Disproportionate lymphoid cell subsets in thalassaemia major: the relative contributions of transfusion and splenectomy.
The relative proportions of T-cell (OKT3-positive, OKT4-positive and OKT8-positive) and B-cell (SIg-positive) populations in peripheral blood obtained from 29 chronically transfused patients with beta-thalassaemia major were compared with those of 17 healthy controls. Changes attributable to blood transfusion and/or splenectomy are described. The percentage of OKT8-positive (T-suppressor) cells found in the thalassaemic patients increased linearly (P less than 0.001) with the number of units transfused, irrespective of splenectomy. The percentage of OKT4-positive (T-helper) cells varied inversely with increasing transfusion in nonsplenectomized patients while in those who were splenectomized no significant correlation was apparent. Thus, in both groups of patients the T4/T8 ratio declined in a transfusion-related manner. The splenectomized patients experienced a marked and persistent lymphocytosis due to an increase in the number of both T- and B-cells. When the results were expressed as percentages, the greatest increase occurred in the number of B-cells, this increase being unrelated to the number of transfusions received. None of the serum parameters usually associated with iron overload or abnormal liver function correlated with the observed increases in T-suppressor and SIg-positive cells. These findings corroborate reports that transfusion of blood products may lead to decreased T4/T8 ratios. However, none of the patients studied manifested clinical signs of acquired immune deficiency syndrome (AIDS). Accordingly, studies which define transfusion related AIDS on the basis of analyses with monoclonal antibodies must be viewed with caution.


Nomogram for overall survival of Japanese patients with bone-metastatic prostate cancer.
We analyzed the relationship between prostate cancer outcomes and pretreatment clinical factors and developed a prognostic nomogram of overall survival (OS) of patients with bone metastasis. From 1993 to 2011, 463 consecutive patients were treated for bone-metastatic prostate cancer. Data sets from 361 patients were used to develop a nomogram (training data), and data sets of 102 patients were used for validation of the nomogram (validation data). Using the external validation data set, the nomogram was assessed for discriminatory ability, and the predictions were assessed for calibration accuracy by plotting actual survival against predicted risk. Of the 361 patients in the training data set, 205 (56.8%) patients died, 169 (46.8%) deaths of which were due to prostate cancer. The median follow-up period was 55.2 months. In the multivariate analysis, patient age, serum prostate-specific antigen level, clinical T stage, extent of disease on bone scan, and biopsy Gleason sum were independent prognostic factors. We developed a prognostic model comprising these five factors for patients with bone-metastatic prostate cancer. This nomogram can be used to estimate 1-, 3-, and 5-year survival probability. External validation of this model using 102 validation data sets showed reasonable accuracy (concordance index, 0.719). Our pretreatment prognostic nomogram might be useful for Japanese patients with bone-metastatic prostate cancer.


Tumor necrosis factor is markedly synergistic with interleukin 1 and interferon-gamma in stimulating the production of nerve growth factor in fibroblasts.
A possible interaction between tumor necrosis factor-alpha (TNF) and other cytokines/growth factors in stimulating the production of nerve growth factor (NGF) in Swiss 3T3 cells was studied. TNF's stimulatory activity on fibroblast NGF production was synergized by interleukin-1 alpha (IL-1 alpha), IL-1 beta and interferon-gamma (IFN-gamma), but was antagonized by transforming growth factor-beta (TGF-beta). The most remarkable synergistic effect was observed between TNF and IL-1 alpha/beta; as little as 0.003 ng/ml of IL-1 beta markedly enhanced TNF's stimulatory activity on NGF production in the cells. These findings reinforce the idea that TNF, in concert with IL-1 alpha/beta, plays an essential role in regulating the regeneration of peripheral nerves following injury through an indirect mechanism by which it stimulates NGF production in fibroblasts.


Cash-Only INcentives to promote insulin DOSE engagement: A protocol paper for the pilot randomized controlled trial of COIN2DOSE.
Most adolescents with type 1 diabetes (T1D) do not achieve a glycated hemoglobin (HbA1c) <7.0%, which is the current clinical target. mHealth can offer a scalable and age-appropriate delivery method for behavioral interventions to lower adolescents' HbA1c levels, while applying established behavior change and behavioral economics theories can enhance scientific rigor. We aim to conduct a pilot randomized clinical trial of a novel mHealth intervention called Coin2Dose (Cash-Only INcentives To promote insulin DOSE engagement), in a sample of youth with T1D: 1) to obtain measures of feasibility and acceptability and 2) to examine preliminary efficacy versus a standard care control group based on differences in youth's daily BOLUS scores, HbA1c levels, and Time in Range (TIR) at post-intervention and 3-month post-intervention follow-up. This pilot RCT is already registered in http://ClinicalTrials.gov (NCT#05280184). Our pilot will recruit youth with T1D 11-17 years-old who use an insulin pump or Bluetooth connected insulin pen and have an average daily BOLUS score ≤2.5. Youth randomized to Coin2Dose will receive the intervention for 12 weeks followed by a 12-week maintenance period. The pilot is scheduled to start July 2022 and to conclude in 2025. At the conclusion of the pilot, we will have information about the feasibility and acceptability of two different behavioral economic incentive structures for improving BOLUS scores. The work is anticipated to progress to final efficacy trial. We will disseminate study results through presentations at local, national, and international conferences and through peer-reviewed diabetes and psychology journals.


Estimates of genetic correlations among clinical measures of the eye.
In a quantitative genetic investigation of seven clinical tests, five pairs were significantly correlated: (1) cover test measures and corneal power, (2) corneal power and divergence recovery points, (3) convergence and divergence recovery points, (4) divergence break and recovery points, and (5) convergence break and recovery points. Common genes may account entirely for the gene influence on cover test measure; thus, parents who are above the population mean for corneal power will have offspring with a tendency toward an exodeviation (phoria). The two convergence amplitudes may depend on the same gene differences, whereas the two divergence amplitudes only partly reflect a common function, suggesting that they may be indexing somewhat different underlying physiologic mechanisms.


Role of pleiotropy in the evolution of a cryptic developmental variation in Caenorhabditis elegans.
Robust biological systems are expected to accumulate cryptic genetic variation that does not affect the system output in standard conditions yet may play an evolutionary role once phenotypically expressed under a strong perturbation. Genetic variation that is cryptic relative to a robust trait may accumulate neutrally as it does not change the phenotype, yet it could also evolve under selection if it affects traits related to fitness in addition to its cryptic effect. Cryptic variation affecting the vulval intercellular signaling network was previously uncovered among wild isolates of Caenorhabditis elegans. Using a quantitative genetic approach, we identify a non-synonymous polymorphism of the previously uncharacterized nath-10 gene that affects the vulval phenotype when the system is sensitized with different mutations, but not in wild-type strains. nath-10 is an essential protein acetyltransferase gene and the homolog of human NAT10. The nath-10 polymorphism also presents non-cryptic effects on life history traits. The nath-10 allele carried by the N2 reference strain leads to a subtle increase in the egg laying rate and in the total number of sperm, a trait affecting the trade-off between fertility and minimal generation time in hermaphrodite individuals. We show that this allele appeared during early laboratory culture of N2, which allowed us to test whether it may have evolved under selection in this novel environment. The derived allele indeed strongly outcompetes the ancestral allele in laboratory conditions. In conclusion, we identified the molecular nature of a cryptic genetic variation and characterized its evolutionary history. These results show that cryptic genetic variation does not necessarily accumulate neutrally at the whole-organism level, but may evolve through selection for pleiotropic effects that alter fitness. In addition, cultivation in the laboratory has led to adaptive evolution of the reference strain N2 to the laboratory environment, which may modify other phenotypes of interest.


Root sepsis associated with insect-dwelling Sebaldella termitidis in a lesser dwarf lemur (Cheirogaleus medius).
Sebaldella termitidis is a rare fastidious microorganism of the Leptotrichiaceae family. A variety of closely related species are associated with severe and even life-threatening disease in humans and animals, such as Streptobacillus moniliformis, the etiological organism of rat-bite fever as well as members of Leptotrichia spp. and Sneathia sanguinegens, which have been reported from cases of septicaemia. In contrast, since its description some 50 years ago, S. ermitidis has so far never been reported as a vertebrate pathogen, nor has it been found aside from its natural termite host. A lesser dwarf lemur was presented with unilateral facial inflammation originating from rotten maxillary teeth and septic root abscess. Surgical intervention and root extraction significantly improved the clinical cause in that a pus-filled cavity underneath the right eye could be drained, sampled and flushed. Bacteria displaying substantial characteristics of S. termitidis were cultured from the sampled pus. Morphological features observed included strictly anaerobic regular Gram-negative rods. Significant shared biochemical properties included negative reactions for cytochrome oxidase, catalase, urease, nitrate reduction and indole production. Furthermore, 16S rRNA gene sequencing revealed 99.9 % sequence homology to the S. termitidis type strain NCTC 11300(T), from which it, nevertheless, differed with respect to rep and rep- and RAPD-PCR profiles. An affiliation of the lemur isolate described in this study with the type strain of S. termitidis as well as a clear discrimination from other members of the Leptotrichiaceae could also be confirmed by matrix-assisted laser desorption/ionization time-of flight mass spectrometry and Fourier transform-infrared spectroscopy. This is the first evidence for clinical disease caused by S. termitidis in a vertebrate species indicating a broader host spectrum of this rarely encountered microorganism.


Patterns of multimorbidity and their effects on adverse outcomes in rheumatoid arthritis: a study of 5658 UK Biobank participants.
To investigate how the type and number of long-term conditions (LTCs) impact on all-cause mortality and major adverse cardiovascular events (MACE) in people with rheumatoid arthritis (RA). Population-based longitudinal cohort study. UK Biobank. UK Biobank participants (n=502 533) aged between 37 and 73 years old. Primary outcome measures were risk of all-cause mortality and MACE. We examined the relationship between LTC count and individual comorbid LTCs (n=42) on adverse clinical outcomes in participants with self-reported RA (n=5658). Risk of all-cause mortality and MACE were compared using Cox's proportional hazard models adjusted for lifestyle factors (smoking, alcohol intake, physical activity), demographic factors (sex, age, socioeconomic status) and rheumatoid factor. 75.7% of participants with RA had multimorbidity and these individuals were at increased risk of all-cause mortality and MACE. RA and >4 LTCs showed a threefold increased risk of all-cause mortality (HR 3.30, 95% CI 2.61 to 4.16), and MACE (HR 3.45, 95% CI 2.66 to 4.49) compared with those without LTCs. Of the comorbid LTCs studied, osteoporosis was most strongly associated with adverse outcomes in participants with RA compared with those without RA or LTCs: twofold increased risk of all-cause mortality (HR 2.20, 95% CI 1.55 to 3.12) and threefold increased risk of MACE (HR 3.17, 95% CI 2.27 to 4.64). These findings remained in a subset (n=3683) with RA diagnosis validated from clinical records or medication reports. Those with RA and other LTCs, particularly comorbid osteoporosis, are at increased risk of adverse outcomes, although the role of corticosteroids could not be evaluated in this study. These results are clinically relevant for the monitoring and management of RA across the healthcare system, and future clinical guidelines for RA should acknowledge the importance of multimorbidity.


Successful treatment of Brucella melitensis end-carditis.
Brucella endocarditis is a rare, but often fatal, complication of brucellosis. A 32 year old man acquired brucellosis while on a visit to his former home in Greece and presented six months later with malaise, fever and aortic regurgitation. Blood cultures grew Brucella melitensis biotype 1. Combined chemotherapy with streptomycin, tetracycline and rifampin sterilized his blood; however, his aortic valve was replaced owing to recurrent emboli and cardiac failure. Over the next 18 months the patient's antibody titer to Brucella fell and his blood reamined sterile. Cure was achieved by resection of the infected aortic valve and 10 weeks of bactericidal therapy for B. melitensis.


Beyond polymer polarity: how the cytoskeleton builds a polarized cell.
Cell polarity relies on the asymmetric organization of cellular components and structures. Actin and microtubules are well suited to provide the structural basis for cell polarization because of their inherent structural polarity along the polymer lattices and intrinsic dynamics that allow them to respond rapidly to polarity cues. In general, the actin cytoskeleton drives the symmetry-breaking process that enables the establishment of a polarized distribution of regulatory molecules, whereas microtubules build on this asymmetry and maintain the stability of the polarized organization. Crosstalk coordinates the functions of the two cytoskeletal systems.


Weyl fermions and spin dynamics of metallic ferromagnet SrRuO3.
Weyl fermions that emerge at band crossings in momentum space caused by the spin-orbit interaction act as magnetic monopoles of the Berry curvature and contribute to a variety of novel transport phenomena such as anomalous Hall effect and magnetoresistance. However, their roles in other physical properties remain mostly unexplored. Here, we provide evidence by neutron Brillouin scattering that the spin dynamics of the metallic ferromagnet SrRuO3 in the very low energy range of milli-electron volts is closely relevant to Weyl fermions near Fermi energy. Although the observed spin wave dispersion is well described by the quadratic momentum dependence, the temperature dependence of the spin wave gap shows a nonmonotonous behaviour, which can be related to that of the anomalous Hall conductivity. This shows that the spin dynamics directly reflects the crucial role of Weyl fermions in the metallic ferromagnet.


1-Benzyl-3-methyl-imidazolium bromide.
The title compound, (BenzMIm)Br (BenzMIm=1-benzyl-3-methyl-imidazo-l-ium), C11H13N2 +·Br-, was obtained as single crystals directly from a pure and liquid sample of the compound over several weeks. The mol-ecular structure of (BenzMIm)Br consists of separated bromide anions and 1-benzyl-3-methyl-imidazolium cations connected via short C-H⋯Br contacts. The compound exhibits a relatively low melting point (m.p. = 72°C) and is a supercooled, highly viscous transparent liquid at ambient conditions. The title compound crystallizes with two unique ion pairs in the asymmetric unit of the orthorhombic unit cell.


In-depth assessment of critical access hospital stewardship program adherence to the CDC Core Elements in Iowa and Nebraska.
In 21 antimicrobial stewardship programs in critical-access hospitals in Nebraska and Iowa that self-reported nonadherence to a CDC Core Element or Elements, in-depth program assessment and feedback revealed that accountability and education most needed improvement. Recommendations included providing physician and pharmacist training, tracking interventions, and providing education. Program barriers included lack of time and/or personnel and antimicrobial stewardship and/or infectious diseases expertise.


Antimicrobial effects of sodium hypochlorite and Er,Cr:YSGG laser against Enterococcus faecalis biofilm.
The aim of this study was to investigate the antimicrobial effect of Erbium, chromium-doped yttrium, scandium, gallium, and garnet (Er,Cr:YSGG) laser with and without NaOCl solution. A total of 81 extracted human mandibular premolar teeth were used. After root canal preparation and sterilization, the samples were inoculated with E. faecalis for 24 hours. The specimens were divided into 4 experimental groups. Group 1 (n = 25) was irradiated with 2 W laser, group 2 (n = 25) was irradiated with 0.75 W laser in combination with 2.5% NaOCl, group 3 (n = 25) was irrigated with 5% NaOCl and group 4 (n = 6) was not treated. Statistical analysis was performed by using Wilcoxon Signed Ranks, Mann-Whitney U and Kruskal-Wallis H tests. The combination of 0.75 W laser with 2.5% NaOCl regime was found to be just as effective at inhibiting the growth of E. faecalis and sterilization of all root canals as 5% NaOCl irrigation (P > 0.001). The 2 W laser had significant bactericidal effect in infected root canals however it did not eradicate all bacteria. The SEM observations were in accordance with the microbiologic findings. Within the limitations of this laboratory study, NaOCl irrigation improved the antimicrobial effect of Er,Cr:YSGG laser irradiation. When the toxic effects of high percentage of NaOCl was considered, the combination of low-powered laser and low concentration of NaOCl can be used as an effective disinfection method in root canal treatment.


Scrapie infection alters the distribution of plasma metabolites in diseased Cheviot sheep indicating a change in energy metabolism.
Nuclear magnetic resonance (NMR) spectroscopy has been used to profile the metabolic status of plasma from; sheep showing clinical signs of scrapie, those known to be infected with scrapie but yet to show clinical signs, and control animals. The NMR measurements have shown that energy metabolism in scrapie infected animals is altered before the onset of clinical symptoms. These metabolic changes may provide the foundation for a pre-clinical diagnostic test for scrapie in sheep.


Recent developments in the classification and functional significance of receptors for ATP and UTP, evidence for nucleotide receptors.
The presence of a nucleotide receptor activated with similar potencies by UTP and ATP is suggested by recent data from a variety of different cell types. This receptor type appears distinct from previously described ATP-sensitive P2-purinoceptor subtypes and, probably, from other UTP-sensitive receptors, however further studies using selective antagonists are necessary to provide a definitive characterisation. Although the functional role of endogenous extracellular ATP has already achieved recognition there are also many diverse examples of cells and tissues which respond to UTP at micromolar or sub-micromolar concentrations. Therefore, the possible physiological importance of UTP is a fertile area for further investigation. The functional significance of ATP/UTP receptors is underlined by recent demonstrations that UTP and ATP modulate chloride ion secretion in human airways epithelium, possibly by activation of a nucleotide receptor, an effect which may have potential clinical utility in the treatment of cystic fibrosis.


[Validity and reliability of the Demirjian distinguish software on estimating dental age].
To study the validity and reliability of the Demirjian distinguish software on dental age estimation. The dental age of 60 children were estimated with both the software three times and manual measures. The correlation coefficients of dental age estimated by the software and by manual measure was 0.974 (female) (P > 0.1) and 0.970 (male) (P > 0.05); the coefficients of interclass correlation of each dental age estimated by the software was 0.977 (female) (P > 0.1) and 0.977 (male) (P > 0.05). Demirjian distinguish software has high validity and reliability in estimating dental age.


Intraoperative imaging of zygomaticomaxillary complex fractures using a 3D C-arm system.
After preclinical studies and evaluation of radiation exposures, intraoperative three-dimensional (3D) C-arm based imaging is now available for the facial skeleton. Fourteen patients admitted for surgical treatment of zygomaticomaxillary complex (ZMC) fractures were included in the study. Preoperative diagnostics and surgical treatment were performed as usual. Intraoperatively, after open reduction, a cone-beam computed tomography (CBCT) dataset was generated using the SIREMOBIL Iso-C3D (Siemens Medical Solutions, Erlangen, Germany). After DICOM-import in eFilm Workstation axial, coronal and sagittal reconstructions were evaluated by five examiners with the help of six defined criteria. In our study, secondary reconstructions were available after 6 min, excluding the time needed for the evaluation of the images. Especially the positioning of the isocentre of the SIREMOBIL Iso-C3D proved to be uncomplicated. Because of the size of the datasets, assessment of the symmetry of the malar projection proved difficult. Best scoring results were found regarding the visualization of the fragment position, bony anchorage of the screws and the fitting of the plates. Remarkable was the low level of metal artefacts in primary and secondary reconstructions. In conclusion, our results demonstrate intraoperative CBCT using the SIREMOBIL Iso-C3D suitable for assessment of postoperative results following ZMC reduction.


Retinoids inhibit phospholipase A2 in human synovial fluid and arachidonic acid release from rat peritoneal macrophages.
Retinoids have demonstrated antiinflammatory activity in certain animal models and human disease states. The mechanism by which retinoids elicit this activity is unknown. Some retinoids are known to inhibit arachidonic acid (AA) release and metabolism in intact cells in vitro. Retinoids may exert their antiinflammatory effects by inhibiting phospholipase A2 (PLA2) and the resultant production of inflammatory AA metabolites. Retinoids were evaluated in vitro as inhibitors of the PLA2 activity in human synovial fluid (HSF-PLA2). Of the naturally occurring, nonaromatic retinoids tested, all-trans-retinal, all-trans-retinoic acid (all-trans-RA) and 13-cis-RA were the most potent inhibitors (IC50 S 6-15 microM), whereas all-trans-retinol was much less potent. Of the synthetic aromatic retinoids and arotinoids examined, the free carboxylic, sulfonic, and sulfinic acid forms were more than 15-fold more potent inhibitors of HSF-PLA2 than their corresponding ethyl esters. These retinoids also were evaluated as inhibitors of calcium ionophore A23187-induced AA release from rat peritoneal macrophages. All-trans-RA and 13-cis-RA were potent inhibitors of AA release from these cells (IC50 S 4 microM), while the other natural retinoids were inactive. Of the aromatic retinoids and arotinoids tested, the free acid forms (IC50 S 2-6 microM) were 5- to 21-fold more potent inhibitors of AA release from the macrophages than their corresponding ethyl esters. The potencies of the arotinoids as inhibitors of HSF-PLA2 appeared to correlate with their potencies as inhibitors of AA release from A23187-stimulated rat peritoneal macrophages. These data support the hypothesis that one possible mechanism for the known antiinflammatory activity of some retinoids may be by inhibition of phospholipase A2.


Live Imaging of the Lung.
Live imaging is critical to determining the dynamics and spatial interactions of cells within the tissue environment. In the lung, this has proven to be difficult due to the motion brought about by ventilation and cardiac contractions. A previous version of this Current Protocols in Cytometry article reported protocols for imaging ex vivo live lung slices and the intact mouse lung. Here, we update those protocols by adding new methodologies, new approaches for quantitative image analysis, and new areas of potential application. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Live imaging of lung slices Support Protocol 1: Staining lung sections with fluorescent antibodies Basic Protocol 2: Live imaging in the mouse lung Support Protocol 2: Intratracheal instillations Support Protocol 3: Intravascular instillations Support Protocol 4: Monitoring vital signs of the mouse during live lung imaging Support Protocol 5: Antibodies Support Protocol 6: Fluorescent reporter mice Basic Protocol 3: Quantification of neutrophil-platelet aggregation in pulmonary vasculature Basic Protocol 4: Quantification of platelet-dependent pulmonary thrombosis Basic Protocol 5: Quantification of pulmonary vascular permeability.


An orbital decompression for severe dysthyroid exophthalmos.
A procedure has been developed for maximizing decompression in patients with nonspecific orbital inflammatory disease or dysthyroid orbitopathy that involves removal of parts of all four orbital walls through a lateral orbitotomy using a 30-mm incision combined with a canthotomy and an inferior cul-de-sac incision. The procedure has been performed on seven orbits of four patients who had greater than 30 mm of forward placement of the eyes as measured by Hertel exophthalmometry and/or a 10-mm difference in the forward placement of the two eyes, indications considered for this procedure. The procedure has resulted in 10-17 mm of reduction in proptosis in these patients, which is consistently greater than that obtained by any other procedure. A complication of spinal fluid leak resulted in the recommendation that the procedure not be performed in patients over 65 whose dura is considered too thin to prevent spontaneous leakage and minimal surgical trauma. In all circumstances, this procedure must be performed by an experienced neuro-ophthalmic orbital surgical team.


Label free and homogeneous histone sensing based on chemiluminescence resonance energy transfer between lucigenin and gold nanoparticles.
In this paper, a label free and homogeneous protocol without recognition elements such as antibody or DNA based on nonenzymatic chemiluminescence resonance energy transfer between lucigenin and gold nanoparticles (AuNPs) is developed for the detection of histone. This chemiluminescence resonance energy transfer process originated from a chemiluminescent donor-acceptor pair in which the chemiluminescence system of the lucigenin-H2O2 as a donor and AuNPs as an acceptor owing to the overlapping of the chemiluminescence spectrum of the lucigenin-H2O2 system and the absorption spectrum of AuNPs, leading to a significant decrease in chemiluminescence signal from the lucigenin-H2O2 system. However, the presence of histone resulted in the aggregation of AuNPs via the electrostatic interaction between negatively charged AuNPs and positively charged histone, which inhibited the chemiluminescence resonance energy transfer process. Thus the chemiluminescence signal of the lucigenin-H2O2 system was restored. This could be used for the detection of histone with a linear range of 30-500ng/mL, and a detection limit of 25ng/mL. This sensitivity increased about 4 orders of magnitude over that of the reported fluorometric method. The proposed strategy for the detection of histone is simple, facile, reliable, and opens a new avenue for the determination of histone.


Procyanidins, from Castanea mollissima Bl. shell, induces autophagy following apoptosis associated with PI3K/AKT/mTOR inhibition in HepG2 cells.
Procyanidins from Castanea mollissima Bl. shell (CSPCs) induced autophagy and apoptosis in HepG2 cells and its mechanism remains to be examined. In this paper, autophagy was measured by the lipid modification of light chain-3 (LC3) and the formation of autophagosomes. Hoechst 33258 staining and flow cytometer analysis were used to measure apoptosis. The western blot analysis was used to examine the effects of CSPCs on the expression of LC3, PI3K, phosphorylation of AKT, mTOR, Bcl-2, Bad, Bax, BID and cleaved caspase 3 in HepG2 cells. The results showed that 3-methyladenine (3-MA) and apoptosis inhibitor (Z-VAD) could inhibited the death of HepG2 induced by CSPCs for 48h (150μg/mL). CSPCs induced the accumulation of autophagosomes and microtubule-associated proteins light chain 3-II (LC3-II, a marker of autophagy). P-AKT, PI3K and mTOR were significantly decreased on CSPCs exposure. However, these phenomena were not observed in the group pretreated with the autophagy inhibitor 3-MA and Z-VAD. CSPCs also induced the expression of Bad, Bax and Beclin-1 proteins and decreased the expression of Bcl-2, which was inhibited by 3-MA and Z-VAD. Moreover the apoptotic cell death could be inhibited by 3-MA. In addition, inhibition of LC3-II by siRNA-dependent knockdown attenuated the cleavage of caspase 3. These results suggested CSPCs could trigger autophagy via inhibition of the PI3K/AKT/mTOR signaling pathway, enhanced apoptosis in HepG2 cells which may be associated with the mitochondria-dependent signaling way.


A novel method of estimating cost of therapy by using patient population characteristics: analysis of fluoroquinolones in various populations with different distributions of renal function.
Formulary decisions regarding a given drug class are often made in the absence of patient outcome and/or sophisticated pharmacoeconomic data. Analyses that consider factors beyond simple acquisition costs may be useful in such situations. For example, the cost implications of using manufacturers' recommendations for dosing in patients with renal dysfunction may be important, depending on the distribution of various levels of renal function within a patient population. Using four 1000-patient populations representing different renal function distributions and a fifth population of our medical center's distribution, we determined the costs of therapy for intravenous and oral levofloxacin, gatifloxacin, and moxifloxacin for a 10-day course of therapy for community-acquired pneumonia. Costs considered were average wholesale prices (AWPs), 50% of AWP, or same daily price, plus intravenous dose preparation and administration costs when applicable. Costs for each renal function distribution were examined for significant differences with an analysis-of-variance test. Also, costs of failing to adjust dosing regimens for decreased renal function were determined. Differences in fluoroquinolone costs (AWP, 50% AWP, or when matched as the same daily price) among the populations were found. When considering same daily prices, differences among populations ranged from about 35,000 dollars with intravenous gatifloxacin to more than 51,000 dollars for intravenous levofloxacin (all fluoroquinolones, p>0.05). Within a population, differences in costs among the intravenous fluoroquinolones ranged from 47,000-99,000 dollars. Rank orders of the drugs and population costs of therapy were affected by the pricing structure used and varied by the specific population and drug. Differences among the fluoroquinolones or populations were much smaller (<2100 dollars) when considering oral regimens. Costs potentially incurred by failing to adjust dosing for renal function were substantial. Formulary decisions can be facilitated by considering factors such as patient characteristics and related dosing in addition to simple acquisition costs. In our example, consideration of the distribution of renal function within a given patient population and related dosing for these fluoroquinolones revealed potentially important differences within the class.


The intergenic region of maize streak virus contains a GC-rich element that activates rightward transcription and binds maize nuclear factors.
Maize streak virus (MSV) is transcribed bidirectionally from an intergenic region and rightward transcription produces an RNA that encodes the coat protein. The intergenic region contains promoter elements required for rightward transcription including an upstream activating sequence (UAS) which endows the promoter with full activity in a maize transient expression system. The UAS contains two GC-rich repeats (GC boxes) and a long inverted repeat or hairpin with a loop harboring a TAATATTAC sequence common to all geminiviruses. Deletions through the UAS demonstrated the presence of an element, called the rightward promoter element (rpe1), which is responsible for transcriptional activation. Rpe1 includes the two GC-rich boxes, which are similar in sequence to Sp1 binding sites in mammalian cells, but not the conserved hairpin loop. Rpe1 binds maize nuclear factors in vitro and the characteristics of the binding interaction have been determined by 1) binding competition with oligonucleotides, 2) methidiumpropyl-EDTA footprinting and 3) methylation interference assays. Binding of maize nuclear factors to the UAS generates two major bands, slow and fast migrating bands, in gel retardation assays. Footprinting and factor titration data suggest that the fast bands arise by the binding of factors to one GC box while the slow bands are generated by factors binding to both boxes. The data further indicate that the factors bind to the two GC-rich boxes with little cooperativity and bind on opposite faces of the DNA helix.


Development of new chromanol antagonists of leukotriene D4.
By addressing the issues of potency and metabolism in 3, a new series of LTD4 antagonists represented by (+)-26 was developed which is equipotent to clinical LTD4 antagonists Zafirlukast (1) and Pranlukast (2).


The effects of the incision types in myringoplasty operations on cosmesis.
There is no study based on objective measurements about the cosmetic results of myringoplasty operation in medical literature. This study aims to show the differences in the auriculomastoid angle between the operations which were done with postauricular and endaural incisions. The auriculomastoid angle of 20 patients having had postauricular myringoplasty operation were measured both preoperative and postoperative periods. The auriculomastoid angles of 17 patients having had endaural myringoplasty operation were also measured in both preoperative and postoperative periods. Also, the patients were asked whether there was a change in the shape of their ears. It was observed that the increase of the auriculomastoid angles of the patients who had myringoplasty by postauricular incision was statistically significant when compared to the ones having had the operation by endaural incision. Also, the patients who had myringoplasty operation by postauricular incision realized the change in their ears significantly when compared to the other group. In this study, esthetically better results were observed in the myringoplasty operations done by endaural incision than the ones done by postauricular incision. When deciding on the incision type, this should be considered.


Bottlenose dolphins perceive object features through echolocation.
How organisms (including people) recognize distant objects is a fundamental question. The correspondence between object characteristics (distal stimuli), like visual shape, and sensory characteristics (proximal stimuli), like retinal projection, is ambiguous. The view that sensory systems are 'designed' to 'pick up' ecologically useful information is vague about how such mechanisms might work. In echolocating dolphins, which are studied as models for object recognition sonar systems, the correspondence between echo characteristics and object characteristics is less clear. Many cognitive scientists assume that object characteristics are extracted from proximal stimuli, but evidence for this remains ambiguous. For example, a dolphin may store 'sound templates' in its brain and identify whole objects by listening for a particular sound. Alternatively, a dolphin's brain may contain algorithms, derived through natural endowments or experience or both, which allow it to identify object characteristics based on sounds. The standard method used to address this question in many species is indirect and has led to equivocal results with dolphins. Here we outline an appropriate method and test it to show that dolphins extract object characteristics directly from echoes.


Distribution characteristics, sources, and risk assessment of organochlorine pesticides in the Fildes Peninsula and Adelaide Island.
This work analyzed the distribution characteristics, sources, and corresponding environmental risks of 10 organochlorine pollutants (OCPs) in soil samples from 13 stations on the Fildes Peninsula and Adelaide Island in Antarctica in 2020. Results showed that the total amount of OCP residues in the study area ranged from N.D. to 2.66 ng·g-1 with an average concentration of 0.90 ng·g-1. Six organochlorine pesticides were detected, and the residue level of DDTs was slightly lower than that of HCHs. The residues of HCHs were mainly γ-HCH and those of DDTs were mainly O, P'-DDT. Source analysis revealed that the residues mainly originated from atmospheric transport and human activities. Environmental risk analysis showed that in the study area, the residues of HCHs and DDTs were present at low levels, and their ecological risk was low.


Low b-value diffusion-weighted imaging: emerging applications in the body.
Thanks to recent advances in magnetic resonance imaging technology, it has become possible to perform intravoxel incoherent motion (IVIM) diffusion-weighted imaging (DWI) in any part of the body. Extracranial applications of DWI are currently under active investigation, especially for oncological imaging. However, the use of non-quantitative low b-value (10-100 s/mm(2)) DWI in the body is still a relatively unexplored field, and its potential is not fully recognized. Non-quantitative low b-value DWI may especially be useful for the evaluation of structures that have an inherently low signal at high b-value DWI, including (but not limited to) the liver, heart, and small bowel. This article will review and discuss the basic principles and potential applications of nonquantitative low b-value DWI in the body.


Cost effectiveness analysis of CAR-T cell therapy for patients with relapsed/refractory multiple myeloma in China.
The landscape of treatment strategies for relapsed/refractory multiple myeloma (RRMM) has dramatically changed due to the emergence of chimeric antigen receptor T (CAR-T) cell therapy. The aim of this study was to evaluate the cost-effectiveness of two CAR-T cell treatments for RRMM patients from the perspective of the Chinese healthcare system. A Markov model was used to compare currently available salvage chemotherapy with Idecabtagene vicleucel (Ide-cel) and Ciltacabtagene autoleucel (Cilta-cel) for treatment of patients with RRMM. The model was developed based on data from three studies: CARTITUDE-1, KarMMa, and MAMMOTH. The healthcare cost and utility of RRMM patients were collected from a provincial clinical center in China. In the base case analysis, 3.4% and 36.6% of RRMM patients were expected to be long-term survivors after 5 years of Ide-cel and Cilta-cel treatment, respectively. Compared to salvage chemotherapy, Ide-cel and Cilta-cel were associated with incremental QALYs of 1.19 and 3.31, and incremental costs of US$140,693 and $119,806, leading to ICERs of $118,229 and $36,195 per QALY, respectively. At an ICER threshold of $37,653/QALY gained, the probability that Ide-cel and Cilta-cel are cost-effective were estimated to be 0% and 72%, respectively. With younger target people entering the model, and a partitioned survival model in scenario analysis, the ICERs of Cilta-cel and Ide-cel changed rather mildly and their cost-effectiveness results were the same as base analysis. Based on the willingness-to-pay of 3-times China's per capita GDP in 2021, Cilta-cel was considered to be a more cost-effective option compared to salvage chemotherapy for RRMM in China, while Ide-cel was not.


Introduction to the Special Issue: Mechanisms of Action in Cognitive-Behavior Therapy.
Research into mechanisms of treatment outcome in cognitive-behavior therapy (CBT), as work evaluating potential mechanisms through mediator and moderator analyses, has been increasing. In recognition of this growth in more refined analyses of functional components of treatment, this special issue highlights a diverse range of research-evaluating mechanisms, as well as mediators and moderators, of outcome. It is anticipated that the research described in these articles will stimulate additional investigations as well as provide important insights for clinicians in refining treatment programs.


Conformationally constrained NO synthase inhibitors: rigid analogs of L-N-iminoethylornithine.
The synthesis of eight rigid analogs of L-N-iminoethylornithine (L-NIO) is described. The compounds have been evaluated for their inhibition of inducible nitric oxide synthase. Preliminary structure-activity relationships are discussed.


Reaction patterns of wheat starch granules substituted with an anionic propylene oxide analog (POA) revealed by confocal microscopy and 3D anaglyph imaging.
A technique was developed for investigating granular reaction patterns of anionic starch derivatives, utilizing the optical sectioning capability of reflectance confocal laser scanning microscopy in combination with 3D anaglyphic imaging (3D glasses required for visualization). The method facilitated improved visualization of internal granular reaction patterns by discarding optical sections comprising the highly-reacted external granule surface (generating 3D image constructs from sections constituting exclusively internal regions of granules) and minimizing confounding background signal associated with non-starch constituents (e.g., proteins). For wheat starch A-type granules substituted with a propylene oxide analog (POA), granular reaction patterns revealed that reagent reached the hilum region of granules through channels, and primarily entered the granule matrix via lateral diffusion from channel surfaces, producing relatively homogeneous reaction patterns. The homogeneous nature of POA granular reaction patterns was attributed to the relatively low reactivity of the reagent (rate of diffusion>rate of reaction).


Efficient synthesis of 3'-glycosylated LacNAc-based oligosaccharides.
LacNAc-based oligosaccharides, including sialyl-(2-->3)-LacNAc, dimeric sialyl-(2-->3)-LacNAc, trimeric sialyl-(2-->3)-LacNAc, beta-glucuronyl-(1-->3)-LacNAc, and 3-sulfo-beta-glucuronyl-(1-->3)-LacNAc, were synthesized efficiently from a single protected LacNAc derivative having both OH-3' and 4' unprotected.


Shorter Incubation Times for Detecting Multi-drug Resistant Bacteria in Patient Samples: Defining Early Imaging Time Points Using Growth Kinetics and Total Laboratory Automation.
The transition from manual processing of patient samples to automated workflows in medical microbiology is challenging. Although automation enables microbiologists to evaluate all samples following the same incubation period, the essential incubation times have yet to be determined. We defined essential incubation times for detecting methicillin-resistant Staphylococcus aureus (MRSA), multi-drug resistant gram-negative bacteria (MDRGN), and vancomycin-resistant enterococci (VRE). We monitored the growth kinetics of MRSA, MDRGN, and VRE between two and 48 hours on chromogenic media to establish the time points of first growth, single colony appearance, and typical morphology for 10², 10⁴, 10⁶, and 10⁸ colony forming units/mL. Subsequently, we imaged plates inoculated with 778 patient samples after 20, 24, and 36 hours. The first growth, single colony appearance, and typical morphology time points were inoculum-dependent. First growth appeared after 6-18 hours, 4-18 hours, and 8-48 hours for MRSA, MDRGN, and VRE, respectively, and single colonies appeared at 12-18 hours, 6-20 hours, and 12-48 hours, respectively. Typical morphology was visible at 14-22 hours and 12-48 hours for MRSA and VRE, but was not determined for MDRGN. By examining patient samples, ≥98% of MRSA and MDRGN were visible 20 hours after the start of incubation. Following 24 hours of incubation, only 79.5% of VRE were clearly visible on the respective plates. An incubation time of 20 hours is sufficient for detecting MRSA and MDRGN. VRE growth is much slower and requires additional imaging after 36 hours.


Influence of beetroot juice supplementation on intermittent exercise performance.
This study tested the hypothesis that nitrate (NO3-) supplementation would improve performance during high-intensity intermittent exercise featuring different work and recovery intervals. Ten male team-sport players completed high-intensity intermittent cycling tests during separate 5-day supplementation periods with NO3 (-)-rich beetroot juice (BR; 8.2 mmol NO3- day(-1)) and NO3 (-)-depleted beetroot juice (PL; 0.08 mmol NO3- day(-1)). Subjects completed: twenty-four 6-s all-out sprints interspersed with 24 s of recovery (24 × 6-s); seven 30-s all-out sprints interspersed with 240 s of recovery (7 × 30-s); and six 60-s self-paced maximal efforts interspersed with 60 s of recovery (6 × 60-s); on days 3, 4, and 5 of supplementation, respectively. Plasma [NO2-] was 237% greater in the BR trials. Mean power output was significantly greater with BR relative to PL in the 24 × 6-s protocol (568 ± 136 vs. 539 ± 136 W; P < 0.05), but not during the 7 × 30-s (558 ± 95 vs. 562 ± 94 W) or 6 × 60-s (374 ± 57 vs. 375 ± 59 W) protocols (P > 0.05). The increase in blood [lactate] across the 24 × 6-s and 7 × 30-s protocols was greater with BR (P < 0.05), but was not different in the 6 × 60-s protocol (P > 0.05). BR might be ergogenic during repeated bouts of short-duration maximal-intensity exercise interspersed with short recovery periods, but not necessarily during longer duration intervals or when a longer recovery duration is applied. These findings suggest that BR might have implications for performance enhancement during some types of intermittent exercise.


Aortic root replacement in 271 Marfan patients: a 24-year experience.
The introduction of composite graft repair of aortic root aneurysm by Hugh Bentall in 1968 promised Marfan patients the choice for a normal life expectancy. We performed our first Bentall composite graft procedure in 1976 and herein report our 24-year experience with 271 Marfan patients. Between September 1976 and August 2000, 232 Marfan patients had a composite graft replacement of the aortic root, 15 patients received a homograft, and 24 had a valve-sparing procedure. Two hundred thirty-five Marfan patients underwent elective aortic root replacement with no 30-day mortality. Two early deaths occurred among 36 patients who underwent urgent or emergent operation. Eighty-three percent of patients in this series are currently alive. The actuarial freedom from thromboembolism, endocarditis, and reoperation on the residual aorta 20 years postoperatively was 93%, 90%, and 74%. Twenty-four patients have undergone valve-sparing procedures with encouraging results. Elective aortic root replacement for Marfan patients can be performed with low operative risk. Elective repair before the aortic root reaches 6 cm in diameter is recommended to minimize risk of dissection and rupture.


Comprehensive analysis of TCGA data reveals correlation between DNA methylation and alternative splicing.
The effect of DNA methylation on the regulation of gene expression has been extensively discussed in the literature. However, the potential association between DNA methylation and alternative splicing is not understood well. In this study, we integrated multiple omics data types from The Cancer Genome Atlas (TCGA) and systematically examined the relationship between DNA methylation and alternative splicing. Using the methylation data and exon expression data, we identified many CpG sites significantly associated with exon expression in various types of cancers. We further observed that the direction and strength of significant CpG-exon correlation tended to be consistent across different cancer contexts, indicating that some CpG-exon correlation patterns reflect fundamental biological mechanisms that transcend tissue- and cancer- types. We also discovered that CpG sites correlated with exon expressions were more likely to be associated with patient survival outcomes compared to CpG sites that did not correlate with exon expressions. Furthermore, we found that CpG sites were more strongly correlated with exon expression than expression of isoforms harboring the corresponding exons. This observation suggests that a major effect of CpG methylation on alternative splicing may be related to the inclusion or exclusion of exons, which subsequently impacts the relative usage of various isoforms. Overall, our study revealed correlation patterns between DNA methylation and alternative splicing, which provides new insights into the role of methylation in the transcriptional process.


Contact of cis-Golgi with ER exit sites executes cargo capture and delivery from the ER.
Protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus is mediated by coat complex II (COPII) vesicles. It has been believed that COPII vesicles containing cargo are released from the ER exit sites (ERES) into the cytosol and then reach and fuse with the first post-ER compartment, cis-Golgi or ER-to-Golgi intermediate compartment (ERGIC). However, it still remains elusive how cargo loading to vesicles, vesicle budding, tethering and fusion are coordinated in vivo. Here we show, using extremely high speed and high resolution confocal microscopy, that the cis-Golgi in the budding yeast Saccharomyces cerevisiae approaches and contacts the ERES. The COPII coat cage then collapses and the cis-Golgi captures cargo. The cis-Golgi, thus loaded with cargo, then leaves the ERES. We propose that this 'hug-and-kiss' behaviour of cis-Golgi ensures efficient and targeted cargo transport from the ERES to cis-Golgi.


CCR5 mimicry by sulfated human anti-HIV-1 antibodies.
HIV-1 envelope glycoprotein interaction with a principal coreceptor, CCR5, requires sulfated tyrosines on the CCR5 amino terminus. In this issue of Cell, Choe et al. identified tyrosine-sulfated, neutralizing antibodies against HIV-1 that apparently mimic CCR5. The results demonstrate for the first time that a natural posttranslational modification of an antibody can contribute substantially to antigen recognition.


Outcomes of Highly Selected Live Donors With a Future Liver Remnant Less Than or Equal to 30%: A Matched Cohort Study.
The main concern with live donor liver transplantation (LDLT) is the risk to the donor. Given the potential risk of liver insufficiency, most centers will only accept candidates with future liver remnants (FLR) >30%. We aimed to compare postoperative outcomes of donors who underwent LDLT with FLR ≤30% and >30%. Adults who underwent right hepatectomy for LDLT between 2000 and 2018 were analyzed. Remnant liver volumes were estimated using hepatic volumetry. To adjust for between-group differences, donors with FLR ≤30% and >30% were matched 1:2 based on baseline characteristics. Postoperative complications including liver dysfunction were compared between the groups. A total of 604 live donors were identified, 28 (4.6%) of whom had a FLR ≤30%. Twenty-eight cases were successfully matched with 56 controls; the matched cohorts were mostly similar in terms of donor and graft characteristics. The calculated median FLR was 29.8 (range, 28.0-30.0) and 35.2 (range, 30.1-68.1) in each respective group. Median follow-up was 36.5 mo (interquartile range, 11.8-66.1). Postoperative outcomes were similar between groups. No difference was observed in overall complication rates (FLR ≤30%: 32.1% versus FLR >30%: 28.6%; odds ratio [OR], 1.22; 95% confidence interval [CI], 0.46-3.27) or major complication rates (FLR ≤30%: 14.3% versus FLR >30%: 14.3%; OR, 1.17; 95% CI, 0.33-4.10). Posthepatectomy liver failure was rare, and no difference was observed (FLR ≤30%: 3.6% versus FLR >30%: 3.6%; OR, 1.09; 95% CI, 0.11-11.1). A calculated FLR between 28% and 30% on its own should not represent a formal contraindication for live donation.


[Clinical evaluation of local application of Merogel after endoscopic sinus surgery].
To study the auxo-action of Merogel (hyaluronic acid) for operative cavity epithelialization after endoscopic sinus surgery (ESS). Sixteen patients (32 sides) were observed, in which all sinuses(maxillary, frontal, ethmoid and sphenoid) were operated on by ESS. For the purpose of control study, the left nasal side was used as Merogel group, and right side contrast group. The patients were followed-up for 12 weeks. In Merogel group, the curative rate was 93.75%, improvement rate 6.25%, and average time of epithelialization was 3.4 weeks. In contrast group, the curative rate was 87.50%, improvement rate 12.50%, and average time of epithelialization was 8.3 weeks. It was distinct that in Merogel group, the epithelialization time was shorter than that in contrast group. Merogel has auxo-action for operative cavity epithelialization after ESS.


Influence of paternal age on ongoing pregnancy rate at eight weeks' gestation in assisted reproduction.
A retrospective cohort study was performed with the followings aims: to evaluate the influence of paternal age on best embryo quality available for embryo transfer on the third day; biochemical pregnancy rate; miscarriage rate and ongoing pregnancy rate at 8 weeks' gestational age, after IVF or intracytoplasmic sperm injection (ICSI) treatment, respectively, including treatment with non-ejaculated spermatozoa. In total, 7051 first IVF/ICSI cycles in Radboud university medical center, between 1 January 2001 and 1 June 2013 were included in this study. A statistical model was used to analyse the effect of paternal age and maternal age. No statistically significant differences between the paternal age groups were found with respect to the probability of an ongoing pregnancy after the first cycle (35-44 years: odds ratio [OR] = 0.97 [95% confidence interval [CI]: 0.86 to 1.10] and ≥45 years: OR = 1.01 [95% CI: 0.82 to 1.26]), respectively, compared with <35 years of age (control). Similar results were found with respect to paternal age and the availability of a top quality embryo for transfer, biochemical pregnancy and miscarriage. However, live birth was not taken into account. In conclusion, paternal age did not affect ongoing pregnancy rates in first IVF/ICSI cycles.


Do children need reminders on the Day-Night task, or simply some way to prevent them from responding too quickly?
We previously reported better performance on the Day-Night task when a ditty was chanted between stimulus presentation and when children could respond (Diamond, Kirkham, & Amso, 2002). Here we investigated competing hypotheses about why the ditty helps. Does it help because it imposes a brief waiting time (the child waits while the ditty is chanted before responding)? Or, does the ditty help because of its content, providing information helpful to performing the task? One-third of the 72 children (age 4) were tested with the ditty previously used which reminds them: "Think about the answer; don't tell me." Another 24 children were tested with a ditty with no task-relevant content: "I hope you have a nice time; I like you." One-third received the standard condition. Performance in both ditty conditions was comparable and better than in the standard condition. That indicates that a factor common to both ditties (that chanting them took time, allowing the prepotent response to subside and the more-considered answer to reach response threshold) likely accounts for their benefit. Whether a ditty reminded children what to do or not did not affect the results. The challenge of the Day-Night task for preschoolers is not its working memory demands but the need to inhibit a dominant response, making a different response instead.


Quantitative phase imaging of living red blood cells combining digital holographic microscopy and deep learning.
Digital holographic microscopy as a non-contacting, non-invasive, and highly accurate measurement technology, is becoming a valuable method for quantitatively investigating cells and tissues. Reconstruction of phases from a digital hologram is a key step in quantitative phase imaging for biological and biomedical research. This study proposes a two-stage deep convolutional neural network named VY-Net, to realize the effective and robust phase reconstruction of living red blood cells. The VY-Net can obtain the phase information of an object directly from a single-shot off-axis digital hologram. We also propose two new indices to evaluate the reconstructed phases. In experiments, the mean of the structural similarity index of reconstructed phases can reach 0.9309, and the mean of the accuracy of reconstructions of reconstructed phases is as high as 91.54%. An unseen phase map of a living human white blood cell is successfully reconstructed by the trained VY-Net, demonstrating its strong generality.


Regulation of renal cytochrome P450s by thyroid hormone in diabetic rats.
Effects of thyroid hormone treatment on renal P450 expression in the diabetic rats were investigated. Diabetes produced by streptozotocin induced CYP4A2 and P450 K-2 (similar form with CYP2C23) but not P450 K-4 (similar form with CYP4A8) and induced lauric acid hydroxylation activity. The serum thyroid hormone level was decreased with diabetes. Treatment of diabetic rats with thyroid hormone (T3) as well as with insulin reversed the increase in the levels of CYP4A2 and P450 K-2. Thyroidectomy also induced CYP4A2 and P450 K-2 in the rat kidney. The increase was reversed by treatment of thyroidectomized rats with T3. These findings suggest that expression of CYP4A2 and P450 K-2 in rat kidney is suppressively regulated by thyroid hormone and the decrease in thyroid hormone level in the diabetic state affects the levels of CYP4A2 and P450 K-2.


Tissue solubility of four volatile anesthetics in fresh and frozen tissue specimens from swine.
To determine tissue solubilities of desflurane, sevoflurane, enflurane, and halothane in swine and to evaluate the effects of freezing specimens on tissue solubility, Arterial blood samples and specimens of brain, heart, liver, kidney, muscle, and subcutaneous fat from 5 healthy female adult Chinese Meishan pigs. Each tissue specimen was divided into 2 parts. One part was used to measure tissue-gas partition coefficients immediately after collection. The other part was frozen at -20 C for 6 days prior to determination of tissue-gas partition coefficients. Tissue-gas and blood-gas partition coefficients were measured by use of gas chromatography, and tissue-blood partition coefficients were calculated. Regression analysis was performed to determine whether fat-gas partition coefficients were correlated with lean tissue-gas partition coefficients. Tissue-gas and blood-gas partition coefficients of halothane were greater than those of enflurane followed by coefficients of sevoflurane and desflurane. However, the order of anesthetic agents with the greatest to smallest tissue-blood partition coefficients was sevoflurane, halothane, enflurane, and desflurane. Muscle-gas partition coefficients of sevoflurane and enflurane, liver-gas partition coefficients of desflurane and halothane, and the kidney-gas partition coefficient of enflurane were significantly greater in frozen specimens, compared with fresh specimens. Lean tissue-gas partition coefficients of all 4 volatile anesthetics correlated directly with fat-gas partition coefficients. The fat content of lean tissue is an important factor in determining the tissue solubility of volatile anesthetics. Freezing specimens before determination of tissue-gas partition coefficients may result in a false increase in tissue solubility.


Initial experience with ACT™ periurethral adjustable balloons to treat urinary incontinence due to intrinsic sphincter deficiency in the pediatric population.
To present our initial experience with periurethral adjustable continence therapy (ACT™) for urinary incontinence due to intrinsic sphincter deficiency (ISD) in children. This is an approved prospective non-randomized pilot study (NCT03351634) aiming to treat children born with spinal dysraphism (SD) or exstrophy epispadias complex (EEC) with ACT™. Endpoints were patient-reported changes in daily pad count, 24-h Pad test and complications. Since April 2018, 13 children (six girls, seven boys) were implanted at the median age of 12 years (5-16). The etiology of incontinence was neurogenic ISD (7/13, 54%) and EEC (6/13, 46%). After ACT™ implantation, continence (no pad or 1 security pad/day) was achieved in 9(69%) patients (5/7 SD, 4/6 EEC). Additionally, two (15%) patients had a significant improvement (decreasing Pad test from 1049 to 310 g at 3 months). One patient (7%) had no improvement. Results were stable at 21 months (6-43) of follow-up. Mean final balloon volume was 2.89 ml (± 0.85) with a median of 3 fillings to obtain continence. We had four revisions due to cutaneous port erosion (n = 3) and balloon migration (n = 1) and two definitive explantations. PinQ score was significantly improved (47 vs 40.5 with balloon, p = ns). Neither degradation of the upper urinary tract nor cystomanometric changes have been observed at 6 and 12 months postoperatively. Urinary incontinence due to ISD owing to EEC or SD can be successfully treated with ACT™ periurethral balloons. Given the minimal invasiveness of this therapy, it might be a first-line option treatment in children with complex stress urinary incontinence.


Clinical Images: Symmetrical Gyriform Restricted Diffusion in Severe Reversible Cerebral Vasoconstriction Syndrome.
A 45-year-old woman with chronic depression and currently receiving dual serotonergic therapy was transferred to our department with sudden dysbasia; she also reported an antecedent thunderclap headache. Magnetic resonance imaging (MRI) revealed a symmetrical bilateral acute infarction in the medial parietal lobe and severe vasoconstriction in the cerebral arteries. We diagnosed reversible cerebral vasoconstriction syndrome (RCVS) and stopped the serotonergic drug regimen. Her symptoms improved after receiving a calcium blocker and magnesium sulfate, and she was discharged after 20 days. Cerebral infarctions developed in 30%-40% of reversible cerebral vasoconstriction syndrome (RCVS) cases, mostly within watershed regions; however, these atypical infarctions were bilateral and symmetrical, potentially because of hypoperfusion caused by long-lasting severe vasoconstriction in the anterior cerebral arteries. Rare bilateral neurologic deficits can be mistaken for hypochondriac delusions, particularly in patients with depression. Since serotonergic drugs are the main cause of RCVS, primary care psychiatrists must be aware of drug-induced RCVS.


Emotional memory for facial expressions in schizophrenia spectrum disorders: The role of encoding method.
Memory is better for emotional rather than neutral events or materials (emotional enhancement of memory; EEM). Evidence suggests that this memory benefit remains intact in schizophrenia, but conflicting findings present the need for further research to understand how and when this process occurs. Here, we examine whether different encoding methods for learning emotional faces may result in different EEM patterns in those with schizophrenia spectrum disorders (SSD). A patient group (n = 28) and healthy comparisons (n = 29) encoded faces in two conditions that manipulated attentional focus to promote direct (emotion judgements) or indirect (sex discrimination) processing of emotional content. Based on literature in schizophrenia showing selective emotion perception deficits on tasks of direct processing but relatively intact emotion perception on indirect processing tasks, we hypothesized that patients would show greater EEM effects when faces were encoded indirectly. This hypothesis was not supported, and the patient group instead showed a similar intact EEM for angry and fearful faces to healthy comparisons in both encoding conditions. Further, using the Remember/Know paradigm, we demonstrated that the EEM in SSD appears selective to recollection-based memory, which helps to explain inconsistencies in past literature that has not differentiated between recognition domains. These findings have important implications for improving emotional memory and functional outcomes in SSD; future research should establish how the EEM for facial expressions may relate to social functioning.


Evaluation of sewage source and fate on southeast Florida coastal reefs.
Water, sponge and coral samples were collected from stations impacted by a variety of pollution sources and screened for human enteric viruses as conservative markers for human sewage. While human enteroviruses and adenoviruses were not detected, noroviruses (NoV; human genogroups I and II) were detected in 31% of samples (especially in sponge tissue). Stations near inlets were the only ones to show multiple sample types positive for NoV. Fecal indicator bacteria and enteric viruses were further evaluated at multiple inlet stations on an outgoing tide. Greatest indicator concentrations and highest prevalence of viruses were found at the mouth of the inlet and offshore in the inlet plume. Results suggest that inlets moving large volumes of water into the coastal zone with tides may be an important source of fecal contaminants. Efforts to reduce run-off or unintended release of water into the Intracoastal Waterway may lower contaminants entering sensitive coastal areas.


Selective conversion of syngas to light olefins.
Although considerable progress has been made in direct synthesis gas (syngas) conversion to light olefins (C2(=)-C4(=)) via Fischer-Tropsch synthesis (FTS), the wide product distribution remains a challenge, with a theoretical limit of only 58% for C2-C4 hydrocarbons. We present a process that reaches C2(=)-C4(=) selectivity as high as 80% and C2-C4 94% at carbon monoxide (CO) conversion of 17%. This is enabled by a bifunctional catalyst affording two types of active sites with complementary properties. The partially reduced oxide surface (ZnCrO(x)) activates CO and H2, and C-C coupling is subsequently manipulated within the confined acidic pores of zeolites. No obvious deactivation is observed within 110 hours. Furthermore, this composite catalyst and the process may allow use of coal- and biomass-derived syngas with a low H2/CO ratio.


Subcutaneous pedicle limberg flap for facial reconstruction.
A residual dog-ear arising from the traditional Limberg flap transposition sometimes influences the cosmetic outcomes, and the Limberg flap with a subcutaneous pedicle is an innovation applicable to a variety of facial defects and free from the dog-ear. To investigate the outcome of the subcutaneous pedicle Limberg flap for the reconstruction of medium-sized facial skin defects. From August 2002 to June 2004, the subcutaneous pedicle Limberg flap was designed to repair facial skin defects in 17 patients (19 flaps), and the size of the lesions ranged from 2.0 x 1.9 cm to 5.0 x 4.5 cm. All flaps survived with primary healing postoperatively. The patients were followed up from 1 to 22 months, and functionally and cosmetically satisfactory outcomes were achieved. This subcutaneous pedicle Limberg flap provides a competitive repair alternative for the treatment of medium-sized skin defects in the face.


Measurement of an isosbestic point in the Raman spectrum of liquid water by use of a backscattering geometry.
Measurements presented here confirm that a temperature-insensitive point occurs in the backscattered Raman spectrum from liquid water. This result, coupled with existing laboratory measurements of Raman scattering from liquid droplets, indicates that a Raman lidar measurement of cloud liquid water is feasible.


Recombinant M-, B- and MB-type isozymes of human phosphoglyceric acid mutase: their large-scale production and preparation of polyclonal antibodies specific to M- and B-type isozymes.
Human phosphoglyceric acid mutase is a dimer comprising M-, B- and MB-type isozymes composed from the combination of the muscle-specific (M) and non-muscle-specific (B) subunits. Human DNAs coding M and B subunits were, respectively, reconstructed at their 5' regions without changing amino acid sequences, and expressed directly in Escherichia coli under the control of the trp promoter. M- and B-type isozymes were over-produced in the bacterial cytoplasm as soluble, active forms, which have been purified and characterized. MB-type was synthesized in vitro by recombining M- and B-type. All three recombinant isozymes thus obtained showed the same properties as the naturally-occurring ones with respect to the properties tested. Polyclonal IgGs specific to the M-type, B-type and MB-type were prepared from rabbits immunized with M- and B-type, using columns bound with M- and B-type. A method for the immunoassay of MB-type which is specifically present in cardiac muscle, is now under development.


Amiodarone in the management of patients with ventricular tachycardia and ventricular fibrillation.
Fifty-eight patients with symptomatic ventricular tachycardia (VT) or ventricular fibrillation (VF) were treated with amiodarone. All had clinical episodes of VT/VF or inducible VT during electropharmacologic testing despite treatment with maximum-tolerated doses of conventional antiarrhythmic agents. Chronic treatment with amiodarone was begun at a dose of 800-1000 mg per day. Thirty-two patients were also treated with a previously ineffective conventional agent. Thirty patients underwent programmed ventricular stimulation after 2.6 +/- 1.7 months (mean +/- S.D.) of treatment with amiodarone at a mean daily dose of 588 +/- 155 mg. VT was induced in 25 patients (sustained in 20, nonsustained in five). Seventeen patients had a recurrence of VT or VF after 0.5-9 months of treatment with amiodarone (fatal in seven, non-fatal in 10). Forty-one patients (71%) had no recurrence of symptomatic VT or VF while being treated with amiodarone (mean follow-up period, 17.1 +/- 12.4 months). Among the 25 patients who had inducible VT with programmed ventricular stimulation while being treated with amiodarone, 19 patients (76%) have had no recurrence of symptomatic VT or VF over a follow-up period of 21.5 +/- 7.3 months. Ambulatory electrocardiographic recordings obtained after one week of treatment with amiodarone were not helpful in predicting clinical response. Twenty-two patients (38%) developed ataxia and/or an intention tremor which improved with a decrease in the amiodarone dose. Amiodarone, either by itself or in combination with conventional antiarrhythmic drugs, has a significant therapeutic effect in high risk patients with refractory VT. The finding of inducible VT during electropharmacologic testing in patients taking amiodarone does not preclude a favorable clinical response. Neurologic toxicity is common in patients treated with 600-800 mg per day of amiodarone.


Haemopericardium in rheumatoid arthritis.
We report two cases of haemopericardium presenting as tamponade in patients with rheumatoid arthritis. Pericardial aspiration produced a good clinical response with no reaccumulation. We suggest that, although rare, haemopericardium can be a feature of rheumatoid disease and may be associated with significant haemodynamic disturbance.


2-Methoxyestradiol, an endogenous estrogen metabolite, induces thyroid cell apoptosis.
The etiology of autoimmune thyroid diseases is unclear; however, the extreme female predominance suggests that sex hormones may have a pathogenic role. 2-Methoxyestradiol (2-ME) is present in the serum of women during the ovulatory and luteal phases of the menstrual cycle, and during pregnancy. We investigated the actions of 2-ME and estrogen on thyroid follicular cells. 2-ME induced dramatic changes in cell morphology and decreased the viability of the cells, as well as disrupted the structural integrity of cultured thyroid follicles. Flow cytometric analysis showed that 2-ME halted cell proliferation by arresting the cells in the G2/M cell-cycle compartment. Prolonged exposure to 2-ME led to apoptosis and to increased release of the autoantigen thyroid peroxidase (TPO). 17beta-estradiol failed to produce a similar effect even in 40-fold molar excess to 2-ME. Co-treatment with estrogen receptor antagonists did not alter the 2-ME effect, indicating that 2-ME was not operating through a classic nuclear estrogen receptor. In conclusion, this study indicates that 2-ME induces G2/M cycle arrest, apoptosis and the disruption of thyroid follicles. This process results in the release of thyroid antigens that may play a role in high incidence of thyroid autoantibodies and autoimmune thyroid disease in women.


Tension Pneumocephalus from Positive Pressure Ventilation Following Endoscopic Skull Base Surgery: Case Series and an Institutional Protocol for the Management of Postoperative Respiratory Distress.
Tension pneumocephalus (TP) is a rare but feared complication of endoscopic endonasal skull base surgery. In contrast to simple pneumocephalus, which is common after endoscopic transnasal approaches and managed conservatively, TP represents a neurosurgical emergency and mandates urgent decompression. Here we present 2 cases of TP as a consequence of positive pressure ventilation following endoscopic endonasal skull base surgery. Both occurred during resuscitation for postoperative hypoxia. These cases prompted the development of an institution-wide protocol to identify and manage patients at risk of TP after extended skull base approaches. To our knowledge, these are the only such cases of postoperative TP following positive pressure ventilation in the literature.


Biogenic ferrihydrite-humin coprecipitate as an electron donor for the enhancement of microbial denitrification by Pseudomonas stutzeri.
Nitrate pollution of groundwater has become an increasingly serious environmental problem that poses a great threat to aquatic ecosystems and to human health. Previous studies have shown that solid-phase humin (HM) can act as an additional electron donor to support microbial denitrification in the bioremediation of nitrate-contaminated groundwater where electron donor is deficient. However, the electron-donating capacities of HMs vary widely. In this study, we introduced ferrihydrite and prepared ferrihydrite-humin (Fh-HM) coprecipitates via biotic means to strengthen their electron-donating capacities. The spectroscopic results showed that the crystal phase of Fh did not change after coprecipitation with HM in the presence of Shewanella oneidensis MR-1, and iron may have complexed with the organic groups of HM. The Fh-HM coprecipitate prepared with an optimal initial Fh-HM mass ratio of 14:1 enhanced the microbial denitrification of Pseudomonas stutzeri with an electron-donating capacity 2.4-fold higher than that of HM alone, and the enhancement was not caused by greater bacterial growth. The alginate bead embedding assay indicated that the oxidation pathway of Fh-HM coprecipitate was mainly through direct contact between P. stutzeri and the coprecipitate. Further analyses suggested that quinone and organic-complexed Fe were the main electron-donating fractions of the coprecipitate. The results of the column experiments demonstrated that the column filled with Fh-HM-coated quartz sand exhibited a higher denitrification rate than the one filled with quartz sand, indicating its potential for practical applications.


[Infant mortality and socioeconomic indicators in Chile: 1985-1999].
The link between economy and some public health indicators like Infant Mortality Rate (IMR) is widely demonstrated. To explore the relationship between some economic indicators and infant mortality in Chile between 1985 and 1999. An ecological study, obtaining information from Demographic Annual Reports edited by the National Institute of Statistics, Health Ministry, Planning Ministry and the Central Bank. There is an inverse association between the Gross National Product, per capita and Real Salary Index and a direct relationship between inflation and unemployment rates with an increased infant mortality. The reduction in infant mortality observed in Chile depends more on socio demographic than on economical factors.


Innate recognition of non-self nucleic acids.
The immune system has evolved a plethora of innate receptors that detect microbial DNA and RNA, including Toll-like receptors in the endosomal compartment and RIG-I-like receptors and Nod-like receptors in the cytosol. Here we discuss the recognition of and responses to non-self nucleic acids via these receptors as well as their involvement in autoimmune diseases.


Localized amyloid of the ureter--case report and review of the literature.
An unusual case of localized amyloidosis of the ureter is described and a review of the 12 previously reported cases is presented. No case had been diagnosed prior to surgery. Loin pain, with or without haematuria, associated with a sterile pyuria and an abnormal pyelogram were the commonest clinical signs. In the case reported an incidental small renal cell carcinoma was also present and the association of amyloidosis and neoplasia is discussed.


Placental transfer and fetal effects of maternal sodium beta-hydroxybutyrate infusion in the baboon.
We examined the effects of maternal sodium beta-hydroxybutyrate (NaBOHB) on the primate fetus to investigate the impact of ketosis not associated with acidosis on fetal metabolism. After a loading dose (600 mg/kg), NaBOHB was infused for 70 min (300 mg/kg.hr) into the maternal femoral vein of eight pregnant baboons, and placental transfer and fetal and maternal metabolic changes were observed during an acute experimental protocol. Maternal arterial levels rose from 0.70 +/- 0.21 to 5.42 +/- 0.93 mM (p less than 0.001), and fetal arterial levels from 0.34 +/- 0.09 to 2.76 +/- 0.64 mM (p less than 0.01). A maternal-fetal gradient of approximately 2:1 was observed in both baseline and steady-state infusion conditions and is similar to the human maternal-fetal ketone gradient. This is in contrast to the sheep where significantly higher gradients have been described. The elevated lactate, from 1.90 +/- 0.34 to 2.88 +/- 0.54 mM (p less than 0.05) and somewhat decreased pO2 values in the fetus from 54.8 +/- 8.9 to 45.0 +/- 3.8 mm Hg (p greater than 0.05 less than 0.1), without change in oxygen consumption (2.00 +/- 0.28 versus 1.73 +/- 0.15 mM/min) are features common to conditions of increased levels of fetal energy substrate. NaBOHB does not appear to contribute to oxidative energy metabolism of the whole fetus but may contribute to lipid stores. The significance of higher levels of BOHB in the primate fetus compared to the sheep fetus remains to be elucidated.


Lateral lymph node dissection reduces local recurrence of locally advanced lower rectal cancer in the absence of preoperative neoadjuvant chemoradiotherapy: a systematic review and meta-analysis.
The impact of lateral lymph node dissection (LLND) in locally advanced lower rectal cancer remains controversial. This study is to compare total mesorectal excision (TME) with or without LLND in lower rectal cancer cases of stage II/III. The electronic databases were systematically searched that compared TME with or without LLND among patients with lower rectal cancer in clinical stage II/III. Subgroup analysis was performed considering neoadjuvant chemoradiotherapy (nCRT). The hazard ratios (HR), relative risk (RR), and weighted mean difference (WMD) were pooled. Twelve studies of 4458 patients of this meta-analysis demonstrate, LLND alone significantly reduced the local recurrence rate of patients who did not receive nCRT (RR 0.71, P = 0.004), while the difference was not significant when combined with nCRT (RR 0.70, P = 0.36). The analysis shows TME with LLND was associated with significantly longer operation time (WMD 90.73 min, P < 0.001), more intraoperative blood loss (WMD 303.20 mL, P < 0.001), and postoperative complications (RR = 1.35, P =0.02). Whereas urinary dysfunction (RR 1.44, P = 0.38), sexual dysfunction (RR 1.41, P = 0.17), and postoperative mortality (RR = 1.52, P = 0.70), were similar between these two groups. Statistically, no significant differences were observed in OS (HR 0.93, P = 0.62), DFS (HR 0.99, P = 0.96), total recurrence (RR 0.98, P = 0.83), lateral recurrence (RR 0.49, P = 0.14), or distal recurrence (RR 0.95, P = 0.78) between these two groups regardless of whether nCRT was performed or not. The study shows LLND alone decreases the local recurrence without using nCRT irrespective of the survival advantage in locally advanced lower rectal cancer. The benefit of controlling local recurrence by LLND alone makes us reconsider the usage of nCRT with LLND. The protocol for this meta-analysis was registered prospectively with PROSPERO ( CRD42020135575 ) on 16 May 2019.


FNCaMP, ratiometric green calcium indicator based on mNeonGreen protein.
Neurobiologists widely use green genetically encoded calcium indicators (GECIs) for visualization of neuronal activity. Among them, ratiometric GECIs allow imaging of both active and non-active neuronal populations. However, they are not popular, since their properties are inferior to intensiometric GCaMP series of GECIs. The most characterized and developed ratiometric green GECI is FGCaMP7. However, the dynamic range and sensitivity of its large Stock's shift green (LSS-Green) form is significantly lower than its Green form and its molecular design is not optimal. To address these drawbacks, we engineered a ratiometric green calcium indicator, called FNCaMP, which is based on bright mNeonGreen protein and calmodulin from A. niger and has optimal NTnC-like design. We compared the properties of the FNCaMP and FGCaMP7 indicators in vitro, in mammalian cells, and in neuronal cultures. Finally, we obtained and analyzed X-ray structure of the FNCaMP indicator.


Ventricular remodeling. Mechanisms and prevention.
The changes that take place in the structure and form of the left ventricle have become closely related to increase in clinical events in patients with heart failure. These changes are included in the term remodeling and occur as a response to the initial loss of left ventricular muscle mass. The mechanisms that initiate and maintain this remodeling process have become of great importance since a variety of effective pharmacologic agents have become available. A more complete understanding of remodeling has the potential of preventing the progression of heart failure. This article considers the various aspects of left ventricular remodeling including the cardiomyocytes, the myocardial interstitium, the importance that apoptosis plays the remodeling process.


The Problem of Vascularization of Precancerous Lesions of the Colon and Development of Hypoxia at the Incipient Stages of Blastomogenesis.
The features of the blood supply in the colon mucosa during the development of hyperplastic and dysplastic processes, and early neoplastic lesions were studied. The study cohort comprised the material of 92 patients of the N. N. Blokhin National Medical Research Center of Oncology collected during 2002-2016. The initial stages of hyperplastic, atypical processes developed against the background of enhanced angiogenesis. During the development of adenomatosis with concomitant dysplasia, the blood supply of newly formed structures decreased, which was associated with overexpression of markers of tissue hypoxia Hif1, CAIX, and Glut1. The expression of these markers was maximal in the foci of highly pronounced dysplasia, but was not detected in the foci of carcinoma in situ and microinvasion, when the cells reach the stroma with abundant vascular supply. It can be hypothesized that hypoxia forces the cells to search the territories with sufficient vascularization and migrate to other structures, which finally manifests in invasion and destruction of adjacent tissues and dissemination of these cells in the body (formation of metastasis).


Randomization with a posteriori constraints: description and properties.
The use of randomization for assigning patients to treatment groups in clinical trials is firmly acknowledged as providing the best quality results. Two standard methods are used in order to achieve well-balanced groups with respect to prognostic factors (i.e. factors influencing the disease outcome): stratification and minimization. Stratification is recommended when the number of strata is not too high--otherwise, minimization is preferred. However, minimization may compromise blinding (since the search for balance is performed a priori) and, furthermore, use of the technique has been questioned by the European Agency for the Evaluation of Medicinal Products. We have developed a new procedure for adaptive randomization, which we have named 'randomization with a posteriori constraints'. By using a search for balance a posteriori, this procedure ensures that patient groups are similar with respect to prognostic factors while being less vulnerable to selection bias. The aim of this work was to describe the new method and to compare it (using simulations) with stratification and minimization. In the case of trials with few prognostic factors, the recourse to minimization or 'randomization with a posteriori constraints' does not appear to be useful. In such a context, stratification has suitable properties and its simplicity of implementation encourages its use. However, when the number of prognostic factors is higher, 'randomization with a posteriori constraints' is less predictable than minimization and the chance of imbalance is lower than for stratification. In conclusion, 'randomization with a posteriori constraints' with an adequate threshold seems to be a good compromise between minimization and stratification.


Assessment of the functional affinity constant of monoclonal antibodies using an improved enzyme-linked immunosorbent assay.
The use of an enzyme-linked immunosorbent assay (ELISA) for the determination of affinity constants implies heterogeneous measurements. Therefore, despite their simplicity, direct solid-phase binding assays are not common. Many investigators have serious, and mostly justified, reservations about the application of solid-phase affinity methods. They refer to problems such as diffusion effects and difficulties in reaching equilibrium due to heterogeneous binding and co-operativity. Accordingly, functional affinity determinations are often described as meaningless. These objections apply to the measurement of the affinity of a monoclonal antibody using the enzyme-linked immunosorbent assay of Beatty et al. (J. Immunol. Methods (1987) 100, 173), which is based on the effect of antibody affinity on the sigmoidal dose response curve. The affinity constant is calculated by mathematical equations, based on the Law of Mass Action and the authors made a number of important assumptions--avoiding the above mentioned problems--in order to justify the use of the Law of Mass Action. By carefully examining these assumptions we have developed an improved ELISA procedure for functional affinity determinations on the basis of a primary coating with the antigen only. the coating conditions were validated by employing gold labelled colloidal particles and physical counting of the bound particles under the scanning electron microscope. Since monovalent binding between human chorionic gonadotropin and its monoclonal antibody could be achieved under equilibrium conditions, the application of the Law of Mass Action and hence of the Beatty formula became possible. We conclude that under these conditions functional affinity determinations are appropriate.


Age-dependent decline in cortisol levels and clinical manifestations in patients with ACTH-independent Cushing's syndrome.
Adrenocortical cells change with age in both subcellular morphology and level of steroidogenesis. This study evaluates the influence of aging on hypercortisolism and clinical manifestations in patients with ACTH-independent Cushing's syndrome due to a cortisol-secreting adrenal adenoma (CS). Restrospective study. Thirty-six (33 females and 3 males) with CS. 31 healthy controls were age- and sex-matched to CS patients. Patient age at diagnosis was compared to the degree of hypercortisolism and clinical manifestations. The degree of hypercortisolism was estimated using serum cortisol levels and urinary free cortisol excretion. Positive clinical manifestations were quantified using 12 symptoms and signs which resulted in a clinical score for each patient. Endogenous creatinine clearance, estimated disease duration and gender were analysed as possible factors affecting adenoma-secreated cortisol. Patient age correlated negatively with serum cortisol level (r = -0.417, P = 0.0107, n = 36) and urinary excretion of free cortisol (r = -0.613, P = 0.0002, n = 31). Analysis of Spearman's rank correlation showed that the clinical score decreased with age (rho = -0.631, P = 0.0004, n = 33). Oedema, weakness/myopathy, hirsutism, striae and psychological changes were infrequent symptoms in the elderly. Using stepwise multiple regression analysis, we estimated the clinical score (Y) by factors of serum cortisol levels (X1:nmol/l), age (X2: years of age) and gender (X3: gender; female = 1, male = 0), i.e. Y = 4.771 + 0.004 X1 - 0.064 X2 + 2.548 X3 (r = 0.743, P < 0.0001, n = 33). Results suggested that aging both suppressed cortisol secretion and repressed manifestations directly as an independent factor, although its direct effect may be weak. Our findings indicated the importance of age as a factor modulating hormonal levels and possible clinical manifestations in patients with cortisol-secreting adenomas.


In vitro and in vivo studies on the antimicrobial effect of lactoferrin against Escherichia coli O157:H7.
Escherichia coli O157:H7 shed by clinically healthy ruminants has been linked to hemorrhagic colitis and the hemolytic uremic syndrome in humans. The bacteria are spread mainly by contaminated food and water, contact with animals carrying the organisms, and person-to-person contact. Although many intervention strategies have been studied to reduce E. coli O157:H7 carriage in ruminants and its spread into the environment, none of the available methods can completely eliminate the infection. Therefore, there is need for new intervention strategies which will effectively reduce E. coli O157:H7 prevalence. Lactoferrin, a member of the transferrin protein family, is an iron-binding glycoprotein that is found in many exocrine secretions, including milk, tears, saliva, and serum. Lactoferrin has a number of biological functions including antimicrobial and immunomodulatory effects. This review summarizes latest data on the antimicrobial effect of lactoferrin against E. coli O157:H7 in in vitro and in vivo studies.


Short-Segment Type B Interrupted Aortic Arch Presenting With Subarachnoid Hemorrhage With Subsequent Primary Percutaneous Repair.
This report describes a young adult man presenting with subarachnoid hemorrhage secondary to an intracranial aneurysm who was found to have a short-segment type B interrupted aortic arch. We describe the clinical presentation, evaluation, and management of this patient and highlight imaging findings and percutaneous repair of the aneurysm and interrupted aortic arch. (Level of Difficulty: Intermediate.).


Osteoporosis is associated with increased marrow fat content and decreased marrow fat unsaturation: a proton MR spectroscopy study.
To use proton magnetic resonance spectroscopy ((1)H-MRS) to evaluate vertebral marrow fat, and to determine whether bone density correlates with fat content and fat unsaturation levels in postmenopausal women. Fifty-three women (mean age = 70 years) underwent dual energy x-ray absorptiometry and (1)H-MRS, and 12 young female controls (mean age = 28 years) underwent (1)H-MRS of the lumber spine. Water and lipid peak amplitudes were measured to calculate fat content and fat unsaturation index. Spearman's correlation tests and a t-test comparison of means were applied. (1)H-MRS was successful in 15 normal, 15 osteopenic, and 20 osteoporotic subjects, and in all controls. Marrow fat content was significantly elevated in osteoporotic (65.5% +/- 10%) and osteopenic (63.5% +/- 9.3%) subjects compared to normal subjects (56.3% +/- 11.2%) and young controls (29% +/- 9.6%). The fat unsaturation index was significantly decreased in osteoporotic (0.091 +/- 0.013) and osteopenic (0.097 +/- 0.014) subjects compared to normal subjects (0.114 +/- 0.016) and young controls (0.127 +/- 0.031). A good inverse correlation was observed between the fat content and the unsaturation index (r(s) = -0.53, P < 0.0001). Osteoporosis is associated with increased marrow fat. As marrow fat increases, saturated lipids appear to increase preferentially to unsaturated lipids.


Simultaneous effect of lead and cadmium on granulosa cells: a cellular model for ovarian toxicity.
Lead (Pb) and cadmium (Cd) are known reproductive toxicants, which accumulate in granulosa cells of the ovary. Female Charles foster rats were treated with sodium acetate (control), lead acetate and cadmium acetate either alone or in combination at a dose 0.05 mg/kg body weight intra-peritoneally for 15 days daily. Animals were killed at proestrous stage and granulosa cells were isolated from the ovaries. Binding of (125)I-luteinizing hormone ((125)I-LH), (125)I-follicle stimulating hormone ((125)I-FSH) and 17beta-hydroxysteroid dehydrogenase activity were measured. As these receptors are localized on the surface of the cell membrane, we also estimated the membrane parameters of these cells. Our results demonstrated that both lead and cadmium caused a significant reduction in gonadotropin binding, which altered steroidogenic enzyme activity of granulosa cells. These changes exhibited a positive correlation with membrane changes of the granulosa cells.


Certificate of need, volume, and percutaneous transluminal coronary angioplasty outcomes.
Florida seeks high hospital volumes for percutaneous transluminal coronary angioplasty (PTCA) by enforcing certificate of need (CON) laws, whereas California has no such laws. This study compares the volume-outcome relation for PTCA in Florida and California. The relation between the number of PTCA procedures performed at hospitals and the rate of inhospital bypass graft surgery and death for 292,457 patients in Florida and 390,880 patients in California between 1988 and 1998 was examined with descriptive statistics and logistic regressions. In 1988, the mean hospital PTCA volumes in Florida (237) and California (218) were not significantly different (P =.44). By 1998, Florida hospital volumes were significantly larger (724 vs 389, P <.001). Logistic regressions indicate that higher log (volume) was associated with lower mortality and urgent bypass grafting rates in both Florida and California during the sample period. Regression estimates indicate that a California hospital with the mean 1998 PTCA volume of 389 procedures had a predicted inpatient mortality rate of 1.4% and urgent bypass grafting rate of 2.2%. If the PTCA volume was raised to the 1998 Florida mean of 724 procedures, the inpatient mortality rate would not fall, although urgent bypass grafting rates were predicted to fall to 2.0%. Florida CON laws were associated with higher average PTCA volumes relative to California hospitals, where no such laws exist. Because a higher PTCA volume was associated with moderately better outcomes, CON may be marginally effective in improving outcomes for PTCA. Future studies should revisit this hypothesis with data from several states.


Simultaneous determination of gypenoside LVI, gypenoside XLVI, 2α-OH-protopanaxadiol and their two metabolites in rat plasma by LC-MS/MS and its application to pharmacokinetic studies.
Gypenoside LVI and gypenoside XLVI are the major bioactive dammarane saponins from Gynostemma pentaphyllum. Gypenoside LVI, gypenoside XLVI, and their metabolite 2α-OH-protopanaxadiol (2α-OH-PPD) possess potent non-small cell lung carcinoma A549 cell inhibitory activity. A sensitive liquid chromatography tandem mass spectrometry method was developed and validated to study the pharmacokinetics of gypenoside LVI and XLVI, 2α-OH-PPD, metabolite 1 (M1), and metabolite 2 (M2) after administration of gypenosides or 2α-OH-PPD. Plasma samples from rats were protein precipitated with methanol. Analytes were detected by triple quadrupole MS/MS with an electrospray ionization source in the positive multiple reaction monitoring mode. The transition m/z 441.4→109.2 was selected to quantify gypenoside LVI and XLVI, and 2α-OH-PPD, because of the extensive conversion of the gypenosides to aglycone in the ionization source. M1 and M2 are isomers that shared the transition m/z 493.4→143.1. To avoid interference, the baseline separation of each analyte was performed on a SunFire C18 column with a gradient of acetonitrile (0.1% formic acid, v/v) and water (0.1% formic acid, v/v). The chromatographic run time was 10min. The linearity was validated over a plasma concentration range from 2.00 to 2000ng/mL for M1 and M2, and from 10.0 to 2000 for gypenosides LVI and XLVI, and 2α-OH-protopanaxadiol. The lower limits of quantification were 10.0, 10.0, 10.0, 2.00, and 2.00ng/mL for gypenoside LVI, gypenoside XLVI, 2α-OH-PPD, M1, and M2, respectively, with acceptable intra-/inter-day precision and accuracy. The extraction recovery rates were >86.9% for each compound. No apparent matrix effect or instability was observed during each step of the bioanalysis. After full validation, this method was proved to be simple, fast, and efficient in analyzing large batches of plasma samples for the analytes.