Uricase-containing coacervate microdroplets as enzyme active membrane-free protocells for detoxification of uric acid in serum.
Based on the unique property of preferential sequestration of guest molecules, coacervate microdroplets are proposed as enzyme active membrane-free protocells, in which uricase is loaded for efficient detoxification of uric acid in serum.


Resolution of four large chromosomes in penicillin-producing filamentous fungi: the penicillin gene cluster is located on chromosome II (9.6 Mb) in Penicillium notatum and chromosome I (10.4 Mb) in Penicillium chrysogenum.
Four chromosomes were resolved by pulsed field gel electrophoresis in Penicillium notatum (10.8, 9.6, 6.3 and 5.4 Mb in size) and in five different strains of Penicillium chrysogenum (10.4, 9.6, 7.3 and 6.8 Mb in the wild type). Small differences in size were found between the four chromosomes of the five P. chrysogenum strains. The penicillin gene cluster was localized by hybridization with a pcbAB probe to chromosome II of P. notatum and to chromosome I of all P. chrysogenum strains except the deletion mutant P. chrysogenum npe10, which lacks this DNA region. The pyrG gene was localized to chromosome I in P. notatum and to chromosome II in all P. chrysogenum strains except in the mutant AS-P-78 where the probe hybridized to chromosome III. A major chromosomal rearrangement seems to have occurred in this high penicillin producing strain. A fast moving DNA band observed in all gels corresponds to mitochondrial DNA. The total genome size has been calculated as 32.1 Mb in P. notatum and 34.1 Mb for the P. chrysogenum strains.


Development and characterization of a glycine biosensor system for fine-tuned metabolic regulation in Escherichia coli.
In vivo biosensors have a wide range of applications, ranging from the detection of metabolites to the regulation of metabolic networks, providing versatile tools for synthetic biology and metabolic engineering. However, in view of the vast array of metabolite molecules, the existing number and performance of biosensors is far from sufficient, limiting their potential applications in metabolic engineering. Therefore, we developed the synthetic glycine-ON and -OFF riboswitches for metabolic regulation and directed evolution of enzyme in Escherichia coli. The results showed that a synthetic glycine-OFF riboswitch (glyOFF6) and an increased-detection-range synthetic glycine-ON riboswitch (glyON14) were successfully screened from a library based on the Bacillus subtilis glycine riboswitch using fluorescence-activated cell sorting (FACS) and tetA-based dual genetic selection. The two synthetic glycine riboswitches were successfully used in tunable regulation of lactate synthesis, dynamic regulation of serine synthesis and directed evolution of alanine-glyoxylate aminotransferase in Escherichia coli, respectively. Mutants AGXT22 and AGXT26 of alanine-glyoxylate aminotransferase with an increase of 58% and 73% enzyme activity were obtained by using a high-throughput screening platform based on the synthetic glycine-OFF riboswitch, and successfully used to increase the 5-aminolevulinic acid yield of engineered Escherichia coli. A synthetic glycine-OFF riboswitch and an increased-detection-range synthetic glycine-ON riboswitch were successfully designed and screened. The developed riboswitches showed broad application in tunable regulation, dynamic regulation and directed evolution of enzyme in E. coli.


Quality of life after stem cell transplantation: a patient, partner and physician perspective.
Background: Although increasing data are available on quality of life (QOL) issues after stem cell transplantation (SCT), many issues are still unresolved. With the increasing use of transplants in non-curative situations, quality of life issues have become especially important. Methods: All patients who had undergone SCT and had no recurrent disease were asked to provide quality of life data using the EORTC QLQ-C30 questionnaire at about 2.5 (n=52) and 4.5 years (n=33) after transplantation. In addition, the patients' partners and physicians were asked to estimate the patients' quality of life. Results: Scores on the functional scales ranged from 65 to 80 and were comparable initially and at the 23-month follow-up. The initial, overall quality of life of the patients was 75. After an additional 23-month follow-up, the overall quality of life was 73. Fatigue, sleeping disturbances, and financial problems scored relatively high. Generally, in contrast to partners, physicians underestimated symptoms and overestimated functions and quality of life significantly. Conclusions: We conclude that SCT has a long-term impact on such quality of life issues as fatigue, sleep disturbances, dyspnea, and financial problems. Physicians tend to overestimate quality of life in their patients. With the increasing numbers of patients undergoing transplantation, more attention should be focussed on these issues.


Short stature in the "hyperimmunoglobulin E syndrome".
A boy presenting the hyperimmunoglobulin E syndrome and short stature is described. Extensive endocrine investigations could not reveal any disturbance. It is suggested that a very active immunological process might cause growth retardation in this syndrome.


Involvement of upper torso stress amplification, tissue compression and distortion in the pathogenesis of keloids.
Keloids are benign tumours composed of fibrous tissue produced during excessive tissue repair triggered by minor injury, trauma or surgical incision. Although it is recognized that keloids have a propensity to form in the upper torso of the body, the predisposing factors responsible for this have not been investigated. It is crucial that the aetiopathoical factors implicated in keloid formation be established to provide guidelines for well-informed more successful treatment. We compared keloid-prone and keloid-protected skin, identified pertinent morphological differences and explored how inherent structural characteristics and intrinsic factors may promote keloid formation. It was determined that keloid prone areas were covered with high tension skin that had low stretch and a low elastic modulus when compared with skin in keloid protected areas where the skin was lax with a high elastic modulus and low pre-stress level. Factors contributing to elevated internal stress in keloid susceptible skin were the protrusion of hard connective tissue such as bony prominences or cartilage into the dermis of skin as well as inherent skin characteristics such as the bundled arrangement of collagen in the reticular dermis, the existent high tension, the low elastic modulus, low stretch ability, contractile forces exerted by wound healing fibroblastic cells and external forces. Stress promotes keloid formation by causing dermal distortion and compression which subsequently stimulate proliferation and enhanced protein synthesis in wound healing fibroblastic cells. The strain caused by stress also compresses and occludes microvessels causing ischaemic effects and reperfusion injury which stimulate growth when blood rich in growth factors returns to the tissue. The growth promoting effects of increased internal stress, primarily, and growth factors released by reperfusing blood, manifest in keloid formation. Other inherent skin characteristics promoting keloid growth during the late stages of wound healing in the upper torso are the thinner epidermis, the presence of vellus hairs, the absence of protective immunoglobulin A (IgA), and the thick fragile quality of upper torso skin. As it is not known why there is a predilection for keloids to form in the upper torso of the body, this hypothesis implicating and associating inherent morphological characteristics and elevated stress in the aetiopathogenesis of keloids is of potential value in terms of prevention, management and treatment of these enigmatic tumours.


Pharmacokinetics-based optimal dose-exploration of mycophenolate mofetil in allogeneic hematopoietic stem cell transplantation.
For better clinical outcomes of mycophenolate mofetil (MMF) in allogeneic hematopoietic stem cell transplantation (alloSCT), higher mycophenolic acid (MPA) plasma levels are proposed to be desirable. Here, we investigate the optimal MMF dosing strategy based on pharmacokinetic studies in 20 Japanese alloSCT patients. The first 11 patients received MMF twice daily at an escalated dose from 15 mg/kg, according to real-time pharmacokinetic monitoring of the total MPA area under the curve (AUC). In the subsequent nine patients, MMF was given at a fixed dose of 1,000 mg three-times daily. The pharmacokinetic data revealed that the dose escalation in each individual did not always increase the AUC. In contrast, the increase of dosing frequency could statistically keep higher MPA plasma levels, as reflected in higher concentration at steady state (C (ss)) or trough value (C (trough)). There was no symptomatic adverse event in both groups. These results suggest that MMF administration of every 8 h after alloSCT would be better to maintain higher MPA plasma levels than that of every 12 h even in the same daily dose. Further studies are necessary to confirm the clinical benefit of MMF to prevent graft failure, as well as severe aGVHD.


Hepatic encephalopathy.
Hepatic encephalopathy (HE) represents a continuum of transient and reversible neurologic and psychiatric dysfunction. It is a reversible state of impaired cognitive function or altered consciousness in patients with liver disease or portosystemic shunting. Over the last several years, high-quality studies have been conducted on various pharmacologic therapies for HE; as more data emerge, it is hoped that HE will become a more easily treated complication of decompensated liver disease. In the interim, it is important that physicians continue to screen for minimal HE and treat patients early in addition to continuing to provide current treatments of overt HE.


Biocatalytic role of potato starch synthase III for α-glucan biosynthesis in Synechocystis sp. PCC6803 mutants.
A potato starch synthase III (PSSIII) was expressed in the Synechocystis mutants deficient in either glycogen synthase I (M1) or II (M2) to replenish α-(1,4) linkage synthesizing activity, resulting in new mutants, PM1 and PM2, respectively. These mutants were applied to study the role of exogenous plant starch synthase for starch/glycogen biosynthesis mechanism established in the cyanobacteria. The remaining glycogen synthase genes in PM1 and PM2 were further disrupted to make the mutants PM12 and PM21 which contained PSSIII as the sole glycogen/starch synthase. Among wild type and mutants, there were no significant differences in the amount of α-glucan produced. All the mutants harboring active PSSIII produced α-glucans with relatively much shorter and less longer α-1,4 chains than wild-type glycogen, which was exactly in accordance with the increase in glycogen branching enzyme activity. In fact, α-glucan structure of PM1 was very similar to those of PM12 and PM21, and PM2 had more intermediate chains than M2. This result suggests PSSIII may have distributive elongation property during α-glucan synthesis. In conclusion, the Synechocystis as an expression model system of plant enzymes can be applied to determine the role of starch synthesizing enzymes and their association during α-glucan synthesis.


Prostaglandin production by guinea-pig placenta and other uterine tissues during mid-pregnancy.
Prostaglandin (PG) output from cultured placenta, sub-placenta, endometrium and fetal membranes of guinea-pigs was measured on days 22, 29 and 36 of pregnancy to establish the source of increased PGF2alpha production during mid-pregnancy. PGF2alpha and 6-keto-PGF1alpha were produced in larger quantities than PGE2 by the placenta, sub-placenta and endometrium; 6-keto-PGF1alpha was in the major prostaglandin produced by the fetal membranes. The initial outputs of PGF2alpha, PGE2 and 6-keto-PGF1alpha from the sub-placenta, fetal membranes and endometrium either decreased or remained fairly constant between days 22 and 36. In contrast, the initial outputs of PGF2alpha, PGE2 and 6-keto-PGF1alpha from the placenta increased 14.7-, 2.5- and 2.0-fold, respectively, between days 22 and 36, indicating that the placenta is the tissue responsible for the increase in PGF2alpha output from the mid-pregnant guinea-pig uterus. Aristolochic acid (a phospholipase A2 inhibitor) inhibited prostaglandin output from the endometrium, but had a more variable effect in prostaglandin output from the other tissues. Thimerosal (an arachidonic acid uptake inhibitor) inhibited PGF2alpha and PGE2 outputs from the endometrium, but generally potentiated 6-keto-PGF1alpha output and prostaglandin output from the other tissues. Arachidonic acid release for prostaglandin synthesis in the endometrium, but not the placenta, sub-placental or fetal membranes, is apparently dependent upon a constant level of phospholipase A2 activity.


Pain, muscle spasms and twitching fingers following brachial plexus avulsion. Report of three cases relieved by dorsal root entry zone coagulation.
Three patients who, following partial brachial plexus avulsion, experienced pain, involuntary finger twitching and muscular spasms are reported. Two exhibited cutaneous trigger zones, stimulation of which exacerbated their pain; changes in emotional tone aggravated both the pain and the spasms. Pain would appear to be due not only to deafferentation and scarring of the dorsal horn, but also to an afferent pathological barrage from partially damaged dorsal roots. The pathological sensory barrage may activate metameric interneuronal circuits and produce involuntary movements. Exacerbation of both the pain and the spasms can be explained on the basis of intrinsic properties of these ectopic pacemakers.


Corrigendum to "Satisfaction with use of telehealth during COVID-19: An integrative review" [International Journal of Nursing Studies Advanced 2 (2020). Article number 100008].
[This corrects the article DOI: 10.1016/j.ijnsa.2020.100008.].


Sympsium on hearing loss--the otolaryngologist's responsibility. Habilitation and rehabilitation of patients with sensorineural hearing losses.
A philosophy of the (re)habilitation of patients with sensorineural hearing losses from a communication point of view is presented. Pediatric and adult populations are discussed separately. Topics of consideration for pediatric patients include the appropriate use of amplification, team management of the hearing handicap, educational placement and the importance of parent counseling. The discussion of the management of adult patients reviews some problems in hearing aid fitting, patient counseling and appropriate case follow-up. The guiding principle behind these (re)habilitation processes is the integration of the hearing-impaired patient into a hearing world.


Advances in developing small molecule SARS 3CLpro inhibitors as potential remedy for corona virus infection.
Originated in China, coronavirus disease 2019 (COVID-19)- the highly contagious and fatal respiratory disease caused by SARS-CoV-2 has already infected more than 29 million people worldwide with a mortality rate of 3.15% (according to World Health Organization's (WHO's) report, September 2020) and the number is exponentially increasing with no remedy whatsoever discovered till date. But it is not the first time this infectious viral disease has appeared, in 2002 SARS-CoV infected more than 8000 individuals of which 9.6% patients died and in 2012 approximately 35% of MERS-CoV infected patients have died. Literature reports indicate that a chymotripsin-like cystein protease (3CLpro) is responsible for the replication of the virus inside the host cell. Therefore, design and synthesis of 3CLpro inhibitor molecules play a great impact in drug development against this COVID-19 pandemic. In this review, we are discussing the anti-SARS effect of some small molecule 3CLpro inhibitors with their various binding modes of interactions to the target protein.


Online training of Covid-19 infection prevention and control for healthcare workers in psychiatric institutes.
This study aimed to develop a unique online infection prevention and control (IPC) training on Covid-19 for healthcare workers in psychiatric institutes in Japan and to examine its efficacy based on its impact on the knowledge, attitude, and confidence about IPC for Covid-19 among the healthcare workers. This quasi-experimental study was conducted using online training on Covid-19 IPC for healthcare workers in various psychiatric institutes from April 2021 to March 2022. An online training video on Covid-19 IPC was developed. Voluntary healthcare workers in psychiatric institutes located in five prefectures in Japan were recruited to participate in this training. The participants then completed 30 min of online training and surveys about knowledge, attitude, and confidence were conducted pre, post, and three months after the training. The video training and surveys were contextually validated by the experts, but not by any previous study. A total of 224 participants were included, of which 108 (54.0%) were men. The mean (standard deviation (SD)) age and the mean occupational experience were 47.4 (9.5) and 18.0 (12.6) years, respectively. Among the participants, 190 (84.8%) completed the post-training, and 131 (58.5%) completed the three-month-later training surveys. The total score on the quizzes in the post-training (+ 31.1%, SD 15.7, p-value < 0.01) and three-month-later training (+ 14.9%, SD 16.8, p-value < 0.01) surveys had significantly increased from that in the pre-training survey. In contrast, the total score in the three-month-later training had significantly decreased from that in the post-training survey (-16.1%, SD 16.7, p-value < 0.01). Thirty minutes of online training about IPC for Covid-19 had improved knowledge, confidence, and attitude among psychiatric healthcare workers. Regular online training would help in preventing the transmission or formation of clusters of Covid-19 in psychiatric healthcare institutes.


Assessment of bilateral filter on 1/2-dose chest-pelvis CT views.
A bilateral filter (BF) is a non-linear filter that has been proved to de-noise images without overrunning edges. Multi-slice computerized tomography (CT) may employ a BF to participate in dose reduction. This paper quantifies the role of the BF in achieving this objective on 1/2-dose CT. Two sets of CT images are acquired for the chest-pelvis at two different radiation doses. The BF was applied on the 1/2-dose CT images by use of various window sizes. Each time, a set of values of the BF range was fixed while the BF domain was modified. The goal was to observe the behavior of the BF on 1/2-dose CT images in comparison with full-dose CT images. The comparison was carried out by use of four co-occurrence matrix descriptors. Additionally, the peak signal-to-noise ratio (PSNR) and the mean square error (MSE) were reported. The study was applied to the sagittal, coronal, and axial CT views. The results showed that the impact of applying a BF varies among different CT views. The BF can retrieve only part of the signal being lost due to reduction of the radiation dose by one half. Yet, the BF improves the appearance of the 1/2-dose chest-pelvis CT examination. Thus, the BF can contribute to a 50% dose reduction. A procedure for employing the BF on CT machines is proposed. The results also showed that texture descriptors are similar to the PSNR and MSE in providing quantities for assessing medical image quality.


Brown Adipose Tissue Thermogenic Capacity Is Regulated by Elovl6.
Although many transcriptional pathways regulating BAT have been identified, the role of lipid biosynthetic enzymes in thermogenesis has been less investigated. Whereas cold exposure causes changes in the fatty acid composition of BAT, the functional consequences of this remains relatively unexplored. In this study, we demonstrate that the enzyme Elongation of Very Long Chain fatty acids 6 (Elovl6) is necessary for the thermogenic action of BAT. Elovl6 is responsible for converting C16 non-essential fatty acids into C18 species. Loss of Elovl6 does not modulate traditional BAT markers; instead, it causes reduced expression of mitochondrial electron transport chain components and lower BAT thermogenic capacity. The reduction in BAT activity appears to be counteracted by increased beiging of scWAT. When beige fat is disabled by thermoneutrality or aging, Elovl6 KO mice gain weight and have increased scWAT mass and impaired carbohydrate metabolism. Overall, our study suggests fatty acid chain length is important for BAT function.


A long distance RT-PCR able to amplify the Pestivirus genome.
A method to amplify long genomic regions (up to approximately 12.3 kb) from pestiviruses in one RT-PCR is described. The difficulty in designing conserved Pestivirus primers for the amplification of genomes from highly divergent isolates simply by means of overlapping segments is demonstrated using new bioinformatic tools. An alternative procedure consisting of optimizing the length of the genomic cDNA fragments and their subsequent amplification by polymerase chain reaction (PCR) using a limited set of specific primers is described. The amplification of long DNA fragments from a variety of sources, including genomic, mitochondrial, and viral DNAs as well as cDNA produced by reverse transcription (RT) has been achieved using this methodology, known as long distance PCR. In the case of viruses, it is necessary to obtain viral particles from infected cells prior to RT procedures. This work provides improvements in four steps of long distance RT-PCR (L-RT-PCR): (i) preparation of a viral stock, (ii) preparation of template RNA, (iii) reverse transcription and (iv) amplification of the cDNA by LD-PCR. The usefulness of L-RT-PCR is discussed in the light of current knowledge on pestivirus diversity. The genomic sequence of Singer_Arg reference strain obtained using this method is presented and characterized.


Inhibition of Pyk2 blocks lung inflammation and injury in a mouse model of acute lung injury.
Proline-rich tyrosine kinase 2 (Pyk2) is essential in neutrophil degranulation and chemotaxis in vitro. However, its effect on the process of lung inflammation and edema formation during LPS induced acute lung injury (ALI) remains unknown. The goal of the present study was to determine the effect of inhibiting Pyk2 on LPS-induced acute lung inflammation and injury in vivo. C57BL6 mice were given either 10 mg/kg LPS or saline intratracheally. Inhibition of Pyk2 was effected by intraperitoneal administration TAT-Pyk2-CT 1 h before challenge. Bronchoalveolar lavage analysis of cell counts, lung histology and protein concentration in BAL were analyzed at 18 h after LPS treatment. KC and MIP-2 concentrations in BAL were measured by a mouse cytokine multiplex kit. The static lung compliance was determined by pressure-volume curve using a computer-controlled small animal ventilator. The extravasated Evans blue concentration in lung homogenate was determined spectrophotometrically. Intratracheal instillation of LPS induced significant neutrophil infiltration into the lung interstitium and alveolar space, which was attenuated by pre-treatment with TAT-Pyk2-CT. TAT-Pyk2-CT pretreatment also attenuated 1) myeloperoxidase content in lung tissues, 2) vascular leakage as measured by Evans blue dye extravasation in the lungs and the increase in protein concentration in bronchoalveolar lavage, and 3) the decrease in lung compliance. In each paradigm, treatment with control protein TAT-GFP had no blocking effect. By contrast, production of neutrophil chemokines MIP-2 and keratinocyte-derived chemokine in the bronchoalveolar lavage was not reduced by TAT-Pyk2-CT. Western blot analysis confirmed that tyrosine phosphorylation of Pyk2 in LPS-challenged lungs was reduced to control levels by TAT-Pyk2-CT pretreatment. These results suggest that Pyk2 plays an important role in the development of acute lung injury in mice and that pharmacological inhibition of Pyk2 might provide a potential therapeutic strategy in the pretreatment for patients at imminent risk of developing acute lung injury.


The effect of endothelin 1 on the retinal microvascular pericyte.
The effect of the highly vasoactive peptide endothelin 1 (ET1) was tested on bovine retinal microvascular pericytes propagated in vitro. Specific binding of 125I-ET1 to retinal pericytes was documented by autoradiography. ET1 caused contraction of pericytes at a concentration of 0.1 nM which was accompanied by increases in inositol phosphates. Exposure of pericytes to 10 nM ET1 resulted in the aggregation and realignment of muscle-specific actins into bundles which were oriented parallel to the long axis of the cell, and ET1 was also mitogenic to pericytes in the presence of low levels of fetal calf serum. These observations suggest that ET1 may play an important role in endothelial cell-pericyte interactions within the microvasculature of the retina and that it may be involved in the autoregulation of retinal blood flow.


Aidi injection altered the activity of CYP2D4, CYP1A2, CYP2C19, CYP3A2, CYP2E1 and CYP2C11 in normal and diethylnitrosamine-induced hepatocellular carcinoma in rats.
Aidi injection (ADI), a traditional chinese medicine preparation, is widely used in combination with chemotherapy for the treatment of various malignant tumors, such as hepatocellular carcinoma (HCC). Studies have shown that changes in cytochrome P450 (CYP450) activity in disease states would affect the metabolism of drugs in vivo, especially liver diseases. However, the changes of Aidi injection on the activities of CYP2D4, CYP1A2, CYP2C19, CYP3A2, CYP2E1 and CYP2C11 in normal and HCC states are still unknown. The cocktail probe drugs method was used to investigate the effects of ADI on the activity of CYP2D4, CYP1A2, CYP2C19, CYP3A2, CYP2E1 and CYP2C11 in normal and HCC rats. The HCC rats was induced by diethylnitrosamine (DEN). Then, both normal and HCC rats were randomly divided into 2 groups (n = 6). They were given saline or ADI (10 mL/kg/d, i.p) for 2 weeks, respectively. On the fifteenth day, cocktail probe mixing solution, including metoprolol (10 mg/kg), caffeine (1.0 mg/kg), omeprazole (2.0 mg/kg), midazolam (2.0 mg/kg), chlorzoxazone (4.0 mg/kg) and tolbutamide (0.5 mg/kg), was injected into tail vein of all rats in each group. The blood sample was obtained at specified time. After the protein is precipitated, six probe drugs are analyzed by ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Compared with control group, the activity of CYP3A2 and CYP2E1 was significantly lower in the ADI group. Compared with the model group, the activities of CYP1A2, CYP3A2, CYP2E1, and CYP2C11 enzymes in the ADI model group were significantly reduced. Additionally, the activity of CYP2D4, CYP1A2, CYP2C19, CYP3A2, CYP2E1 and CYP2C11 enzymes in model group was significantly lower than control group. ADI can inhibit a lot of CYP450 enzyme, so it may reduce the dosage of chemotherapeutic drugs to reach the required plasma concentration of chemotherapeutic drugs, which is of great significance for the combination of anti-tumor chemotherapeutic drugs and is worthy of further in-depth study and clinical attention.


A TIR domain receptor-associated protein (TIRAP) variant SNP (rs8177374) confers protection against premature birth.
To investigate whether single nucleotide polymorphisms (SNPs) in genes encoding the Toll-like receptor (TLR) signaling pathway modulate susceptibility to preterm birth (PTB). Prospective case-control study examining the contribution of nine TLR SNPs to PTB (<37 weeks) and PTB <32 weeks. Genotyping was done on neonatal blood using a multiplexed single-base extension assay. Chi-square test, Fischer's exact test and classification trees were used for data analysis. Preterm infants (n=177) were more likely to be African American (P=0.02), and were more likely to be born to mothers who smoked (P=0.007), had pregnancy-induced hypertension (PIH; P=0.002) and placental abruption (P=0.0004) when compared with term infants (n=146). The TLR2, TLR4, TLR5, TLR9, nuclear factor-kappa B1 (NFκB1), NFκBIA and IRAK1 variants were not associated with PTB whereas the TIR domain receptor-associated protein (TIRAP) variant was more prevalent in term infants when compared with preterm infants born <32 weeks (P=0.004). PTB <32 weeks was more prevalent in infants without the TIRAP variant whose mothers had PIH and did not smoke (P=0.001). Presence of the TIRAP variant protected against PTB <32 weeks (P=0.015) in Caucasian infants. In our study, a TLR pathway adapter variant (TIRAP (rs8177374)) protected against PTB<32 weeks, supporting our hypothesis that genetic variation in the innate immune signaling pathway contributes to altered risk of PTB.


Field evaluation of a fast anti-Leishmania antibody detection assay in Ethiopia.
A fast agglutination screening test (FAST) for the detection of Leishmania antibodies in human serum samples was evaluated under harsh field conditions in northern Ethiopia. Test performance was compared with a standard serological test, namely the direct agglutination test (DAT), and with parasitology. In total, 103 suspected cases were recruited for the study. Based on parasitological examination, 49 patients were confirmed of having visceral leishmaniasis (VL) and the other 54 suspected cases were parasitologically negative. Field evaluation of FAST was possible in blood samples of 89 patients. FAST had 4 false negative results and 13 false positive results. DAT had 2 false negative results and 20 false positive results. A good degree of agreement (86.9%) was observed between FAST and DAT (kappa value 0.73). In this field-based evalauation, the sensitivity and specificity of FAST were found to be 91.1% (95% CI 77.9-97.1) and 70.5% (95% CI 54.6-82.8), respectively, compared with 95.3% (95% CI 82.9-99.2) and 62.3% (95% CI 47.9-74.9) for DAT. FAST had a high predictive value of a negative test, demonstrating that FAST could be utilised to exclude rapidly non-VL patients from a large population of suspects with fever and splenomegaly in endemic areas.


Cartilage-on-cartilage versus metal-on-cartilage impact characteristics and responses.
A common in vitro model for studying acute mechanical damage in cartilage is to impact an isolated osteochondral or cartilage specimen with a metallic impactor. The mechanics of a cartilage-on-cartilage (COC) impact, as encountered in vivo, are likely different than those of a metal-on-cartilage (MOC) impact. The hypothesis of this study was that impacted in vitro COC and MOC specimens would differ in their impact behavior, mechanical properties, chondrocyte viability, cell metabolism, and histologic structural damage. Osteochondral specimens were impacted with either an osteochondral plug or a metallic cylinder at the same delivered impact energy per unit area, and processed after 14 days in culture. The COC impacts resulted in about half of the impact maximum stress and a quarter of the impact maximum stress rate of change, as compared to the MOC impacts. The impacted COC specimens had smaller changes in mechanical properties, smaller decreases in chondrocyte viability, higher total proteoglycan content, and less histologic structural damage, as compared to the impacted MOC specimens. If MOC impact conditions are to be used for modeling of articular injuries and post-traumatic osteoarthritis, the differences between COC and MOC impacts must be kept in mind.


Hepatocyte growth factor mediates a novel form of hepatic stem/progenitor cell-induced tolerance in a rat xenogeneic liver rejection model.
We have previously identified novel neural/glial antigen 2-expressing hepatic stem/progenitor cells (NG2+ HSPs) that are beneficial for tissue repair by inhibiting the immune cell response. In this in vivo study, we investigated the use of hepatocyte growth factor (HGF)-secreting NG2+ HSPs as a tolerogen in the well-established Syrian golden hamster (SGH) to Lewis (LEW) xenogeneic rat acute liver rejection (ARJ) model. Liver and blood cells were collected for histology and functional analyses using immunofluorescence staining, western blot, ELISA, and TUNEL assays. All recipient rats were randomly divided into 5 groups (n = 14 rats/group) and treated with: (1) ARJ + PBS: (2) ARJ + NG2: tail vein injection of NG2+ HSPs; (3) ARJ + tacrolimus (FK506, oral administration); (4) ARJ + an anti-cMet functional blocking antibody (a-cMet-Ab, I.V) 24 h before the injection of NG2+ HSPs; (5) ARJ + cHGF (clinically used HGF). LEW to LEW syngeneic rats were considered "normal" (n = 14, namely Syn). Significantly prolonged mean survival times (MSTs) and improved graft functions were observed after NG2+ HSP transplantation. An anti-cMet Ab significantly blocked the effect of NG2+ HSPs, suggesting that the effects were likely associated with HGF secreted from NG2+ HSPs. Notably, when intravenously injected into the xenogeneic rat model, the injected cHGF not only prolonged the MST of recipient rats but also increased the number of TUNEL-expressing xenoreactive cytotoxic T lymphocytes (CD8+ T cells). Based on these results, HGF-secreting NG2+ HSPs may specifically target recipient CD8+ T cells by inducing their apoptosis.


Immune privilege for stem cells: not as simple as it looked.
A recent paper in PNAS (Swijnenburg et al., 2008) reveals that human embryonic stem cells are not immune privileged. Potential therapeutic use of these cells will thus depend on the development of new strategies to counter foreign graft rejection by patient immune systems.


Cochlear implantation in a 16-month-old with Chudley-McCullough Syndrome.
The purpose of this report is to describe a case of bilateral cochlear implantation (CI) in a pediatric patient with Chudley-McCullough Syndrome (CMS). By reviewing the literature, we hope to describe common clinical presentations to aid in early diagnosis and management of pediatric patients with CMS. Case report with literature review. We present a case of a 16-month-old female with CMS who presented to clinic after a failed newborn hearing screen and was found to have bilateral sensorineural hearing loss. After a failed trial of hearing amplification, the patient underwent successful bilateral CI. The patient had no surgical complications, and her follow up visit showed satisfactory speech and language development. This case validates that cochlear implantation in pediatric patients who present with CMS is both safe and efficacious. It also demonstrates the importance of early detection and treatment of sensorineural hearing loss in CMS to prevent speech and language delay.


Configurational entropy modulates the mechanical stability of protein GB1.
Configurational entropy plays important roles in defining the thermodynamic stability as well as the folding/unfolding kinetics of proteins. Here we combine single-molecule atomic force microscopy and protein engineering techniques to directly examine the role of configurational entropy in the mechanical unfolding kinetics and mechanical stability of proteins. We used a small protein, GB1, as a model system and constructed four mutants that elongate loop 2 of GB1 by 2, 5, 24 and 46 flexible residues, respectively. These loop elongation mutants fold properly as determined by far-UV circular dichroism spectroscopy, suggesting that loop 2 is well tolerant of loop insertions without affecting GB1's native structure. Our single-molecule atomic force microscopy results reveal that loop elongation decreases the mechanical stability of GB1 and accelerates the mechanical unfolding kinetics. These results can be explained by the loss of configurational entropy upon closing an unstructured flexible loop using classical polymer theory, highlighting the important role of loop regions in the mechanical unfolding of proteins. This study not only demonstrates a general approach to investigating the structural deformation of the loop regions in mechanical unfolding transition state, but also provides the foundation to use configurational entropy as an effective means to modulate the mechanical stability of proteins, which is of critical importance towards engineering artificial elastomeric proteins with tailored nanomechanical properties.


Characterization of virus-like particle assembly for DNA delivery using asymmetrical flow field-flow fractionation and light scattering.
This study illustrates the application of asymmetrical flow field-flow fractionation (AF4) and light scattering analysis during the development of a gene delivery vehicle based on virus-like particles (VLPs) derived from the human polyoma JC virus. The analytical system was created by connecting an AF4 apparatus to the following detectors: diode array, fluorescence, multiangle light scattering, dynamic light scattering, and refractometer. From a single analysis, the molar mass, root mean square and hydrodynamic radii, composition, and purity of the sample could be determined. The VLPs were purified from baculovirus-infected Sf158 insect cells overexpressing the recombinant VP1 protein using weak anion exchange chromatography. The VLPs were dissociated to VP1 pentamers, and the contaminating DNA and proteins were removed using strong anion exchange chromatography. The gene delivery vehicle was created by reassembling the VP1 pentamers in the presence of the desired DNA. The newly formed VLPs encapsulated the DNA and were shown to be capable of delivering the gene of interest to target cells where it was translated into protein. This paper describes the scalable process that was derived to produce the VLPs and demonstrates how the AF4-based analytical characterization was indispensable during the development process.


Clinician Response to a Child Who Completes Suicide.
Although suicide is a leading cause of death for children and adolescents, there is a dearth of literature on clinician responses to suicides in that age group. However, most psychiatrists experience the death of a patient by suicide, with resulting grief reactions including shock, isolation, rumination, self-doubt, and impact on clinical decision making. The impact is more pronounced in trainee clinicians. Postvention is the clinical, administrative, legal, and emotional processes following a suicide. These processes are discussed in detail, with recommendations for policies and training that assist clinicians with this tragic, but common, professional crisis.


Portable fizzy extraction ion-mobility spectrometry system.
Fizzy extraction (FE) is a technique that utilizes effervescence phenomenon to extract volatile organic compounds (VOCs) from liquid matrices for subsequent analysis. To induce effervescence, a liquid sample is first pressurized (at ∼ 150 kPa) with an extractant gas (here, nitrogen), and then rapidly depressurized. In this work, we combine an in-house-built FE system with a commercial ion-mobility spectrometry (IMS) module in order to develop a portable analytical platform for in-situ analysis of VOCs in liquid samples. The size and shape of the FE-IMS platform are similar to those of a typical airline catering trolley. Its operation is enabled by several electronic and electromechanical components (a single-board computer, two microcontroller boards, a relay board, six DC-DC converters, a pressure regulator, a solenoid valve, and a pinch valve). The platform can carry out the extraction procedure as well as acquire, process, and transmit the data to a cloud-storage service. A custom-designed graphical user interface allows the user to select one of the available operation modes: full spectrum mode, ion current profile mode, and cleaning mode. The interface also allows one to follow the analysis progress, display the final result, and upload it to the Internet cloud. The platform has been characterized using three standards: ethyl acetate, ethyl propanoate, and butanone; and their limits of detection are 4.51 × 10-8 M, 2.74 × 10-8 M, and 1.26 × 10-7 M, respectively. Furthermore, its ability to analyze real samples (alcoholic and non-alcoholic beverages) has been demonstrated.


Use of Clinical Faculty Input in Development of an Error and Near-Miss Reporting Form.
Nonreporting of medical errors is an impediment to error prevention and safety in the clinical environment. Many prelicensure nursing education programs do not have policies, procedures, or forms to support student reporting of errors or near misses during their clinical experiences. The purpose of this study was to describe the experiences of clinical faculty with prelicensure nursing student errors or near misses in the clinical environment and to use this information to create a reporting form. Focus groups were used to obtain information about the experiences of clinical faculty (n = 28) with errors or near misses of students. Clinical faculty reported students having safety issues with medication administration, being too task oriented, and modeling shortcut behaviors they had witnessed in clinical practice rather than performing skills as they had learned them. Using information obtained, a clinical error and near-miss reporting form was developed. This form has the potential to increase patient safety in the clinical environment by improving reporting.


MAC1, a nuclear regulatory protein related to Cu-dependent transcription factors is involved in Cu/Fe utilization and stress resistance in yeast.
The related transcription factors ACE1 of Saccharomyces cerevisiae and AMT1 of Candida glabrata are involved in copper metabolism by activating the transcription of copper metallothionein genes. ACE1 and AMT1 are 'copper-fist' transcription factors which possess a conserved cysteine-rich copper binding domain required for DNA binding. Here we report the identification of a nuclear protein from S. cerevisiae, MAC1, whose N-terminal region is highly similar to the copper and DNA binding domains of ACE1 and AMT1. Loss-of-function mutants of MAC1 have a defect in the plasma membrane Cu(II) and Fe(III) reductase activity, are slow growing, respiratory deficient, and hypersensitive to heat and exposure to cadmium, zinc, lead and H2O2. Conversely, a dominant gain-of-function mutant of MAC1 shows an elevated reductase activity and is hypersensitive to copper. We have identified two target genes of MAC1 whose altered expression in mutants of MAC1 can account for some of the observed mutant phenotypes. First, MAC1 is involved in basal level transcription of FRE1, encoding a plasma membrane component associated with both Cu(II) and Fe(III) reduction. Second, MAC1 is involved in the H2O2-induced transcription of CTT1, encoding the cytosolic catalase. This suggests that MAC1 may encode a novel metal-fist transcription factor required for both basal and regulated transcription of genes involved in Cu/Fe utilization and the stress response.


Mesenteric fibromatosis: cytologic, histologic, and ultrastructural findings in a case.
The cytologic findings in a case of mesenteric fibromatosis initially suggested the diagnosis of a benign spindle-cell soft-tissue tumor. Subsequent histology and electron microscopy were performed on the resected mass, and the definitive diagnosis was established. The patient had a history of previous abdominal surgery, but no features of Gardner's syndrome. The difficulties associated with the diagnosis of mesenteric fibromatosis and the cytologic diagnosis of benign and spindle-cell soft-tissue tumors and low-grade sarcomas in general are discussed.


Preclinical exploration of combined glucagon inhibition and liver-preferential insulin for treatment of diabetes using in vitro assays and rat and mouse models.
Normalisation of blood glucose in individuals with diabetes is recommended to reduce development of diabetic complications. However, risk of severe hypoglycaemia with intensive insulin therapy is a major obstacle that prevents many individuals with diabetes from obtaining the recommended reduction in HbA1c. Inhibition of glucagon receptor signalling and liver-preferential insulin action have been shown individually to have beneficial effects in preclinical models and individuals with diabetes (i.e. improved glycaemic control), but also have effects that are potential safety risks (i.e. alpha cell hyperplasia in response to glucagon receptor antagonists and increased levels of liver triacylglycerols and plasma alanine aminotransferase activity in response to glucagon receptor antagonists and liver-preferential insulin). We hypothesised that a combination of glucagon inhibition and liver-preferential insulin action in a dual-acting molecule would widen the therapeutic window. By correcting two pathogenic mechanisms (dysregulated glucagon signalling and non-physiological distribution of conventional insulin administered s.c.), we hypothesised that lower doses of each component would be required to obtain sufficient reduction of hyperglycaemia, and that the undesirable effects that have previously been observed for monotreatment with glucagon antagonists and liver-preferential insulin could be avoided. A dual-acting glucagon receptor inhibitor and liver-preferential insulin molecule was designed and tested in rodent models (normal rats, rats with streptozotocin-induced hyperglycaemia, db/db mice and mice with diet-induced obesity and streptozotocin-induced hyperglycaemia), allowing detailed characterisation of the pharmacokinetic and pharmacodynamic properties of the dual-acting molecule and relevant control compounds, as well as exploration of how the dual-acting molecule influenced glucagon-induced recovery and spontaneous recovery from acute hypoglycaemia. This molecule normalised blood glucose in diabetic models, and was markedly less prone to induce hypoglycaemia than conventional insulin treatment (approximately 4.6-fold less potent under hypoglycaemic conditions than under normoglycaemic conditions). However, compared to treatment with conventional long-acting insulin, this dual-acting molecule also increased triacylglycerol levels in the liver (approximately 60%), plasma alanine aminotransferase levels (approximately twofold) and alpha cell mass (approximately twofold). While the dual-acting glucagon receptor inhibitor and liver-preferential insulin molecule showed markedly improved regulation of blood glucose, effects that are potential safety concerns persisted in the pharmacologically relevant dose range.


Development of a Reverse Transcription Loop - Mediated Isothermal Amplification [RT-LAMP] as a early rapid detection assay for Crimean Congo Hemorrhagic Fever virus.
Presently diagnosis of Crimean Congo Hemorrhagic Fever virus (CCHFV) infection relies on real-time and end-point RT-PCR, and serodiagnostic assay. These assays are time consuming and cannot be used as a routine screening test. The objective of this study was to develop a rapid diagnostic test that could be completed in < 60 minutes. Rapid detection of CCHFV infection is important for faster delivery of appropriate therapeutics, clinical management of patient and also important to contain the outbreak. In the present study, we have developed a rapid and sensitive single tube reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for detection of CCHFV. The limit of detection of RT-LAMP vis-a-vis Real-time RT-PCR assay is 10 RNA copies. Further, CCHFV specific RT-LAMP assay was successfully evaluated with human and tick samples. The assay correctly picked up diverse CCHFV isolates indicating its applicability for different strains. A comparative evaluation of the RT-LAMP assay vis-à-vis with the real-time RT-PCR revealed 100% concordance with 100 % sensitivity and specificity respectively. No cross reactivity with related Flaviviruses and hemorrhagic fever viruses was observed. The assay is a rapid, isothermal, simple to perform molecular diagnostic, which can be performed in a portable heating block device. CCHF RT-LAMP assay can be used in low resource laboratories for monitoring of CCHFV outbreaks in remote rural regions in affected countries.


Rehabilitation of Patients with Post-COVID-19 Syndrome: A Narrative Review.
: We aimed to review the most recent articles on the rehabilitation of patients after coronavirus disease 2019 (COVID-19) and to identify the methods and effects of rehabilitation on such patients. : A literature search was conducted using PubMed and Web of Science from study inception to October 2022 using the following search terms to identify meta-analyses and randomized controlled studies with abstracts written in English: ["COVID-19" or "COVID 19" or "2019-nCoV" or "SARS-CoV" or "novel coronavirus" or "SARS-CoV-2"] and ["rehabilitation"]. Publications investigating the effects of pulmonary and physical rehabilitation on patients with COVID-19 were extracted. The extraction process selected four meta-analyses, two systematic reviews, two literature reviews, and two randomized controlled trials. Pulmonary rehabilitation recovered forced vital capacity (FVC), 6-min walk distance (6MWD), health-related quality of life (HRQOL), and dyspnea. Pulmonary rehabilitation increased predicted FVC, distance in the 6MWD test, and HRQOL score compared with baseline values. Physical rehabilitation, comprising aerobic exercises and resistance training, effectively improved fatigue, functional capacity, and quality of life with no adverse events. Telerehabilitation was an effective tool to provide rehabilitation for patients with COVID-19. Our study suggests that rehabilitation after COVID-19 should be considered an effective therapeutic strategy to improve the functional capacity and quality of life of patients with COVID-19.


[Quantification of plasma DNA as a tumor marker in patients with pancreatic cancer].
The plasma DNA concentration of patients with cancer is known to be higher than normal controls. Increased DNA concentration and tumor-specific genes in plasma can be used as tumor markers in some cancers. This study was designed to evaluate whether quantification of plasma DNA concentration by using real-time PCR is useful as a tumor marker in the diagnosis of pancreatic cancer. Twenty-four patients (M:F=16:8, mean age; 60.5+/-11.5 years) with pancreatic cancer were recruited for this study. Fifteen patients with chronic pancreatitis and fifteen healthy persons were selected as controls (M:F=26:4, 53.5+/-11.2 years). The concentration of plasma DNA was determined by real-time PCR for telomerase reverse transcriptase gene. Plasma DNA concentration in patients with pancreatic cancer (46.4+/-63.2 ng/mL) was higher than that of chronic pancreatitis (p=0.041) and normal controls (p=0.030). The sensitivity and specificity in detecting pancreatic cancer were 75% and 70% respectively when the cut-off value of plasma DNA concentration was set at 46.9 ng/mL. Plasma DNA concentration in patients with pancreatic cancer was higher than that of controls. However, its sensitivity and specificity is not high enough to be used as a tumor marker for pancreatic cancer.


Disappearance of intraperitoneal gas following gynaecological laparoscopy.
We investigated 25 day case gynaecological laparoscopy patients to determine the duration of the pneumoperitoneum, the rate at which residual gas disappeared and the relative contribution of this gas to pain in the early postoperative period. The volume of gas was calculated from measurements of the subdiaphragmatic gas bubble obtained from an erect chest X-ray. Each patient was X-rayed twice, either immediately prior to discharge and at 24 h post laparoscopy, or at 24 h and 48 h post surgery. Patients were contacted by telephone at 24 and 48 h to confirm fitness to return for the X-ray and at the same time verbal pain and activity scores were recorded. We found that residual gas was almost entirely gone by 48 h following surgery and that it appeared to reduce in an exponential manner. The contribution of this gas to postoperative pain was significant in the first 24 h, but by 48 h it was considerably reduced. We conclude that in day case gynaecological laparoscopy patients, postoperative pneumoperitoneum is short-lived, and that by 48 h its volume and contribution to postoperative pain should be minimal.


N-dihydrogalactochitosan reduces mortality in a lethal mouse model of SARS-CoV-2.
The rapid emergence and global dissemination of SARS-CoV-2 that causes COVID-19 continues to cause an unprecedented global health burden resulting in nearly 7 million deaths. While multiple vaccine countermeasures have been approved for emergency use, additional treatments are still needed due to sluggish vaccine rollout, vaccine hesitancy, and inefficient vaccine-mediated protection. Immunoadjuvant compounds delivered intranasally can guide non-specific innate immune responses during the critical early stages of viral replication, reducing morbidity and mortality. N-dihydrogalactochitosan (GC) is a novel mucoadhesive immunostimulatory polymer of β-0-4-linked N-acetylglucosamine that is solubilized by the conjugation of galactose glycans with current applications as a cancer immunotherapeutic. We tested GC as a potential countermeasure for COVID-19. GC was well-tolerated and did not produce histopathologic lesions in the mouse lung. GC administered intranasally before and after SARS-CoV-2 exposure diminished morbidity and mortality in humanized ACE2 receptor expressing mice by up to 75% and reduced infectious virus levels in the upper airway. Fluorescent labeling of GC shows that it is confined to the lumen or superficial mucosa of the nasal cavity, without involvement of adjacent or deeper tissues. Our findings demonstrate a new application for soluble immunoadjuvants such as GC for preventing disease associated with SARS-CoV-2 and may be particularly attractive to persons who are needle-averse.


Modification of regression of virally xenogenized tumor cells by cyclophosphamide and busulfan.
Rat fibrosarcoma cells infected with Friend leukemia virus (FV-KMT-17) grow for a short time and then regress spontaneously in syngeneic hosts. This regression mechanism was examined by analyzing the immunomodulating action of the antitumor drugs busulfan (BU) and cyclophosphamide (CY). In preliminary experiments, the optimum dosages of BU and CY for the enhancement of DTH responses to SRBC were 10 mg/kg and 40 mg/kg respectively. Treatment of rats with BU (10 mg/kg) on day 5 induced the regression of KMT-17 cells, while in contrast, the same drug delayed the spontaneous regression of FV-KMT-17 cells. Pretreatment with CY (40 mg/kg) on day 5 did not affect the growth of KMT-17 or FV-KMT-17 cells. After the same treatment schedule, BU inhibited humoral antibody formation against SRBC and against virus-associated antigen (VAA), NK cell activity, and ADCC effector cell activity. On the other hand, CY did not affect the activities of NK cells or ADCC effector cells, although it significantly augmented the DTH responses to SRBC and the production of antibody to VAA but had no effect on production of antibodies to SRBC. These results suggest that NK cells and ADCC may play an important role in the initial stage of the spontaneous regression of FV-KMT-17 cells.


Pharmacogenetic approach for capecitabine or 5-fluorouracil selection to be combined with oxaliplatin as first-line chemotherapy in advanced colorectal cancer.
We studied the role of TS (5'VNTR, 5'SNP and 3'UTR), XRCC1-399, XPD-751, ERCC1-118 and XRCC3-241 genetic polymorphisms in tailoring fluroropyrimidine/oxaliplatin treatment. For this purpose, 110 XELOX (capecitabine/oxaliplatin)- or FUOX (fluorouracil/oxaliplatin)-treated metastatic colorectal cancer patients were selected prospectively for genotyping. In the FUOX group, TS-3'UTR +6bp/+6bp (hazards ratio, HR=2.62, p=0.007) and ERCC1-118C/T or C/C (HR=1.96, p=0.050) genotypes correlated with a shorter progression-free survival (PFS). When analysed jointly, the higher the number of favourable genotypes (FG) the longer the PFS (6.8m, 9.6m and 25.8m for 0, 1 or 2 FG; p=0.005). Disease-control rate was 100% in patients with 2 FG (87% and 38.5% for 1 or 0 FG; p=0.001). In the multivariate analysis, ERCC1-118 (HR=2.12, p=0.0037) and TS-3'UTR (HR=2.68, p=0.006) were strong independent prognostic factors. According to this, patients harbouring TS-3'UTR +6bp/+6bp and ERCC1-118C/T or C/C genotypes may better receive capecitabine instead of 5FU in an oxaliplatin-based first-line treatment.


Placenta-specific gene manipulation using lentiviral vector and its application.
The placenta is an essential organ for embryo development in the uterus of eutherian mammals. Large contributions in unveiling molecular mechanisms and physiological functions underlying placental formation were made by analyzing mutant and transgenic animals. However, it had been difficult to elucidate whether the placental defects observed in such animals originate from the placenta itself or from the fetus, as both placental and fetal genomes are modified. Therefore strategies to modify the placental genome without affecting the "fetal genome" had been needed. Through the ingenious use of lentiviral (LV) vectors, placenta-specific modification is now possible. Lentivirus is a genus of retroviruses that use reverse-transcriptase to convert its single-strand RNA genome to double-strand DNA and integrate into the host genome. Previous studies showed that when LV vectors were used to transduce embryos at the 2-cell stage, the viral genome is systemically introduced into host genome. Interestingly, by delaying the timing of transduction to the blastocyst stage, the transgene is expressed specifically in the placenta as a consequence of trophectoderm-specific viral transduction. This review summarizes the development of the LV vector-mediated placenta-specific gene manipulation technology and its application in placental research over the past decade. A perspective for future application of LV vectors to further placenta research, especially in combination with next generation genome editing technologies, is also presented.


Circulating long noncoding RNA ZNFX1 antisense RNA negatively correlates with disease risk, severity, inflammatory markers, and predicts poor prognosis in sepsis patients.
This study aimed to investigate the correlation of long noncoding RNA (lncRNA) ZNFX1 antisense RNA (ZFAS1) with disease risk, severity, inflammation markers, and prognosis in sepsis patients.A total of 202 sepsis patients were consecutively enrolled, and 200 healthy volunteers were also recruited as healthy controls (HCs). Plasma samples of all patients and HCs were collected. LncRNA ZFAS1 expression was determined by quantitative polymerase chain reaction assay and inflammatory cytokines levels were detected by enzyme-linked immunosorbent assay.The median value of lncRNA ZFAS1 expression in sepsis patients was (0.639 [0.325-1.071]), which was lower compared to HCs (1.957 [0.876-3.245], P < .001], and receiver operating characteristics (ROC) curve revealed that lncRNA ZFAS1 expression had a good diagnostic value for sepsis with area under curve (AUC) of 0.814 (95% confidence interval [CI]: 0.771-0.857). Spearman test disclosed that lncRNA ZFAS1 expression was negatively correlated with Acute Physiology and Chronic Health Evaluation (APACHE) II score (r = -0.505, P < .001), and it was negatively associated with levels of C-creative protein (r = -0.241, P = .001), tumor necrosis factor-α (r = -0.253, P < .001), and interleukin (IL)-6 (r = -0.177, P = .012) while positively associated with IL-10 level (r = 0.173, P = .014). Also, lncRNA ZFAS1 expression was lower in survivor group compared to nonsurvivor group (P < .001), and it presented with a good predictive value on distinguishing nonsurvivors from survivors in sepsis patients with AUC of 0.628 (95% CI: 0.538-0.717).Circulating lncRNA ZFAS1 expression negatively correlates with disease risk, severity, and inflammatory markers levels, and might predict worse survival in sepsis patients.


A Prospective Study Evaluating the Feasibility and Accuracy of Post-operative Laryngeal Ultrasonography (LUSG) in Assessment of Vocal Cord Function After Esophagectomy.
Vocal cord paresis (VCP) is a serious complication after esophagectomy. Conventional diagnosis of VCP relies on flexible laryngoscopy (FL), which is invasive. Laryngeal ultrasonography (LUSG) is non-invasive and convenient. It has provided accurate VC evaluation after thyroidectomy but it is unclear if it is just as accurate following esophagectomy. This prospective study evaluated the feasibility and accuracy of LUSG in VC assessment on day-1 after esophagectomy. Consecutive patients from a tertiary teaching hospital who underwent elective esophagectomy were prospectively recruited. All received pre-operative FL, and post-operative LUSG and FL on Day-1, each performed by a blinded, independent assessor. The primary outcomes were feasibility and accuracy of LUSG in the diagnosis of VCP on Day-1 post-esophagectomy. The accuracy of voice assessment (VA) was analyzed. Twenty-six patients were eligible for analysis. The median age was 70 years (66-73). Majority were male (84.6%). Twenty-five (96.2%) received three-phase esophagectomy. Twenty-four (96%) had same-stage anastomosis at the neck. Three (11.5%) developed temporary and one (3.8%) developed permanent unilateral VCP. Overall VC visualization rate by LUSG was 100%; sensitivity, specificity, positive predictive value, negative predictive value (NPV) and accuracy of LUSG were 75.0%, 100%, 100%, 98.0%, 98.1% respectively, and superior to VA. Combining LUSG with VA findings could pick up all VCPs i.e. improved sensitivity and NPV to 100%. LUSG is a highly feasible, accurate and non-invasive method to evaluate VC function early after esophagectomy. Post-operative FL may be avoided in patients with both normal LUSG and voice.


Variational Hilbert Quantitative Phase Imaging.
Utilizing the refractive index as the endogenous contrast agent to noninvasively study transparent cells is a working principle of emerging quantitative phase imaging (QPI). In this contribution, we propose the Variational Hilbert Quantitative Phase Imaging (VHQPI)-end-to-end purely computational add-on module able to improve performance of a QPI-unit without hardware modifications. The VHQPI, deploying unique merger of tailored variational image decomposition and enhanced Hilbert spiral transform, adaptively provides high quality map of sample-induced phase delay, accepting particularly wide range of input single-shot interferograms (from off-axis to quasi on-axis configurations). It especially promotes high space-bandwidth-product QPI configurations alleviating the spectral overlapping problem. The VHQPI is tailored to deal with cumbersome interference patterns related to detailed locally varying biological objects with possibly high dynamic range of phase and relatively low carrier. In post-processing, the slowly varying phase-term associated with the instrumental optical aberrations is eliminated upon variational analysis to further boost the phase-imaging capabilities. The VHQPI is thoroughly studied employing numerical simulations and successfully validated using static and dynamic cells phase-analysis. It compares favorably with other single-shot phase reconstruction techniques based on the Fourier and Hilbert-Huang transforms, both in terms of visual inspection and quantitative evaluation, potentially opening up new possibilities in QPI.


[Cost analysis of the diagnosis and treatment of soft tissue sarcomas in reference centres].
To determine the diagnostic and treatment costs in patients referred to a reference centre with a suspected soft tissue sarcoma. The study consisted of a historic cohort of 48 consecutive patients who were diagnosed with soft tissue sarcomas and treated in our centres between the years 2009 and 2011, both prior to and after performing the biopsy. The cost per procedure was taken from the official list of prices published in the year 2009. A comparative study of the data was performed using the Mann-Whitney U and Wilcoxon tests. The mean cost per patient was 14,427.58€. In those referred before the biopsy, the overall mean cost was 11,818.67€, and in those referred afterwards, it was 6,456.74€ (p=.0073). There were no significant differences in the diagnostic costs between the groups. However, the mean cosy of the treatment per patient was higher in the second group (p=.0121). The referral to centres with experienced multidisciplinary teams in this disease is a common fact highlighted in many articles, where the financial savings have also been demonstrated. This study shows that, as well as improving the care aspect of the patients when they are referred to a specialist centre prior to the biopsy, there is also a lower health care cost.


Sudden unexplained death in children with epilepsy: a cohort study with an eighteen-year follow-up.
Sudden unexplained death is a significant cause of mortality in adults with epilepsy. Only a few data exist about this risk in childhood. Cases of sudden unexplained death in epilepsy (SUDEP) up to the age of 18 y occurring at our hospital between 1984 and 2001 were identified. The incidence rate was calculated on the basis of diagnosed epileptics registered with a statutory disability insurance scheme. Four cases of SUDEP were identified during the 18-y period. The incidence of SUDEP was 4.3 per 10,000 patient-years. All children showed polytherapy-refractory epilepsy, developmental retardation and early-onset epilepsy. Two witnessed cases had shown no previous signs of seizure. SUDEP is rare in childhood. Children with uncomplicated epilepsy seem not to be at risk.


Associations among palatal impaction of canine, sella turcica bridging, and ponticulus posticus (atlas arcuate foramen).
The palatal impaction of canine (PIC) can be predicted by some head and neck skeletal anomalies or variants. Since studies on this regard (especially vertebral anomalies) are scarce, this study was conducted. This case-control study was done on 46 PIC orthodontic patients (34 females, 12 males) and 46 control orthodontic patients (36 females, 10 males). The diagnosis of PIC was done on lateral cephalographs and panoramic radiographs. On cephalographs, sella turcica bridging (occurrence and severity) and ponticulus posticus (occurrence and severity) were assessed. Associations between PIC, sella bridging, and ponticulus posticus were examined statistically (α = 0.05, β ≤ 0.2). Cases' and controls' mean ages were 17.7 ± 4.0 and 17.4 ± 3.5, respectively. Of the case subjects, 22, 22, and 2 had respectively types I (normal), II, and III of sella bridging, while these numbers were 34, 12, and 0 in controls (chi-square P = 0.023 for severity, 0.010 for occurrence). Ponticulus posticus was observed in 28 cases (7 completed) and 17 controls (6 completed, P = 0.022 for occurrence, 0.056 for severity). Sella bridging was not associated with ponticulus posticus (Spearman P = 0.150). According to binary logistic regression, sella bridging can increase the odds of palatal canine impaction for OR = 2.8 times, while ponticulus posticus for OR = 2.6. Age and sex did not affect sella bridging or ponticulus posticus. Both sella bridging and ponticulus posticus can predict an increased rate of PIC for more than 2.5 times.


The fine structure of the feline superior vestibular nucleus: identification and synaptology of the primary vestibular afferents.
The superior vestibular nucleus of the cat and its primary vestibular efferents were examined by light and electron microscopy. The primary vestibular afferents branch within the nucleus in a sheet-like pattern, in the transverse plane. The dendritic fields of many secondary neurons are shaped like discs and are also oriented in the transverse plane. This relation between the primary afferents and dendritic fields may be relevant to the convergence of primary afferents innervating particular endorgans onto secondary neurons. Synaptic boutons in the SV were divided into 3 putative types on the basis of the size and shape of their synaptic vesicles. The primary afferent bouton was identified by comparing the SV of the two sides after unilateral lesions of the vestibular ganglion. Its boutons contain round vesicles of 40 nm average diameter and are associated with prominent postsynaptic densities; the two other putative bouton types contain smaller, round vesicles, and pleomorphic vesicles. The primary afferent boutons largely contact proximal dendrites, their appendages, and cell somata of the secondary neurons. In animals receiving unilateral lesions of the vestibular ganglion and allowed to survive long enough for the primary afferent boutons to disappear (5--6 days), there occurs in the denervated as compared to normal SV: (1) a decrease in the fraction of the somal surface of the secondary neurons covered by boutons with small round vesicles; and (2) a decrease in the ratio: number of boutons with small round vesicles to number of boutons with pleomorphic vesicles. In addition, there appears on the lesioned side a new group of boutons with pleomorphic vesicles smaller than those in boutons from the control side. These observations suggest plastic changes in response to deafferentation, and may be related to the marked behavioral recovery which occurs within a few days after lesion of the vestibular ganglion.


Soil habitat function after innovative nanoagriproducts application: Effect of ageing on the avoidance behaviour of the soil invertebrates Enchytraeus crypticus and Folsomia candida.
Research on nanotechnology with applications in agriculture has been gathering attention because it may achieve a good balance between agricultural production and environmental integrity. Among the vast nanomaterials, layered double hydroxides (LDHs) are a promising solution for supplying crops with macro- and/or micronutrients. Still, little is known about their safety implications for non-target organisms, such as soil invertebrates. The habitat function of soils might be impacted by potential stressors, which can be assessed through avoidance behaviour tests. This study aimed to assess the effect of two innovative agriproducts, Zn-Al-NO3 LDH and Mg-Al-NO3 LDH, on the avoidance behaviour of the enchytraeid Enchytraeus crypticus and the collembolan Folsomia candida, over time. Simultaneously, Zn and Mg potential release from LDHs to soil was evaluated. Overall, the behaviour of soil invertebrates differed between species, with enchytraeids being more sensitive to LDHs-treated soils than collembolans, possibly explained by their different physiological traits. The behaviour of soil organisms also depended on the LDH structural composition and was time-variable. Soil treated with Zn-Al-NO3 LDH was perceived as less favourable compared to Mg-Al-NO3 LDH, which was preferred to clean soil at most tested concentrations. LDHs toxicity was partly, but not exclusively, related to Zn and Mg release. Cations release over time was demonstrated in the chemical assessment. Still, LDHs toxicity to soil invertebrates decreased as increasing AC50 values were derived over time. Slower dissolution over time might explain the decrease in toxicity. Our study demonstrates that both soil invertebrates could sense LDHs in soil and eventually adapt their behaviour by avoiding or preferring, according to the type and level of LDH present.


[Rapid detection of Streptococcus mutans and streptococcus sobrinus in human saliva by nested polymerase chain reaction].
To establish a simple and rapid method to detect Streptococcus mutans and streptococcus sobrinus simultaneously in human saliva. Chromosomal DNA from the bacteria was obtained by the extraction method with phenol-chloroform. A nested PCR method with two sets of primers specific for portions of the glucosyltransferase genes (gtfB of S. mutans and gtfI of S. sobrinus), was optimized to detect S. mutans and S. sobrinus from standard strains, clinical strains and directly in human saliva. The first process of nested PCR was capable of amplifying DNA fragments specific for these species from chromosomal DNA extracted from 10(5) CFU cells of standard and clinical strains, or from 1 ml clinical saliva samples containing 10(5) CFU cells of either species. a second process of nested PCR, using the first PCR product as a template with new internal primers to detect 10(3) CFU of either streptococcal species in 1ml saliva samples. Nested PCR could detect S. mutans and S. sobrinus rapidly and simply in human saliva. This finding would be important to studies of elucidation the role of these two streptococcal species in the etiology of dental caries.


A locally aggressive solitary fibrous tumor of the leg: Case report and literature review.
The solitary fibrous tumor (SFT) is a rare soft tissue tumor with a substantially benign clinical behavior. However, malignant neoplasms with local recurrence or distant metastases have been reported. The authors present a case of an aggressive SFT of the leg, in a 55 years old Caucasian man. Radiological, histological and molecular findings are reported. The differential diagnosis, therapy and outcome of this rare tumor are also discussed. An extensive review of literature showed SFT's clinical behavior as substantially benign, anyway aggressive or malignant neoplasms have been described. The potential risk of local recurrence and distant metastasis thus suggests wide surgical resection and careful long-term follow-up. Differential diagnosis may be quite laborious as SFT can mimic a variety of benign and malignant mesenchymal tumors; immunohistochemical analysis for CD34, CD99, vimentin and bcl-2 is then mandatory. Our clinical experience confirmed that SFT may have an aggressive behavior, however, conservative surgical treatment may be successful in the long term.


First-Principles Study on Interlayer Spacing and Structure Stability of NiAl-Layered Double Hydroxides.
Interlayer spacing and structure stability of layered double hydroxides (LDHs) on their application performance in adsorption, ion exchange, catalysis, carrier, and energy storage is important. The effect of different interlayer anions on the interlayer spacing and structure stability of LDHs has been less studied, but it is of great significance. Therefore, based on density functional theory (DFT), the computational model with 10 kinds of anions intercalated Ni3Al-A-LDHs (A = Cl-, Br-, I-, OH-, NO3 -, CO3 2-, SO4 2-, HCOO-, C6H5SO3 -, C12H25SO3 -) and four Ni R Al-Cl-LDH models with different Ni2+/Al3+ ratios (R = 2, 3, 5, 8) were constructed to calculate and analyze interlayer spacing, structural stability, and their influence factors. It was found that the interlayer spacing order of Ni3Al-A-LDHs intercalated with different anions is OH- < CO3 2- < Cl- < Br- < I- < HCOO- < SO4 2- < NO3 - < C6H5SO3 - < C12H25SO3 -. The hydrogen bond network between the base layer and the interlayer anions affects the arrangement structure of the interlayer anions, which affects the interlayer spacing. For interlayer monatomic anions Cl-, Br-, and I- and the anion of comparable size in each direction SO4 2-, the interlayer spacing is positively correlated with the interlayer anion diameter. The larger difference between the long-axis and short-axis dimensions of the polyatomic anions results in the long axis of the anion being perpendicular to the basal layer, increasing interlayer spacing. The long-chain anion C12H25SO3 - intercalation system exhibits the largest layer spacing of 24.262 Å. As R value increases from 2 to 8, the interlayer spacing of Ni R Al-Cl-LDHs gradually increases from 7.964 to 8.124 Å. The binding energy order between the interlayer anion and basal layer is CO3 2- > SO4 2- > OH- > Cl- > Br- > I- > HCOO- > NO3 - > C12H25SO3 - > C6H5SO3 -. The smaller the interlayer spacing, the higher the binding energy and the stronger the structural stability of LDHs. The factors affecting structural stability mainly include the bond length and bond angle of the hydrogen bond and the charge interaction between the basal layer and interlayer anion. In the CO3 2- intercalated system, the hydrogen bond length exhibits the shortest of 1.95 Å and the largest bond angle of 163.68°. The density of states and energy band analysis show that the higher the number of charges carried by the anion, the stronger its ability to provide electrons to the basal layer.


Glucocorticoids increase extracellular [3H]D-aspartate overflow in hippocampal cultures during cyanide-induced ischemia.
Glucocorticoids (GCs), the adrenal steroid hormones secreted during stress, exacerbate neuronal death in the hippocampus during ischemia. Since ischemia brain damage is ascribed to an elevated level of extracellular excitatory amino acids (EAAs), this study was undertaken to investigate the effect of GCs on EAA homeostasis in hippocampal cell cultures during the insult of cyanide exposure. Using D-[2,3-3H]aspartic acid ([3H]D-Asp) as a tracer, we found that corticosterone (CORT, the physiological GC in rats) increased the accumulation of extracellular [3H]D-Asp by 25% in hippocampal cultures during cyanide-induced ischemia. CORT had no effect on the release of [3H]D-Asp. Instead, analysis of [3H]D-Asp uptake kinetics indicates that CORT decreased the maximum uptake rate and the Michaelis constant by 44% and 50%, respectively, in cells treated with cyanide. It is concluded that, during cyanide-induced ischemia, CORT might enhance extracellular overflow of [3H]D-Asp by decreasing its uptake, thereby endangering neurons.


The HLA-B7 allele confers susceptibility to breast cancer in Spanish women.
Several HLA alleles are associated with susceptibility or protection in breast cancer. The particular allele varies depending on the geographical region. A study in a small group of Spanish patients using serological methods found an association with HLA-B7. We undertook a larger study in southern Spain using molecular biology techniques. Genotype variants of HLA class I and II were typed by PCR-SSP in 132 breast cancer patients and 382 healthy controls. The frequency of the HLA-B7 allele was increased in the patients compared to the controls (P=0.0019; 95% confidence interval, 1.337-3.409, relative risk=2.135). Bonferroni correction of the P showed it was still significant (P(c)=0.0285). These results support previous suggestions that HLA-B7 is associated with the development of breast cancer in our area.


Incidence of allo-immunization and allergic reactions to cryoprecipitate in haemophilia.
38 haemophiliacs who had received frequent cryoprecipitate infusions, were investigated for the presence of antibodies directed against red cell, HLA, Gm, Inv and platelet-specific antigens. 15 had detectable antibodies against one or more antigen systems. 11 experienced allergic reactions following infusions of cryoprecipitate. There was no correlation between allergic reactions and the presence of detectable antibodies.


Diurnal changes in melatonin binding sites of hamster and rat brains. Correlations with neuroendocrine responsiveness to melatonin.
A diurnal variation in neuroendocrine sensitivity to melatonin is known to occur in hamsters and rats. The present experiments were carried out to examine the possibility that affinity and/or number of melatonin binding sites in brain could change accordingly at the two times when exogenous melatonin is known to be ineffective or effective to produce neuroendocrine changes, i.e., at 07 : 00 or 20 : 00 h (lights on from 07 : 00 to 21 : 00 h daily). The number of melatonin binding sites in hamster and rat brains was at 20 : 00 h 34--56% higher than at 07 : 00 h, without changing their affinity towards [3H]melatonin (hamster: Kd = 53 nM; rat: Kd = 73--77 nM). These alterations in melatonin receptor density may play a role in daily changes in sensitivity to the exogenous methoxyindole.


Relative hepatotoxicity of ortho and meta mono substituted thiobenzamides in the rat.
Thiobenzamide is known to be hepatotoxic in the rat and the relative hepatotoxicity of para-substituted thiobenzamides has previously been shown to depend strictly on the electronic character of the para substituent. We have now extended this study to include ortho and meta monosubstituted thiobenzamides. Among the meta-substituted compounds, hepatotoxicity varies in strict accordance with the electronic character of the substituent, whereas the ortho-substituted compounds show no toxicity at comparable doses regardless of the nature of the substituent. Explanations for these substituent effects are provided in terms of the chemical reactivity of the compounds and their corresponding S-oxide and S,S-dioxide metabolites.


Site-specific analysis of drug interactions and damage in DNA using sequencing techniques.
DNA-sequencing techniques can be adapted to provide powerful analytical tools for pinpointing the sites at which DNA is modified by either radiations or chemicals. Base modifications, covalent adducts, crosslinks, and noncovalent binding can all be detected. This review outlines the adaptions of the Maxam-Gilbert and Sanger dideoxy sequencing techniques which make such studies possible. Practical aspects and limitations of the various methods are given. Assays which test the ability of DNA polymerase to bypass damage and misincorporate bases are also discussed. It is concluded that these techniques constitute the most powerful method currently available for the study of sequence-specific DNA interactions.


Reuse of used paper egg carton boxes as a source to produce hybrid AgNPs- carboxyl nanocellulose through bio-synthesis and its application in active food packaging.
The proper disposal of disposable synthetic plastic food packaging materials presents a significant challenge for both the environment and the solid waste management community. To address this issue, an antibacterial-based high-strength bio-composite serves as the optimal alternative to conventional packaging materials. This study aims to produce a hybrid material of AgNPs-carboxyl cellulose nanocrystals (AgNPs-CCNCs), obtained from used egg carton boxes (UECBs), through bio acid hydrolysis and an in-situ generation process. Furthermore, AgNPs- carboxyl cellulose nanofibers (AgNPs-CCNFs) will be synthesized through a combination of bio acid hydrolysis and ball milling, followed by an additional in-situ generation step. The AgNPs-carboxyl nanocellulose (AgNPs-CCNCs, and AgNPs-CCNFs) exhibited excellent crystallinity index, morphology, thermal, and antibacterial properties. The morphological analysis was performed by electron microscopy, and the results showed the uniform distribution and spherical form of AgNPs appearing over the carboxyl nanocellulose through the in-situ generation process, which was confirmed through XRD analysis. The study further explores the impact of AgNPs-carboxyl nanocellulose on the mechanical, chemical, antibacterial, and thermal properties of the PVA matrix. The results demonstrate that the bio-nanocomposite film offers opportunities for utilization in active packaging applications.


Associations between Health Behaviors, Health Self-Efficacy, and Long-Term Outcomes in Survivors of Childhood Cancer: A Cross-Sectional Study.
Many survivors of childhood cancer experience multiple long-term chronic health conditions. Health behaviors are important because they contribute to chronic disease yet are highly modifiable. With growing pressure on cancer services, alternative models of care are required to address survivorship care needs. The authors sought to inform the development of a community-based model of cancer survivorship care for young people. This exploratory cross-sectional study aimed to assess the feasibility of study measures and processes, as well as investigate associations between various modifiable health behaviors, health self-efficacy, quality of life, and persistent symptoms. Participants were recruited from a long-term follow-up clinic for childhood cancer survivors. A self-report survey was completed, and participants received an activity tracker. Bivariate regression analyses were used to explore the relationship between variables. The study measures and processes were deemed feasible with >70% of eligible survivors enrolling in the study and completing >70% of study measures. Thirty participants (mean age 22 ± 4.4 years) were enrolled; 83.3% finished treatment ≥5 years previously and 36.7% were overweight or obese. Bivariate regression identified those with higher scores of health self-efficacy were more likely to meet physical activity guidelines, as were those who achieved more sleep and consumed greater servings of vegetables. Meeting the physical activity guidelines, was significantly positively associated with higher quality of life and self-efficacy. Interventions that target health self-efficacy have the potential to improve a range of health behaviors and long-term outcomes for survivors of childhood cancer. Nurses are ideally placed to use this knowledge to support patients with recommendations to optimise their recovery and rehabilitation.


Abnormal urodynamic findings after radical hysterectomy or pelvic irradiation for cervical cancer.
To assess urodynamic study results in patients with cervical cancer who had received radical hysterectomy or pelvic irradiation or radical hysterectomy with pelvic irradiation. Forty-two patients with stage IB cervical cancer after radical hysterectomy (group A), 11 patients at stage IB or IIA after pelvic irradiation (group B), 15 patients at stage IB or IIA after both radical hysterectomy and pelvic irradiation (group C) and 17 patients at stage IB before treatment (group D) as control were recruited for urodynamic examination. The evaluations for each case included a 20-min pad test, uroflowmetry, both filling and voiding cystometry, and stress urethral pressure profile. ANOVA method with Bonferroni test and Pearson chi2-test were utilized for statistical analysis. The mean ages in sequential groups A, B, C and D were 52.9 +/- 10.2, 62.5 +/- 13.5, 49.8 +/- 11.7 and 49.4 +/- 12.5 years (P = 0.02), respectively. The occurring frequency of either detrusor instability or low bladder compliance was 57%, 45%, 80% and 24%, respectively. Each group revealed decreased bladder capacity as 268.4 +/- 102.8, 164.1 +/- 62.9, 233.5 +/- 73.9 and 293.0 +/- 47.2 ml (P < 0.0001). However, the frequency of abdominal strain voiding was 100% in groups A, B and C as compared to 0% in group D (P < 0.01), and the frequency of abnormal residual urine (> 50 ml) was 41%, 27%, 40% and 24%. Although each case showed a poor pressure transmission ratio (< 100%), the frequency of positive pad test in each group was 81%, 46%, 100% and 18% (P < 0.001). The functional urethral length decreased in each group and was 2.6 +/- 0.8, 2.3 +/- 0.8, 2.5 +/- 0.8 and 2.9 +/- 0.6 cm, but there were no significant differences in maximal urethral pressure or urethral closure pressure among the four groups. Our data show that abnormal urodynamic findings pre-exist in patients with cervical cancer before treatment especially in bladder storing function, and that these findings may worsen, or that new abnormal findings may happen after radical hysterectomy or pelvic irradiation, or both.


Comparative potency and pharmacology of isomers of leukotriene D4 on guinea-pig trachea: requirement for a 5(S)6(R) configuration.
The relative contractile activity of C5 and C6 diastereomers of Leukotriene D4 (LTD4), as well as 11-trans stereoisomers were evaluated in guinea-pig tracheal smooth muscle. 5(S)6(R) LTD4 was 1000 times more potent than histamine as a contractile agent. While a change of the 11-ethylenic bond from cis to trans resulted in a four fold decrease in potency, a change in configuration of the 5-hydroxyl group and/or the 6-peptide adduct resulted in a decrease in potency of at least 2 to 3 orders of magnitude. The contractile activity of all LTD4 isomers was inhibited by FPL 55712, whereas indomethacin markedly enhanced the contractile activity of 5(S)6(R) LTD4, but appeared to have less of an effect on the other diastereomers. The results demonstrate the critical nature of configuration of the 5-hydroxyl and the 6-peptide adduct of eicosatetraenoic acid for maintenance of high affinity for receptors.


4-Kallikrein Test and Kallikrein Markers in Prostate Cancer Screening.
A preponderance of clinical evidence supports a significant public health benefit for prostate-specific antigen (PSA)-based screening and early detection of prostate cancer in appropriately counseled and selected men. Population-based screening with PSA decreases prostate cancer mortality; however, because of relatively poor specificity, PSA-based screening may also increase the detection of clinically insignificant cancers that would otherwise never require treatment. Use of newer biomarkers that increase the specificity for prostate cancer detection may aid in risk stratification and the appropriate identification of men for prostate biopsy. The authors review the 4-kallikrein panel and 4K probability score.


Exploiting intracellular iron and iron-rich compounds to effect tumor cell lysis.
The regimen outlined in this brief paper focuses on therapeutically utilizing iron and iron compounds in transferrin receptor-rich tumor cells in ways that inhibit and lyse same.


MR-optical imaging of cardiovascular molecular targets.
Our understanding of the intricate inflammation biology underlying atherosclerosis is rapidly progressing. Molecular imaging strategies, harnessing this body of knowledge, have been developed to visualize some key cellular and molecular events in plaque evolution and vulnerability. Here, we discuss recent advances in magnetic resonance and fluorescence imaging of key biomarkers including adhesion molecules, inflammatory cells, and enzyme activity. We discuss strengths and limitations of respective imaging technologies, and comment on the potential of multi-modality imaging approaches.


High-resolution computed tomography in hypersensitivity pneumonitis - correlation with pulmonary function.
In 20 patients with chronic form of hypersensitivity pneumonitis (HP) pulmonary function tests (FEV(1), FVC, DCO) and high resolution computed tomography (HRCT) scans were obtained. Extent of pathological changes, i.e. nodularity, lines, honeycombing but not ground-glass estimated by HRCT and expressed as a score correlated significantly with lung function abnormalities.


Fentanyl trauma analgesia use in air medical scene transports.
This study assessed frequency, safety and efficacy of prehospital fentanyl analgesia during 6 months' adult and pediatric helicopter trauma scene transports (213 doses in 177 patients). We reviewed flight records for pain assessment and analgesia provision, effect, and complications. Analgesia was administered to 46/49 (93.9%) intubated patients. In non-intubated patients, pain assessment was documented in 112 of 128 (87.5%), and analgesia was offered, or there was no pain, in 97/128 (75.8%). Of the 67 non-intubated patients to whom analgesia was administered, post-analgesia pain assessment was documented in 62 (92.5%) and pain improved in 53 (79.1% of 67). Post-analgesia blood pressure dropped below 90 torr in 2/177 cases (1.1%, 95% confidence interval [CI] 0.1-4.0%). Post-analgesia S(p)O(2) did not drop below 90% in any patients (95% CI 0-2.3%). In this study, prehospital providers performed well with respect to pain assessment and treatment. Fentanyl was provided frequently, with good effect and minimal cardiorespiratory consequence.


Exosomes originating from infection with the cytoplasmic single-stranded RNA virus Rift Valley fever virus (RVFV) protect recipient cells by inducing RIG-I mediated IFN-B response that leads to activation of autophagy.
Although multiple studies have demonstrated a role for exosomes during virus infections, our understanding of the mechanisms by which exosome exchange regulates immune response during viral infections and affects viral pathogenesis is still in its infancy. In particular, very little is known for cytoplasmic single-stranded RNA viruses such as SARS-CoV-2 and Rift Valley fever virus (RVFV). We have used RVFV infection as a model for cytoplasmic single-stranded RNA viruses to address this gap in knowledge. RVFV is a highly pathogenic agent that causes RVF, a zoonotic disease for which no effective therapeutic or approved human vaccine exist. We show here that exosomes released from cells infected with RVFV (designated as EXi-RVFV) serve a protective role for the host and provide a mechanistic model for these effects. Our results show that treatment of both naïve immune cells (U937 monocytes) and naïve non-immune cells (HSAECs) with EXi-RVFV induces a strong RIG-I dependent activation of IFN-B. We also demonstrate that this strong anti-viral response leads to activation of autophagy in treated cells and correlates with resistance to subsequent viral infection. Since we have shown that viral RNA genome is associated with EXi-RVFV, RIG-I activation might be mediated by the presence of packaged viral RNA sequences. Using RVFV infection as a model for cytoplasmic single-stranded RNA viruses, our results show a novel mechanism of host protection by exosomes released from infected cells (EXi) whereby the EXi activate RIG-I to induce IFN-dependent activation of autophagy in naïve recipient cells including monocytes. Because monocytes serve as reservoirs for RVFV replication, this EXi-RVFV-induced activation of autophagy in monocytes may work to slow down or halt viral dissemination in the infected organism. These findings offer novel mechanistic insights that may aid in future development of effective vaccines or therapeutics, and that may be applicable for a better molecular understanding of how exosome release regulates innate immune response to other cytoplasmic single-stranded RNA viruses.


Levels and trends of HBCD and BDEs in the European and Asian environments, with some information for other BFRs.
In this paper, we review recent data for brominated flame retardants (particularly BDEs, HBCD and TBBP-A) in samples from the European and Asian environments, including recent temporal trend studies. Research is active and we cite over 100 studies published during 2005-2007. Environmental compartments studied comprise the atmosphere, indoor and outdoor air, sewage sludges, soils and sediments and a variety of biological samples and food chains. Findings include that the lack of reference materials for use in analytical quality control and method validation identified earlier has been addressed, and certified concentrations are now available for a number of BDE congeners in six pre-existing and one new reference materials. BDE209 was certified in three samples of indoor dust. The analysis of BDE209 remains problematic, but suitable methods for its determination are available and need to be applied. The contamination of indoor environments with BFRs has been recognised as representing a significant uptake pathway, particularly via dust. Additional data for TBBP-A are needed from areas, where it is produced and used, primarily Asia, as the worst-case scenario. As a reactive flame retardant it is less likely to leach from finished products. Also, issues regarding the importance of e-waste recycling as a source of BFRs to the local populations and the local environment require urgent study, certainly in China and possibly elsewhere. Generally, trends show a levelling in concentrations of BDEs and increases in concentrations of HBCD wherever determined and BDE209 in Asia.


Utilization of a natural tooth in acid-etch bridging.
This report reviews nine cases where despite the wide range of treatments applied to the dentition after a traumatic episode, tooth loss was inevitable and the crown of the damaged tooth was used as a pontic for an immediate bridge. Advantages and disadvantages are reviewed.


Temperature gradient focusing of bio-analyte in a microfluidic channel dealing with non-Newtonian electrolyte considering temperature-dependent zeta potential.
Temperature gradient focusing (TGF) relies on establishing a precise balance between the electrophoretic motility of a target analyte and the advective flow of the background electrolyte (BGE) to locally concentrate the analyte in a microfluidic configuration. This paper presents a finite-element-based numerical analysis where the coupled electric field and the transport equations are solved to describe the effects of the shear-dependent apparent viscosity of a non-Newtonian BGE on the localized concentration buildup of a charged bio-sample inside a microchannel by TGF via Joule heating. Effects of the temperature-dependent nature of the wall zeta potential and the flow behavior index (n) of BGE on the flow, thermal, and species concentration profiles inside the microchannel have been investigated. Study using a fluorescein-Na analyte sample shows that the maximum normalized analyte concentration (Cmax /C0 ) reduces as the zeta potential increases linearly with temperature. The maximum concentration enhancement is achieved when the BGE displays the Newtonian rheology. For example, Cmax /C0 increases 134- to 280-fold when n is increased from 0.8 to 1 (pseudoplastic regime) and again reduces to 190-fold when n increases further from 1 to 1.2 (dilatant regime).


Regulation and possible function of axl expression in immature human mast cells.
The receptor tyrosine kinase Axl which expresses extracellular domains reminiscent of cell adhesion molecules, is involved in homotypic binding as well as in intracellular signaling of myeloid progenitor cells. In order to investigate factors which might influence differentiation pathways through changes of the adhesive properties of cells, we analyzed the expression of axl in immature basophil and mast cell lines and in cultured basophil and mast cell precursors. Axl expression was induced by interferon-alpha in the human leukemic mast cell line HMC-1 and in cultured mast cells derived from CD34+ peripheral blood cells. Axl induction was dose dependent, appeared within 1 h, and was independent of de novo protein synthesis. IFNalpha-treated HMC-1 cells expressing axl formed large cell aggregates within 40 h while untreated cells did not. HMC-1 cells also expressed gas6, the putative ligand of axl, which has been shown to induce axl-mediated homotypic binding. Gas6 expression was independent of interferon treatment in HMC-1 cells. The present results suggest that axl-mediated changes of cellular adhesive properties in mast cells may be important in mast cell differentiation as well as in mast cell-associated inflammation.