors in the control of meal size.
The adiposity hormone leptin has been implicated in the regulation of behavioral and metabolic controls of body weight. Leptin receptors are found in multiple peripheral and central tissues, particularly within hypothalamic and brainstem neuronal populations. Central leptinergic signaling acts as an indirect control to modulate the feeding inhibitory potency of the direct controls of meal size. Mouse models of neuronal leptin loss and gain of function have helped to identify and characterize how central leptin contributes to the central control of food intake.


Hydroxylation and dechlorination of 3,3',4,4'-tetrachlorobiphenyl (CB77) by rat and human CYP1A1s and critical roles of amino acids composing their substrate-binding cavity.
Cytochrome P450 (CYP) monooxygenases play critical roles in determining the toxicity of polychlorinated biphenyls (PCBs) in mammals. Hydroxylation of PCBs by these enzymes leads to increased water solubility, promoting the elimination of PCBs from the body. The CYP1 family is mainly responsible for metabolizing PCBs that exhibit a dioxin-like toxicity. Although the dioxin-like PCB 3,3',4,4'-tetrachlorobiphenyl (CB77) is abundant in the environment and accumulates in organisms, information on CB77 metabolism by CYP1A1s is limited. In this study, recombinant rat CYP1A1 metabolized CB77 to 4'-hydroxy (OH)-3,3',4,5'-tetrachlorobiphenyl (CB79) and 4'-OH-3,3',4-trichlorobiphenyl (CB35), whereas human CYP1A1 produced only 4'-OH-CB79. Rat CYP1A1 exhibited much higher metabolizing activity than human CYP1A1 because CB77 was stably accommodated in the substrate-binding cavity of rat CYP1A1 and was close to its heme. In a rat CYP1A1 mutant with two human-type amino acids, the production of 4'-OH-CB79 decreased, whereas that of the dechlorinated metabolite 4'-OH-CB35 increased. These results are explained by a shift in the CB77 positions toward the heme. This study provides insight into the development of enzymes with high metabolizing activity and clarifies the structural basis of PCB metabolism, as dechlorination contributes to a drastic decrease in dioxin-like toxicity.


Assessment of tracer 99mTc(V)-DMSA uptake as a measure of tumor cell proliferation in vitro.
To examine whether (99m)Tc(V)-DMSA could be used as a non-invasive measure of cancer cell proliferation. Human breast cancer MCF-7, MDA-MB-231 and pII, and prostate cancer PC-3 cell lines were grown to 30, 50 and 100% confluency and pulsed with (99m)Tc(V)-DMSA in media for 60 min at 37°C. DNA synthesis was analysed by quantification of the S phase using flow cytometry, [methyl-(3)H]thymidine incorporation and expression of proliferation markers PCNA and Ki-67 using realtime PCR. One way ANOVA was used to compare groups. In all cell lines rates of (99m)Tc(V)-DMSA uptake were inversely related to cell density. This was paralleled by similar trends in S phase proportions, [methyl-(3)H]thymidine incorporation and expression of PCNA and Ki-67. Rates of (99m)Tc(V)-DMSA uptake into different types of tumour cells correlate well with cell density that is useful as a non-invasive measure of tumour cellular proliferation in vivo.


Laparoscopic Transabdominal Cerclage for Cervical Incompetence: A Feasible and Effective Treatment in 10 Steps.
To demonstrate the surgical technique of laparoscopic cerclage (LAC) in nonpregnant women with a clinical diagnosis of cervical incompetence. In this video, the authors describe the complete procedure in 10 steps to standardize and facilitate the comprehension and performance of the procedure in a simple and safe way. Step-by-step video demonstration of the surgical technique. Private hospital in Curitiba, Paraná, Brazil. The patient was 32 years old (gravidity and parity, G3A3; late progressive miscarriage), had no comorbidities, and had a radiologic diagnosis of cervical incompetence. The main steps of LAC are described in detail. A complete laparoscopic approach was performed. Under general anesthesia, the patient was placed in the 0-degree supine decubitus position with arms alongside her body. The operative setup included a 15-mm Hg pneumoperitoneum created using the closed Veress technique and 4 trocars: a 10-mm trocar at the umbilicus for a 0-degree laparoscope; a 5-mm trocar in the right iliac fossa; a 5-mm trocar in the left iliac fossa; and a 5-mm trocar in the suprapubic area. After systematic exploration of the pelvic and abdominal cavities, the procedure began. Step 1 involved identification of anatomic key landmarks and exposure of the operation field. Step 2 involved opening of the anterior peritoneum. The anterior peritoneal reflection was opened over the peritoneum uterovesicalis and then extended laterally until the uterine artery could be clearly identified on both sides. Step 3 involved dissection of the avascular space on each side of the uterus. The vesical-cervical avascular space was created, and the bladder was pushed down, away from the isthmus area. Step 4 involved preparation for a perfect stitch placement. A 5-mm Mersilene suture (Ethicon, Somerville, NJ) with a straight needle was introduced by a suprapubic trocar into the abdominal cavity before a complete identification of uterine vessels at both the sides using atraumatic graspers. Step 5 involved identification of the perfect space in the posterior aspect for Mersilene suture placement. Step 6 was to make a perfect anterior stitch. For this, the needle was grasped at the proximal portion in a 90-degree angle. In posterior position and when helped by a cranial and posterior uterine mobilization, the needle passed through the right, broad ligament in the avascular space created on the anterior leaf and medially from the uterine artery until the tip of the needle was seen on the posterior face above the uterosacral ligament. All steps were possible by synchronic uterine mobilization. Step 7 was to make a perfect posterior stitch. The procedure was then repeated contralaterally following the same anatomic and technical precepts but from posteriorly to anteriorly. Step 8 involved correct positioning and orientation of the Mersilene suture far away from the ureter and medial to the uterine arteries 2 cm over the uterosacral ligaments. Step 9 involved fixation of the Mersilene suture with an adequate blocking sequence. Step 10 involved fixation of the Mersilene suture and reperitonealization. The tape was knotted with an adequate blocking intracorporeal suturing sequence at the cervicoisthmic junction, and a Monocryl 2-0 stitch (Ethicon, Somerville, NJ) was made to fix the knot and left it horizontally. Finally, the procedure was ended with anterior reperitonealization, covering all the plica uterovesicalis and mesh, leaving it completely extraperitoneal. The surgery ended without any intraoperative complications and within 30 minutes. Patient was discharged on the first day postoperatively and became pregnant 6 months after surgery, with a C-section delivery of a healthy term newborn at 39 weeks of gestational age. LAC in nonpregnant women with a diagnosis of cervical incompetence is safe and feasible in experienced hands, adding all the intrinsic advantages of minimally invasive surgery and providing better obstetric outcomes. In this patient, the procedure was performed without any intra- or postoperative complications, and the patient had an uneventful term pregnancy in the follow-up period. We must remember that adequate standardization of surgical procedures will help reduce the learning curve.


[Predictive factors for pain in technical dermatological procedures].
Technical procedures in dermatology are painful. The aim of the study was to determine predictive factors for pain in such procedures. This observational study evaluated pain by means of a numerical pain rating scale and a questionnaire about the circumstances of the pain and the use of analgesic methods. Data were analysed using Excel and SAS software. Five hundred and forty-six questionnaires were analysed. Among the patients, 45.4% had mild pain, 34.5% moderate pain and 20.1% severe pain. The least painful procedures were cryotherapy, surgical excision and biopsy, debridement and curettage. Procedures causing moderate to severe pain were treatments involving laser and lights as well as injections (hyaluronic acid, intralesional corticosteroids, botulinum toxin). Pain scores were higher for procedures involving fingers, toes and armpits, while the least painful areas were the trunk and limbs. Among the patients, 62.5% had no preventive analgesia before the procedure, 37.5% had preventive analgesia and 34.2% underwent local or topical anaesthesia (3.7% by cold, 2.0% by oral premedication, 0.5% by hypnosis and 0.4% by nitrous oxide). Awareness of predictive factors for pain could allow better use of analgesic methods to offer the patient relief and improve the quality of the procedure.


Trade-off study and the influence of different fuel injection strategies to enhance the engine characteristics powered with ternary fuel in CRDI diesel engine.
The effects of a diesel engine combustion chamber geometries (PB1, PB2, PB3) and fuel injection pressure (FIP) (500, 750, and 1000 bar) with ternary fuel are discussed in this paper. It is observed that at maximum load as the FIP increased, the PB3 surpasses the PB1 and PB2 in brake thermal efficiency (4.86%) and hydrocarbon (7.2% decrease). However, a minor reduction in NOx (3%) is seen with increased FIP in the PB2. The NHRR (net heat release rate), peak pressure (in-cylinder), and ROPR (rate of pressure increases) all considerably increase with an enhancement in FIP in the case of PB3 by 2.96%, 3.86%, and 1.98%, respectively. It is discovered that altering the fuel injection pressure and piston shape concurrently offers a viable substitute for raising engine output and reducing emissions.


Partial characteristics of hemolytic factors secreted from airborne Aspergillus and Penicillium, and an enhancement of hemolysis by Aspergillus micronesiensis CAMP-like factor via Staphylococcus aureus-sphingomyelinase.
One of the advantages for initial survival of inhaled fungal spores in the respiratory tract is the ability for iron acquisition via hemolytic factor-production. To examine the ability of indoor Aspergillus and Penicillium affecting hemolysis, the secreted factors during the growth of thirteen strains from eight species were characterized in vitro for their hemolytic activity (HA) and CAMP-like reaction. The hemolytic index of HA on human blood agar of Aspergillus micronesiensis, Aspergillus wentii, Aspergillus westerdijkiae, Penicillium citrinum, Penicillium copticola, Penicillium paxilli, Penicillium steckii, and Penicillium sumatrense were 1.72 ± 0.34, 1.61 ± 0.41, 1.69 ± 0.16, 1.58 ± 0.46, 3.10 ± 0.51, 1.22 ± 0.19, 2.55 ± 0.22, and 1.90 ± 0.14, respectively. The secreted factors of an Aspergillus wentii showed high HA when grown in undernourished broth at 25°C at an exponential phase and were heat sensitive. Its secreted proteins have an estimated relative molecular weight over 50 kDa. Whereas, the factors of Penicillium steckii were secreted in a similar condition at a late exponential phase but showed low HA and heat tolerance. In a CAMP-like test with sheep blood, the synergistic hemolytic reactions between most tested mold strains and Staphylococcus aureus were identified. Moreover, the enhancement of α-hemolysis of Staphylococcus aureus could occur through the interaction of Staphylococcus aureus-sphingomyelinase and CAMP-like factors secreted from Aspergillus micronesiensis. Further studies on the characterization of purified hemolytic- and CAMP-like-factors secreted from Aspergillus wentii and Aspergillus micronesiensis may lead to more understanding of their involvement of hemolysis and cytolysis for fungal survival prior to pathogenesis.


Cardiac performance in children with pectus excavatum.
Fourteen children with pectus excavatum and 14 normal control patients underwent graded exercise testing using a cycle ergometer and the James protocol. All the subjects were preoperative. The children were exercised to exhaustion during the test. The pectus and control groups were broken down into subgroups consisting of subjects less than or equal to 10 years of age and subjects greater than or equal to 11 years of age (pectus and controls less than or equal to 10 years old, PI and CI, respectively; pectus and controls greater than or equal to 11 years old, PII and CII, respectively). Maximal workload, oxygen consumption, cardiac output, and stroke volume were not significantly different when comparing the total groups or when each of the respective subgroups were compared. However, maximal diastolic BP was significantly elevated when the entire pectus and control groups were compared. When the subgroups were compared, maximal diastolic BP was elevated only in the older pectus patients (PII) and remained so until five minutes after exercise. Additionally, left ventricular systolic time intervals were measured immediately after exercise in all the children. The ratio of preejection period to left ventricular ejection time (P/L) was significantly shortened in the total pectus group. When the subgroups were compared, the P/L ratio was significantly decreased only in the older patients. Also, the preejection period (PEP) was significantly shortened in the older pectus patients. It appears that children with pectus excavatum have a normal exercise tolerance and oxygen transport. However, older pectus children develop an increased diastolic BP and a shortened P/L and PEP in response to exercise.(ABSTRACT TRUNCATED AT 250 WORDS)


En Face OCT Detects Preretinal Abnormal Tissues Before and After Internal Limiting Membrane Peeling in Eyes with Macular Hole.
To assess preretinal abnormal tissue (PAT) using en face OCT in eyes with idiopathic macular holes (MHs). Retrospective, observational study. Patients with MH who received 6 × 6-mm spectral-domain-OCT scans. Preretinal abnormal tissue was detected by en face OCT images with a custom slab, defined with an anterior boundary of 6 μm anterior to the internal limiting membrane (ILM) and a posterior boundary at 3 μm posterior to the ILM. The PAT was defined as any abnormal tissues observed in en face OCT. Characteristics of preoperative and postoperative PAT. Sixty eyes with MH from 60 patients were included. Fifty-one eyes underwent preoperative analysis, and 46 eyes underwent postoperative examination. Before surgery, 48 (94%) eyes had a mean (standard deviation [SD]) PAT of 6.6 (9.8) mm2. The corresponding cross-sectional OCT showed PAT consisting of epiretinal proliferation, epivascular glia, and a preretinal hyperreflective band. Preretinal abnormal tissue was contiguous to the hole (peri-MH PAT) in 12 eyes (24%), whereas others (36 eyes, 76%) had a scattered distribution (extrafoveal PAT). The area of PAT was greater in eyes with a peri-MH PAT than in eyes with an extrafoveal PAT (P < 0.0001). Compared to the eyes with the extrafoveal PAT, the eyes with the peri-MH PAT were associated with men (P = 0.0059) and worse baseline visual acuity (VA) (P = 0.0002). In eyes with ILM peeling (42 eyes), postoperative PAT proliferation was observed from the edge of the ILM peeling toward the periphery over a 1-year follow-up. The mean (SD) area of PAT at 2 weeks after surgery was 3.4 (3.6) mm2 and increased to 12.1 (6.4) mm2 at 12 months (P < 0.0001). However, no PAT increase was observed in 4 eyes that underwent vitrectomy without ILM peeling. The postoperative PAT size was not associated with the postoperative VA. En face OCT revealed PAT in most eyes with MHs. Peri-MH PAT was associated with worse VA and was seen more frequently in male patients. Postoperative PAT proliferation may be triggered by ILM peeling, but it is not associated with worse VA.


A Molecular Electron Density Theory Study of the Chemoselectivity, Regioselectivity, and Diastereofacial Selectivity in the Synthesis of an Anticancer Spiroisoxazoline derived from α-Santonin.
The [3 + 2] cycloaddition (32CA) reaction of an α-santonin derivative, which has an exocyclic C⁻C double bond, with p-bromophenyl nitrile oxide yielding only one spiroisoxazoline, has been studied within the molecular electron density theory (MEDT) at the MPWB1K/6-311G(d,p) computational level. Analysis of the conceptual density functional theory (CDFT) reactivity indices and the global electron density transfer (GEDT) account for the non-polar character of this zwitterionic-type 32CA reaction, which presents an activation enthalpy of 13.3 kcal·mol-1. This 32CA reaction takes place with total ortho regioselectivity and syn diastereofacial selectivity involving the exocyclic C⁻C double bond, which is in complete agreement with the experimental outcomes. While the C⁻C bond formation involving the β-conjugated carbon of α-santonin derivative is more favorable than the C⁻O one, which is responsible for the ortho regioselectivity, the favorable electronic interactions taking place between the oxygen of the nitrile oxide and two axial hydrogen atoms of the α-santonin derivative are responsible for the syn diastereofacial selectivity.


Results of pericardiectomy for constrictive pericarditis : Single-center experience.
We evaluated our early and late outcomes after pericardiectomy in patients with constrictive pericarditis (CP). We retrospectively reviewed 31 patients who underwent pericardiectomy for CP from 1997 to 2015. Their mean age was 49.2 ± 18.5 years and 74.2 % of them were male. The vast majority had severe functional impairment (NYHA class III-IV) with a mean duration of symptoms of 14.2 ± 10.1 months. Early mortality was 9.7 %: n = 3; multiorgan failure (MOF) in 1, respiratory failure in 1, and left heart failure in 1. Preoperative systolic pulmonary artery pressure over 60 mmHg (p = 0.038, odds ratio [OR] = 0.12) and postoperative low cardiac output syndrome (p = 0.005, OR = 13.5) were significant predictors of early mortality in univariate analysis. Mean follow-up time was 57.8 ± 61.9 months (4-216 months). Late mortality was 6.8 % (2/28 patients) and the cause was MOF secondary to end-stage right heart failure. In Kaplan-Meier analyses, actuarial (including early mortality) and event-free survival rates were 83.9 and 51.1 % at 216 months, respectively. At the end of follow-up, the majority of patients (23/26, 92.9 %) were in good functional status (NYHA class I-II). There were fewer patients under diuretic therapy in the postoperative than in the preoperative period; however, the difference was not statistically significant (12/31 vs. 4/26, p = 0.76). There was no significant difference between the preoperative and follow-up tricuspid annular plane systolic excursion values (15.5 ± 2.2 and 16.6 ± 2.2 mm, respectively, p = 0.088). Left ventricular systolic function was preserved in all patients postoperatively. Although early mortality after pericardiectomy remains high, the procedure provides significant improvement in functional status in the long term.


Nucleosomes inhibit the initiation of transcription but allow chain elongation with the displacement of histones.
Promoters were assembled in nucleosomes or ligated to nucleosomes and transcribed with SP6 RNA polymerase or with mammalian RNA polymerase II and accessory factors. Neither polymerase would initiate transcription at a promoter in a nucleosome, but once engaged in transcription, both polymerases were capable of reading through a nucleosome. In the course of readthrough transcription, the histones were displaced from the DNA, as shown by the exposure of restriction sites and by a shift of the template to the position of naked DNA in a gel. It may be true, in general, that processive enzymes will traverse regions of DNA organized in nucleosomes and displace histones.


TR-FRET-based duplex immunoassay reveals an inverse correlation of soluble and aggregated mutant huntingtin in huntington's disease.
Huntington's disease (HD) is an inherited neurodegenerative disorder caused by the amplification of a polyglutamine stretch at the N terminus of the huntingtin protein. N-terminal fragments of the mutant huntingtin (mHtt) aggregate and form intracellular inclusions in brain and peripheral tissues. Aggregates are an important hallmark of the disease, translating into a high need to quantify them in vitro and in vivo. We developed a one-step TR-FRET-based immunoassay to quantify soluble and aggregated mHtt in cell and tissue homogenates. Strikingly, quantification revealed a decrease of soluble mHtt correlating with an increase of aggregated protein in primary neuronal cell cultures, transgenic R6/2, and HdhQ150 knock-in HD mice. These results emphasize the assay's efficiency for highly sensitive and quantitative detection of soluble and aggregated mHtt and its application in high-throughput screening and characterization of HD models.


Molecular insights into quorum sensing in Acidithiobacillus ferrooxidans bacteria via molecular modelling of the transcriptional regulator AfeR and of the binding mode of long-chain acyl homoserine lactones.
Amino acid sequence alignments of the transcriptional regulator AfeR, which is involved in type 1 quorum sensing (QS) in Acidithiobacillus ferrooxidans bacteria, with other acyl homoserine lactone (AHL)-dependent QS regulators, revealed the presence of strictly or highly conserved residues located in the active site of these proteins. As a consequence, a model of AfeR was constructed to study the binding mode of long-chain AHLs using molecular dynamics and subsequent rigid ligand docking. This study, performed on the tetradecanoyl homoserine lactone C14-AHL, showed that the binding mode involved a curved conformation. Based on these results, the binding mode of tetradec-7-Z enoyl homoserine lactone, an AHL that is conformationally constrained due to the presence of the cis double bond, was investigated. This mono-unsaturated AHL with its preferential curved shape conformation was found to be particularly well adapted to the active site of AfeR. These results should be helpful in the rational design of QS modulators with potential biotechnological applications and especially in the improvement of industrial bioleaching from ores.


Age and sexual culture among homosexually oriented males.
It is here argued that the thesis has been overgeneralized that the culture of homosexually oriented males is heavily youth oriented. It is suggested that any "youth orientedness" is due to their largely single "marital status" rather than to sexual orientation. Utilizing data from 243 males from the Detroit area, several qualifications to the thesis of youth orientedness are indicated. It was found that a majority of males aged 18-24 prefer a male partner who is older, a majority of those aged 25-34 years prefer a same-age person, and 50% of theose over 35 prefer a younger partner. These relationships were found to be especially strong among those segments of the sample who attend gay bars infrequently or who are of lower occupational levels. Also, higher occupational status was found to be associated with preference for the younger, independent of age. It is suggested that, because of the heterogeneity of the male homosexual community, the degree of emphasis of youth will be found to vary with social setting.


Determinants of early sexual initiation among female youth in Ethiopia: a multilevel analysis of 2016 Ethiopian Demographic and Health Survey.
Evidences on determinants of early sexual initiation among female youth is still limited especially; community-level factors are not investigated in Ethiopia. Therefore, the aim of this study was to assess individual and community-level factors associated with early sexual initiation among female youth in Ethiopia. The 2016 Ethiopian Demographic and Health Survey (EDHS) dataset were used and a total of 6143 participants (female youth) were included. Multi-level mixed-effect logistic regression was done to identify individual and community-level factors. Adjusted odds ratio along with 95% confidence interval was used to show the strength and direction of the association. Finally, the level of statistical significance was declared at P value less than 0.05. Individual-level factors significantly associated with early sexual initiation among female youth were; age group from 19 to 24 years [AOR = 5.8, 95% CI = (4.6, 7.3)], not attending school [AOR = 14.1, 95% CI = (8.1, 24.7)], ever chewing Chat [AOR = 2.0, 95% CI = (1.3, 3.0)]. From community-level factors: living in Addis Ababa [AOR = 0.3, 95% CI = (0.2, 0.5)], living in Gambella [AOR = 2.7, 95% CI = (1.7, 4.3)] and live in a low proportion of poor communities [AOR = 0.7, 95% CI = (0.5, 0.9)] were significantly associated with early sexual initiation among female youth in Ethiopia. Age, low educational status, ever chewing Chat, region and live in a high proportion of poor community had a statistical association with early sexual initiation among female youth in Ethiopia. Improving educational coverage and community-level of wealth status are important intervention areas to delay the age of early sexual initiation.


Determination of dodecylbenzenesulphonates and ethoxylated alkylphenols in liquid pesticide formulations by high-performance liquid chromatography.
A method is presented for determining mixtures of calcium dodecylbenzene-sulphonate (CaDBS) and ethoxylated alkylphenols in pesticidal emulsifiable concentrates. The pesticide formulations are adsorbed on a pre-concentration column. The active ingredients, the solvents and the emulsifiers are eluted successively by using solvents with increasing polarities. The various homologues of CaDBS and the ethoxylated alkylphenols are separated by reversed-phase high performance liquid chromatography using methanol-water containing tetramethylammonium bromide. The oligomers of the ethoxylated phenols are separated on an aminopropyl-modified column using a solvent gradient (propan-2-ol-water and hexane-tetrahydrofuran). Quantitative results are given for synthetic samples. The degree of ethoxylation of the ethoxylated alkylphenols has been determined and the CaDBS components characterized.


Boundary-directed epitaxy of block copolymers.
Directed self-assembly of block copolymers (BCPs) enables nanofabrication at sub-10 nm dimensions, beyond the resolution of conventional lithography. However, directing the position, orientation, and long-range lateral order of BCP domains to produce technologically-useful patterns is a challenge. Here, we present a promising approach to direct assembly using spatial boundaries between planar, low-resolution regions on a surface with different composition. Pairs of boundaries are formed at the edges of isolated stripes on a background substrate. Vertical lamellae nucleate at and are pinned by chemical contrast at each stripe/substrate boundary, align parallel to boundaries, selectively propagate from boundaries into stripe interiors (whereas horizontal lamellae form on the background), and register to wide stripes to multiply the feature density. Ordered BCP line arrays with half-pitch of 6.4 nm are demonstrated on stripes >80 nm wide. Boundary-directed epitaxy provides an attractive path towards assembling, creating, and lithographically defining materials on sub-10 nm scales.


Survey of anesthesia practice in spine surgery patients in the United States.
There is a wide range of anesthetic practice in spine surgery with respect to anesthetic choice, blood conservation, and monitoring. There is no ideal technique with respect to each of these choices. This study was designed to determine the usual practice for members of the Society of Neurosurgical Anesthesia and Critical Care (SNACC), with regard to the use of anesthetic technique, the type of monitoring used (and the person responsible for its application and interpretation), and the blood conservation strategies most often utilized for the management of patients undergoing spinal surgery. A total of 415 questionnaires were mailed to all current members of the SNACC. Of these, 85 (23%) were completed and returned. The majority of the respondents administer general anesthesia for lumbar laminectomy. Somatosensory evoked potential (SSEP) monitoring is available at most of the institutions (94%) but only utilized in 75% of patients undergoing a Harrington rod placement. Motor evoked potential monitoring is available at 50% of the institutions, but utilized 25% of the time for these surgeries. The two most frequent blood conservation strategies utilized are intraoperative salvage and autologous donation. The most frequent monitoring utilized for major spinal surgeries is SSEP. Autologous donation and intraoperative salvage are the most frequent blood conservation methods utilized.


Gut microbiota changes in patients with autism spectrum disorders.
Autism spectrum disorder (ASD) has a high incidence of intestinal comorbidity, indicating a strong association with gut microbiota. The purpose of this study was to characterize gut microbiota profiles in children with ASD. Seventy-seven children with ASD [33 with mild ASD and 44 with severe ASD according to the Childhood Autism Rating Scale score] and 50 age-matched healthy children were enrolled. Compared with children in the healthy control (HC) group, those in the ASD group showed higher biomass, richness, and biodiversity of gut microbiota, and an altered microbial community structure. At the genus level, there was a significant increase in the relative abundance of unidentified Lachnospiraceae, Clostridiales, Erysipelotrichaceae, Dorea, Collinsella, and Lachnoclostridium, whereas Bacteroides, Faecalibacterium, Parasutterella, and Paraprevotella were significantly lower in the ASD group than in the control group. The presence of unidentified Erysipelotrichaceae, Faecalibacterium, and Lachnospiraceae was positively correlated with ASD severity. Notably, three microbial markers (Faecalitalea, Caproiciproducens and Collinsella) were identified in a random forest model with an area under the curve (AUC) of 0.94 for differentiation between HCs and ASD patients. Furthermore, the validation model was consistent with the discovery set (AUC = 0.98, 95% CI: 97.9%-100%). The training and testing sets were more effective when the number of bacteria was increased. In addition, the functional properties (such as galactose metabolism, glycosyltransferase activity, and glutathione metabolism) displayed significant differences between the ASD and HC groups. The current study provides evidence for the relationship between gut microbiota and ASD, with the findings suggesting that gut microbiota could contribute to symptomology. Thus, modulation of gut microbiota may be a new therapeutic strategy for ASD.


Securing the future of ground water resources in the Great Lakes basin.
Ground water is a vital, but underappreciated, natural resource in the Great Lakes basin. It meets many human needs and contributes significantly to the hydrology of the Great Lakes and the health of ecosystems. This paper provides an overview of ground water in the Great Lakes and the institutional and legal setting that governs the use, protection, diversion, and removal of water from the basin and proposes a citizen-centered vision for management of ground water in the 21st century.


Patient-reported migraine-specific quality of life, activity impairment and headache impact with once-daily atogepant for preventive treatment of migraine in a randomized, 52-week trial.
Atogepant is an oral, small-molecule, calcitonin gene-related peptide receptor antagonist for the preventive treatment of episodic migraine. In this 52-week, multicenter, randomized, open-label trial, adults with 4-14 monthly migraine days received atogepant 60 mg once-daily or standard care. Health outcome endpoints collected from participants randomized to atogepant included change from baseline in Migraine-Specific Quality of Life Questionnaire version 2.1 (MSQ v2.1) Role Function-Restrictive (RFR), Role Function-Preventive (RFP) and Emotional Function (EF) domain scores, change in Activity Impairment in Migraine-Diary (AIM-D) Performance of Daily Activities (PDA) and Physical Impairment (PI) domain scores, and change in Headache Impact Test-6 (HIT-6) total score. Of 744 randomized participants, 521 received atogepant 60 mg in the modified intent-to-treat population. Least-squares mean changes from baseline in MSQ-RFR score were 30.02 (95% confidence interval = 28.16-31.87) at week 12 and 34.70 (95% confidence interval = 32.74-36.66) at week 52. Improvements were also observed in other MSQ domains, AIM-D PDA, PI and HIT-6 total scores. A ≥5-point improvement from baseline in HIT-6 score was observed in 59.9% of participants at week 4 and 80.8% of participants at week 52. Over 52 weeks, atogepant 60 mg once-daily was associated with sustained improvements in quality of life and reductions in activity impairment and headache impact.Trial Registration: NCT03700320.


Modulation of human B cell responses by a monoclonal antibody to an activation antigen 4F2.
Antigen specific human antibody responses can be modulated in vitro by the addition of 4F2 antibody, a monoclonal antibody (MoAb) which recognizes an antigen on activated T cells and B cells. Specific antibody responses induced with the antigen are suppressed by the addition of 4F2. However, specific antibody responses induced with the polyclonal activator, pokeweed mitogen (PWM), are significantly enhanced by the addition of 4F2. Proliferative responses to both antigen and PWM are suppressed by the addition of 4F2. The enhancement of PWM stimulated responses by 4F2 is mediated by T cells. However, in the absence of T cells, 4F2 can directly inhibit antigen specific B cells. Polyclonal Ig production stimulated by PWM was also enhanced by 4F2. Thus, the immunomodulating effects of the 4F2 MoAb are the result of a balance of enhancement and suppression mediated at the T cell and the B cell level, respectively.


Classification of acute myocardial ischemia by artificial neural network using echocardiographic strain waveforms.
Echocardiographic strain waveforms are highly variable, so their interpretation is experience-dependent and subjective. We tested whether an artificial neural network (ANN) can distinguish between strain waveforms obtained at baseline and during experimentally induced acute ischemia. An open-chest model of coronary occlusion and acute ischemia was used in 14 adult pigs. Strain waveforms were obtained using a GE Vivid 7 ultrasound system. An ANN design was implemented in MATLAB, and backpropagation and "leave-one-out" processes were used to train and test it. Specificity of 86% and sensitivity of 87% suggest that ANNs could aid in diagnostic prescreening of echocardiographic strain waveforms.


Broad spectrum antiviral nucleosides-Our best hope for the future.
The current focus for many researchers has turned to the development of therapeutics that have the potential for serving as broad-spectrum inhibitors that can target numerous viruses, both within a particular family, as well as to span across multiple viral families. This will allow us to build an arsenal of therapeutics that could be used for the next outbreak. In that regard, nucleosides have served as the cornerstone for antiviral therapy for many decades. As detailed herein, many nucleosides have been shown to inhibit multiple viruses due to the conserved nature of many viral enzyme binding sites. Thus, it is somewhat surprising that up until very recently, many researchers focused more on "one bug one drug," rather than trying to target multiple viruses given those similarities. This attitude is now changing due to the realization that we need to be proactive rather than reactive when it comes to combating emerging and reemerging infectious diseases. A brief summary of prominent nucleoside analogues that previously exhibited broad-spectrum activity and are now under renewed interest, as well as new analogues, that are currently under investigation against SARS-CoV-2 and other viruses is discussed herein.


Functional role of PKC in contraction of cultured human prostatic stromal cells.
The contractile activity of prostatic stromal cells contributes to symptoms of benign prostatic hyperplasia (BPH). However, the mechanisms for this contraction have not yet been fully elucidated. In this study, we investigated the role of protein kinase C (PKC) in prostatic contraction by measuring the isometric tension development of cultured human prostatic stromal cells (CHPSCs) derived from BPH patients. Fresh human BPH tissue was used only in a Western blot analysis. A ring preparation made of CHPSCs and collagen gel could develop an isometric tension during activation with various agonists. Phorbol 12,13 dibutyrate (PDBu), a PKC activator, induced a relaxation. A Western blot analysis revealed the expression of PKC-potentiated protein phosphatase-1 inhibitory protein (CPI-17) in both CHPSCs and fresh human BPH tissue to be much lower than that in the rabbit aorta. When CPI-17 was over-expressed, PDBu induced a large contraction, but the agonist-induced contraction did not become larger than expected. In alpha-toxin permeabilized preparations, PDBu induced a relaxation in control CHPSCs, while it induced a contraction at a constant [Ca2+]i in CPI-17 over-expressing CHPSCs. These results indicated that the activation of PKC in CHPSCs induces a relaxation probably due to low expression level of CPI-17 and also that the PKC-CPI-17 pathway does not appear to play a major role in the agonist-induced contraction even when CPI-17 was over-expressed.


Characterization of two novel lytic bacteriophages having lysis potential against MDR avian pathogenic Escherichia coli strains of zoonotic potential.
Avian pathogenic E. coli (APEC) is associated with local and systemic infections in poultry, ducks, turkeys, and many other avian species, leading to heavy economical losses. These APEC strains are presumed to possess zoonotic potential due to common virulence markers that can cause urinary tract infections in humans. The prophylactic use of antibiotics in the poultry sector has led to the rapid emergence of Multiple Drug Resistant (MDR) APEC strains that act as reservoirs and put human populations at risk. This calls for consideration of alternative strategies to decrease the bacterial load. Here, we report isolation, preliminary characterization, and genome analysis of two novel lytic phage species (Escherichia phage SKA49 and Escherichia phage SKA64) against MDR strain of APEC, QZJM25. Both phages were able to keep QZJM25 growth significantly less than the untreated bacterial control for approximately 18 h. The host range was tested against Escherichia coli strains of poultry and human UTI infections. SKA49 had a broader host range in contrast to SKA64. Both phages were stable at 37 °C only. Their genome analysis indicated their safety as no recombination, integration and host virulence genes were identified. Both these phages can be good candidates for control of APEC strains based on their lysis potential.


Distribution and ecological risks of pharmaceuticals and personal care products with different anthropogenic stresses in a coastal watershed of China.
The occurrences of pharmaceutical and personal care products (PPCPs) in both freshwater and sea have been widely reported. However, pollution control requires further information on riverine discharges with influence of land-based activities and associated risks to estuarine ecosystems. This study investigated the spatial occurrences and the relationship to sociodemographic parameters of 30 PPCPs in 67 rivers along the Bohai coastal region. The results showed that PPCPs were mainly deposited in aquatic phase, and the partitioning coefficient between water and sediment was highly determined by chemical properties. The levels of 30 PPCPs in rivers ranged from 8.33 to 894.48 ng/L, showing a large variance among regions. Caffeine, sulfamethoxazole, sulfamethazine, ofloxacin, anhydro-erythromycin, and trimethoprim were found to be the major pollutants. Multivariable analysis method was used to assess the correlation of PPCPs markers to socio-economic parameters. The results indicated that domestic emissions contributed most to the occurrences of PPCPs in the riverine water. Risk assessment result indicated that sulfamethoxazole, caffeine, tetracycline, and carbamazepine ranked top four with the highest risks to the most sensitive aquatic organisms. The results identified caffeine and carbamazepine with high detection frequency and concentration as the priority chemicals, while sulfamethoxazole and erythromycin should also be concerned due to their potential threats in specific rivers. This study provides valuable information for pollution control over PPCPs riverine discharges in estuarine regions.


Chemoenzymatic synthesis of iminocyclitol derivatives: a useful library strategy for the development of selective fucosyltransfer enzymes inhibitors.
A chemoenzymatic strategy has been developed for the synthesis of libraries of iminocyclitol derivatives for the discovery of new and selective fucosidase inhibitors.


An ultrabright and monochromatic electron point source made of a LaB6 nanowire.
Electron sources in the form of one-dimensional nanotubes and nanowires are an essential tool for investigations in a variety of fields, such as X-ray computed tomography, flexible displays, chemical sensors and electron optics applications. However, field emission instability and the need to work under high-vacuum or high-temperature conditions have imposed stringent requirements that are currently limiting the range of application of electron sources. Here we report the fabrication of a LaB6 nanowire with only a few La atoms bonded on the tip that emits collimated electrons from a single point with high monochromaticity. The nanostructured tip has a low work function of 2.07 eV (lower than that of Cs) while remaining chemically inert, two properties usually regarded as mutually exclusive. Installed in a scanning electron microscope (SEM) field emission gun, our tip shows a current density gain that is about 1,000 times greater than that achievable with W(310) tips, and no emission decay for tens of hours of operation. Using this new SEM, we acquired very low-noise, high-resolution images together with rapid chemical compositional mapping using a tip operated at room temperature and at 10-times higher residual gas pressure than that required for W tips.


Human intestinal brush border peptidases.
Hydrolysis of small peptides, like disaccharide hydrolysis, is an important function of the intestinal brush border, but little is known of the individual human peptidases. The purposes of this study were to detect all human brush border enzymes hydrolyzing dipeptides and tripeptides, identify the most discriminating substrate for each enzyme in order to permit assays in crude mixtures, and begin biochemical characterization of each enzyme. Four brush border peptidases were identified. Enzymes I (aspartate aminopeptidase, E.C. 184.108.40.206) and III (amino-oligopeptidase, E.C. 184.108.40.206) are known brush border enzymes. Enzymes II (membrane Gly-Leu peptidase) and IV (zinc stable Asp-Lys peptidase) have not been identified in human brush border previously. They are distinct from dipeptidyl aminopeptidase IV, carboxypeptidase, and gamma-glutamyl transferase. The substrate most discriminating for each enzyme is alpha-Glu-beta-naphthylamide for I (100% of the brush border activity for this substrate is due to enzyme I), glycylleucine for II (80%), leucyl-beta-naphthylamide for III (91%), and aspartyl-lysine in 5 mM Zn2+ for IV (63%). The enzymes are immunologically distinct and antibodies to each one localize to the brush border on immunohistochemical staining. Purification of 142-, 79-, 158-, and 46-fold was achieved for enzymes I through IV, respectively. Biochemical characteristics include slightly alkaline pH optima, molecular weights of 91,000-190,000, and evidence of metal ion involvement in activity. These studies provide necessary information for determining the role of brush border peptidase deficiencies in human disease.


Clinico-pathologic characteristics and prognostic factors of ovarian carcinoma with different histologic subtypes - A benchmark analysis of 482 cases.
Ovarian carcinomas (OCX) have traditionally been thought to arise from the ovarian surface epithelium. However, recent (immuno-) histopathological and molecular analyses suggest that OCX consist of morphological subtypes with different epidemiologic features and a varying prognosis. The data of 482 OCX from the Clinical Cancer Registry of Leipzig who were surgically treated between 2000 and 2019 and were evaluated regarding incidence, clinico-pathologic characteristics and prognostic factors. Cases were separated into high-grade and non-high-grade serous tumors. Both groups were analyzed regarding the tumor stage, lymph node involvement, site of origin and prognosis. The overall incidence for OCX was 17.9. The most common histological subtype was high-grade serous OCX (57.9%; 279/482). Patients with high-grade were significantly older than those with a non-high-grade serous OCX (63.9 versus 58.5 years; p < 0.001), more frequently diagnosed at an advanced stage >pT3 (78.5% (219/279) versus 42.8% (87/203); p < 0.001) and showed a 2.4-fold higher frequency of lymph node metastases (53.4% vs. 21.2%; p < 0.02) with a 4.6-fold higher rate of > 1 cm metastatic deposits (pN1b) within the lymph nodes (14.8% vs. 4.6%; p < 0.02). Irrespective of tumor stage and morphological subtype, the 1- and 5-year overall survival (OAS) was 72.9% and 40.8%, respectively. Patients with high-grade serous OCX showed a shorter 5-year OAS compared to non-high-grade serous OCX (34.1% vs. 57.0%; p 0.001). This association was reproducible in patients with an advanced tumor stage irrespective of the histopathologic tumor type serous OCX (pT3: 32.4% vs. pT1: 75.1%; p 0.001) as well as within high-grade (pT3: 28.7% vs. pT1: 55.5%; p = 0.003) and non-high-grade serous OCX (pT3: 43.0% vs. 80.0%; p 0.001). There were no differences in OAS depending on the site of origin (fallopian tube, ovary, peritoneum) within the two histologic subgroups. OCX cases from a single institution with uniform surgical treatment and a standardized histopathological workup were evaluated. The poor prognostic outcome of patients with high-grade serous compared non-high-grade serous OCX as well as an advanced stage of the disease was confirmed. This study demonstrates for the first time that the histopathological distinction into high-grade serous and non-high-grade serous tumors may be much more prognostically relevant than the site of origin.


Protein-polymer conjugation-moving beyond PEGylation.
In this review, we summarize-from a materials science perspective-the current state of the field of polymer conjugates of peptide and protein drugs, with a focus on polymers that have been developed as alternatives to the current gold standard, poly(ethylene glycol) (PEG). PEGylation, or the covalent conjugation of PEG to biological therapeutics to improve their therapeutic efficacy by increasing their circulation half-lives and stability, has been the gold standard in the pharmaceutical industry for several decades. After years of research and development, the limitations of PEG, specifically its non-degradability and immunogenicity have become increasingly apparent. While PEG is still currently the best polymer available with the longest clinical track record, extensive research is underway to develop alternative materials in an effort to address these limitations of PEG. Many of these alternative materials have shown promise, though most of them are still in an early stage of development and their in vivo distribution, mechanism of degradation, route of elimination and immunogenicity have not been investigated to a similar extent as for PEG. Thus, further in-depth in vivo testing is essential to validate whether any of the alternative materials discussed in this review qualify as a replacement for PEG.


The soft tissue cover of the mandibular condyle. Differentiation in histological forms and age-related changes of aggrecan- and versican-like proteoglycans.
Age-related changes of the composition of the extracellular matrix of the soft tissue cover of the mandibular condyle (STC), especially of the large proteoglycans, have been investigated. Proteoglycans were extracted from the STC of neonatal, juvenile and adult domestic pigs, fractionated by density gradient centrifugation and analyzed by electrophoresis/Western blotting. Experiments revealed firstly that a large CS/KS proteoglycan (aggrecan) is an essential constituent of the STC at all ages. This proteoglycan is required for nutrition of avascular tissues, and age-related changes in its average size and substitution with KS (keratan sulfate) may be a response to altered functional loading and tissue architecture of the STC. Secondly it was shown that a large CS/DS (chondroitin sulfate/dermatan sulfate) proteoglycan characterized by a doublet of core proteins at 200 and 250 kDa, thereby resembling perlecan, is present in the tissue of adults, but not of neonates and juveniles. Thirdly a large CS/DS proteoglycan characterized by core proteins at 350, 450 and 550 kDa, thereby resembling versican, was present in juveniles. It was detectable only weakly in neonates and not in adults. Results of core protein analysis were confirmed by results of agarose gel electrophoresis/Western blotting of the undigested proteoglycans isolated directly from the tissue extracts. Versican is believed to destabilize cell-matrix interactions required for cell proliferation and differentiation. In this context, presence of versican-like proteoglycans in the STC of growing individuals and its disappearance in adults appears to be related to the growth potential of the mandibular condyle.


S100 family proteins in inflammation and beyond.
The S100 family proteins possess a variety of intracellular and extracellular functions. They interact with multiple receptors and signal transducers to regulate pathways that govern inflammation, cell differentiation, proliferation, energy metabolism, apoptosis, calcium homeostasis, cell cytoskeleton and microbial resistance. S100 proteins are also emerging as novel diagnostic markers for identifying and monitoring various diseases. Strategies aimed at targeting S100-mediated signaling pathways hold a great potential in developing novel therapeutics for multiple diseases. In this chapter, we aim to summarize the current knowledge about the role of S100 family proteins in health and disease with a major focus on their role in inflammatory conditions.


Serotonin transporter and saitohin genes in risk of Alzheimer's disease and frontotemporal lobar dementia: preliminary findings.
Serotonergic transmission impairment and abnormal phosphorylation of tau protein have been implicated in the physiopathology of Alzheimer's disease (AD) and frontotemporal lobar dementia (FTLD). Associations between a functional polymorphism (5-HTTLPR), in the promoter region of the serotonin transporter gene, and susceptibility to sporadic AD and FTLD have been reported. A polymorphism (Q7R) in saitohin gene inside the microtubule-associated protein tau gene has also been related to dementia. To determine the possible role of the two polymorphisms in susceptibility to AD and FTLD, we performed a case-control study collecting 218 Italian sporadic dementia patients and 54 controls. We found a significant excess of 5-HTTLPR short alleles and an interaction between 5-HTTLPR and Q7R polymorphisms in demented subjects. Our study confirms the role of 5-HTTLPR as a potential susceptibility factor for sporadic dementia in the Italian population, and suggests a possible interaction between 5-HTTLPR and Q7R polymorphisms in neurodegenerative diseases.


The effect of antidepressants on aversive periaqueductal gray stimulation in rats.
The selective serotonin (5-HT) uptake inhibitors fluvoxamine and sertraline had anti-aversive effects when administered acutely. Imipramine and clomipramine, which combine noradrenaline (NA) and 5-HT uptake blocking properties, did not have significant effects, whereas the mixed dopamine (DA)/NA uptake blocker nomifensine decreased the thresholds for escape from aversive periaqueductal gray stimulation. These results suggest that indirect 5-HT receptor activation suppresses periaqueductal gray aversion. Conversely, indirect DA and perhaps NA receptor activation appears to enhance periaqueductal gray aversion in rats.


Siamon Gordon: A half-century fascination with macrophages.
Siamon Gordon is a Glaxo Wellcome Professor Emeritus of Cellular Pathology at the University of Oxford and a fellow of the Royal Society. Throughout his career, Siamon has focused on macrophages, and his work led to the identification of the pan-macrophage marker F4/80 and the description of a role for Dectin-1 in the innate recognition of β-glucans. I caught up with Siamon to discuss his career path and his thoughts on macrophages.


Evaluation of frictional forces during dental alignment: an experimental model with 3 nonleveled brackets.
The aim of this in-vitro study was to evaluate the frictional forces generated by various combinations of brackets and orthodontic wires by using an experimental model with 3 nonleveled brackets to gain a better understanding of the resistance to sliding during dental alignment and leveling. Two kinds of orthodontic brackets were tested: passive self-ligating brackets and conventional twin stainless steel brackets. The following wires were tested: 3 nickel-titanium (.014, .016, and .016 x .022 in), 2 stainless steel (multistranded .0155 and .016 in), and 1 beta-titanium alloy (.016 in). The ligatures used with conventional brackets were elastomeric modules (power 'O' 110) and preformed stainless steel ligature wire (.010). Each of the 10 bracket-archwire combinations was tested 10 times. Kinetic frictional forces were measured on a specially designed testing machine. The wires tested were pulled through a set of multiple nonleveled brackets at a speed of 4 mm per minute over a distance of 5 mm. All data were statistically analyzed. The sliding of the wire in the 3-bracket nonaligned system was significantly influenced by wire cross-section dimension (P < 0.001), wire material (P < 0.001), number of wire strands (P < 0.001), and type of ligation (P < 0.001). Frictional forces can be reduced during alignment by using self-ligating brackets, small dimensions, and less stiff wires, thereby inducing the wire to slide in the slots. Under such conditions, the force required by the orthodontic wire to overcome resistance to sliding is reduced. This allows the wire to exploit its mechanical characteristics more efficiently.


Significant Improvement in Outcomes of Patients with Intrahepatic Cholangiocarcinoma after Surgery.
The prognosis of intrahepatic cholangiocarcinoma (ICC) remains poor despite improvements in treatment and post-operative clinical management. We review our experiences and evaluate our current surgical approaches by comparing patients from two consecutive treatment periods. One hundred forty-four patients who underwent hepatectomy for ICC between 1993 and 2014 were divided into groups that received treatment before (n = 65, first period) and after 2006 (n = 79, second period), when new treatment options such as adjuvant chemotherapy and multimodal therapy for recurrence were introduced. Clinicopathological characteristics and survival outcomes were compared between the groups. First-period patients exhibited more advanced tumor characteristics, including larger tumors, higher serum carbohydrate antigen 19-9 levels, and vascular invasion. Median overall survival (OS) durations of the first- and second-period groups were 21.4 and 57.7 months, respectively (p < 0.001); corresponding median disease-free survival (DFS) durations were 12.2 and 16.6 months, respectively (p = 0.027). Multivariate analysis found an independent association of the treatment time period with OS and DFS. Notably, second-period patients with N1 disease achieved a longer OS and DFS (median OS time: 12.4 and 26.0 months, p = 0.0018, and median DFS: 4.7 and 10.7 months p = 0.019, respectively). Among recurrent patients (first, n = 50 and second, n = 44), second-period patients had a significantly longer survival after recurrence (8.0 vs. 22.3 months, p < 0.001). ICC patients, particularly those with N1 disease, achieved significant survival improvements that were partly attributable to patient selection, adjuvant chemotherapy, and multimodal treatment after recurrence.