[Akt activation mediates estrogen-induced neuroprotection against beta amyloid].
To investigate the effects of estrogen 17beta-estradiol on beta-amyloid protein 25-35 (Abeta25-35) -induced neurotoxicity and possible mechanism thereof. Primary cortical neurons were obtained from the brain of a SD rat and cultured and treated with Abeta25-35 and 17beta-estradiol. I order to investigate the possible mechanism of antagonism of estrogen against the neurotoxicity of Abeta25-35, Akt- I, a specific Akt inhibitor and ICI-182780, an inhibitor of estrogen receptor were added before the addition of estrogen. The Abeta32-35 induced cell viability and lactate dehydrogenase (LDH) release into the cell media was detected with spectrophotometer. Western blotting was used to detect the level of phosphorylated Akt, a cell-protecting factor, and level of nonphosphorylated Akt in different conditions. 20 micromol/L Abeta25-35 decreased the cell viability of the rat cortical neurons to 40.4% (P < 0.01) as compared with the control group and the cell viability of the group with the addition of 17beta-estradiol was 84.2%, significantly higher than that of the Abeta25-35 group (P < 0.01). The LDH secretion level of the Abeta25-35 was 172.5% as high as that of the control group (P < 0.01), while that of the 17beta-estradiol preincubation group was only 118.5% that of the control group (P < 0.01). Both Akt inhibitor and estrogen receptor antagonist partially antagonized estrogen's protective effects against Abeta25-35 in cell viability. Abeta25-35 decreased the phosphorylated Akt level to 69.5% (P < 0.01), while estrogen 17beta-estradiol pretreatment antagonized this effect to 94.7% (P < 0.01), and the addition of Akt inhibitor partially blocked the estrogen's rescue effect (75.4%, P < 0.05). Estrogen treatment alone increased Akt phosphorylation, but this phosphorylation could be partially blocked by the estrogen receptor antagonist, the latter alone had no effect on Akt phosphorylation. Under all circumstances, the amount of nonphosphorylated Akt didn't change significantly. The estrogen 17beta-estradiol partially alleviates the Abeta25-35-induced neurotoxicity and Akt activation may be involved in estrogen's neuroprotective effect. Estrogen receptor may play a role in estrogen induced Akt activation.


Use of the Hollywood Impact Base and standard stationary base to reduce sliding and base-running injuries in baseball and softball.
To determine if the use of the Hollywood Impact Base would significantly reduce the frequency of sliding and base-running injuries to baseball and softball players, we recorded injury information for several teams who used these bases and standard stationary bases. The investigators, under the direction of the Central Michigan University team physician, verified each recorded incident by contacting the injured participant. The sample included interscholastic, intercollegiate, recreational, and intramural softball and baseball teams. There were 33,153 athlete-exposures with the Hollywood Impact Base in use and 3999 with the standard stationary base over a 2-year period (1990 and 1991). There were four injuries that could be directly attributed to the standard stationary base and only one injury that could be attributed to the Hollywood Impact Base. The results (t = -1.94, P < 0.05) verify that using the Hollywood Impact Base significantly reduced the frequency of sliding and base-running injuries. The Hollywood Impact Base injury rate of 0.08% per game compares favorably with the reported breakaway base injury rate. The results of our study verify that the Hollywood Impact Base is similarly effective in reducing the risk of base-related injuries.


The Polymorphisms of Ser49Gly and Gly389Arg in Beta-1-Adrenergic Receptor Gene in Major Depression.
It was reported that the genetic susceptibility of major depressive disorder (MDD) is related with genetic polymorphisms. The aim of this study was to investigate the possible association of the genotype and allele frequencies of Ser49Gly and Arg389Gly polymorphisms in MDD by comparing them with healthy subjects. A total of 144 patients with MDD diagnosed according to Diagnostic and Statistical Manual of Mental Disorders, 4th Edition (DSM-IV) criteria and 105 healthy controls were included in the study. Polymerase chain reaction (PCR) with restriction fragment length polymorphism (RFLP) was used for genotyping. Of the 144 participants in the MDD group, 77 (53.5%) had homozygous wild type (AA), 57 (39.6%) had heterozygous type (AG), and 10 (6.9%) had mutant (GG) genotype for Ser49Gly, whereas 75 (52.1%) had homozygous wild type (GG), 59 (41.0%) had heterozygous (GC) type, and 10 (6.9%) had mutant homozygous (CC) genotype for Gly386Arg. There were no significant difference in the allele and genotype frequencies of the beta-1-adrenergic receptor (ADRB1) gene for Ser49Gly and Arg389Gly polymorphisms after comparing with healthy controls (p=0.626; p=0.863 and p=0.625; p=0.914). The results of our study did not reveal a major effect of the polymorphism of Ser49Gly and Gly389Arg in the ADRB1 gene in MDD. Further studies with larger sample size are required to elucidate the role of other beta-1 adrenergic gene polymorphisms in MDD.


Spatial rearrangement of Purkinje cell subsets forms the transverse and longitudinal compartmentalization in the mouse embryonic cerebellum.
Transversely oriented lobules and longitudinally arrayed stripes of Purkinje cell subsets subdivide the cerebellar cortex into multiple compartments that are involved in diverse functions. In the mammalian cerebellum, anterior, and posterior lobules, which are involved in somatosensorimotor function, show an alternation of aldolase C (zebrin II) -positive and -negative stripes, whereas the central lobules (lobules VIb-VII and crus I), which are implicated in nonmotor functions, show a laterally expanded arrangement solely of aldolase C-positive stripes. To understand the developmental process of this compartmental pattern, we identified groups of Purkinje cell subsets in the entire mouse cerebellum at embryonic day (E) 14.5 by staining Purkinje cell subset markers. We then tracked four major domains of Protocadherin 10 (Pcdh10)-positive Purkinje cell subsets (medial, dorsal, central, and mid-lateral subsets), which were clearly demarcated during E14.5-17.5. These domains of Purkinje cell subsets shifted predominantly in the longitudinal direction to be positioned in the anterior and posterior lobules. However, a particular portion of the medial and mid-lateral domains, and the whole of the central domain shift in the lateral direction to be positioned in the central lobules. The results indicate that while the longitudinal shift of domains of Purkinje cell subsets forms the longitudinally striped compartments in the anterior and posterior cerebellum, the lateral shift of particular domains of Purkinje cell subsets underlies the laterally expanded arrangement of stripes in central lobules. Thus, the rearrangement of Purkinje cell subsets in the embryonic cerebellum is critically related to the compartmental organization in the mammalian cerebellum.


A single intranasal administration of virus-like particle vaccine induces an efficient protection for mice against human respiratory syncytial virus.
Human respiratory syncytial virus (RSV) is an important pediatric pathogen causing acute viral respiratory disease in infants and young children. However, no licensed vaccines are currently available. Virus-like particles (VLPs) may bring new hope to producing RSV VLP vaccine with high immunogenicity and safety. Here, we constructed the recombinants of matrix protein (M) and fusion glycoprotein (F) of RSV, respectively into a replication-deficient first-generation adenoviral vector (FGAd), which were used to co-infect Vero cells to assemble RSV VLPs successfully. The resulting VLPs showed similar immunoreactivity and function to RSV virion in vitro. Moreover, Th1 polarized response, and effective mucosal virus-neutralizing antibody and CD8+ T-cell responses were induced by a single intranasal (i.n.) administration of RSV VLPs rather than intramuscular (i.m.) inoculation, although the comparable RSV F-specific serum IgG and long-lasting RSV-specific neutralizing antibody were detected in the mice immunized by both routes. Upon RSV challenge, VLP-immunized mice showed increased viral clearance but decreased signs of enhanced lung pathology and fewer eosinophils compared to mice immunized with formalin-inactivated RSV (FI-RSV). In addition, a single i.n. RSV VLP vaccine has the capability to induce RSV-specific long-lasting neutralizing antibody responses observable up to 15 months. Our results demonstrate that the long-term and memory immune responses in mice against RSV were induced by a single i.n. administration of RSV VLP vaccine, suggesting a successful approach of RSV VLPs as an effective and safe mucosal vaccine against RSV infection, and an applicable and qualified platform of FGAd-infected Vero cells for VLP production.


In situ high-pressure study of sodium amide by Raman and infrared spectroscopies.
Here we report the first in situ high-pressure study of sodium amide (NaNH(2)) as an agent in potential hydrogen storage applications by using combined Raman and infrared (IR) spectroscopies at room temperature and pressures up to ~16 GPa. Starting with an orthorhombic crystal structure at ambient pressure, sodium amide was found to transform to two new phases upon compression as evidenced by the changes in the characteristic Raman and IR modes as well as by examining the pressure dependences of these modes. Raman and IR measurements on NaNH(2) collectively provided consistent information about the structural evolutions of NaNH(2) under compression. Upon decompression, all Raman and IR modes were completely recovered, indicating the reversibility of the pressure-induced transformations in the entire pressure region. The combined Raman and IR spectroscopic data together allowed for the analysis of possible structures of the new high-pressure phases of NaNH(2).


Effect of sodium-glucose transporter 2 inhibitors on sarcopenia in patients with type 2 diabetes mellitus: a systematic review and meta-analysis.
Sarcopenia has been recognized as the third category of disabling complications in patients with type 2 diabetes mellitus(T2DM), in addition to micro- and macrovascular complications. Sodium-glucose co-transporter 2 (SGLT2) inhibitors are innovative glucose-lowering treatments that have been shown to reduce body weight and enhance cardiovascular and renal outcomes. However, there is vigilance that SGLT2 inhibitors should be taken cautiously because they target skeletal muscle and may raise the risk of sarcopenia. Herein, we conducted a meta-analysis of randomized controlled trials to evaluate the effects of SGLT2 inhibitors on sarcopenia in patients with T2DM. Relevant studies were obtained from PubMed, Embase, Medicine, Cochrane, and Web of Science databases to determine eligible studies until February 2023, without any language restrictions. A random effects model was utilized irrespective of heterogeneity, and the I2 statistic was used to evaluate study heterogeneity. The differences in results were measured using the weighted average difference (WMD) of the continuous data, along with a 95% confidence interval (CI). A total of 25 randomized controlled trials with 2,286 participants were included. SGLT2 inhibitors significantly reduced weight-related changes and fat-related changes, including body weight(BW) (WMD= -2.74, 95% CI: -3.26 to -2.23, P<0.01), body mass index(BMI) (WMD= -0.72, 95% CI: -0.95 to -0.49, P<0.01), waist circumference(WC) (WMD= -1.60, 95% CI: -2.99 to -0.22, P=0.02), fat mass(FM)(WMD= -1.49, 95% CI: -2.18 to -0.80, P<0.01), percentage body fat(PBF) (WMD= -1.28, 95% CI: -1.83 to -0.74, P<0.01), visceral fat area(VFA)(WMD= -19.52, 95% CI: -25.90 to -13.14, P<0.01), subcutaneous fat area(SFA)(WMD= -19.11, 95% CI: -31.18 to -7.03, P=0.002), In terms of muscle-related changes, lean mass(LM)(WMD= -0.80, 95% CI: -1.43 to -0.16, P=0.01), and skeletal muscle mass(SMM) (WMD= -0.38, 95% CI: -0.65 to -0.10, P=0.007), skeletal muscle index(SMI) (WMD= -0.12, 95% CI: -0.22 to -0.02, P=0.02)were also significantly reduced. In addition, body water likewise decreased significantly (WMD=-0.96, 95% CI: -1.68 to -0.23, P=0.009). As one of the most widely used hypoglycemic, SGLT2 inhibitors have beneficial effects on FM and BW weight loss in T2DM, such as BW, BMI, WC, FM, PBF, VFA, and SFA. However, the negative influence on muscle mass paralleled the reduction in FM and BW, and the consequent increased risk of sarcopenia warrants high attention, especially as patients are already predisposed to physical frailty. https://www.crd.york.ac.uk/prospero/#myprospero, identifier PROSPERO (No.CRD 42023396278).


Curcumin-induced histone acetylation inhibition improves stress-induced gastric ulcer disease in rats.
Curcumin is known to possess anti‑inflammatory properties. Despite the fact that curcumin is known to be a strong inhibitor of H+, K+‑ATPase activity, the mechanism underlying the curcumin‑induced inhibition of the transcription of the H+, K+‑ATPase α subunit in gastric mucosal parietal cells remains unclear. The present study investigated the possible mechanism by which curcumin inhibits stomach H+, K+‑ATPase activity during the acute phase of gastric ulcer disease. A rat model of stress‑induced gastric ulcers was produced, in which the anti‑ulcer effects of curcumin were examined. Curcumin‑induced inhibition of the H+, K+‑ATPase promoter via histone acetylation, was verified using a chromatin immunoprecipitation assay. The results showed that curcumin improved stress‑induced gastric ulcer disease in rats, as demonstrated by increased pH values and reduced gastric mucosal hemorrhage and ulcer index. These effects were accompanied by a significant reduction in the level of histone H3 acetylation at the site of the H+, K+‑ATPase promoter and in the expression of the gastric H+,K+‑ATPase α subunit gene and protein. In conclusion, curcumin downregulated the acetylation of histone H3 at the site of the H+, K+‑ATPase promoter gene, thereby inhibiting the transcription and expression of the H+, K+‑ATPase gene. Curcumin was shown to have a preventive and therapeutic effect in gastric ulcer disease.


Effects of clonidine and dopamine on gastric tone.
The effects of clonidine and dopamine, both alone and together, on gastric tone were studied using an electronic barostat. This enabled volume changes to be measured in an intragastric bag with a constant preset pressure. Nine healthy male volunteers were each studied on two occasions in a randomized order. During each study period, a continuous infusion of dopamine was given, starting with a dose of 2.5 microg kg(-1) min(-1), and then increasing at 15-min intervals to 5.0 and 7.5 microg kg(-1) min(-1). Clonidine 150 microg intravenously was given on one occasion during the infusion of dopamine (7.5 microgkg(-1) min(-1)) and on the other occasion 15 min before the dopamine infusion started. During dopamine infusion, the intragastric bag volume increased (gastric tone therefore decreasing) in a dose-related manner (total increase 290 +/- 114 mL). Clonidine given either during or before dopamine infusion did not influence the bag volume. When the dopamine infusion started 15 min after clonidine, the bag volume did not change until the infusion of dopamine reached 7.5 microg kg(-1) min(-1) (total increase 205 +/- 156 mL). Dopamine reduced gastric tone in a dose-related manner, and clonidine did not influence gastric tone per se. If clonidine is given before dopamine, the effects of dopamine are reduced.


Infection with enterovirus 71 or expression of its 2A protease induces apoptotic cell death.
Enterovirus 71 (EV71) is the causative agent of human diseases with distinct severity, from mild hand-foot-and-mouth disease to severe neurological syndromes, such as encephalitis and meningitis. Infection of several different cell lines with EV71 causes extensive cytopathic effect, leading to destruction of the entire monolayer and the death of infected cells. In this study, cell death processes during EV71 infection and the underlying mechanisms of them were investigated. The hallmarks of apoptosis, nuclear condensation and fragmentation, were observed 24 h after infection. Apoptosis in infected cells was also confirmed by detectable cleavage of cellular DNA and degradation of poly(ADP-ribose) polymerase. Transient expression of EV71 2A protease (2A(pro)) alone resulted in the induction of apoptotic change. Infection of EV71 or expression of EV71 2A(pro) leads to cleavage of the eukaryotic initiation factor 4GI, a key factor for host protein synthesis. This study added one more example to the growing list of human viruses that induce apoptosis by a virus-encoded protein.


The spleen in liver cirrhosis: revisiting an old enemy with novel targets.
The spleen is a secondary lymphoid organ which can influence the progression of multiple diseases, notably liver cirrhosis. In chronic liver diseases, splenomegaly and hypersplenism can manifest following the development of portal hypertension. These splenic abnormalities correlate with and have been postulated to facilitate the progression of liver fibrosis to cirrhosis, although precise mechanisms remain poorly understood. In this review, we summarize the literature to highlight the mechanistic contributions of splenomegaly and hypersplenism to the development of liver cirrhosis, focusing on three key aspects: hepatic fibrogenesis, hepatic immune microenvironment dysregulation and liver regeneration. We conclude with a discussion of the possible therapeutic strategies for modulating splenic abnormalities, including the novel potential usage of nanomedicine in non-surgically targetting splenic disorders for the treatment of liver cirrhosis.


In vitro differentiation of chicken embryo skin cells transformed by Rous sarcoma virus.
The epidermal cells isolated from 14-day chicken embryo shank skin epidermis were infected in vitro with Rous sarcoma virus (RSV). Within a few weeks, rapidly growing colonies of epithelial cells appeared among the sea of transformed fibroblastic cells. When isolated and subcultured, these cells were found to possess typical markers of skin epidermis. The presence of major keratin and typical epithelial cell type morphology strongly suggested that these cells were transformed epidermal cells retaining their differentiated characteristics but having the capacity to propagate in cell culture. If RSV tsNY68, an RSV mutant having a temperature lesion in the src gene, was used, similar transformed epidermal cells were obtained at 36 degrees C (permissive temperature). At the nonpermissive temperature (41 degrees C) the growth rate of these cells decreased and additional keratin species appeared. At 41 degrees C the cells were flattened and lost the refractivity in their peripheries. All the keratins which are synthesized at the nonpermissive temperature were present in normal differentiated shank skin of 19-day old chick embryo. These cells also had "cornified envelop," indicating extensive differentiation. Viral production was as efficient as transformed fibroblasts during the rapid growth phase, while it declined significantly after the cells reached confluency, exhibiting the differentiated characteristics. Since no normal epidermal cells could be cultured under our experimental conditions, these results represent examples in which the src gene is essential for propagation of differentiated cells in cell culture while it abolishes only a part of differentiated characteristics.


Loss of FHIT protein expression is a marker of adverse evolution in good prognosis localized breast cancer.
The FHIT tumor suppressor gene, which encompasses the fragile site FRA3B at 3p14.2, is altered frequently in many types of human cancers. To determine its importance as a prognostic marker in breast cancer, the expression of the FHIT protein was studied in a series of 452 breast carcinomas by using immunohistochemistry on sections of tissue microarrays. Three distinct levels of FHIT expression were observed: in 154 cases (34.1%) expression was unchanged as compared to normal level; in 78 (17.2%) no expression was found; in the remaining 220 cases (48.7%), expression was intermediate. Overall, two-thirds of the cases had abnormal levels of the protein. Absence of FHIT was significantly associated with a higher grade (p < 0.01) and absence of hormone receptors (p < 0.001). The patients were separated into Group I (153 node-negative good prognosis patients who did not receive adjuvant chemotherapy) and Group II (226 high-risk patients treated by adjuvant chemotherapy) according to the St.-Gallen conference consensus. The median follow-up was 48 months. Among Group I but not Group II patients, a multivariate analysis showed that FHIT expression was significantly associated with disease-free survival. The relative risk of recurrence for FHIT-negative Group I patients was 2.37 (1.21-4.64; p = 0.03). Thus, among the patients who present with tumors of apparent good prognosis, FHIT is an independent prognostic factor that distinguishes a subgroup of patients who could benefit from adjuvant treatment.


Polymorphism of HLA class I genes in the Brazilian population from the Northeastern State of Pernambuco corroborates anthropological evidence of its origin.
The allelic distribution of human leukocyte antigen (HLA) class I genes (HLA-A, HLA-B, and HLA-Cw) of the population from the State of Pernambuco in Northeastern Brazil was studied in a sample of 101 healthy unrelated individuals. Low to medium resolution HLA class I typing was performed using polymerase chain reaction-amplified DNA hybridized to sequence specific primers (PCR-SSPs). Twenty allele groups were detected for HLA-A, 28 for HLA-B, and 14 for HLA-Cw. The most frequent alleles were HLA-A*02(0.2871), HLA-B*15(0.1238), and HLA-Cw*04(0.2277), and the most frequent genotypes were A*02/A*02(0.0990), B*15/B*15(0.0594), and Cw*04/Cw*04 and Cw*07/Cw*07, both with a frequency of 0.0792. The observed heterozygosity for the studied loci was 79.21% for HLA-A, 87.13% for HLA-B, and 77.23% for HLA-Cw. The most frequent haplotype was A*02-Cw*04-B*35(0.0485), which is also present in Western European, Amerindian, and Brazilian Mulatto populations, but absent in African populations. Taken together, these data corroborate the historic anthropological evidences of the origin of the Northeastern Brazilian population from Pernambuco.


The Increasing Role of Short-Term Sperm Storage and Cryopreservation in Conserving Threatened Amphibian Species.
Multidisciplinary approaches to conserve threatened species are required to curb biodiversity loss. Globally, amphibians are facing the most severe declines of any vertebrate class. In response, conservation breeding programs have been established in a growing number of amphibian species as a safeguard against further extinction. One of the main challenges to the long-term success of conservation breeding programs is the maintenance of genetic diversity, which, if lost, poses threats to the viability and adaptive potential of at-risk populations. Integrating reproductive technologies into conservation breeding programs can greatly assist genetic management and facilitate genetic exchange between captive and wild populations, as well as reinvigorate genetic diversity from expired genotypes. The generation of offspring produced via assisted fertilisation using frozen-thawed sperm has been achieved in a small but growing number of amphibian species and is poised to be a valuable tool for the genetic management of many more threatened species globally. This review discusses the role of sperm storage in amphibian conservation, presents the state of current technologies for the short-term cold storage and cryopreservation of amphibian sperm, and discusses the generation of cryo-derived offspring.


[Experimental testing of Pflüger's reflex hypothesis of menstruation in late 19th century].
Pflüger's hypothesis of a nerve reflex as the cause of menstruation published in 1865 and accepted by many, nonetheless did not lead to experimental investigations for 25 years. According to this hypothesis the nerve reflex starts in the ovary by an increase of the intraovarian pressure by the growing follicles. In 1884 Adolph Kehrer proposed a program to test the nerve reflex, but only in 1890, Cohnstein artificially increased the intraovarian pressure in women by bimanual compression from the outside and the vagina. His results were not convincing. Six years later, Strassmann injected fluids into ovaries of animals and obtained changes in the uterus resembling those of oestrus. His results seemed to verify a prognosis derived from Pflüger's hypothesis. Thus, after a long interval, that hypothesis had become a paradigma. Though reasons can be given for the delay, it is little understood, why experimental testing started so late.


The Role of miRNA for the Treatment of MGMT Unmethylated Glioblastoma Multiforme.
Glioblastoma multiforme (GBM) is the most common high-grade intracranial tumor in adults. It is characterized by uncontrolled proliferation, diffuse infiltration due to high invasive and migratory capacities, as well as intense resistance to chemo- and radiotherapy. With a five-year survival of less than 3% and an average survival rate of 12 months after diagnosis, GBM has become a focus of current research to urgently develop new therapeutic approaches in order to prolong survival of GBM patients. The methylation status of the promoter region of the O6-methylguanine-DNA methyltransferase (MGMT) is nowadays routinely analyzed since a methylated promoter region is beneficial for an effective response to temozolomide-based chemotherapy. Furthermore, several miRNAs were identified regulating MGMT expression, apart from promoter methylation, by degrading MGMT mRNA before protein translation. These miRNAs could be a promising innovative treatment approach to enhance Temozolomide (TMZ) sensitivity in MGMT unmethylated patients and to increase progression-free survival as well as long-term survival. In this review, the relevant miRNAs are systematically reviewed.


The occurrence of auto-antibodies in patients with gastro-duodenal lesions.
The occurrence of auto-antibodies in patients with the autoimmune disease pernicious anaemia and in patients with active duodenal ulcers was investigated. In order to characterize antigenic structures, various cellular and subcellular fractions were prepared from pig gastric mucosa and from a homogenate of duodenal mucosa. By means of an enzyme-linked immunosorbent assay and immunoblotting, both the H+,K(+)-ATPase and pepsinogen/pepsin were shown to constitute the major antigens. All of the seven pernicious-anaemia sera that were tested contained auto-antibodies against both antigens, and the epitopes of the H+,K(+)-ATPase were shown to be localized on its cytoplasmic face. In 75% (18/24) of the sera from patients with duodenal ulcers, auto-antibodies were detected when using purified antigens. Six sera reacted with H+,K(+)-ATPase and twelve reacted with pepsinogen, one reacted with both antigens, and four sera reacted with the duodenal mucosal antigen. The occurrence of auto-antibodies indicates that there is a mucosal lesion and that immunological factors may be involved in the pathogenesis of the disease in some patients.


Artificial intelligence-based decision-making for age-related macular degeneration.
Artificial intelligence (AI) based on convolutional neural networks (CNNs) has a great potential to enhance medical workflow and improve health care quality. Of particular interest is practical implementation of such AI-based software as a cloud-based tool aimed for telemedicine, the practice of providing medical care from a distance using electronic interfaces. Methods: In this study, we used a dataset of labeled 35,900 optical coherence tomography (OCT) images obtained from age-related macular degeneration (AMD) patients and used them to train three types of CNNs to perform AMD diagnosis. Results: Here, we present an AI- and cloud-based telemedicine interaction tool for diagnosis and proposed treatment of AMD. Through deep learning process based on the analysis of preprocessed optical coherence tomography (OCT) imaging data, our AI-based system achieved the same image discrimination rate as that of retinal specialists in our hospital. The AI platform's detection accuracy was generally higher than 90% and was significantly superior (p < 0.001) to that of medical students (69.4% and 68.9%) and equal (p = 0.99) to that of retinal specialists (92.73% and 91.90%). Furthermore, it provided appropriate treatment recommendations comparable to those of retinal specialists. Conclusions: We therefore developed a website for realistic cloud computing based on this AI platform, available at https://www.ym.edu.tw/~AI-OCT/. Patients can upload their OCT images to the website to verify whether they have AMD and require treatment. Using an AI-based cloud service represents a real solution for medical imaging diagnostics and telemedicine.


Real-time kinetics of high-mobility group box 1 (HMGB1) oxidation in extracellular fluids studied by in situ protein NMR spectroscopy.
Some extracellular proteins are initially secreted in reduced forms via a non-canonical pathway bypassing the endoplasmic reticulum and become oxidized in the extracellular space. One such protein is HMGB1 (high-mobility group box 1). Extracellular HMGB1 has different redox states that play distinct roles in inflammation. Using a unique NMR-based approach, we have investigated the kinetics of HMGB1 oxidation and the half-lives of all-thiol and disulfide HMGB1 species in serum, saliva, and cell culture medium. In this approach, salt-free lyophilized (15)N-labeled all-thiol HMGB1 was dissolved in actual extracellular fluids, and the oxidation and clearance kinetics were monitored in situ by recording a series of heteronuclear (1)H-(15)N correlation spectra. We found that the half-life depends significantly on the extracellular environment. For example, the half-life of all-thiol HMGB1 ranged from ~17 min (in human serum and saliva) to 3 h (in prostate cancer cell culture medium). Furthermore, the binding of ligands (glycyrrhizin and heparin) to HMGB1 significantly modulated the oxidation kinetics. Thus, the balance between the roles of all-thiol and disulfide HMGB1 proteins depends significantly on the extracellular environment and can also be artificially modulated by ligands. This is important because extracellular HMGB1 has been suggested as a therapeutic target for inflammatory diseases and cancer. Our work demonstrates that the in situ protein NMR approach is powerful for investigating the behavior of proteins in actual extracellular fluids containing an enormous number of different molecules.


Maternal diet and cord blood leptin and adiponectin concentrations at birth.
The purpose of this study was to determine the effects of total energy intake, macronutrient intake, and maternal adherence to Mediterranean diet or Alternative Healthy Eating Index (AHEI) on cord blood leptin and adiponectin levels, which have been associated with childhood adiposity. We used multivariable linear regression to assess associations of maternal diet, averaged over 1st and 2nd trimesters, with cord blood adipokines of 780 women from the prospective cohort study Project Viva. Mean (SD) energy intake during pregnancy was 2135 (596) kcal. Mean (SD) cord blood levels of leptin and adiponectin were 9.0 (6.6) ng/ml and 28.6 (6.7) μg/ml, respectively. Neither closer adherence to a Mediterranean/AHEI pattern diet nor energy intake was associated with either cord blood leptin or adiponectin. Protein intake was associated with both marginally lower leptin (-0.22 ng/ml [95% CI -0.41, -0.02] for each 1% of energy) and adiponectin (-0.25 μg/ml [95% CI -0.48, -0.02]). Closer adherence to a Mediterranean/AHEI pattern diet during pregnancy was not associated with cord blood leptin or adiponectin. Maternal protein intake was weakly but significantly associated with lower cord blood leptin and adiponectin.


Poor sequestration of toxic host plant cardenolides and their rapid loss in the milkweed butterfly Danaus chrysippus (Lepidoptera: Nymphalidae: Danainae: Danaini).
Butterflies of the genus Danaus are known to sequester toxic cardenolides from milkweed host plants (Apocynaceae). In particular, Danaus plexippus efficiently sequesters and stores these compounds, whereas D. chrysippus, is considered to poorly sequester cardenolides. To estimate its sequestration capability compared with that of D. plexippus, larvae of both species were jointly reared on Asclepias curassavica and the major cardenolides of the host plant, calotropin and calactin, were analyzed in adults sampled at different time intervals after eclosion. Both cardenolides were detected in body and wings of D. plexippus. Whereas the calotropin-concentration remained constant over a period of 24 days, that of calactin steadily decreased. In the body, but not in the wings of D. chrysippus, calactin only was detected in low amounts, which was then almost completely lost during the following 8 days after eclosion, suggesting that in contrast to D. plexippus, cardenolides seem to be less important for that butterfly's defence against predators.


Indomethacin lowers optic nerve oxygen tension and reduces the effect of carbonic anhydrase inhibition and carbon dioxide breathing.
Prostaglandins are important in blood flow regulation. Carbon dioxide (CO(2)) breathing and carbonic anhydrase inhibition increase the oxygen tension in the retina and optic nerve. To study the mechanism of this effect and the role of cyclo-oxygenase in the regulation of optic nerve oxygen tension (ONPO(2)), the authors investigated how indomethacin affects ONPO(2) and the ONPO(2) increases caused by CO(2) breathing and carbonic anhydrase inhibition in the pig. Optic nerve oxygen tension was measured in 11 pigs with a polarographic oxygen electrode. The tip of the electrode was placed 0.5 mm above the optic disc. The effects of indomethacin, CO(2) breathing (3%) before and after indomethacin treatment, and carbonic anhydrase inhibition with or without indomethacin treatment were investigated. Administration of 300 mg indomethacin decreased optic nerve oxygen tension significantly. Carbonic anhydrase inhibition and CO(2) breathing increased ONPO(2) significantly. After indomethacin had been given, the rise in ONPO(2) caused by CO(2) breathing and carbonic anhydrase inhibition was significantly reduced. Systemic administration of indomethacin decreases the optic nerve oxygen tension; this is probably the result of decreased blood flow through vasoconstriction of vessels in the optic nerve. Additionally, indomethacin diminishes the ONPO(2) increasing effect of CO(2) breathing and carbonic anhydrase inhibition, thus affecting the reactivity of vessels in the optic nerve.


[Changes in occupational morbidity in major economic branches of Bashkortostan Republic over 2002-2006].
The authors analyzed occupational morbidity in major economic branches of Bashkortostan Republic over 2002-2006, in comparison with Russian Federation, performed ranking of the economic branches according to the share of accumulated occupational morbidity, gave brief hygienic evaluation of work conditions.


Reconstitution of an insulin signaling pathway in Xenopus laevis oocytes: coexpression of a mammalian insulin receptor and three different mammalian hexose transporters.
We report the functional expression of the mammalian muscle-adipocyte insulin-sensitive hexose transporter in Xenopus laevis oocytes. Oocytes microinjected with RNA synthesized in vitro showed enhanced hexose transport activity compared with uninjected controls. However, like the endogenous oocyte hexose transporter, activity was stimulated only twofold by 1 microM insulin. X. laevis oocytes injected with in vitro-synthesized RNA encoding the human insulin proreceptor expressed a functionally active insulin receptor that enhanced the insulin sensitivity of injected oocytes. This increase was not observed in oocytes expressing a mutant insulin receptor that lacked protein tyrosine kinase activity. In the presence of the coexpressed human insulin receptor, insulin induced a two- to threefold increase in hexose transport. The muscle-, brain-, and liver-type hexose carriers normally expressed in tissues with different responses to insulin exhibited the same insulin sensitivity when expressed in oocytes. This was observed whether or not the insulin signal was transduced through a coexpressed human insulin receptor or the endogenous oocyte insulin-like growth factor I receptor. We conclude that the expressed human insulin receptor is able to couple efficiently with preexisting postreceptor regulatory pathways in oocytes and that the regulation of hexose transport in these cells can be mediated through the combined actions of the expressed human insulin receptor and the endogenous oocyte insulin-like growth factor I receptor.


Agrobacterium infection and plant defense-transformation success hangs by a thread.
The value of Agrobacterium tumefaciens for plant molecular biologists cannot be appreciated enough. This soil-borne pathogen has the unique capability to transfer DNA (T-DNA) into plant systems. Gene transfer involves both bacterial and host factors, and it is the orchestration of these factors that determines the success of transformation. Some plant species readily accept integration of foreign DNA, while others are recalcitrant. The timing and intensity of the microbially activated host defense repertoire sets the switch to "yes" or "no." This repertoire is comprised of the specific induction of mitogen-activated protein kinases (MAPKs), defense gene expression, production of reactive oxygen species (ROS) and hormonal adjustments. Agrobacterium tumefaciens abuses components of the host immunity system it mimics plant protein functions and manipulates hormone levels to bypass or override plant defenses. A better understanding of the ongoing molecular battle between agrobacteria and attacked hosts paves the way toward developing transformation protocols for recalcitrant plant species. This review highlights recent findings in agrobacterial transformation research conducted in diverse plant species. Efficiency-limiting factors, both of plant and bacterial origin, are summarized and discussed in a thought-provoking manner.


Behavioral abnormalities precede neuropathological markers in rats transgenic for Huntington's disease.
Huntington's disease (HD) is caused by an expanded CAG repeat leading to the synthesis of an aberrant protein and to the formation of polyglutamine (polyQ)-containing inclusions and aggregates. Limited information is available concerning the association of neuropathological markers with the development of behavioral markers in HD. Using a previously generated transgenic rat model of HD (tgHD rat), we performed association studies on the time-course of behavioral symptoms (motor function, learning, anxiety) and the appearance of striatal atrophy, 1C2 immunopositive aggregates and polyQ recruitment sites, a precursor to these aggregates. At the age of 1 month, tgHD rats exhibited reduced anxiety and improved motor performance, while at 6 months motor impairments and at 9 months cognitive decline occurred. In contrast, polyQ recruitment sites appeared at around 6-9 months of age, indicating that HD-like behavioral markers preceded the appearance of currently detectable neuropathological markers. Interestingly, numerous punctate sites containing polyQ aggregates were also seen in areas receiving afferents from the densely recruiting regions suggesting either transport of recruitment-competent aggregates to terminal projections where initially 1C2 positive aggregates were formed or different internal properties of neurons in different regions. Furthermore, striatal atrophy was observed at the age of 12 months. Taken together, our findings support the hypothesis of a dynamic process leading to region- and age-specific polyQ recruitment and aggregation. The dissociation of onset between behavioral and neuropathological markers is suggestive of as yet undetected processes, which contribute to the early phenotype of these HD transgenic rats.


Effects of morphine and respiratory depression on sulfobromophthalein disposition in rats.
Morphine, 20 mg X kg-1, sc, halved the plasma clearance of sulfobromophthalein (BSP) while tripling hepatic tissue levels of this dye. Since narcotics depress respiration, effects of hypoxia, hypercapnia, and acidosis on BSP disposition were studied. Ambient gases breathed by rats were adjusted to achieve blood gas levels identical to those of morphine-induced respiratory depression. Saline-treated rats breathing room air had PAO2 of 87 +/- 3 mmHg (mean +/- SE) and PaCO2 of 40 +/- 2 mmHg. After intraarterial injection of BSP, 100 mg X kg-1, plasma clearance of this dye was 7.1 +/- 1.1 ml X min-1 and BSP levels in the liver at 40 min after injection were 163.3 +/- 19.8 micrograms X g-1. After morphine, 20 mg X kg-1, PaO2 decreased to 47 +/- 4 mmHg and PaCO2 increased to 89 +/- 5 mmHg. In these rats BSP clearance dropped to 3.5 +/- 0.4 ml X min-1, and 40-min liver dye levels were increased to 596.4 +/- 60.4 micrograms X g-1. Similar hypoxia and hypercapnia caused by breathing 9% O2 and 8% CO2 in the absence of morphine caused plasma BSP clearance to be decreased to 4.4 +/- 0.2 ml X min-1 and 40-min hepatic BSP to be increased to 292.5 +/- 31.8 micrograms X g-1. Hypercapnia and acidosis alone did not affect BSP disposition, while hypoxia without hypercapnia decreased its plasma clearance to 5.5 +/- 0.3 ml X min-1 and increased liver levels to 339.1 +/- 35.1 micrograms X g-1. Hypoxia was reversed completely in morphine-treated rats by placing them in 40% O2. In these animals, despite normal oxygen, plasma BSP clearance was decreased to 4.4 +/- 0.6 ml X min-1, and liver BSP was increased to 497.9 +/- 65.6 micrograms X g-1. Thus, respiratory depression with hypoxia may contribute to morphine-induced effects on BSP disposition, but altered blood gases cannot account fully for these narcotic effects.


A carbon-centered radical as a reaction species in the DNA strand-breakage by D-glucosamine.
Electron spin resonance(ESR) studies using 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and sodium 3,5-dibromo-4-nitrosobenzensulfonate (DBNBS) as a spin-trapping agent revealed the formation of both hydroxyl and carbon-centered radical-derived spin adducts in Cu2+-containing 50 mM Tris-HCl buffered solutions (pH 7.1) of D-glucosamine, D-mannosamine, and D-galactosamine, which were previously shown to have the ability to break the single-strand of plasmid pBR322 DNA in a nucleotide sequence-specific manner. HCl-free D-glucosamine has higher DNA breaking activity, and this activity is promoted more by the presence of Cu2+ than the original D-glucosamine hydrochloride, exhibits stronger radical signals in the ESR spectrum. It is suggested that D-glucosamine is unstable around neutral pH, being converted into certain intermediate(s) such as a dihydropyrazine compound, which generate(s) carbon-centered radicals, and that, besides the hydroxyl radical, the intermediate(s) is/are responsible for DNA strand breakage.


An Alternative Technique for Monitoring the Live Interaction of Monocytes and Tumor Cells with Nanoparticles in the Mouse Lung.
Nanomaterials are increasingly used for the diagnosis and treatment of cancer, including lung cancer. For the clinical translation of nano-based theranostics, it is vital to detect and monitor their accumulation in the tumor, as well as their interaction with tumor, immune cells, and the tumor microenvironment (TME). While high resolution microscopy of fixed tumor specimens can provide some of this information from individual thin slices, it cannot capture cellular events over time and lacks 3D information of the tumor tissue. On the other hand, in vivo optical procedures either fall short of providing the necessary cellular resolution, as in the case of epifluorescence optical imaging, or are very demanding, as for instance intravital lung microscopy. We describe an alternative approach to investigate nanoparticle-cell interactions in entire mouse lung lobes, by longitudinal live cell confocal microscopy at nanometer resolution. By filling the lung ex vivo with 1% agarose, we were able to stabilize the lung lobes and visualize the interaction of fluorescent cells and nanoparticles for at least 4 hours post mortem. This high resolution ex vivo live cell imaging approach is an easy 4D tool for assessing several dynamic processes in tumor tissue, such as the traffic of cells, shedding of extracellular vesicles (EVs), and the accumulation of nanoparticles in tumor tissue. Graphic abstract: Schematic of the workflow for live cell imaging in the mouse lung.


Pharmacogenetics of capecitabine in advanced breast cancer patients.
Germinal gene polymorphisms can explain a part of the interpatient pharmacodynamic variability of anticancer drugs, particularly fluoropyrimidines. Genes for which polymorphisms may potentially influence pharmacodynamics of fluoropyrimidines, including capecitabine, are thymidylate synthase (TS), methylenetetrahydrofolate reductase (MTHFR), and dihydropyrimidine dehydrogenase (DPD). The aim of this prospective pilot study was to analyze the effect of TS, MTHFR, and DPD gene polymorphisms on toxicity and efficacy in advanced breast cancer patients receiving capecitabine as monotherapy. Germinal polymorphisms of TS (6 bp deletion in the 3' region and 28 bp repeats, including G>C mutation in the 5' region), MTHFR (677C>T and 1298A>C), and DPD (IVS14+1G>A) were determined in 105 consecutive patients. A trend toward a higher global toxicity grade 3 and 4 was observed in patients homozygous for the TS 3RG allele compared with patients heterozygous for the 3RG allele or patients not carrying the 3RG allele (50% versus 19% versus 13% respectively, P=0.064). The sole patient bearing the DPD IVS14+1G>A mutation (heterozygous) deceased from hematologic toxicity. The median response duration was 5.8 months (95% confidence interval, 4.3-7.2). Duration of response was significantly shortened in patients homozygous for the 3RG allele compared with others (P=0.037). The present data suggest that 3RG3RG breast cancer patients are not good candidates for capecitabine therapy. In addition, attention should be paid to DPD deficiency in breast cancer patients receiving capecitabine. These preliminary data require further confirmation on a larger number of patients.


Eldecalcitol prevented OVX-induced osteoporosis through inhibiting BMSCs senescence by regulating the SIRT1-Nrf2 signal.
Background: Aging and oxidative stress are considered to be the proximal culprits of postmenopausal osteoporosis. Eldecalcitol (ED-71), a new active vitamin D derivative, has shown a good therapeutic effect on different types of osteoporosis, but the mechanism is unclear. This study focused on exploring whether ED-71 could prevent bone loss in postmenopausal osteoporosis by regulating the cell senescence of bone mesenchymal stem cells (BMSCs), and explaining its specific mechanism of action. Materials and methods: An ovariectomized (OVX) rat model was established and 30 ng/kg ED-71 was administered orally once a day. The weight of rats was recorded regularly. Micro-computed tomography (CT) and histochemical staining were used to evaluate bone mass, histological parameters, and aging-related factors. Rat bone mesenchymal stem cells were extracted and cultivated in vitro. Aging cells were marked with senescence-associated β-gal (SA-β-gal) dyeing. The mRNA and protein levels of aging-related factors and SIRT1-Nrf2 signal were detected by RT-PCR, Western blot, and immunofluorescence staining. The reactive oxygen species (ROS) levels were detected by DCFH-DA staining. Results: Compared with the Sham group, the bone volume of the ovariectomized group rats decreased while their weight increased significantly. ED-71 prevented bone loss and inhibited weight gain in ovariectomized rats. More importantly, although the expression of aging-related factors in the bone tissue increased in the ovariectomized group, the addition of ED-71 reversed changes in these factors. After extracting and in vitro culturing bone mesenchymal stem cells, the proportion of aging bone mesenchymal stem cells was higher in the ovariectomized group than in the Sham group, accompanied by a significant decrease in the osteogenic capacity. ED-71 significantly improved the bone mesenchymal stem cells senescence caused by ovariectomized. In addition, ED-71 increased the expression of SIRT1 and Nrf2 in ovariectomized rat bone mesenchymal stem cells. Inhibition of SIRT1 or Nrf2 decreased the inhibitory effect of ED-71 on bone mesenchymal stem cells senescence. ED-71 also showed a suppression effect on the reactive oxygen species level in bone mesenchymal stem cells. Conclusion: Our results demonstrated that ED-71 could inhibit the cell senescence of bone mesenchymal stem cells in ovariectomized rats by regulating the SIRT1-Nrf2 signal, thereby preventing bone loss caused by osteoporosis.


Frequent use of household cleaning products is associated with rhinitis in Chinese children.
Despite the popular use of household cleaning products worldwide, there is no published study investigating the health effects of these products on rhinitis in children. We sought to investigate the household use of cleaning products and rhinitis patterns in Chinese children. A total of 2299 children were recruited from 21 primary schools with wide geographic coverage in Hong Kong. Self-administered questionnaires were completed by parents/guardians to collect detailed information on respiratory symptoms and household use of 14 types of chemical cleaning products, as well as clean water. Students were categorized into 4 mutually exclusive rhinitis patterns (never, occasional, frequent, and persistent). The total chemical burden (TCB) score was used as the exposure indicator by calculating the total time of exposure to the 14 cleaning products. Multinomial logistic regression was used to assess the relationship between rhinitis patterns and the use of household cleaning products. Every 10-unit increment of TCB score was associated with an increase in the odds of occasional (odds ratio [OR], 1.21; 95% CI, 1.05-1.41), frequent (OR, 1.36; 95% CI, 1.13-1.60), and persistent (OR, 1.21; 95% CI, 1.01-1.56) rhinitis after adjustment for a wide range of potential confounders. Compared with the children within the lowest tertile of TCB scores, the adjusted ORs of occasional, frequent, and persistent rhinitis in children within the highest tertile were 1.29 (95% CI, 1.01-1.65), 1.97 (95% CI, 1.40-2.76), and 1.67 (95% CI, 1.10-2.54), respectively. Frequent use of chemical cleaning products at home is associated with an increase in the odds of rhinitis in Chinese primary school children.


A Randomized, Open-Label, Comparative Study of Efficacy and Safety of Tolterodine Combined with Tamsulosin or Doxazosin in Patients with Benign Prostatic Hyperplasia.
BACKGROUND Benign prostatic hyperplasia (BPH), a common disease in men over age 50 years, often causes bladder outlet obstruction and lower urinary tract symptoms (LUTS). Alpha blockers in combination with muscarinic receptor antagonists may have the potential to improve symptoms. This study aimed to assess the efficacy and safety of doxazosin or tamsulosin combined with tolterodine extend release (ER) in patients with BPH and LUTS. MATERIAL AND METHODS In a prospective, randomized, open-label study (ChiCTR-IPR-15005763), 220 consecutive men with BPH and LUTS were allocated to receive doxazosin 4 mg and tolterodine ER 4 mg per day (doxazosin group) or tamsulosin 0.2 mg and tolterodine ER 4 mg per day (tamsulosin group). Treatment lasted 12 weeks. The primary endpoint was the international prostatic symptom score (IPSS). Secondary endpoints were quality of life (QoL) and maximum flow rate (Qmax), which were evaluated at 0, 6, and 12 weeks, and urodynamic parameters assessed at 0 and 12 weeks. RESULTS A total of 192 patients completed the trial. Baseline measurements showed no differences between the groups. After 6 weeks, IPSS improved in both groups and QoL was significantly better in the doxazosin group (P=0.01). After 12 weeks, Qmax, IPSS, QoL, intravesical pressure (Pves), and bladder compliance (BC) in the doxazosin group were significantly better than in the tamsulosin group (P=0.03, P<0.001, P<0.001, P=0.027, and P=0.044, respectively). CONCLUSIONS Administration of alpha blockers combined with muscarinic receptor blocker for 12 weeks improved LUTS in men with BPH.


Long-Term Health Impact of Early Nutrition: The Power of Programming.
The Power of Programming conference 2016 at Ludwig-Maximilians-Universität Munich brought together about 600 researchers and other stakeholders from around the world who reviewed the recent evidence on the lasting health impact of environment and nutrition during early life, from pre-pregnancy to early childhood. The conference was hosted by the Early Nutrition Project, a multidisciplinary research collaboration funded by the European Commission with collaborating researchers from 35 institutions in 15 countries in Europe, the United States and Australia. The project explores the early origins of obesity, adiposity and associated non-communicable diseases, underlying mechanisms and opportunities for prevention. The project also proactively supports translational application of research findings. In fact, some existing evidence has already been rapidly adopted into policy, regulatory standards and practice. Further, broad dissemination of findings is achieved through the established digital eLearning platform of the Early Nutrition eAcademy, video clip-based learning and graphically supported messaging to consumers. The project demonstrated powerful effects of early metabolic programming on later health. Compared to other common prevention strategies, modifying risk trajectories in early life can achieve a much larger risk reduction and be more cost-effective. While some effective prevention strategies have been promptly implemented in policy and guidelines, legislation and practice, in other areas, the uptake is limited by a paucity of quality human intervention trials and insufficient evaluation of the feasibility of implementation and econometric impact. This needs to be strengthened by future collaborative research work.


Introduction to Focus Issue: Dynamical disease: A translational approach.
The concept of Dynamical Diseases provides a framework to understand physiological control systems in pathological states due to their operating in an abnormal range of control parameters: this allows for the possibility of a return to normal condition by a redress of the values of the governing parameters. The analogy with bifurcations in dynamical systems opens the possibility of mathematically modeling clinical conditions and investigating possible parameter changes that lead to avoidance of their pathological states. Since its introduction, this concept has been applied to a number of physiological systems, most notably cardiac, hematological, and neurological. A quarter century after the inaugural meeting on dynamical diseases held in Mont Tremblant, Québec [Bélair et al., Dynamical Diseases: Mathematical Analysis of Human Illness (American Institute of Physics, Woodbury, NY, 1995)], this Focus Issue offers an opportunity to reflect on the evolution of the field in traditional areas as well as contemporary data-based methods.


Implant-supported prosthesis in a child with hereditary mandibular anodontia: the use of ball attachments.
This report describes the provision of an implant-supported hybrid overdenture in an 8-year-old child with mandibular anodontia. An interdisciplinary approach to treatment planning was employed. Two implant fixtures were placed in the canine regions and ball attachments connected to provide support and retention for the prosthesis. There was only one implant-related complication during the first follow-up year. Close follow-up during the child's growing years is essential for the maintenance of the peri-implant tissues and to replace the prosthesis as mandibular growth occurs.


A new model for pancreatitis.
Pancreatitis induced by ligation of the pancreatic duct produces morphologic similarities to human pancreatitis. This model is easily performed in big animals, but it is very difficult to perform pancreatic duct ligation in small animals. Many experimental studies of pharmaceutical treatments for pancreatitis used pancreatic duct-ligation models, but it is also difficult to evaluate the efficacy of the drugs used, because the animals used are of different species with individual differences. To overcome these problems, we ligated the main pancreatic duct of the splenic lobe by a 5.0 absorbable suture by using a surgical microscope and left the gastroduodenal lobe intact in the same rats. This model produced damaged pancreatic tissue in one part and normal pancreatic tissue in another part of the pancreas in the same animals, biochemically and histologically. We evaluated the effect of a new protease inhibitor (ONO-3404) on this preliminary model and found this new protease inhibitor demonstrated a hypertrophic effect on the damaged pancreatic tissue and the normal pancreatic tissue in the same animals. This model is also useful to study pharmaceutic treatment for pancreatic insufficiency and to study chemically induced pancreatic carcinogenesis in the damaged pancreatic tissue and the normal pancreatic tissue in the same animals.


[Changes in HDL subfractions in patients with type I diabetes mellitus before and after metabolic control].
In order to further investigate the behaviour of high density lipoproteins in diabetes mellitus, we studied HDL subclasses, HDL2 and HDL3, in 10 patients with newly detected, untreated insulin-deficient diabetes before starting insulin treatment and after getting a good metabolic control. We used the extractive method of Abell to determine HDL-cholesterol after LDL and VLDL precipitation with polyanions and HDL3-cholesterol after HDL2 precipitation with dextransulphate 15,000 m.w. After insulin therapy, we observed a significant increase in HDL-cholesterol and a decrease in serum triglycerides. Only HDL2-cholesterol, but not HDL3-cholesterol, raised; moreover, we found a significant inverse relationship between HDL-cholesterol (and also HDL2-cholesterol) and triglycerides. So, we think that an increase of lipoprotein lipase activity, owing to insulin treatment, could account for our results.


Peptides and anxiety: a dose-response evaluation of pentagastrin in healthy volunteers.
A large body of data suggest that brain cholecystokinin (CCK) systems are involved in the regulation of anxiety, and numerous studies have demonstrated that CCK-4, a CCKB agonist, reliably induces panic attacks in patients with panic disorder. Recently, pentagastrin, a commercially available CCKB agonist, has been reported to have similar anxiogenic properties. To further explore the utility of pentagastrin as a challenge agent and to determine whether its effects are dose-related, a dose-response study was conducted in ten healthy volunteers. Pentagastrin (0.2 microgram/kg, 0.6 microgram/kg and 1.0 microgram/kg) and inactive placebo were infused over one minute on four separate challenge days in a double-blind fashion. Subjects received pentagastrin while participating in a structured social interaction task. Repeated measures of anxiety, blood pressure, pulse, ACTH, and cortisol were taken at baseline and postinfusion. Pentagastrin administration led to increases in anxiety, pulse, ACTH, cortisol and physical symptoms of panic, in a dose-related manner. Participation in the social interaction task led to increases in measures of anxiety as well as increases in pulse and blood pressure. Few differences were found between the 0.2 microgram/kg dose of pentagastrin and placebo, or between the 0.6 microgram/kg and the 1.0 microgram/kg doses of pentagastrin. These findings support the notion that CCK systems are involved in the regulation of anxiety, and suggest that the 0.6 microgram/kg dose may be optimal for increasing symptoms of anxiety while minimizing unpleasant side effects. The powerful anxiogenic effects of the social interaction task underscore the importance of contextual variables in challenge studies.


[Pentamidine inhalation for Pneumocystis carinii pneumonia in AIDS].
Pneumocystis carinii pneumonia (PCP) is the major cause of death in AIDS. The use of standard drugs for PCP is associated with a remarkably high rate of adverse reactions in AIDS patients. We treated a 35-year-old homosexual with a first episode of PCP with intravenous trimethoprim, 20 mg/kg, and sulfamethoxazole, 100 mg/kg and later with pentamidine injections. However medication had to be stopped because of severe side effects. A new method of therapy using aerosolized pentamidine (300 mg per inhalation, daily) was then applied successfully. The patient recovered and has been having prophylactic inhalations and getting azothymidine for the past 8 months. In studies on relatively small numbers of patients this treatment has been reported to give good results with only minor side effects. However, further data are needed, especially on the optimal characteristics of the generated aerosol and the best delivery system.


Lung Cancer Screening with Low-Dose CT in Female Never Smokers: Retrospective Cohort Study with Long-term National Data Follow-up.
Because of growing concerns about lung cancer in female never smokers, chest low-dose computed tomography (LDCT) screening is often performed although it has never shown clinical benefits. We examinewhether or not female never smokers really need annual LDCT screening when the initial LDCT showed negative findings. This retrospective cohort study included 4,365 female never smokers aged 40 to 79 years who performed initial LDCT from Aug 2002 to Dec 2007. Lung cancer diagnosis was identified from the Korea Central Cancer Registry Database registered until December 31, 2013. We calculated the incidence, cumulative probability, and standardized incidence ratio (SIR) of lung cancer by Lung Imaging Reporting and Data System (Lung-RADS) categories showed on initial LDCT. After median follow-up of 9.69 years, 22 (0.5%) had lung cancer. Lung cancer incidence for Lung-RADS category 4 was 1,848.4 (95% confidence interval [CI], 1,132.4 to 3,017.2) per 100,000 person-years and 16.4 (95% CI, 7.4 to 36.4) for categories 1, 2, and 3 combined. The cumulative probability of lung cancer for category 4 was 10.6% at 5 years and 14.8% at 10 years while they were 0.07% and 0.17% when categories 1, 2, and 3 were combined. The SIR for subjects with category 4 was 43.80 (95% CI, 25.03 to 71.14), which was much higher than 0.47 (95% CI, 0.17 to 1.02) for categories 1, 2, and 3 combined. Considering the low risk of lung cancer development in female never smokers, it seems unnecessary to repeat annual LDCT screening for at least 5 years or even longer unless the initial LDCT showed Lung-RADS category 4 findings.


Influence of free drug samples on prescribing by physicians: A cross sectional survey.
Free drug samples are distributed among doctors as a promotional tool. We investigated the effects of dispensing samples on prescriptions by the doctors and their opinion about samples through a cross-sectional survey. A questionnaire was distributed among the doctors in the Department of Medicine, Nishtar Hospital Multan. It contained drug choices for two hypothetical scenarios, options and reasons for using free samples and doctors' views about them. Response rate remained 83% (166/198). In scenario 1 (rheumatoid arthritis), 100 % and in scenario 2 (acid peptic disease), 13% of sample users dispensed against their preference while in both cases 78 % prescribed the same brand afterwards. Trainees used samples more frequently in both vignettes (p value 0.24 and 0.001 respectively). Mainly, samples were used as a cost-effective measure and were considered a source of medication for poor, significantly. (p value 0.007). But in this process, physicians ignored their first choice and inadvertently increased the total expenditure.


Premature expression of a muscle fibrosis axis in chronic HIV infection.
Despite the success of highly active antiretroviral therapy (HAART), HIV infected individuals remain at increased risk for frailty and declines in physical function that are more often observed in older uninfected individuals. This may reflect premature or accelerated muscle aging. Skeletal muscle gene expression profiles were evaluated in three uninfected independent microarray datasets including young (19 to 29 years old), middle aged (40 to 45 years old) and older (65 to 85 years old) subjects, and a muscle dataset from HIV infected subjects (36 to 51 years old). Using Bayesian analysis, a ten gene muscle aging signature was identified that distinguished young from old uninfected muscle and included the senescence and cell cycle arrest gene p21/Cip1 (CDKN1A). This ten gene signature was then evaluated in muscle specimens from a cohort of middle aged (30 to 55 years old) HIV infected individuals. Expression of p21/Cip1 and related pathways were validated and further analyzed in a rodent model for HIV infection. We identify and replicate the expression of a set of muscle aging genes that were prematurely expressed in HIV infected, but not uninfected, middle aged subjects. We validated select genes in a rodent model of chronic HIV infection. Because the signature included p21/Cip1, a cell cycle arrest gene previously associated with muscle aging and fibrosis, we explored pathways related to senescence and fibrosis. In addition to p21/Cip1, we observed HIV associated upregulation of the senescence factor p16INK4a (CDKN2A) and fibrosis associated TGFβ1, CTGF, COL1A1 and COL1A2. Fibrosis in muscle tissue was quantified based on collagen deposition and confirmed to be elevated in association with infection status. Fiber type composition was also measured and displayed a significant increase in slow twitch fibers associated with infection. The expression of genes associated with a muscle aging signature is prematurely upregulated in HIV infection, with a prominent role for fibrotic pathways. Based on these data, therapeutic interventions that promote muscle function and attenuate pro-fibrotic gene expression should be considered in future studies.


[Nonpenetrating brachiocephalic arterial injury].
The injury of brachiocephalic artery is uncommon in the blunt trauma and sometimes it was accompanied by other organ damages. We reported a case and reviewed 47 cases in literature. A 28-year-old woman was transferred to our hospital with the blunt chest trauma and the cerebral infarction due to the traffic accident. The cineangiogram showed complete obstruction at the middle portion of brachiocephalic artery and the subclavian steal-carotid recovery phenomenon. Thirty days after trauma, the operation was performed under the monitoring of right superficial temporal arterial pressure. The vessel with intimal defect was plugged by a clot and it was replaced with a Gore-Tex graft during simple occlusion of the right carotid, subclavian and brachiocephalic arteries without any monitoring pressure change. The postoperative course was uneventful and the symptom improved. The review decided that the main cause of the injury is traffic accidents and the cineangiogram is important to find the arterial damage and the monitoring of superficial temporal arterial pressure is helpful to avoid the brain ischemia during operation.


[Myocardial metabolism as affected by propofol in geriatric patients. A comparison with etomidate].
This study was designed to compare the effects of propofol and etomidate on myocardial metabolism in elderly patients without clinical manifestations of heart failure or coronary artery disease. Twenty geriatric patients (age 65-82 years) scheduled to undergo elective major upper-abdominal surgery were studied and randomly allocated to two equal groups (propofol and etomidate). All patients were premedicated with piritramide, 7.5 mg, and promethazine, 25 mg, intramuscularly 1 h before arrival in the anesthesia room. Ten patients received propofol (1.5 mg/kg) for induction of anesthesia, followed by 10-min infusion of an induction dose; thereafter, anesthesia was maintained with a continuous infusion of 0.1 mg/kg per min. Ten patients received etomidate, 18 mg, for induction, followed by 2.4 mg/min for maintenance. Vecuronium was used for neuromuscular blockade. Cardiovascular dynamics were recorded while the patients were awake, 1-2 min after induction during apnoea, and 1, 5 and 30 min after tracheal intubation without surgical stimulation. Coronary blood flow (argon wash-in technique with sampling of blood from the coronary sinus), myocardial oxygen consumption and myocardial uptake of glucose, free fatty acids and lactate were determined in the awake state and 5 and 30 min after intubation. Arterial plasma concentrations of propofol (high-pressure liquid chromatography with fluorescence detection) and etomidate (gas chromatography) were measured every 5 min throughout the investigation period, which lasted 45 min. Overall mean plasma concentrations of propofol were 3.69 +/- 0.16 micrograms/ml and of etomidate 1.1 +/- 0.16 microgram/ml.(ABSTRACT TRUNCATED AT 250 WORDS)


Dispersion frailty models and HGLMs.
In medical research recurrent event times can be analysed using a frailty model in which the frailties for different individuals are independent and identically distributed. However, such a homogeneous assumption about frailties could sometimes be suspect. For modelling heterogeneity in frailties we describe dispersion frailty models arising from a new class of models, namely hierarchical generalized linear models. Using the kidney infection data we illustrate how to detect and model heterogeneity among frailties. Stratification of frailty models is also investigated.


Progenitor cells in the kidney: biology and therapeutic perspectives.
The stem cell may be viewed as an engineer who can read the blue print and become the building. The role of this fascinating cell in physiology and pathophysiology has recently attracted a great deal of interest. The archetype of stem cells is the zygote: one cell capable of endless proliferation and differentiation into all tissue types in the human body. Historically, the differentiation of embryonic stem cells is seen as an irreversible process with restricting possibilities for differentiation leading finally to a terminally differentiated cell type. Stem cells have also been described in the adult. They were first defined in tissues with a high cell turnover like skin and gut. Today, stem cells have also been shown in tissues with no or low regenerative potential and turnover, like the kidney. Traditionally, adult stem cells were thought to be restricted in their differentiative and regenerative potential to the tissues in which they reside. However, the stem cell concept is changing rapidly as evidence is mounting that adult stem cells not only reside locally in specific niches, but may also be recruited from the circulation to actively participate in the regeneration of various tissues. Furthermore, reverse differentiation has been demonstrated. This means that highly specialized cell types are able to dedifferentiate and engage in stem cell like activities. Moreover, transdifferentiation of mature cells into different cell types has been reported. This paper will review our current knowledge on renal stem cells and progenitor cells. Specifically, it will discuss the role of progenitor cells and transdifferentiation in renal repair and maintenance. Finally, the potential clinical implications of these findings will be discussed.


Long-lived alpha MUPA transgenic mice exhibit increased mitochondrion-mediated apoptotic capacity.
Caloric restriction (CR) is currently the only therapeutic intervention known to attenuate aging in mammals, but the mechanisms underlying this phenomenon are still poorly understood. To study this issue, the transgenic model of alpha MUPA mice, which previously were reported to spontaneously eat less and live longer compared with their wild-type (WT) control mice, were used. Currently, two transgenic lines that eat less are available, thus implicating the transgenic enzyme, that is, the urokinase-type plasminogen activator (uPA), in causing the reduced appetite. Recently, several changes in the alpha MUPA liver were noted, at the mitochondrial and cellular level, which consistently pointed to an enhanced capacity to induce apoptosis. In addition, alpha MUPA mice showed a reduced level of serum IGF-1 and a reduced incidence of spontaneously occurring or carcinogen-induced tumors in several tissues. Overall, the alpha MUPA model suggests that long-lasting, moderately increased apoptotic capacity, possibly linked in part to modulation of serum IGF-1 and mitochondrial functions, could play a role in the attenuation of aging in calorically restricted mice.


Different classification systems yield different dementia occurrence among nonagenarians and centenarians.
Literature data consistently show different prevalence estimates of dementia when different classification systems are used in the same population. Very few data are available for the oldest old of the elderly. We investigated the occurrence of dementia among 34 nonagenarians and centenarians according to four classification systems: the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders, third edition revised (DSM-III-R) and fourth edition (DSM-IV), the World Health Organization's International Classification of Diseases, 10th revision (ICD-10), and the Cambridge Examination for Mental Disorders of the Elderly (CAMDEX). Cognitive functioning, work, social function and independence in activities of daily living were evaluated by using an extensive neuropsychological examination. The prevalence (95% CI) of dementia was the following: 47.1% (95% CI 30.3-63.8) with the DSM-III-R criteria, 41.2% (95% CI 24.6-57.7) with the DSM-IV criteria, 29.4% (95% CI 14.1-44.7) with the ICD-10 criteria and 38.2% (95% CI 21.9-54.6) with the CAMDEX. The factors that best predicted disagreement between DSM-III-R and DSM-IV were calculation impairment and the presence or absence of personality changes. DSM-III-R and ICD-10 were differentiated by the weight given to executive functions that all have to be impaired according to ICD-10, whereas progressive deterioration differentiated CAMDEX from DSM-III-R. It should be noted that although the DSM-III-R diagnoses differ by a factor of 1.6 times from the ICD-10 and 1.2 times from the CAMDEX diagnoses, we are speaking about dementia, which is very frequent in nonagenarians and centenarians. Moreover, with regard to public health, an estimation of the number of subjects who will lose their autonomy is rather more useful and informative than simple prevalence figures of dementia by itself. In this light, classification systems, such as the ICD-10, that do not include impairment of social function as a criterion for assessing dementia become less adequate.


Inhibition of leukotriene formation in human leukocytes by halothane.
The effects of an inhalation anesthetic, halothane (2-bromo-2-chloro-1,1,1-trifluoroethane) on the formation of 5-lipoxygenase metabolites such as leukotriene B4, 5(S)-hydroxyeicosatetraenoic acid (5-HETE), 6-trans-isomers of leukotriene B4 and leukotriene C4 were studied in human leukocytes stimulated with calcium ionophore A23187. Halothane inhibited the formation of all these metabolites dose dependently and the formation was restored by removal of the drug. The anesthetic also reversibly inhibited the release of [3H]arachidonic acid from neutrophils with a half-inhibition concentration of less than 0.19 mM. The formation of 5-lipoxygenase metabolites was not inhibited by the anesthetic when leukocytes were stimulated with the ionophore in the presence of exogenous arachidonic acid. These observations indicate that the inhibitory effect of halothane on the formation of 5-lipoxygenase metabolites in leukocytes is mainly due to the inhibition of arachidonic acid release.


Briarane-type diterpenoids from the Vietnamese gorgonian Junceella fragilis.
Ten briarane-type diterpenoids (1-10), including one new stereoisomer 17-epi-junceellolide B (1), were isolated from the MeOH extract of the Vietnamese gorgonian Junceella fragilis. Their structures were elucidated by spectroscopic experiments including 1D and 2D NMR, and HR-QTOF-MS. In addition, the in vitro cytotoxic activity against eight human cancer cell lines (LNCaP, HepG2, KB, MCF-7, SK-Mel2, HL-60, LU-1 and SW480) of all isolated compounds was evaluated by SRB assays.


Mechanisms of disease in hemorrhagic fever with renal syndrome.
Hemorrhagic fever with renal syndrome (HFRS) is a dramatic illness characterized by fever, and variable degrees of circulatory failure, hemorrhage, and renal insufficiency. Hantaviruses are the cause of this syndrome. In its severe form, it is associated with significant mortality. Vascular dysfunction is the key physiologic derangement in this disorder. There is a growing body of evidence that suggests that immune mechanisms account for this derangement.


On the origin of the regeneration factor.
The data presented indicate a complexity of the process through which the hepatocyte replication is controled after partial hepatectomy. It can be said that several factors may be working together, including hepatic inhibitor, splenic inhibitor, intestine associated stimulator and pancreatic factor.


Anatomical Study of the Hepatic Veins in Segment 4 of the Liver Using Three-Dimensional Visualization.
Objective: The current study aimed to examine the anatomical structure of the hepatic vein of segment IV liver (S4) of the liver using three-dimensional (3D) visualization technology in order to explore the surgical value of the middle hepatic vein (MHV) manipulation and highlight the importance of current research in hepatic surgery. Methods: Between January 2014 and December 2019, 52 patients with abdominal diseases(not including hepatic disease) were selected for multiphasic computed tomography-enhanced scans of the upper abdomen. A 3D visualization system was utilized to display the structural details of the hepatic veins in S4 of their livers. Couinaud's eight-segment classification system was used to denote the liver' sections. Results: The constructed 3D model clearly displayed vascular morphological characteristics and their location in the liver, hepatic artery and vein system, and portal vein system. Of the 52 patients, 43 had an umbilical fissure vein (UFV) (82.7%), 19 had an accessory S4 liver vein (36.5%), 16 had both a UFV (30.8%) and an accessory S4 liver vein, and 6 had neither (11.5%). A total of 79% of the patients with a UFV and 74.2% of those with an accessory S4 liver vein had venous blood returning into the left hepatic vein. Conclusion: 3D visualization technology was used to determine hepatic venous return of S4 hepatic veins and was found to improve the safety of evaluation in hepatic surgery.


Method to eliminate the zero spectra in Fourier transform profilometry based on a cost function.
To increase the accuracy, speed, and robustness of 3D measurements in Fourier transform profilometry (FTP), this paper introduces a cost function according to the intrinsic features of the amplitude and frequency modulated (AF/M) signal and proposes two new algorithms to eliminate the background components of the fringe pattern based on the proposed cost function. Finally, the standard Fourier transform (FT) is used to calculate the phase of the pattern, which no longer contains background components. The two proposed methods are both data-driven and require no parameter adjustments in advance. Theoretical analysis and 80 experimental results show that the proposed cost function is valid. The results of more than 80 experiments with different types of fringe patterns, different carrier frequencies, and different noise variances with frequency overlap and sudden phase variation show that the proposed two methods are more accurate and robust than the 2D Gabor wavelet transform, the 2D Fan wavelet transform, and the 1D complex Morlet wavelet transform profilometry, and they are approximately 70 times faster than the 1D complex Morlet wavelet transform profilometry.


Hanstruepera crassostreae sp. nov., a novel marine bacterium of the family Flavobacteriaceae isolated from an oyster.
A Gram-stain-negative, rod-shaped, non-motile, aerobic and orange-pigmented marine bacterium, designated strain L53T, was isolated from an oyster sample collected from the coast of Weihai, China (122.0° E 37.5° N). Growth of strain L53T occurred at 4-40 °C (optimum, 33 °C), at pH 6.5-8.0 (optimum, pH 7.5) and with 1.0-7.0 % (w/v) NaCl (optimum, 3.0 %). Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain L53T was closely related to Bizionia echini KCTC 22015T (96.9 %) and Hanstruepera neustonica JCM 19743T (96.1 %). Strain L53T was located in a distinct phyletic lineage in a discrete clade associated with H. neustonica JCM 19743T. The DNA G+C content of strain L53T was 33.5 mol%. The sole menaquinone was MK-6. The polar lipids comprised one phosphatidylethanolamine, four unidentified aminolipids and four unidentified lipids. The major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH and iso-C15 : 0 G. Based on morphological, physiological and molecular properties as well as on phylogenetic distinctiveness, strain L53T should be placed in the genus Hanstruepera as representing a novel species, for which the name Hanstruepera crassostreae sp. nov. is proposed. The type strain is L53T (=KCTC 62247T=MCCC 1H00246T).


Investigations of brain tumours with 99Tcm-HMPAO SPECT.
The cerebral uptake of 99Tcm-hexamethylpropyleneamine oxime (99Tcm-HMPAO) as measured with the use of single photon emission computed tomography (SPECT) was studied in 66 patients with various types of brain tumours and quantified by tumour-to-cerebellum ratios. The uptake of 99Tcm-HMPAO by gliomas and meningiomas showed wide ranges of values. There were no significant differences among primary malignant gliomas (0.75 +/- 0.27, n = 25), recurrent malignant gliomas (0.81 +/- 0.25, n = 14) and benign gliomas (0.77 +/- 0.21, n = 9). Compared to gliomas, meningiomas exhibited a significantly higher 99Tcm-HMPAO uptake (1.14 +/- 0.31, n = 13, p less than 0.001) while the remaining four patients with tumours of various histopathology showed a low 99Tcm-HMPAO uptake. Three of the 66 patients were scanned immediately and again 2 h after injection and they revealed a decrease in tumour activity. No changes in the pattern of uptake were observed in two patients with gliomas which were studied before and after intra-arterial chemotherapy, but a decrease in tumour uptake was found in the glioma patient who was studied before and after radiotherapy. The results obtained with 99Tcm-HMPAO SPECT are in agreement with those on regional cerebral blood flow (rCBF) in brain tumours reported in the literature.


P-Stereogenic β-Aminophosphines: Preparation and Applications in Enantioselective Organocatalysis.
The synthesis of stereodefined β-aminophosphines having both carbon- and phosphorus-based chirality centers is described. The method involves resolution of a mixture of β-aminophosphine oxide diastereomers accessed by ring-opening of an amino alcohol-derived cyclic sulfamidate. A stereospecific, borane-promoted reduction of β-aminophosphine oxides, which occurs under mild conditions and with inversion of configuration at phosphorus, is a key step in this process. The products obtained are new building blocks for the synthesis of P-chiral ligands and organocatalysts. In a proof-of-concept application in organocatalysis, the diastereomeric P-chiral β-aminophosphine-based bifunctional thioureas displayed significantly different activities in the Morita-Baylis-Hillman reaction of methyl acrylate with benzaldehyde derivatives.


[Mechanisms of metastasis and factors in antimetastatic resistance].
The present notions of pathogenetic mechanisms of metastatic spreading are discussed, and results of the investigations devoted to an increase of antimetastatic resistance of the body, application of different immunomodulators, neurotropic and antiangiogenic influences during surgery, chemotherapy, and irradiation, as well as individualization of antimetastatic therapy aimed at prevention, diagnosis and treatment of metastases of human malignant tumours are described.


Genetic differences in Pneumocystis isolates recovered from immunocompetent infants and from adults with AIDS: Epidemiological Implications.
Polymerase chain reaction analysis, direct DNA sequencing, and histological staining were used to determine whether Pneumocystis jirovecii was present in lung tissue specimens obtained, at autopsy, from 58 infants without identifiable immunodeficiency. The results of genotyping of these specimens were compared with the results of genotyping of specimens obtained from 384 human immunodeficiency virus (HIV)-infected adults with Pneumocystis pneumonia. P. jirovecii DNA was detected at the mitochondrial large subunit rRNA and dihydropteroate synthase loci in 100% and 53%, respectively, of the specimens obtained from infants. All specimens obtained from adults tested positive for P. jirovecii at both loci. Genotype distributions at both loci were significantly different in the 2 populations (P < .0001). The observation of different strains circulating in immunocompetent infants and HIV-infected adults suggests independent transmission cycles that warrant further study.


Ultrasound-mediated delivery enhances therapeutic efficacy of MMP sensitive liposomes.
To improve therapeutic efficacy of nanocarrier drug delivery systems, it is essential to improve their uptake and penetration in tumour tissue, enhance cellular uptake and ensure efficient drug release at the tumour site. Here we introduce a tumour targeting drug delivery system based on the ultrasound-mediated delivery of enzyme sensitive liposomes. These enzyme sensitive liposomes are coated with cleavable poly(ethylene glycol) (PEG) which will be cleaved by two members of the enzyme matrix metalloproteinase family (MMP-2 and MMP-9). Cleavage of the PEG coat can increase cellular uptake and will destabilize the liposomal membrane which can result in accelerated drug release. The main aim of the work was to study the effect of focused ultrasound and microbubbles on the delivery and therapeutic efficacy of the MMP sensitive liposome. The performance of the MMP sensitive liposome was compared to a non-MMP sensitive version and Doxil-like liposomes. In vitro, the cellular uptake and cytotoxicity of the liposomes were studied, while in vivo the effect of ultrasound and microbubbles on the tumour accumulation, biodistribution, microdistribution, and therapeutic efficacy were investigated. For all tested liposomes, ultrasound and microbubble treatment resulted in an improved tumour accumulation, increased extravasation, and increased penetration of the liposomes from blood vessels into the extracellular matrix. Surprisingly, penetration depth was independent of the ultrasound intensity used. Ultrasound-mediated delivery of free doxorubicin and the Doxil-like and MMP sensitive liposome resulted in a significant reduction in tumour volume 28 days post the first treatment and increased median survival. The MMP sensitive liposome showed better therapeutic efficacy than the non-MMP sensitive version indicating that cleaving the PEG-layer is important. However, the Doxil-like liposome outcompeted the MMP and non-MMP sensitive liposome, both with and without the use of ultrasound and microbubbles.


Solid tumor risks after high doses of ionizing radiation.
There is increasing concern regarding radiation-related second-cancer risks in long-term radiotherapy survivors and a corresponding need to be able to predict cancer risks at high radiation doses. Although cancer risks at moderately low radiation doses are reasonably understood from atomic bomb survivor studies, there is much more uncertainty at the high doses used in radiotherapy. It has generally been assumed that cancer induction decreases rapidly at high doses due to cell killing. However, recent studies of radiation-induced second cancers in the lung and breast, covering a very wide range of doses, contradict this assumption. A likely resolution of this disagreement comes from considering cellular repopulation during and after radiation exposure. Such repopulation tends to counteract cell killing and accounts for the large discrepancies between the current standard model for cancer induction at high doses and recent second-cancer data. We describe and apply a biologically based minimally parameterized model of dose-dependent cancer risks, incorporating carcinogenic effects, cell killing, and, additionally, proliferation/repopulation effects. Including stem-cell repopulation leads to risk estimates consistent with high-dose second-cancer data. A simplified version of the model provides a practical and parameter-free approach to predicting high-dose cancer risks, based only on data for atomic bomb survivors (who were exposed to lower total doses) and the demographic variables of the population of interest. Incorporating repopulation effects provides both a mechanistic understanding of cancer risks at high doses and a practical methodology for predicting cancer risks in organs exposed to high radiation doses, such as during radiotherapy.


Different endogenous opioid effects on delta- and mu-receptor subtypes in antral and duodenal motility of conscious dogs.
To evaluate the role of endogenous opioids in regulation of upper gastrointestinal motility in unanesthetized dogs and to differentiate the actions on mu- and delta-opioid receptors, seven strain gauges and five platinum electrodes were chronically implanted at the serosa along the antrum and duodenum and connected to a plug in the neck of the dog. Signals were processed by a Hellige AC amplifier, a rectilinear recorder, and a data aquisition system. A motility index (MI) was calculated and together with the electrical data a contractile activity percentage (CAP) was determined for consecutive 30 min periods. The delta-opioid antagonist ICI 741 864 and the mu-receptor blocker naloxone were injected intraarterially through a chronic Groshong catheter placed in the pyloric region and connected to a subcutaneous port. After a meal of solid food, ICI 174 864 increased motility relative to controls in the antrum averaged over 5 hr by 144.4%+/-26 for the MI and 73%+/-26 for CAP; after naloxone MI increased by 222%+/-60 and CAP 121%+/-76. In the duodenum, ICI 174 864 decreased MI over a range of 57% to 22% (P < 0.05). Naloxone increased MI and CAP significantly after 2.5 hr. We interpreted the results of ICI 174 864 in the duodenum to reflect suppression of a tonic opioid influence at the delta-receptor mainly at the prevertebral ganglion. In the interdigestive state in 56% of the dogs, naloxone delayed the occurrence of phase III of the migrating myoelectric complex (MMC) for up to 370 min, while under ICI 174 864, normal interdigestive cycles were present. Disturbance of the timing of the interdigestive cycles at central mu-opioid receptors may by involved in the effect.


Interferon lambda 1-3 expression in infants hospitalized for RSV or HRV associated bronchiolitis.
The airway expression of type III interferons (IFNs) was evaluated in infants hospitalized for respiratory syncytial virus (RSV) or rhinovirus (HRV) bronchiolitis. As an additional objective we sought to determine whether a different expression of IFN lambda 1-3 was associated with different harboring viruses, the clinical course of bronchiolitis or with the levels of well established IFN stimulated genes (ISGs), such as mixovirus resistance A (MxA) and ISG56. The analysis was undertaken in 118 infants with RSV or HRV bronchiolitis. Nasopharyngeal washes were collected for virological studies and molecular analysis of type III IFN responses. RSV elicited higher levels of IFN lambda subtypes when compared with HRV. A similar expression of type III IFN was found in RSVA or RSVB infected infants and in those infected with HRVA or HRVC viruses. Results also indicate that IFN lambda 1 and IFN lambda 2-3 levels were correlated with each other and with MxA and ISG56-mRNAs. In addition, a positive correlation exists between the IFN lambda1 levels and the clinical score index during RSV infection. In particular, higher IFN lambda 1 levels are associated to an increase of respiratory rate. These findings show that differences in the IFN lambda 1-3 levels in infants with RSV or HRV infections are present and that the expression of IFN lambda 1 correlates with the severity of RSV bronchiolitis.


Engineered porcine pepsinogen exhibits dominant unimolecular activation.
An engineered pepsinogen, which was a fusion protein of thioredoxin and pepsinogen, exhibited dominant self-activation (unimolecular reaction; intramolecular activation) in contrast to recombinant pepsinogen which exhibited both unimolecular and bimolecular reactions (intermolecular activation mediated by pepsin released during activation). At pH values of 1.1, 2.0, and 3.0, activation curves for the engineered pepsinogen were hyperbolic rather than sigmoidal, indicating that self-activation was the dominant activation mechanism in comparison to the slower bimolecular activation. To confirm which activation mechanism was dominant, an equal mole of pepsin was added to accelerate the bimolecular reaction during activation. The addition of exogenous pepsin did not affect the activation rate of the engineered pepsinogen but accelerated pepsinogen activation through the bimolecular reaction. The above results indicated that the engineered pepsinogen exhibited, primarily, a self-activation mechanism and that bimolecular activation was negligible.


Kinetics and mechanisms of the oxidation of iodide and bromide in aqueous solutions by a trans-dioxoruthenium(VI) complex.
The kinetics and mechanisms of the oxidation of I (-) and Br (-) by trans-[Ru (VI)(N 2O 2)(O) 2] (2+) have been investigated in aqueous solutions. The reactions have the following stoichiometry: trans-[Ru (VI)(N 2O 2)(O) 2] (2+) + 3X (-) + 2H (+) --> trans-[Ru (IV)(N 2O 2)(O)(OH 2)] (2+) + X 3 (-) (X = Br, I). In the oxidation of I (-) the I 3 (-)is produced in two distinct phases. The first phase produces 45% of I 3 (-) with the rate law d[I 3 (-)]/dt = ( k a + k b[H (+)])[Ru (VI)][I (-)]. The remaining I 3 (-) is produced in the second phase which is much slower, and it follows first-order kinetics but the rate constant is independent of [I (-)], [H (+)], and ionic strength. In the proposed mechanism the first phase involves formation of a charge-transfer complex between Ru (VI) and I (-), which then undergoes a parallel acid-catalyzed oxygen atom transfer to produce [Ru (IV)(N 2O 2)(O)(OHI)] (2+), and a one electron transfer to give [Ru (V)(N 2O 2)(O)(OH)] (2+) and I (*). [Ru (V)(N 2O 2)(O)(OH)] (2+) is a stronger oxidant than [Ru (VI)(N 2O 2)(O) 2] (2+) and will rapidly oxidize another I (-) to I (*). In the second phase the [Ru (IV)(N 2O 2)(O)(OHI)] (2+) undergoes rate-limiting aquation to produce HOI which reacts rapidly with I (-) to produce I 2. In the oxidation of Br (-) the rate law is -d[Ru (VI)]/d t = {( k a2 + k b2[H (+)]) + ( k a3 + k b3[H (+)]) [Br (-)]}[Ru (VI)][Br (-)]. At 298.0 K and I = 0.1 M, k a2 = (2.03 +/- 0.03) x 10 (-2) M (-1) s (-1), k b2 = (1.50 +/- 0.07) x 10 (-1) M (-2) s (-1), k a3 = (7.22 +/- 2.19) x 10 (-1) M (-2) s (-1) and k b3 = (4.85 +/- 0.04) x 10 (2) M (-3) s (-1). The proposed mechanism involves initial oxygen atom transfer from trans-[Ru (VI)(N 2O 2)(O) 2] (2+) to Br (-) to give trans-[Ru (IV)(N 2O 2)(O)(OBr)] (+), which then undergoes parallel aquation and oxidation of Br (-), and both reactions are acid-catalyzed.


Indications and Predictors for Reconstructive Surgery After Hand Burns.
The objective of this study was to analyze the prevalence, indications, and type of reconstructive surgery and predictors of the outcomes of reconstructive surgery after hand burns. A retrospective cohort study was conducted that included all patients admitted with acute hand burns in the Dutch burn centers from January 1998 through December 2002. The details of reconstruction including frequency, timing, indication, and techniques were collected over a 10-year follow-up period. Hand burns were seen in 42% (n = 562 of 1,334) of all patients admitted with acute burns. Reconstructive surgery during the 10-year follow-up period was required in 15%. Contractures, especially of the first web space and little finger, were the most frequent indications for reconstructive surgery. Web spaces 1 to 3 and the little finger were the location most frequently operated on. The most frequently performed surgical technique was release of the contractures and the use of a random flap. Eighty percent of the reconstructive surgery patients required more than 1 reconstructive procedure, most often within 2 years of the initial injury. Secondary operations at the same location were required in 12%. In 40% of the patients, the first reconstructive surgery was performed within the first postburn year. Significant independent factors related to the need for reconstructive hand surgery were a larger area of full-thickness burns and surgical treatment of the hand during the acute phase. Reconstructive surgery was required in 15% of patients who sustained hand burns. The majority of the patients requiring reconstructive surgery of the hand needed 2 or more operations to correct the contractures of the hand. Contractures of the little finger and first web space were the locations most frequently operated on. Patients with more extensive burns and who required hand surgery during the acute phase were more likely to need reconstructive surgery. Prognostic IV.