Genetic Diversity of Pseudomonas syringae pv. actinidiae Strains from Different Geographic Regions in China.
Pseudomonas syringae pv. actinidiae causes kiwifruit bacterial canker, with severe infection of the kiwifruit plant resulting in heavy economic losses. Little is known regarding the biodiversity and genetic variation of populations of P. syringae pv. actinidiae in China. A collection of 269 strains of P. syringae pv. actinidiae was identified from 300 isolates obtained from eight sampling sites in five provinces in China. The profiles of 50 strains of P. syringae pv. actinidiae and one strain of P. syringae pv. actinidifoliorum were characterized by Rep-, insertion sequences 50, and randomly amplified polymorphic DNA polymerase chain reaction (PCR). Discriminant analysis of principal coordinates, principal component analysis, and hierarchical cluster analysis were used to analyze the combined fingerprints of the different PCR assays. The results revealed that all isolates belonged to the Psa3 group, that strains of P. syringae pv. actinidiae from China have broad genetic variability that was related to source geographic region, and that Chinese strains can be readily differentiated from strains from France but are very similar to those from Italy. Multilocus sequence typing of 24 representative isolates using the concatenated sequences of five housekeeping genes (cts, gapA, gyrB, pfk, and rpoD) demonstrated that strain Jzhy2 from China formed an independent clade compared with the other biovars, which possessed the hopH1 effector gene but lacked the hopA1 effector gene. A constellation analysis based on the presence or absence of the four loci coding for phytotoxins and a cluster analysis based on the 11 effector genes showed that strains from China formed two distinct clades. All of the strains, including K3 isolated in 1997 from Jeju, Korea, lacked the cfl gene coding for coronatine. In contrast, the tox-argK gene cluster coding for phaseolotoxin was detected in K3 and in the biovar 1 strains (K3, Kw30, and Psa92), and produced a false-positive amplicon for the hopAM1-like gene in this study. To date, only one biovar (biovar 3) is represented by the strains of P. syringae pv. actinidiae from China, despite China being the center of origin for kiwifruit.


Gastric remnant cancer and long-term survival in Central Norway 2001 to 2016 - A population-based study.
Gastric remnant cancer (GRC) has been defined as a distinct clinical entity and is reported to account for 1-8% of all gastric cancers. We aimed to characterize GRC patients and assess survival in a Western population. Retrospective population-based cohort study including 1217 patients diagnosed with gastric adenocarcinoma in Central Norway 2001-2016. GRCs (n = 78) defined as adenocarcinomas arising in the residual stomach after distal gastrectomy were compared to non-GRC (n = 1139) and to proximal non-GRC (n = 595). 78 (6.4 %) gastric cancers were GRC. The annual number and proportion of GRC declined during the study period (p = 0.003). Median latency from distal gastrectomy to GRC diagnosis was 37.6 years (15.7-68.0) and previous Billroth II reconstruction was most common (87.7%). Compared to controls, GRC patients were more frequently males (83.3%), diagnosed in earlier TNM stages and were older at diagnosis. A smaller proportion of GRC patients received perioperative or palliative chemotherapy, but the R0/R1resection rate of 41.0% was no different from non-GRC patients. Overall median survival for GRC patients irrespective of treatment was 7.0 months, which did not differ from non-GRCs or proximal non-GRC. In multivariate analyses TNM stage and age were independently associated with mortality, whereas GRC per se was not. Numbers of GRCs declined during the study period, but the latency between distal gastrectomy and GRC diagnosis was long. GRC patients were more frequently male and older than other gastric cancer patients. GRC was not independently associated with survival after adjusting for TNM stage and tumor location.


Histone H3S10 phosphorylation by the JIL-1 kinase in pericentric heterochromatin and on the fourth chromosome creates a composite H3S10phK9me2 epigenetic mark.
The JIL-1 kinase mainly localizes to euchromatic interband regions of polytene chromosomes and is the kinase responsible for histone H3S10 phosphorylation at interphase in Drosophila. However, recent findings raised the possibility that the binding of some H3S10ph antibodies may be occluded by the H3K9me2 mark obscuring some H3S10 phosphorylation sites. Therefore, we have characterized an antibody to the epigenetic H3S10phK9me2 double mark as well as three commercially available H3S10ph antibodies. The results showed that for some H3S10ph antibodies their labeling indeed can be occluded by the concomitant presence of the H3K9me2 mark. Furthermore, we demonstrate that the double H3S10phK9me2 mark is present in pericentric heterochromatin as well as on the fourth chromosome of wild-type polytene chromosomes but not in preparations from JIL-1 or Su(var)3-9 null larvae. Su(var)3-9 is a methyltransferase mediating H3K9 dimethylation. Furthermore, the H3S10phK9me2 labeling overlapped with that of the non-occluded H3S10ph antibodies as well as with H3K9me2 antibody labeling. Interestingly, when a Lac-I-Su(var)3-9 transgene is overexpressed, it upregulates H3K9me2 dimethylation on the chromosome arms creating extensive ectopic H3S10phK9me2 marks suggesting that the H3K9 dimethylation occurred at euchromatic H3S10ph sites. This is further supported by the finding that under these conditions euchromatic H3S10ph labeling by the occluded antibodies was abolished. Thus, our findings indicate a novel role for the JIL-1 kinase in epigenetic regulation of heterochromatin in the context of the chromocenter and fourth chromosome by creating a composite H3S10phK9me2 mark together with the Su(var)3-9 methyltransferase.


Lesion load quantification in serial MR of early relapsing multiple sclerosis patients in azathioprine treatment. A retrospective study.
Azathioprine (AZA) has a slight but consistent effect on clinical outcome in multiple sclerosis (MS), but very few data are available on magnetic resonance imaging (MRI) changes. We performed a retrospective study aimed to quantify changes of lesion load in two serial proton density weighted MRI sequences (TR 2500, TE 30, 1.5 T) at a mean interval of 2.5 years in 36 relapsing-remitting (RR) MS patients: 19 had been treated with AZA, beside steroids after relapses (AZA group), and 17 had been treated with steroids only (control group). All but 3 patients were in the early phase of the disease. Total lesion area (TLA) was measured by manual outlining method and the arbitrary score proposed by Ormerod (total score) was also calculated from the number and diameter of lesions. Lesion load was the same at baseline, but median percentage difference of TLA between first and second scan was + 15.6% in control, -43.7% in the AZA group (p < 0.05, Mann-Whitney test). The distribution of patients according to TLA change, assuming that an increase or decrease was significant if larger than 50%, was found to be significantly different in favor of AZA-treated patients (chi(2) = 35.92, p < 0.001). These results suggest an effect of AZA treatment on MRI lesion load in early RR MS: a larger prospective study is worthwhile.


Birth Defects in Northern Iran (2008-2013).
Congenital anomalies are important medical and public health conditions. The pattern and prevalence of birth defects may vary over time or with geographical location. We investigated the live birth prevalence and occurrence pattern of birth defects in Golestan Province, northern Iran. This cross-sectional descriptive study was carried out on 144920 live newborns in 13 hospitals in Golestan Province, northern Iran, from 21 Jan 2008 to 19 Mar 2013. The newborns were examined for the presence of birth defects and mothers were interviewed for variables such as maternal age and ethnicity. In addition, data for each newborn was filed in a questionnaire and the coding of birth defects was translated to the International Classification of Diseases 10th revision-clinical modification (ICD-10-CM). Overall, 1690 infants were diagnosed as having birth defects among 144920 live newborns. The prevalence rate of birth defects was 11.66 per 1000 live births, the prevalence of birth defects per 1000 was11.62 in males and 11.42 in females. The prevalence of congenital anomalies among native Fars, Turkmen and Sistani were 13.03, 11.16 and 13.07, respectively, per 1000 live births. Anomalies of the cardiovascular system were the most common defects; the prevalence rate of cardiovascular system was 8.34 per 1000 live birth. The prevalence rate of birth defects in this area was lower than in the other regions in Iran (20.3 per 1000 live births) but higher than in some parts of Asia (7.33 per 1000 live births).


Pneumococcal arthritis complicating gout. Case report and literature review.
The simultaneous occurrence of septic arthritis and gout is quite uncommon. We describe a patient with chronic gout who developed pneumococcal pneumonia. Twenty-four hours later, his previously involved joints became acutely inflamed and were found to contain both pneumococcal organisms and monosodium urate crystals intracellularly. The mechanisms which may confer upon a gouty joint selective resistance or susceptibility to colonization by a blood borne organism are discussed. Septic arthritis must remain a consideration in patients with chronic gouty arthritis.


Interictal networks in magnetoencephalography.
Epileptic networks involve complex relationships across several brain areas. Such networks have been shown on intracerebral EEG (stereotaxic EEG, SEEG), an invasive technique. Magnetoencephalography (MEG) is a noninvasive tool, which was recently proven to be efficient for localizing the generators of epileptiform discharges. However, despite the importance of characterizing non-invasively network aspects in partial epilepsies, only few studies have attempted to retrieve fine spatiotemporal dynamics of interictal discharges with MEG. Our goal was to assess the relevance of magnetoencephalography for detecting and characterizing the brain networks involved in interictal epileptic discharges. We propose here a semi-automatic method based on independent component analysis (ICA) and on co-occurrence of events across components. The method was evaluated in a series of seven patients by comparing its results with networks identified in SEEG. On both MEG and SEEG, we found that interictal discharges can involve remote regions which are acting in synchrony. More regions were identified in SEEG (38 in total) than in MEG (20). All MEG regions were confirmed by SEEG when an electrode was present in the vicinity. In all patients, at least one region could be identified as leading according to our criteria. A majority (71%) of MEG leaders were confirmed by SEEG. We have therefore shown that MEG measurements can extract a significant proportion of the networks visible in SEEG. This suggests that MEG can be a useful tool for defining noninvasively interictal epileptic networks, in terms of regions and patterns of connectivity, in search for a "primary irritative zone".


[Vntr-genotyping of Francisella tularensis strains isolated in the former USSR territory and some European countries during epizootics in 1988 - 1989].
Retrospective VNTR-analysis of 159 Francisella tularensis subsp. holarctica strains isolated in December 1988 - February 1989 in former USSR and some European countries was carried out. Analysis of heterogenic genotypes of strains allow to subdivide them into 30 groups of variants by individual genotypes, while cluster analysis--to subdivide them in 7 clusters with different number of compositions. The predominance of genotype C1 strains isolated on the Rostov and Archangelsk regions and the Crimea was established. F. tularensis strains isolated in winter time 1988 - 1989 in different geographic regions were supposed to be resident cultures typical for their biotope in natural focus of disease.


Hyperbaric oxygen therapy in acute necrotizing infections with a special reference to the effects on tissue gas tensions.
Clostridial gas gangrene and perineal necrotizing fasciitis or Fournier's gangrene are rare but serious infections with an acute onset, rapid progression, systemic toxemia and a high mortality rate. The aim of this study was to investigate the efficacy of surgery, antibiotic treatment, surgical intensive care and in particular the role of hyperbaric oxygen (HBO) in the management of these infections. An experimental rat model was used to investigate the possibilities for measuring tissue oxygen and carbon dioxide tensions during hyperbaric oxygen treatment. In addition to this preliminary experimental study, Silastic tube tonometer and capillary sampling techniques were tested to measure the effect of hyperbaric oxygen treatment on subcutaneous oxygen and carbon dioxide tensions in patients with necrotizing fasciitis and healthy controls. Between January 1971 and April 1997, 53 patients with Clostridial gas gangrene were treated in the Department of Surgery, University of Turku. The patients underwent surgical debridement, broad spectrum antibiotic therapy and a series of hyperbaric oxygen treatments at 2.5 atmospheres absolute pressure (ATA). Twelve patients died (22.6%). Hyperbaric oxygen therapy in gas gangrene seems to be life-, limb- and tissue saving. Early diagnosis remains essential. Patient survival can be improved if the disease is recognized early and appropriate therapy instituted promptly. Between February 1971 and September 1996, 33 patients with perineal necrotizing fasciitis were treated in the Department of Surgery, University of Turku. The management included surgical debridement of the necrotic tissue with incisions and drainage of the involved areas, antibiotic therapy, hyperbaric oxygen treatment at 2.5 ATA pressure and surgical intensive care. Three patients died giving a mortality rate of 9.1%. The survivors received hyperbaric oxygen therapy for 2-12 times. Our results indicate that hyperbaric oxygenation is an important therapeutic adjunct in the treatment of Fournier's gangrene. Electrical equipment should not be used unsheltered in a hyperbaric chamber due to the increased risk of fire. The subcutaneous tissue gas tensions of rats were therefore measured using a subcutaneously implanted Silastic tube tonometer and a capillary sampling technique. The method was successfully adapted to hyperbaric conditions. The subcutaneous oxygen tension levels increased five fold and the carbon dioxide tension levels two fold compared to initial levels. The PO2 and PCO2 of subcutaneous tissue and arterial blood were measured directly in six patients with necrotizing fasciitis and three healthy volunteers in normobaric conditions and during hyperbaric oxygen exposure at 2.5 ATA pressure. The measurements were carried out in healthy tissue and at the same time in the vicinity of the infected area of the patients. During HBO at 2.5 ATA subcutaneous oxygen tensions increased several fold from baseline values and carbon dioxide tensions also increased, but to a lesser degree in both healthy and infected tissues. When examining the subcutaneous PO2 levels measured from patients with necrotizing fasciitis, the PO2 was regularly higher in the vicinity of the infected area than in healthy tissue. In general, HBO treatment resulted in a marked increase in tissue oxygenation in both healthy tissue and in the vicinity of infected tissue. The hyper-oxygenated tissue zone surrounding the infected area may be of significance in preventing the extension of invading microorganisms.


Compromise of brain tissue caused by cortical venous reflux of intracranial dural arteriovenous fistulas: assessment with diffusion-weighted magnetic resonance imaging.
Cortical venous reflux (CVR) is a high risk factor for aggressive behavior of intracranial dural arteriovenous fistulas (DAVF). The pathological conditions in brain tissue affected by CVR were investigated by diffusion-weighted magnetic resonance imaging. A retrospective review identified 56 patients with DAVFs who underwent diffusion-weighted imaging before treatment. Twenty patients had neurological symptoms corresponding to the brain area affected by CVR (Group I), 21 patients with CVR had no focal brain dysfunctions (Group II), and 15 patients had no CVR (Group III). Apparent diffusion coefficient (ADC) was measured for 11 brain areas predefined based on normal venous drainage patterns in the 56 patients and in 21 normal volunteers. The mean ADC ratio was calculated for each area by dividing the ADC value of patients by that of normal volunteers. Areas affected by CVR in Group I showed a mean ADC-to-control ratio of 0.72, which was significantly lower than that of Group II (0.96, P<0.01). Follow-up studies demonstrated significantly increased ADC ratios in brain areas affected by CVR after the DAVFs were treated successfully. The mean ADC ratio of an affected area remained low, with persistent symptoms in 1 patient who underwent palliative treatment. Decreased ADC was observed in the brain parenchyma affected by CVR and was associated with regional brain dysfunction. Successful treatment of the DAVF increased the ADC toward normal levels. The ADC may be a useful indicator of the severity of CVR.


Binding of norbinaltorphimine (norBNI) congeners to wild-type and mutant mu and kappa opioid receptors: molecular recognition loci for the pharmacophore and address components of kappa antagonists.
Molecular modifications of both the kappa opioid antagonist norbinaltorphimine (norBNI, 1) and the kappa receptor have provided evidence that the selectivity of this ligand is conferred through ionic interaction if its N17' protonated amine group (an "address") with a nonconserved acidic residue (Glu297) on the kappa receptor. In the present study, we have examined the effect of structural modifications on the affinity of norBNI analogues for wild-type and mutant kappa and mu opioid receptors expressed in COS-7 cells. Compounds 2, 3, and 7, which have an antagonist pharmacophore and basic N17' group in common with norBNI, retained high affinity for the wild-type kappa but exhibited greatly reduced affinity for mutant kappa receptors (E297K and E297A). Modification of the phenolic or N-substituent groups of the antagonist pharmacophore (4 and 5) or removal of basicity at the address N17' center (6) led to greatly reduced affinity for the wild-type and mutant receptors. The reduced affinity upon modification of the kappa receptor is consistent with the ionic interaction of the protonated N17' group of kappa antagonists (1-3, 7) with the carboxylate group of E297 at the top of TM6. This was supported by the greatly enhanced affinity of compounds 1-3 for the mutant mu receptor (K303E), as compared to the wild-type mu receptor, given that residue K303 occupies a position equivalent to that of E297 in the kappa receptor. In view of the high degree of homology of the seven TM domains of the kappa and mu opioid receptors, it is suggested that the antagonist pharmacophore is bound within this highly conserved region of the kappa or mutant mu receptor and that an anionic residue at the top of TM6 (E297 or K303E, respectively) provides additional binding affinity.


Changing Landscape of Dravet Syndrome Management: An Overview.
Dravet syndrome (DS), previously known as severe myoclonic epilepsy of infancy, is a severe developmental and epileptic encephalopathy caused by loss-of-function mutations in one copy of SCN1A (haploinsufficiency), located on chromosome 2q24, with decreased function of Nav1.1 sodium channels in GABAergic inhibitory interneurons. Pharmacoresistant seizures in DS start in the infancy in the form of hemiclonic febrile status epilepticus. Later, other intractable seizure types develop including myoclonic seizures. Early normal development in infancy evolves into moderate to severe intellectual impairment, motor impairment, behavioral abnormalities, and later a characteristic crouching gait. Clobazam, valproate, levetiracetam, topiramate, zonisamide, ketogenic diet, and vagus nerve stimulation had been shown to be effective, but even with polytherapy, only 10% of patients get adequate seizure control. The author provides a narrative review of the current treatment paradigm as well as recent advances in the management of DS based on a comprehensive literature review (MEDLINE using PubMed and OvidSP vendors with appropriate keywords to incorporate recent evidence), personal practice, and experience. In recent years, the treatment paradigm of DS is changing with the approval of pharmaceutical-grade cannabidiol oil and stiripentol. Another novel antiepileptic drug (AED), fenfluramine, had also shown excellent efficacy in phase 3 studies of DS. However, these AEDs primarily control seizures without addressing the underlying pathogenesis and other important common comorbidities such as cognitive impairment, autistic behavior, neuropsychiatric abnormalities, and motor impairment including crouching gait. Several agents targeted for DS are in the developmental stage: TAK935, lorcaserin, clemizole, huperzine analog, ataluren, selective sodium channel modulators and activators, antisense oligonucleotide therapy, and adenoviral vector therapy. As DS is associated with a high risk of sudden unexpected death in epilepsy, seizure detection devices can be used in this population for testing and clinical validation of these devices.


Maternal smoking and low family income during pregnancy as predictors of the relationship between depression and adiposity in young adults.
There is an increasing incidence of overweight/obesity and mental health disorders in young adults and the two conditions often coexist. We aimed to investigate the influence of antenatal and postnatal factors that may underlie this association with a focus on maternal prenatal smoking, socio-economic status and gender. Data from the Western Australian Pregnancy Cohort (Raine) Study (women enrolled 1989-1991) including 1056 offspring aged 20 years (cohort recalled 2010-2012) were analyzed (2015-2016) using multivariable models for associations between offspring depression scores (DASS-21 Depression-scale) and body mass index (BMI), adjusting for pregnancy and early life factors and offspring behaviours. There was a significant positive relationship between offspring depression-score and BMI independent of gender and other psychosocial covariates. There was a significant interaction between maternal prenatal smoking and depression-score (interaction coefficient=0.096; 95% CI: 0.006, 0.19, P=0.037), indicating the relationship between depression-score and BMI differed according to maternal prenatal smoking status. In offspring of maternal prenatal smokers, a positive association between BMI and depression-score (coefficient=0.133; 95% CI: 0.05, 0.21, P=0.001) equated to 1.1 kg/m2 increase in BMI for every 1standard deviation (8 units) increase in depression-score. Substituting low family income during pregnancy for maternal prenatal smoking in the interaction (interaction coefficient=0.091; 95% CI: 0.01, 0.17, P=0.027) showed a positive association between BMI and depression score only among offspring of mothers with a low family income during pregnancy (coefficient=0.118; 95% CI: 0.06, 0.18, P<0.001). There were no significant effects of gender on these associations. Whilst further studies are needed to determine whether these associations are supported in other populations, they suggest potentially important maternal behavioural and socio-economic factors that identify individuals vulnerable to the coexistence of obesity and depression in early adulthood.


Monocyte chemoattractant protein-1 derived from biliary innate immunity contributes to hepatic fibrogenesis.
Monocyte chemoattractant protein-1 (MCP-1) is a major chemotactic factor for hepatic stellate cells (HSCs) associated with hepatic fibrosis. In this study, among several fibrogenetic factors derived from biliary epithelial cells (BECs), MCP-1 produced by the biliary innate immune system was found to be most critical in the histogenesis of hepatic fibrogenesis. Using cultured human BECs, the expression of five fibrogenetic factors including MCP-1 on stimulation with Toll-like receptor ligands, inflammatory cytokines or bile acids was examined. Moreover, in situ detection of MCP-1 and α-smooth muscle actin proteins was performed using sections from normal and diseased livers by immunohistochemistry. All fibrogenetic factors were detected in BECs, but only MCP-1 expression was upregulated, by all the Toll-like receptor ligands, IL-1β, and tumour necrosis factor-alpha. Proliferating bile ductules in interface areas expressed MCP-1 in diseased livers accompanying α-smooth muscle actin-positive activated HSCs. Bile ductules proliferate in various hepatobiliary diseases, and its significance is still unknown. This study demonstrated that BECs in bile ductules could produce MCP-1, particularly, via biliary innate immunity, suggesting that MCP-1 derived from BECs plays an important role in the recruitment of HSCs to interface areas and the activation of HSCs resulting in the progression of periportal fibrosis.


Hormone replacement therapy and physical function in healthy older men. Time to talk hormones?
Improving physical function and mobility in a continuously expanding elderly population emerges as a high priority of medicine today. Muscle mass, strength/power, and maximal exercise capacity are major determinants of physical function, and all decline with aging. This contributes to the incidence of frailty and disability observed in older men. Furthermore, it facilitates the accumulation of body fat and development of insulin resistance. Muscle adaptation to exercise is strongly influenced by anabolic endocrine hormones and local load-sensitive autocrine/paracrine growth factors. GH, IGF-I, and testosterone (T) are directly involved in muscle adaptation to exercise because they promote muscle protein synthesis, whereas T and locally expressed IGF-I have been reported to activate muscle stem cells. Although exercise programs improve physical function, in the long-term most older men fail to comply. The GH/IGF-I axis and T levels decline markedly with aging, whereas accumulating evidence supports their indispensable role in maintaining physical function integrity. Several studies have reported that the administration of T improves lean body mass and maximal voluntary strength in healthy older men. On the other hand, most studies have shown that administration of GH alone failed to improve muscle strength despite amelioration of the detrimental somatic changes of aging. Both GH and T are anabolic agents that promote muscle protein synthesis and hypertrophy but work through separate mechanisms, and the combined administration of GH and T, albeit in only a few studies, has resulted in greater efficacy than either hormone alone. Although it is clear that this combined approach is effective, this review concludes that further studies are needed to assess the long-term efficacy and safety of combined hormone replacement therapy in older men before the medical rationale of prescribing hormone replacement therapy for combating the sarcopenia of aging can be established.


Methods for analyzing data from probabilistic linkage strategies based on partially identifying variables.
In record linkage studies, unique identifiers are often not available, and therefore, the linkage procedure depends on combinations of partially identifying variables with low discriminating power. As a consequence, wrongly linked covariate and outcome pairs will be created and bias further analysis of the linked data. In this article, we investigated two estimators that correct for linkage error in regression analysis. We extended the estimators developed by Lahiri and Larsen and also suggested a weighted least squares approach to deal with linkage error. We considered both linear and logistic regression problems and evaluated the performance of both methods with simulations. Our results show that all wrong covariate and outcome pairs need to be removed from the analysis in order to calculate unbiased regression coefficients in both approaches. This removal requires strong assumptions on the structure of the data. In addition, the bias significantly increases when the assumptions do not hold and wrongly linked records influence the coefficient estimation. Our simulations showed that both methods had similar performance in linear regression problems. With logistic regression problems, the weighted least squares method showed less bias. Because the specific structure of the data in record linkage problems often leads to different assumptions, it is necessary that the analyst has prior knowledge on the nature of the data. These assumptions are more easily introduced in the weighted least squares approach than in the Lahiri and Larsen estimator.


In vivo imaging of nanoparticle delivery and tumor microvasculature with multimodal optical coherence tomography.
Current imaging techniques capable of tracking nanoparticles in vivo supply either a large field of view or cellular resolution, but not both. Here, we demonstrate a multimodality imaging platform of optical coherence tomography (OCT) techniques for high resolution, wide field of view in vivo imaging of nanoparticles. This platform includes the first in vivo images of nanoparticle pharmacokinetics acquired with photothermal OCT (PTOCT), along with overlaying images of microvascular and tissue morphology. Gold nanorods (51.8 ± 8.1 nm by 15.2 ± 3.3 nm) were intravenously injected into mice, and their accumulation into mammary tumors was non-invasively imaged in vivo in three dimensions over 24 hours using PTOCT. Spatial frequency analysis of PTOCT images indicated that gold nanorods reached peak distribution throughout the tumors by 16 hours, and remained well-dispersed up to 24 hours post-injection. In contrast, the overall accumulation of gold nanorods within the tumors peaked around 16 hours post-injection. The accumulation of gold nanorods within the tumors was validated post-mortem with multiphoton microscopy. This shows the utility of PTOCT as part of a powerful multimodality imaging platform for the development of nanomedicines and drug delivery technologies.


A Rapid Robust Method for Subgrouping Non-NF2 Meningiomas According to Genotype and Detection of Lower Levels of M2 Macrophages in AKT1 E17K Mutated Tumours.
The majority of meningiomas are grade I, but some grade I tumours are clinically more aggressive. Recent advances in the genetic study of meningiomas has allowed investigation into the influence of genetics on the tumour microenvironment, which is important for tumorigenesis. We have established that the endpoint genotyping method Kompetitive Allele Specific PCR (KASP™) is a fast, reliable method for the screening of meningioma samples into different non-NF2 mutational groups using a standard real-time PCR instrument. This genotyping method and four-colour flow cytometry has enabled us to assess the variability in the largest immune cell infiltrate population, M2 macrophages (CD45+HLA-DR+CD14+CD163+) in 42 meningioma samples, and to suggest that underlying genetics is relevant. Further immunohistochemistry analysis comparing AKT1 E17K mutants to WHO grade I NF2-negative samples showed significantly lower levels of CD163-positive activated M2 macrophages in meningiomas with mutated AKT1 E17K, signifying a more immunosuppressive tumour microenvironment in NF2 meningiomas. Our data suggested that underlying tumour genetics play a part in the development of the immune composition of the tumour microenvironment. Stratifying meningiomas by mutational status and correlating this with their cellular composition will aid in the development of new immunotherapies for patients.


Cheiloscopy, Palatoscopy and Odontometrics in Sex Prediction and Dis-crimination - a Comparative Study.
Human identification is the forensic odontologist's primary duty in the fields like violent crime, child abuse, elder abuse, missing persons and mass disaster scenarios. In each context, dental traits may produce compelling evidence to aid victim identity, suspect identity and narrow down the outcome of investigative casework. Sometimes it becomes necessary to apply some least known and less popular techniques in identification procedure where lip prints, rugae patterns and canine odontometrics can give us comparatively valid conclusions pertaining to person's identification. This study elucidates the significance of cheiloscopy, palatoscopy and canine odontometry in sex prediction and discrimination. A cross- sectional study involving a total of 60 subjects, 30 males and 30 females were selected from the outpatient department of oral medicine and radiology. Lip prints were recorded using lipstick, palatal impressions were taken with alginate and odontometric measurements were taken with digital vernier calipers from every subject. All the obtained records were analyzed by two observers. Reliability of lip prints was assessed using Kappa coefficient. Comparison of rugae patterns was done using Chi-square test. Mean canine and inter canine width was compared using t test. A p-value of <0.05 was considered significant. Statistically significant difference was found with the different lip print patterns analyzed in males and females, while no significant difference was observed in the rugae patterns but a significant difference in the mesio-distal width of mandibular canines in males and females was found with right mandibular canine(3.73%) showing greater sexual dimorphism compared to left mandibular canine(3.06%). This study shows the uniqueness of the lip prints and rugae patterns with the lip prints showing sensitivity of 81.7% giving reliable prediction of sex over palatoscopy. Hence, cheiloscopy along with the canine odontometrics aid in sex determination and can be considered as an ancilliary forensic tool in identification.


Large-scale genomics unveils the genetic architecture of psychiatric disorders.
Family study results are consistent with genetic effects making substantial contributions to risk of psychiatric disorders such as schizophrenia, yet robust identification of specific genetic variants that explain variation in population risk had been disappointing until the advent of technologies that assay the entire genome in large samples. We highlight recent progress that has led to a better understanding of the number of risk variants in the population and the interaction of allele frequency and effect size. The emerging genetic architecture implies a large number of contributing loci (that is, a high genome-wide mutational target) and suggests that genetic risk of psychiatric disorders involves the combined effects of many common variants of small effect, as well as rare and de novo variants of large effect. The capture of a substantial proportion of genetic risk facilitates new study designs to investigate the combined effects of genes and the environment.


Efficient DNA knock-in using AAV-mediated delivery with 2-cell embryo CRISPR-Cas9 electroporation.
Recent advances in CRISPR-Cas genome editing technology have been instrumental in improving the efficiency to produce genetically modified animal models. In this study we have combined four very promising approaches to come up with a highly effective pipeline to produce knock-in mouse and rat models. The four combined methods include: AAV-mediated DNA delivery, single-stranded DNA donor templates, 2-cell embryo modification, and CRISPR-Cas ribonucleoprotein (RNP) electroporation. Using this new combined approach, we were able to produce successfully targeted knock-in rat models containing either Cre or Flp recombinase sequences with knock-in efficiencies over 90%. Furthermore, we were able to produce a knock-in mouse model containing a Cre recombinase targeted insertion with over 50% knock-in efficiency directly comparing efficiencies to other commonly used approaches. Our modified AAV-mediated DNA delivery with 2-cell embryo CRISPR-Cas9 RNP electroporation technique has proven to be highly effective for generating both knock-in mouse and knock-in rat models.


A tale of two tissues: stem cells in cartilage and corneal tissue engineering.
Laboratory investigations of stems cells in regenerative medicine have generated considerable interest within recent years, however some of this excitement is yet to be matched in the clinical arena. Two fields that are well poised to make significant clinical impact in the coming years are those of cartilage and corneal regeneration. In the case of cornea, it is widely acknowledged that corneal epithelium is derived from an adult stem cell type resident within the cornea. These cells, known as limbal stem cells (LSC's), have been widely investigated for their ex-vivo culture and subsequent transplantation efficacy, with some techniques already enjoying limited clinical application. Thus far however, only preliminary evidence currently exists to suggest that there is a population of adult stem cells which gives rise to stromal keratocytes or to the corneal endothelium. A handful of reports have discussed studies in which non-LSC adult stem cells such as mesenchymal stems cells (MSCs) or embryonic stem cells (ESCs) are being applied to corneal regeneration. Though adult stem cells have been shown to exist in articular cartilage, they have proven elusive, which corroborates the limited ability of this tissue to self-repair. Rather, MSCs, ESCs as well as adipose-derived, periosteum-derived, muscle-derived and synovium-derived stem cells (ADCs, PDCs, MDCs and SDCs respectively) are being extensively explored for cartilage regeneration. This review discusses emerging trends in the applications of both adult and embryonic stem cells to cartilage and corneal regeneration, with an emphasis on those techniques that have been applied clinically or which show significant potential for clinical translation.


[Satisfaction with life and functionality among elderly patients in a geriatric outpatient clinic].
Aging is often accompanied by functional limitations that affect self esteem, lowering the level of satisfaction with life. Studies highlight satisfaction with life as a predictor of quality of life and has a marked correlation with a decrease in functionality. Therefore, this study sought to examine the relationship between satisfaction with life and functional independence and performance of the lower limbs (muscle strength, gait speed and balance) among the elderly in outpatient care with respect to age groups and genders. A total of 125 elderly men and women aged 60 years and above, attending a geriatric outpatient clinic. The instruments used were: 1) Functional Independence Measure (FIM) to evaluate functional dependence; 2) Short Physical Performance Battery (SPPB) to measure physical performance; 3) Satisfaction with life. The sample was random, with a predominance of females, in which the older participants had greater functional impairment. In univariate logistic regression analysis and multivariate analysis, overall self reported and comparative satisfaction with life was more satisfactory among the oldest elderly. The results suggest that older individuals have better satisfaction with life even though they have greater functional impairment.


Spatial coherence measurements of a 13.2 nm transient nickel-like cadmium soft x-ray laser pumped at grazing incidence.
The spatial coherence of a 13.2 nm transient collisional Ni-like Cd soft X-ray laser pumped at 23 degrees grazing incidence was measured in a series of Young's double-slit experiments. We observed pronounced fringe visibility variations associated with microstructures in the beam's intensity profile. The transverse coherence length was measured to be about 1/20 of the beam diameter and did not significantly improve with longer plasma columns. The equivalent incoherent source size is determined to be 10 mum and the laser's peak spectral brightness ~ 3 x 10(23) photons/sec/mm(2)/mrad(2) within less than 0.01% spectral bandwidth.


Impact of income inequalities on subjective perception of older adult health: An analysis of healthy life expectancy in China from 1997 to 2006.
Driven by rapid socioeconomic development over recent decades, there are widening income inequalities and subjective health disparities among older adults in China. This study explored the relationship between income inequalities and self-rated healthy life expectancy (HLE) considering potential sex-specific differences. From a cohort of 1760 individuals aged ≥60 years from the China Health and Nutrition Survey (1997-2006), we calculated age-specific life expectancy (LE), HLE and the proportion of HLE to LE (HLE/LE) by sex using the IMaCh software, incorporated time-varying income levels of older adults. Although longevity has significantly improved, the well-being of Chinese older adults could be further enhanced by assessing HLE, as the results showed that at age 60 years, ~20% of their LE was characterized by unhealthiness. Discriminated by economic status, lower-income individuals experienced worse self-rated health compared with the general population and affluent counterparts. For instance, at age 60 years, the LE, HLE and HLE/LE of low-income men were 19.8 (95% CI 18.4-21.1), 16.2 years (95% CI 15.0-17.5) and 81.8%, respectively, lower than their general (21.7, 95% CI: 20.4-23.0; 18.0, 95% CI 16.8-19.2; and 82.9%) and high-income counterparts (25.1, 95% CI 23.1-27.2; 21.4, 95% CI 19.5-23.3; and 85.3%). Sex disparities were noticeable among low-income individuals in HLE/LE, with low-income women showing the most significant disadvantage. Income inequalities exacerbated subjective health disparities among older adults, particularly among lower-income individuals and women. Our findings carry significant implications for formulating public health and social welfare strategies, especially in nations grappling with an aging population and undergoing parallel socioeconomic development. Geriatr Gerontol Int 2023; ••: ••-••.


TNF-α and IGF1 modify the microRNA signature in skeletal muscle cell differentiation.
Elevated levels of the inflammatory cytokine TNF-α are common in chronic diseases or inherited or degenerative muscle disorders and can lead to muscle wasting. By contrast, IGF1 has a growth promoting effect on skeletal muscle. The molecular mechanisms mediating the effect of TNF-α and IGF1 on muscle cell differentiation are not completely understood. Muscle cell proliferation and differentiation are regulated by microRNAs (miRNAs) which play a dominant role in this process. This study aims at elucidating how TNF-α or IGF1 regulate microRNA expression to affect myoblast differentiation and myotube formation. In this study, we analyzed the impact of TNF-α or IGF1 treatment on miRNA expression in myogenic cells. Results reveal that i) TNF-α and IGF1 regulate miRNA expression during skeletal muscle cell differentiation in vitro, ii) microRNA targets can mediate the negative effect of TNF-α on fusion capacity of skeletal myoblasts by targeting genes associated with axon guidance, MAPK signalling, focal adhesion, and neurotrophin signalling pathway, iii) inhibition of miR-155 in combination with overexpression of miR-503 partially abrogates the inhibitory effect of TNF-α on myotube formation, and iv) MAPK/ERK inhibition might participate in modulating the effect of TNF-α and IGF1 on miRNA abundance. The inhibitory effects of TNF-α or the growth promoting effects of IGF1 on skeletal muscle differentiation include the deregulation of known muscle-regulatory miRNAs as well as miRNAs which have not yet been associated with skeletal muscle differentiation or response to TNF-α or IGF1. This study indicates that miRNAs are mediators of the inhibitory effect of TNF-α on myoblast differentiation. We show that intervention at the miRNA level can ameliorate the negative effect of TNF-α by promoting myoblast differentiation. Moreover, we cautiously suggest that TNF-α or IGF1 modulate the miRNA biogenesis of some miRNAs via MAPK/ERK signalling. Finally, this study identifies indicative biomarkers of myoblast differentiation and cytokine influence and points to novel RNA targets.


[Aquaculture and public health. The emergence of diphyllobothriasis in Chile and the world].
Recent clinical and epidemiológical information, an analysis of the literature, and study of the technical aspects of Chilean salmon aquaculture indicate that this activity has the ability to expand the range of diphyllobothriasis caused by the fish tapeworm Diphyllobothrium latum. Evidence for expansion of the range of the fish tapeworm includes the emergences of clinical cases in Brazil related to consumption of salmon produced in Chile. Expansion of the range of this parasite is also suggested by an increase of its geographical range in Chile, beyond its previously endemic foci in the lakes of Regions IX and X. Prevention of further dissemination of this parasitic disease rests on an improvement of sanitation and sewage disposal around the lakes of Regions IX, X and XI in Chile, improvement in aquaculture methods including curtailing the use of fish tapeworm-contaminated lakes to grow juveniles forms of salmonids and more measures to decrease the number of salmonid escapees from marine pens to prevent their return to rivers and lakes carrying the infestation. Moreover, tracking the origin of juveniles in marketed salmon, determining the presence of plerocercoids in them, and increased education of the public regarding the potential dangers of eating raw fish should also be implemented. Only by stimulating the dialogue between the industry, consumers and state regulators will it be possible to implement appropriate measures to prevent further expansion of this parasitic disease by salmon aquaculture.


Functional Foods for Management of Diarrhea and Malnutrition in Rats Emphasizing on Nucleotides Role.
Diarrhea and malnutrition are major health problems in developing countries. Inflammation, high oxidative stress, poor nutritional status, and fatty liver were encountered during such diseases. Patents for diarrhea and malnutrition management (WO2007/130882A2, WO00/37106A1, WO2014/152420, and CA2987364A1) were published. The objective was to introduce anti-diarrhea functional foods with a preventive effect on malnutrition. Two processing techniques were applied for preparing functional foods (formula 1 ingredients were made into cookies followed by grinding; formula 2 ingredients were pre-cooked, dried, and mixed in powder form) that were evaluated in a rat model of diarrhea with malnutrition (DM). Formula 2 was also assessed when mixed with nucleotides. The ingredients were edible plants that possess an anti-diarrheal effect with high protein sources (legumes and casein). Induction of diarrhea with malnutrition, high oxidative stress, inflammation, accumulation of liver fat, and histopathological changes were demonstrated in DM control compared to normal control. The functional foods produced variable improvement in growth curves, food efficiency ratio, hemoglobin, hematocrit and plasma zinc, protein, albumin, globulin, lipase activity, and MDA. Formula 1 was superior in improving intestinal histopathology while formula 2 was more efficient in elevating plasma iron. Formula 2 with nucleotides was the best in improving growth curves, alkaline phosphatase, and reducing liver fat. Intestinal mucosa reduced glutathione and nitrite showed an efficient significant reduction on treatment with formula 2 with or without nucleotides. The formulas showed an anti-diarrheal effect by improving feces weight and moisture content. Studied functional foods showed an anti-diarrheal effect and malnutrition improvement with different degrees.


Biosurfactants from yeasts: characteristics, production and application.
Biosurfactants are surface-active compounds from biological sources, usually extracellular, produced by bacteria, yeast or fungi. Research on biological surfactant production has grown significantly due to the advantages they present over synthetic compounds such as biodegradability, low toxicity, diversity of applications and functionality under extreme conditions. Although the majority of microbial surfactants have been reported in bacteria, the pathogenic nature of some producers restricts the wide application of these compounds. A growing number of aspects related to the production of biosurfactants from yeasts have been the topic of research during the last decade. Given the industrial importance of yeasts and their potential to biosurfactant production, the goal of this chapter is to review the biosurfactants identified up to present, focusing the relevant parameters that influence biosurfactant production by yeasts and its characteristics, revealing the potential of application of such compounds in the industrial field and presenting some directions for the future development of this area, taking into account the production costs.


[Effect of specific physiotherapy on chronic pain, functional level and quality of life in osteoporosis. A prospective randomized single-blind placebo-controlled study].
Patients suffering from osteoporotic vertebral fractures are handicapped by pain and reduced quality of life. Our aim was to investigate the effect of a short training program for osteoporotic patients with regard to pain level, use of analgetics and quality of life. We performed a prospective randomized single-blinded placebo-controlled study. The training program included general training of balance and muscle strength and stabilization of the back. The participants were randomised to 10 weeks of ambulatory training. Controls and training participants were tested weekly by registration of pain level and analgetic intake. Questionnaires on daily level of function and quality of life were given at the start and after five and 10 weeks. After three months both groups filled out the questionnaires at home. The training group had a significant reduction in pain score and use of analgetics. The distribution of functional score improved during training. Quality of life score improved significantly throughout the study and after three months. In conclusion, this ambulatory training program is effective for training osteoporotic patients with moderately severe pain and the training should be continued.


Community perceptions of genomic research: implications for addressing health disparities.
Increasing the engagement of racial and ethnic minorities in genomic research may help alleviate health disparities. This paper examines community perceptions of the relationships between race, genes, environment, and health disparities, and it discusses how such perceptions may influence participation in genomic research. We conducted semi-structured interviews with 91 African American, Latino, and white lay community members and community leaders in North Carolina. Using constant comparison methods, we identified, compared, and developed linkages between conceptual categories and respondent groups. Participants described gene-environment interactions as contributing to group differences in health outcomes, expressed the belief that genetic predisposition to disease differs across groups, and said that social conditions trigger group-level genetic differences and create poorer health outcomes among African Americans. Given the regional presence of major research institutions and the relatively high education level of many participants, this sample may not reflect the perspectives of those most disparately affected by health disparities. Members from multiple community sectors share perceptions and may respond to similar approaches when attempts are made to increase participation in genomic research. Researchers may inadvertently fuel the perception that health disparities experienced by minorities are rooted in the shared genomes of a particular group as distinct from those of other groups. The way researchers use race and ethnicity in recruitment, analysis, and communication of research findings inaccurately implies that there are genetic differences between races, when categories of social experience or ancestry may more accurately characterize health differences. Understanding these issues is crucial to designing effective community engagement strategies, recruitment plans, and messages about genomic research, which could ultimately help to lessen health disparities.


Perceived work environment and cognitive style.
Differences in perceived work environment are examined by age, gender, and occupation type, along with whether cognitive style predicts perceived work environment. The sample consists of 2185 men and women ranging in age from 21 to 93 years. Perceived work environment was measured using Moos's Work Environment Scale (WES) for the subscales of Autonomy, Control, and Innovation. The Test of Behavioral Rigidity (TBR) was used to measure cognitive style. Findings indicate significant mean level differences in perceived work environment based on gender, age, and occupation type. Hierarchical regression analyses indicated that perceived work environment predicted concurrent cognitive style beyond demographic variables.


DNA-vaccine platform development against H1N1 subtype of swine influenza A viruses.
Swine influenza virus (SIV) is an important viral pathogen in pig populations. However, commercial vaccines cannot provide complete protection with induced humoral immunity only, and require frequent updates to fight against current isolates. DNA vaccination is an effective means of eliciting both arms of the immune system, the humoral and cellular immune responses. In this study, DNA vector pcDNA3.1 was inserted with a chimeric intron downstream of the CMV promoter region followed by a Kozak sequence to enhance the expression of gene inserts. The C-terminal of the VP22 gene (VP22c), encoding the tegument protein of bovine herpesvirus-1, was fused separately to the N-terminal of four quadruplicated epitopes: two B-cell epitopes (HA91-108 and M2e), and two T-cell epitopes (NP366-374 and NP380-393), which were conserved, at least among the three SIV subtypes prevailing in pig populations in North America. Linker -KK- was used to space between each copy of the two B-cell epitopes, and -RVKR- was used for the two T-cell epitopes, in order to enhance the presentation of epitopes to the immune system. The expression of epitopes was confirmed in in vitro transfection of 293FT cells, and higher percentages of epitope-positive cells were achieved from the plasmids containing VP22c than those without. After the DNA plasmids were administered to mice intramuscularly in combination or separately, or boosted with recombinant proteins of quadruplicated epitopes fused to VP22c, the vaccine stimulated the desired epitope-specific humoral immunity to the two B-cell epitopes, and cellular immunity to the epitope NP380-393. Our results indicate that plasmids with quadruplicated epitopes fused to the VP22c may be a potential vehicle in developing epitopes as vaccines against SIV.


Catheter ablation related mitral valve injury: the importance of early recognition and rescue mitral valve repair.
An increasing number of catheter ablations involve the mitral annular region and valve apparatus, increasing the risk of catheter interaction with the mitral valve (MV) complex. We review our experience with catheter ablation-related MV injury resulting in severe mitral regurgitation (MR) to delineate mechanisms of injury and outcomes. We searched the Mayo Clinic MV surgical database over a 19-year period (1993-2012) and the electrophysiologic procedures database over a 23-year period (1990-2013) and identified 9 patients with catheter ablation related MV injury requiring clinical intervention. Indications for ablation included atrial fibrillation (AF) [n = 4], ventricular tachycardia (VT) [n = 3], and left-sided accessory pathways [n = 2]. In all 4 AF patients, a circular mapping catheter entrapped in the MV apparatus was responsible for severe MR. In all 3 VT patients, radiofrequency energy delivery led to direct injury to the MV apparatus. In the 2 patients with accessory pathways, both mechanisms were involved (1 per patient). Six patients required surgical intervention (5 MV repair, 1 catheter removal). One patient developed severe functional MR upon successful endovascular catheter disentanglement that improved spontaneously. Two VT patients with persistent severe postablation MR were managed nonsurgically, 1 of whom died 3 months postprocedure. Circular mapping catheter entrapment and ablation at the mitral annulus are the most common etiologies of MV injury during catheter ablation. Close surveillance of the MV is needed during such procedures and early surgical repair is important for successful salvage if significant injury occurs.


Amelioration of nerve Na(+)-K(+)-ATPase activity independently of myo-inositol level by PGE1 analogue OP-1206.alpha-CD in streptozocin-induced diabetic rats.
An oral prostaglandin E1 (PGE1) analogue, OP-1206.alpha-CD, was given to rats with streptozocin (STZ)-induced diabetes to examine the therapeutic effects of OP-1206 on short-term and long-term diabetic neuropathy and its action mechanism with special reference to nerve Na(+)-K(+)-ATPase activity. In the short-term experiment, OP-1206 was administered daily to diabetic rats in 3- and 30-mg/kg doses for 4 wk from the day of STZ injection. In the long-term study, 10 micrograms/kg OP-1206 was also given daily for 8 wk from 7 mo after induction of diabetes. The compound improved decreased sciatic motor nerve conduction velocity in both short-term and long-term diabetic rats. The nerve Na(+)-K(+)-ATPase activity of diabetic rats, reduced by 40% compared with controls, was reversed to the level of controls in both experiments, whereas weight loss and hyperglycemia were unchanged, and neither nerve sorbitol accumulation nor myo-inositol depletion was corrected. In a morphometric analysis of myelinated nerve fibers (MNFs) in long-term diabetes, the mean diameter of the largest 10% of MNFs was significantly reduced in untreated diabetic compared with control rats, but OP-1206 completely reversed this reduction. The results suggest that OP-1206 ameliorates a decrease in nerve Na(+)-K(+)-ATPase activity without any effect on nerve myo-inositol level and that the compound may be not only a potent therapeutic agent for the treatment of diabetic neuropathy but also a useful research tool to investigate the mechanism of nerve Na(+)-K(+)-ATPase activity regulation.


Hypothyroidism and depression. Evidence from complete thyroid function evaluation.
To evaluate the relationship between hypothyroidism and depression, thyroid function was evaluated in 250 consecutive patients referred to a psychiatric hospital for treatment of depression or anergia. Twenty of the 250 patients had some degree of hypothyroidism. Two patients (less than 1%) were identified with grade 1 (overt); nine patients (3.6%), grade 2 (mild); and ten patients (4%), grade 3 (subclinical) hypothyroidism. These results suggest that a significant proportion of patients with depression and anergia may have early hypothyroidism, the cases of about half of which are detected only by thyrotropin-releasing hormone (TRH) testing. Because hypothyroidism can produce signs and symptoms of depression and can coexist as a second illness in depressed patients, patients with early hypothyroidism may be candidates for thyroid replacement therapy. Clinical examination and measurement of triiodothyronine resin uptake thyroxine and baseline thyroid-stimulating hormone (TSH) levels, and TSH response to TRH are necessary to identify candidates for thyroid replacement among cases diagnosed by descriptive criteria as having either major or minor depression, particularly those that are atypical or treatment resistant.


What do people living with HIV/AIDS expect from their physicians? Professional expertise and the doctor-patient relationship.
This qualitative study identifies the types of professional expertise that physicians are seen to possess in clinical encounters from the perspective of people living with HIV/AIDS (PLWHA). Respondents looked to their physicians for expert knowledge in 3 key areas: medical/clinical; legal/statutory; and ethical/moral. Physicians were seen to be authorities in each of these areas and their judgments, though not always agreed with, were taken seriously and influenced the health care decisions made by PLWHA. The authority that comes with professional expertise in each of the areas identified was experienced both positively and negatively by PLWHA. Understanding the expectations of patients in the medical encounter can assist physicians in providing optimal care in the management of HIV/AIDS.


Dynamics of conformational transitions in polymers.
Conformational transitions in polymers involve large angle rotations about bonds. The process must proceed in a way that does not require gross movements of the macromolecules. The dynamics have been investigated by computer simulation and kinetic theory. The rate-determining step in the transition is found to occur in a mode which is kept local by distortion of nearby parts of the molecule. One especially important type of cooperativity, crank-like counterrotation of second-neighbor bonds, is identified. Experiments which provide evidence about the dynamics of conformational transitions are discussed.


Further characterization of Escherichia coli brain microvascular endothelial cell invasion gene ibeA by deletion, complementation, and protein expression.
The ibeA gene (ibe10) previously identified by TnphoA mutagenesis is part of a 50-kDa full-length open-reading frame (ORF) encoded by a 1.37-kb DNA fragment. An isogenic in-frame deletion mutant of ibeA (ZD1) was constructed by chromosomal gene replacement with a suicide plasmid pCVD442 carrying a 2.1-kb DNA fragment with an ibeA deletion. Similar to the previously described TnphoA insertion mutant of ibeA, the isogenic ibeA deletion mutant ZD1 was significantly less invasive in human brain microvascular endothelial cells (BMECs) than the parent strain. The mutant ZD1 was fully complemented by the ibeA ORF. The ibeA gene was subcloned into pET28a(+) and was expressed as a recombinant protein with an N-terminal histidine tag. The recombinant IbeA protein had much greater activity (50 times) in blocking the invasion of BMECs by Escherichia coli K1 than did the partial protein fragment, which provides further evidence that ibeA is an important determinant for E. coli K1 invasion of BMECs.


Ubiquitin metabolism affects cellular response to volatile anesthetics in yeast.
To investigate the mechanism of action of volatile anesthetics, we are studying mutants of the yeast Saccharomyces cerevisiae that have altered sensitivity to isoflurane, a widely used clinical anesthetic. Several lines of evidence from these studies implicate a role for ubiquitin metabolism in cellular response to volatile anesthetics: (i) mutations in the ZZZ1 gene render cells resistant to isoflurane, and the ZZZ1 gene is identical to BUL1 (binds ubiquitin ligase), which appears to be involved in the ubiquitination pathway; (ii) ZZZ4, which we previously found is involved in anesthetic response, is identical to the DOA1/UFD3 gene, which was identified based on altered degradation of ubiquitinated proteins; (iii) analysis of zzz1Delta zzz4Delta double mutants suggests that these genes encode products involved in the same pathway for anesthetic response since the double mutant is no more resistant to anesthetic than either of the single mutant parents; (iv) ubiquitin ligase (MDP1/RSP5) mutants are altered in their response to isoflurane; and (v) mutants with decreased proteasome activity are resistant to isoflurane. The ZZZ1 and MDP1/RSP5 gene products appear to play important roles in determining effective anesthetic dose in yeast since increased levels of either gene increases isoflurane sensitivity whereas decreased activity decreases sensitivity. Like zzz4 strains, zzz1 mutants are resistant to all five volatile anesthetics tested, suggesting there are similarities in the mechanisms of action of a variety of volatile anesthetics in yeast and that ubiquitin metabolism affects response to all the agents examined.


Bilateral subthalamic stimulation for Parkinson's disease by using three-dimensional stereotactic magnetic resonance imaging and electrophysiological guidance.
Several methods are used for stereotactically guided implantation of electrodes into the subthalamic nucleus (STN) for continuous high-frequency stimulation in the treatment of Parkinson's disease (PD). The authors present a stereotactic magnetic resonance (MR) method relying on three-dimensional (3D) T1-weighted images for surgical planning and multiplanar T2-weighted images for direct visualization of the STN, coupled with electrophysiological recording and stimulation guidance. Twelve patients with advanced PD were enrolled in this study of bilateral STN implantation. Both STNs were visible as 3D ovoid biconvex hypointense structures located in the upper mesencephalon. The coordinates of the centers of the STNs were determined with reference to the patient's anterior commissure-posterior commissure line by using a new landmark, the anterior border of the red nucleus. Electrophysiological monitoring through five parallel tracks was performed simultaneously to define the functional target accurately. Microelectrode recording identified high-frequency, spontaneous, movement-related activity and tremor-related cells within the STNs. Acute STN macrostimulation improved contralateral rigidity and akinesia, suppressed tremor when present, and could induce dyskinesias. The central track, which was directed at the predetermined target by using MR imaging, was selected for implantation of 19 of 24 electrodes. No surgical complications were noted. At evaluation 6 months after surgery, continuous STN stimulation was shown to have improved parkinsonian motor disability by 64% and 78% in the "off' and "on" medication states, respectively. Antiparkinsonian drug treatment was reduced by 70% in 10 patients and withdrawn in two patients. The severity of levodopa-induced dyskinesias was reduced by 83% and motor fluctuations by 88%. Continuous high-frequency stimulation of the STN applied through electrodes implanted with the aid of 3D MR imaging and electrophysiological guidance is a safe and effective therapy for patients suffering from severe, advanced levodopa-responsive PD.


Second messengers, ion channels and pharmacology of airway smooth muscle.
The airway smooth muscle cell is the chief effector cell governing the control of airway calibre in the human lung. The contractile state of the airway smooth muscle cell is predominantly influenced by the balance of constrictor and relaxant stimuli. Agents such as histamine and acetylcholine cause airway smooth muscle cells to contract through activation of specific cell surface receptors and engagement of signal transduction pathways and/or ion channels. The predominant pathway mediating constriction is activation of phospholipase C, with release of inositol 1,4,5-triphosphate and elevation of intracellular calcium levels. Relaxation is brought about predominantly by stimulation of adenylyl cyclase-coupled receptors (e.g. the beta2-adrenoceptor) resulting in elevation of cell cyclic adenosine monophosphate content. Complex crosstalk occurs between both of these pathways and also ion channels expressed on the airway smooth muscle cell membrane, leading to careful regulation of airway smooth muscle tone. A greater understanding of the mechanisms governing control of these pathways will lead to the identification of novel therapeutic targets which will in turn lead to new agents for the treatment of asthma.


Cerebral malaria: clinical features, pathophysiology and treatment.
Herbert Gilles played an important role in the establishment of the Wellcome-Mahidol University, Oxford Tropical Medicine Research Programme in Thailand in 1979. The randomized, placebo-controlled trial of dexamethasone in cerebral malaria that was carried out in Chantaburi in 1980 yielded results which led to the abandonment of ancillary corticosteroid therapy in this disease and contributed to a rejection of the 'permeability hypothesis'. The clinical manifestations of strictly defined cerebral malaria have not been described both in non-immune adults in Thailand and Vietnam and in African children. Clinical and histopathological studies in human patients, together with laboratory studies of cyto-adherence, malaria 'toxin' and cytokine production have provided some evidence for both the 'mechanical' and 'toxin-cytokine' hypotheses to explain the pathophysiology of this condition. Chemotherapy is challenged by the continuing evolution of antimalarial resistance. Recently, the most powerful studies ever carried out with antimalarial drugs have demonstrated that artemether and quinine achieve similar case fatalities, in the range 11%-21%, and that both drugs have some advantages and disadvantages. Further studies are needed to define the efficacy and safety of prophylactic anticonvulsants and exchange transfusion in cerebral malaria. Cerebral malaria remains a major cause of mortality and, in African children, morbidity.


Bacterial RecA Protein Promotes Adenoviral Recombination during In Vitro Infection.
Adenovirus infections in humans are common and sometimes lethal. Adenovirus-derived vectors are also commonly chosen for gene therapy in human clinical trials. We have shown in previous work that homologous recombination between adenoviral genomes of human adenovirus species D (HAdV-D), the largest and fastest growing HAdV species, is responsible for the rapid evolution of this species. Because adenovirus infection initiates in mucosal epithelia, particularly at the gastrointestinal, respiratory, genitourinary, and ocular surfaces, we sought to determine a possible role for mucosal microbiota in adenovirus genome diversity. By analysis of known recombination hot spots across 38 human adenovirus genomes in species D (HAdV-D), we identified nucleotide sequence motifs similar to bacterial Chi sequences, which facilitate homologous recombination in the presence of bacterial Rec enzymes. These motifs, referred to here as ChiAD, were identified immediately 5' to the sequence encoding penton base hypervariable loop 2, which expresses the arginine-glycine-aspartate moiety critical to adenoviral cellular entry. Coinfection with two HAdV-Ds in the presence of an Escherichia coli lysate increased recombination; this was blocked in a RecA mutant strain, E. coli DH5α, or upon RecA depletion. Recombination increased in the presence of E. coli lysate despite a general reduction in viral replication. RecA colocalized with viral DNA in HAdV-D-infected cell nuclei and was shown to bind specifically to ChiAD sequences. These results indicate that adenoviruses may repurpose bacterial recombination machinery, a sharing of evolutionary mechanisms across a diverse microbiota, and unique example of viral commensalism.IMPORTANCE Adenoviruses are common human mucosal pathogens of the gastrointestinal, respiratory, and genitourinary tracts and ocular surface. Here, we report finding Chi-like sequences in adenovirus recombination hot spots. Adenovirus coinfection in the presence of bacterial RecA protein facilitated homologous recombination between viruses. Genetic recombination led to evolution of an important external feature on the adenoviral capsid, namely, the penton base protein hypervariable loop 2, which contains the arginine-glycine-aspartic acid motif critical to viral internalization. We speculate that free Rec proteins present in gastrointestinal secretions upon bacterial cell death facilitate the evolution of human adenoviruses through homologous recombination, an example of viral commensalism and the complexity of virus-host interactions, including regional microbiota.


[Changes in the human fetal membranes in late pregnancy toxicoses].
Changes of a destructive and adaptational character have been revealed in the extraplacental coats in late toxicoses of pregnant women by histological, histochemical and spectrophotometrical methods. The intesity of the apotomical way of the glucose oxidation decreased in the aminiotic epithelium. While the activity of enzymes converting the lipid decomposition products into glycolysis increased. The glucose utilization in the chorial epithelium occurs mainly due to anaerobic glycolysis. Histochemical methods have established a high content of acid glucoaminoglycanes in the smooth chorion under these pathological conditions and the appearance of fibrinoid deposits on the border with the decidual tissue. The number of polyploid cells containing a considerable amount of RNA has been found to increase in the amniotic and chorial epithelium by spectrophotometric methods. The possible role of the extraplacental coats of the fetus in pathogenesis of late toxicoses is discussed.


Fluorescence In Situ Hybridization and Polymerase Chain Reaction to Detect Infections in Patients With Left Ventricular Assist Devices.
The development of driveline infections following left ventricular assist device (LVAD) implantation remains a major problem. We investigated the impact of fluorescence in situ hybridization (FISH) combined with 16S rRNA gene sequencing on the diagnosis of driveline infections. LVAD drivelines (n = 61) from 60 consecutive patients were obtained during LVAD explantation and subjected to FISH analysis. 16S rRNA gene polymerase chain reaction (PCR) and sequencing to identify the microorganisms were performed. Results were compared with those of a standard microbiological culture. The reasons for pump removal were heart transplantation (n = 22), weaning (n = 14), pump exchange due to pump thrombosis (n = 12), technical problems (n = 7), or death (n = 5). Of the 60 patients, 26 exhibited clinical signs of a VAD-specific infection, while 34 (with 35 drivelines) showed no clinical signs of infection before explantation. The spectrum of identified pathogens differed between FISH/PCR and conventional microbiological diagnostics. In general, the bacterial spectrum was more diverse in FISH/PCR as compared with conventional microbiology, which more often showed only typical skin flora (coagulase-negative staphylococci and Corynebacteriaceae). In addition to identifying the species, FISH/PCR provided information about the spatial distribution and invasiveness of the microorganisms. Cultures usually represent the only source of microbiological information for clinicians and often prove to be unsatisfactory in complex LVAD cases. FISH/PCR not only identified a greater number and variety of microorganisms than standard culture did, but it also provided information about the number, localization, and biofilm state of the pathogens, making it a useful tool for diagnosing the specific cause of LVAD driveline infections.


Microtubule polymerization dynamics.
The polymerization dynamics of microtubules are central to their biological functions. Polymerization dynamics allow microtubules to adopt spatial arrangements that can change rapidly in response to cellular needs and, in some cases, to perform mechanical work. Microtubules utilize the energy of GTP hydrolysis to fuel a unique polymerization mechanism termed dynamic instability. In this review, we first describe progress toward understanding the mechanism of dynamic instability of pure tubulin and then discuss the function and regulation of microtubule dynamic instability in living cells.


Insulating state in tetralayers reveals an even-odd interaction effect in multilayer graphene.
Close to charge neutrality, the electronic properties of graphene and its multilayers are sensitive to electron-electron interactions. In bilayers, for instance, interactions are predicted to open a gap between valence and conduction bands, turning the system into an insulator. In mono and (Bernal-stacked) trilayers, which remain conducting at low temperature, interactions do not have equally drastic consequences. It is expected that interaction effects become weaker for thicker multilayers, whose behaviour should converge to that of graphite. Here we show that this expectation does not correspond to reality by revealing the occurrence of an insulating state close to charge neutrality in Bernal-stacked tetralayer graphene. The phenomenology-incompatible with the behaviour expected from the single-particle band structure-resembles that observed in bilayers, but the insulating state in tetralayers is visible at higher temperature. We explain our findings, and the systematic even-odd effect of interactions in Bernal-stacked layers of different thickness that emerges from experiments, in terms of a generalization of the interaction-driven, symmetry-broken states proposed for bilayers.


[Establishment of time-resolved fluorescence immunochromatographic assay for detection of carbohydrate antigen 19-9].
To establish a time-resolved fluorescence immunochromatographic assay for quantitative determination of carbohydrate antigen 19-9 (CA19-9) in serum, we prepared CA19-9 test strips by integrating double-antibody sandwich method and fluorescence immunochromatography technique. Carboxy fluorescent microspheres and nitrocellulose membrane were used as carriers for labeling and coating CA19-9 pairing antibodies. We optimized the process by adjusting the amount of labeling and coating antibody. According to the linear range, lowest detection limit and precision, We evaluated the time-resolved fluorescence immunochromatographic assay of CA19-9. When the amount of labeled antibody was 80 μg for 20 μL fluorescent microspheres, and the concentration of coated antibody on the test line was 1.5 mg/mL, the optimal reaction time was 15 minutes. Assay linear range was 12.5 to 800 U/mL and the minimum detection limit was 6.32 U/mL. The Within-run and between-run coefficient of variation were less than 15%. Average recovery rate was 101%. By detecting 50 clinical samples in parallel with Roche electrochemical luminescence detection kit, correlation coefficient was 0.980 6. The experiment, initially established a fluorescence immunochromatographic detection method to quantitative detection of serum CA19-9, which has a good clinical application prospect.


Molecular architecture of a miRNA-regulated 3' UTR.
Animal genomes contain hundreds of microRNAs (miRNAs), small regulatory RNAs that control gene expression by binding to complementary sites in target mRNAs. Some rules that govern miRNA/target interaction have been elucidated but their general applicability awaits further experimentation on a case-by-case basis. We use here an assay system in transgenic nematodes to analyze the interaction of the Caenorhabditis elegans lsy-6 miRNA with 3' UTR sequences. In contrast to many previously described assay systems used to analyze miRNA/target interactions, our assay system operates within the cellular context in which lsy-6 normally functions, a single neuron in the nervous system of C. elegans. Through extensive mutational analysis, we define features in the known and experimentally validated target of lsy-6, the 3' UTR of the cog-1 homeobox gene, that are required for a functional miRNA/target interaction. We describe that both in the context of the cog-1 3' UTR and in the context of heterologous 3' UTRs, one or more seed matches are not a reliable predictor for a functional miRNA/target interaction. We rather find that two nonsequence specific contextual features beyond miRNA target sites are critical determinants of miRNA-mediated 3' UTR regulation. The contextual features reside 3' of lsy-6 binding sites in the 3' UTR and act in a combinatorial manner; mutation of each results in limited defects in 3' UTR regulation, but a combinatorial deletion results in complete loss of 3' UTR regulation. Together with two lsy-6 sites, these two contextual features are capable of imparting regulation on a heterologous 3' UTR. Moreover, the contextual features need to be present in a specific configuration relative to miRNA binding sites and could either represent protein binding sites or provide an appropriate structural context. We conclude that a given target site resides in a 3' UTR context that evolved beyond target site complementarity to support regulation by a specific miRNA. The large number of 3' UTRs that we analyzed in this study will also be useful to computational biologists in designing the next generation of miRNA/target prediction algorithms.


Confidentiality on the information highway: balancing the needs of individual patients and society.
State and federal agencies, professional associations and business coalitions are actively promoting the development of regional or statewide health database organizations (HDOs). One of the key missions of HDOs is the public release of aggregated healthcare data and analyses to facilitate improved patient care. In 1992 and 1993 a committee appointed by the National Academy of Sciences' Institute of Medicine studied the implications of HDO formation in a landmark report. The committee's recommendations on how to assure accuracy and completeness of data, minimize potential harm from released data and guarantee appropriate protections for individually identifiable data are summarized and interpreted in the following article.


The effect of inhaled nitric oxide on the carrageenan-induced paw edema.
Inhaled nitric oxide therapy reaches not only pulmonary vessels, but also other vasculatures, presenting anti-inflammatory effects. Therefore, this study investigated the effects of inhaled nitric oxide on a mice model of carrageenan-induced paw edema. Paw edema was induced in male Swiss mice (20-30 g) by subplantar injection of carrageenan (0.05 ml of a 1% suspension in 0.9% saline). The evaluation of time-course edema (mililiter) was measured by plethysmometry until 12 h following carrageenan administration. Thirty minutes after carrageenan injection, some groups received inhaled nitric oxide (300 ppm at variable doses and times) or Indometacin (INDO 5 mg/Kg, v.o), while others received sildenafil (1 mg/Kg, i.p) or rolipram (3 mg/Kg, i.p.) with or without inhaled nitric oxide. Paws were assessed for edema levels by plethysmometry, mieloperoxidase activity and histological analysis. Inhaled nitric oxide significantly reduced carrageenan-induced paw edema, mieloperoxidase activity and inflammatory infiltrate, although similar results were also observed in sildenafil and rolipram treated groups. In addition, significant effects between inhaled nitric oxide with pharmacological therapy was observed. Inhaled nitric oxide presents anti-inflammatory effects on carrageenan-induce paw edema, as observed through reduced edema, mieloperoxidase activity and neutrophil infiltration, indicating that inhaled nitric oxide therapy goes beyond lung vascular effects.


Inflow and infiltration assessment of a prototype sanitary sewer network in a coastal city in China.
A 16-month monitoring program was conducted on a prototype sanitary system in a coastal city in China. The groundwater infiltration (GWI) on dry weather days and the rain-derived inflow and infiltration (RDII) on wet weather days were quantified and analyzed. The proportion of monthly averaged GWI to total flow can be as high as 70% during the observation period mainly due to the high groundwater level. The results also show that the ratio of RDII volume to total rainfall volume (defined as R-value) reaches a limited value of approximately 10% for the studied system when the total rainfall depth increases. A reference indicator Rlim for the limited R-value was proposed for assessing the conditions of sewer systems in terms of RDII. The Rlim value depends on local sewer conditions and in general, a lower Rlim value represents a better performance on RDII and vice versa. This study enriches the case studies on the performance of a specific sanitary sewer system on inflow and infiltration in a typical coastal city with exceptionally high groundwater levels, excess rainfall events in the monitoring season and possible typhoon events, which addresses the unique locational and hydrological properties of a representative coastal city.


Quantitative interpretation of bone marrow biopsies in MPN-What's the point in a molecular age?
The diagnosis of myeloproliferative neoplasms (MPN) requires the integration of clinical, morphological, genetic and immunophenotypic findings. Recently, there has been a transformation in our understanding of the cellular and molecular mechanisms underlying disease initiation and progression in MPN. This has been accompanied by the widespread application of high-resolution quantitative molecular techniques. By contrast, microscopic interpretation of bone marrow biopsies by haematologists/haematopathologists remains subjective and qualitative. However, advances in tissue image analysis and artificial intelligence (AI) promise to transform haematopathology. Pioneering studies in bone marrow image analysis offer to refine our understanding of the boundaries between reactive samples and MPN subtypes and better capture the morphological correlates of high-risk disease. They also demonstrate potential to improve the evaluation of current and novel therapeutics for MPN and other blood cancers. With increased therapeutic targeting of diverse molecular, cellular and extra-cellular components of the marrow, these approaches can address the unmet need for improved objective and quantitative measures of disease modification in the context of clinical trials. This review focuses on the state-of-the-art in image analysis/AI of bone marrow tissue, with an emphasis on its potential to complement and inform future clinical studies and research in MPN.


Morphokinetics and in vitro developmental potential of monopronucleated ICSI zygotes until the blastocyst stage.
The aim of this study was to provide a more comprehensive understanding of 1PN intracytoplasmic sperm injection (ICSI) zygotes. To achieve this objective, we assessed whether all 1PN-derived embryos showed a similar morphokinetic pattern, and if the morphokinetic behaviour of 1PN-derived embryos was comparable with that of 2PN-derived embryos. In total, 149 1PN ICSI zygotes (study group) and 195 2PN ICSI zygotes (control group) were included in the study. Embryo development potential was evaluated in terms of blastocyst rate. Morphokinetic parameters, including the pronucleus diameter and kinetics of in vitro development, were also analyzed. Embryos derived from 1PN ICSI zygotes showed impaired development compared with 2PN-derived embryos, with blastocyst rates of 28.9% and 67.2%, respectively. The diameter of the pronucleus of 1PN zygotes was larger than that of 2PN zygotes. When compared with 2PN-derived embryos, those derived from 1PN zygotes had a visible pronucleus for a shorter time, in addition to a longer syngamy time and slower kinetic behaviour from two to nine cells. When 1PN-derived blastocysts and 2PN-derived blastocysts were compared, the developmental kinetics were similar in both groups, except for a delayed and longer duration of the compaction phase in 1PN-derived embryos. In conclusion, monopronucleated ICSI zygotes present differences in developmental capacity and morphokinetic behaviour compared with 2PN ICSI zygotes, showing particular morphokinetic parameters related to pronucleus formation. Only the 1PN ICSI-derived embryos that reached the blastocyst stage have similar morphokinetic development to blastocysts from 2PN zygotes.


Identification of substrate-specific inhibitors of cathepsin K through high-throughput screening.
Cathepsin K (CatK) is a cysteine protease and drug target for skeletal disorders that is known for its potent collagenase and elastase activity. The formation of oligomeric complexes of CatK in the presence of glycosaminoglycans has been associated with its collagenase activity. Inhibitors that disrupt these complexes can selectively block the collagenase activity without interfering with the other regulatory proteolytic activities of the enzyme. Here, we have developed a fluorescence polarization (FP) assay to screen 4761 compounds for substrate-specific ectosteric collagenase inhibitors of CatK. A total of 38 compounds were identified that block the collagenase activity without interfering with the hydrolysis of active site substrates such as the synthetic peptide substrate, benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin, and gelatin. The identified inhibitors can be divided into two main classes, negatively charged and polyaromatic compounds which suggest the binding to different ectosteric sites. Two of the inhibitors were highly effective in preventing the bone-resorption activity of CatK in osteoclasts. Interestingly, some of the ectosteric inhibitors were capable of differentiating between the collagenase and elastase activity of CatK depending on the ectosteric site utilized by the compound. Owing to their substrate-specific selectivity, ectosteric inhibitors represent a viable alternative to side effect-prone active site-directed inhibitors.


Dynamic Survival Analysis for EHR Data with Personalized Parametric Distributions.
The widespread availability of high-dimensional electronic healthcare record (EHR) datasets has led to significant interest in using such data to derive clinical insights and make risk predictions. More specifically, techniques from machine learning are being increasingly applied to the problem of dynamic survival analysis, where updated time-to-event risk predictions are learned as a function of the full covariate trajectory from EHR datasets. EHR data presents unique challenges in the context of dynamic survival analysis, involving a variety of decisions about data representation, modeling, interpretability, and clinically meaningful evaluation. In this paper we propose a new approach to dynamic survival analysis which addresses some of these challenges. Our modeling approach is based on learning a global parametric distribution to represent population characteristics and then dynamically locating individuals on the time-axis of this distribution conditioned on their histories. For evaluation we also propose a new version of the dynamic C-Index for clinically meaningful evaluation of dynamic survival models. To validate our approach we conduct dynamic risk prediction on three real-world datasets, involving COVID-19 severe outcomes, cardiovascular disease (CVD) onset, and primary biliary cirrhosis (PBC) time-to-transplant. We find that our proposed modeling approach is competitive with other well-known statistical and machine learning approaches for dynamic risk prediction, while offering potential advantages in terms of interepretability of predictions at the individual level.


Novel genetic and peptide-based strategies targeting the Bcl-2 family, an update.
By controlling mitochondrial membrane permeability, the Bcl-2 family of proteins plays a crucial role in the regulation of cell death. Members of this family are functionally categorized as either pro- or anti-apoptotic based on their ability to promote or inhibit mitochondrial-dependent cell death signals. The critical role of Bcl-2 family proteins in human diseases has made them the focus of intense research by the pharmaceutical industry and academia. Anti-apoptotic Bcl-2 homologues have been widely explored as potential therapeutic targets for anticancer therapy through the development of nucleic-acid and peptide-based inhibitory molecules. Recently, strategies for the development of peptides or polypeptides derived from specific Bcl-2 family members have also been devised. This paper provides a brief review of the most recent patents that have been published relating to this area.


Human Cytomegalovirus Glycoprotein-Initiated Signaling Mediates the Aberrant Activation of Akt.
Human cytomegalovirus (HCMV) is a major cause of morbidity and mortality among immunocompromised and immunonaive individuals. HCMV-induced signaling initiated during viral entry stimulates a rapid noncanonical activation of Akt to drive the differentiation of short-lived monocytes into long-lived macrophages, which is essential for viral dissemination and persistence. We found that HCMV glycoproteins gB and gH directly bind and activate cellular epidermal growth factor receptor (EGFR) and integrin β1, respectively, to reshape canonical Akt signaling within monocytes. The remodeling of the Akt signaling network was due to the recruitment of nontraditional Akt activators to either the gB- or gH-generated receptor signaling complexes. Phosphoinositide 3-kinase (PI3K) comprised of the p110β catalytic subunit was recruited to the gB/EGFR complex despite p110δ being the primary PI3K isoform found within monocytes. Concomitantly, SH2 domain-containing inositol 5-phosphatase 1 (SHIP1) was recruited to the gH/integrin β1 complex, which is critical to aberrant Akt activation, as SHIP1 diverts PI3K signaling toward a noncanonical pathway. Although integrin β1 was required for SHIP1 recruitment, gB-activated EGFR mediated SHIP1 activation, underscoring the importance of the interplay between gB- and gH-mediated signaling to the unique activation of Akt during HCMV infection. Indeed, SHIP1 activation mediated the increased expression of Mcl-1 and HSP27, two Akt-dependent antiapoptotic proteins specifically upregulated during HCMV infection but not during growth factor treatment. Overall, our data indicate that HCMV glycoproteins gB and gH work in concert to initiate an HCMV-specific signalosome responsible for the atypical activation of Akt required for infected monocyte survival and ultimately viral persistence.IMPORTANCE Human cytomegalovirus (HCMV) infection is endemic throughout the world regardless of socioeconomic conditions and geographic locations with a seroprevalence reaching up to 100% in some developing countries. Although asymptomatic in healthy individuals, HCMV can cause severe multiorgan disease in immunocompromised or immunonaive patients. HCMV disease is a direct consequence of monocyte-mediated systematic spread of the virus following infection. Because monocytes are short-lived cells, HCMV must subvert the natural short life-span of these blood cells by inducing a distinct activation of Akt, a serine/theonine protein kinase. In this work, we demonstrate that HCMV glycoproteins gB and gH work in tandem to reroute classical host cellular receptor signaling to aberrantly activate Akt and drive survival of infected monocytes. Deciphering how HCMV modulates the cellular pathway to induce monocyte survival is important to develop a new class of anti-HCMV drugs that could target and prevent spread of the virus by eliminating infected monocytes.


G6PC2 Modulates the Effects of Dexamethasone on Fasting Blood Glucose and Glucose Tolerance.
The glucose-6-phosphatase catalytic subunit 2 (G6PC2) gene encodes an islet-specific glucose-6-phosphatase catalytic subunit. G6PC2 forms a substrate cycle with glucokinase that determines the glucose sensitivity of insulin secretion. Consequently, deletion of G6pc2 lowers fasting blood glucose (FBG) without affecting fasting plasma insulin. Although chronic elevation of FBG is detrimental to health, glucocorticoids induce G6PC2 expression, suggesting that G6PC2 evolved to transiently modulate FBG under conditions of glucocorticoid-related stress. We show, using competition and mutagenesis experiments, that the synthetic glucocorticoid dexamethasone (Dex) induces G6PC2 promoter activity through a mechanism involving displacement of the islet-enriched transcription factor MafA by the glucocorticoid receptor. The induction of G6PC2 promoter activity by Dex is modulated by a single nucleotide polymorphism, previously linked to altered FBG in humans, that affects FOXA2 binding. A 5-day repeated injection paradigm was used to examine the chronic effect of Dex on FBG and glucose tolerance in wild-type (WT) and G6pc2 knockout mice. Acute Dex treatment only induces G6pc2 expression in 129SvEv but not C57BL/6J mice, but this chronic treatment induced G6pc2 expression in both. In 6-hour fasted C57BL/6J WT mice, Dex treatment lowered FBG and improved glucose tolerance, with G6pc2 deletion exacerbating the decrease in FBG and enhancing the improvement in glucose tolerance. In contrast, in 24-hour fasted C57BL/6J WT mice, Dex treatment raised FBG but still improved glucose tolerance, with G6pc2 deletion limiting the increase in FBG and enhancing the improvement in glucose tolerance. These observations demonstrate that G6pc2 modulates the complex effects of Dex on both FBG and glucose tolerance.


Pathological diagnosis of bone sarcoma.
The diagnosis of bone lesions should be established in every case by combined clinical, radiologic and pathologic investigations. For pathologic investigation, a surgical biopsy or needle biopsy should be carried out. Before attempting to make a diagnosis, the pathologist must determine that the tissue to be studied is representative. If clinical and radiologic information is not readily available, the pathologist must insist on its submission before rendering a diagnosis on a slide. It is said that to merely "read slides" without full comprehension of the clinical setting and the radiologic clues of the biologic behavior of the tumor can easily lead to an erroneous diagnosis. A specimen from aspiration biopsy is generally adequate for immediate diagnosis for some bone tumors, but in many cases it is not sufficient for accurate classification of either benign or malignant tumors.


Characterization of ampicillin resistance plasmids from Haemophilus ducreyi.
Seven strains of Haemophilus ducreyi from diverse geographic origins were analyzed for their plasmid content. All strains were multiply resistant, but only resistance to ampicillin was transferred to Escherichia coli by transformation. The H. ducreyi plasmids encoding for ampicillin resistance were 7.4, 5.7, and 3.6 megadaltons and encoded for part or all of TnA, and ampicillin transposon. The relatedness of these plasmids was examined by restriction endonuclease digestion and DNA-DNA homology with isolated DNA fragments from TnA.