Levels of C-peptide, body mass index and age, and their usefulness in classification of diabetes in relation to autoimmunity, in adults with newly diagnosed diabetes in Kronoberg, Sweden.
C-peptide is a main outcome measure in treatment trials of diabetes. C-peptide also has a role in the classification of diabetes, which is often difficult in adults and this is also increasingly recognised in adolescents and elders. We aimed to describe the levels of C-peptide in relation to age and body mass index (BMI) in a large population-based cohort of adults with newly diagnosed diabetes and compare the capabilities of C-peptide, age and BMI to discriminate between autoimmune and non-autoimmune diabetes. Blood samples from 1180 patients were analysed regarding islet cell antibody, glutamic acid decarboxylase antibody and fasting C-peptide (FCP). Receiver operating characteristics (ROC) curves were analysed to check the ability of age, BMI and C-peptide to discriminate between autoantibody-positive (Ab(+)) and -negative (Ab(-)) diabetes. Mean FCP was 0.73±0.5 (range 0.13-1.80) nmol/l in the Ab(+) and 1.42±0.9 (range 0.13-8.30) nmol/l in the Ab(-). FCP was 0.02 nmol/l higher per year increase in age at diagnosis of diabetes. Mean BMI was 26.0±4.8 (range 18.0-39.0) kg/m(2) in the Ab(+) and 28.9±5.3 (range 15.5-62.6) kg/m(2) in the Ab(-). FCP increased with age also within each BMI group. The highest area under the curve (AUC) in the ROC analysis was found for C-peptide, followed by age and BMI (0.78, 0.68 and 0.66 respectively). At diagnosis of diabetes, C-peptide was superior to age and BMI in discriminating between autoimmune and non-autoimmune diabetes. C-peptide increased significantly with BMI and age, latter also within each BMI group. Most of the adults had normal or high levels of C-peptide at presentation of diabetes among the autoimmune patients.


Immune aspects of endometriosis: relevance of the uterine mucosal immune system.
Endometriosis is classically defined as the growth of endometrial cells at sites outside the uterus. It is a common disease characterized by infertility, chronic pain and adhesion formation. Immune dysregulation, evidenced by decreased clearance of endometrial cells and aberrant production of cytokines by peritoneal fluid leukocytes, has been proposed as a mechanism which allows implantation and growth of ectopic endometrium. Cytokines are primary components of intercellular signaling between uterine epithelial and stromal cells, leukocytes, and the developing conceptus. Because their production is regulated by sex hormones, cytokines are well-placed to play a key role in the extensive tissue remodeling required to accommodate menstruation, implantation and pregnancy. Understanding this specialized hormonally-responsive mucosal immune system within the uterus will be critical to understanding the potential importance of the immune system in the pathogenesis of endometriosis. In this review, highlights of studies describing leukocyte populations, cytokines and cytokine receptors in uterine and ectopic endometrium and their proposed role in the regulation of immune processes and endometrial growth are presented, followed by a review of current data on immune aspects of endometriosis. Studies directed at investigating the hormonal regulation of cytokine secretion by uterine and peritoneal cell populations, and the effect of cytokines on endometrial proliferation, should provide a more complete understanding of their potential role in normal uterine growth and in the pathogenesis of endometriosis.


Characterization of nuclear localization signals in the type III effectors HsvG and HsvB of the gall-forming bacterium Pantoea agglomerans.
HsvG and HsvB, two paralogous type III effectors of the gall-forming bacteria Pantoea agglomerans pv. gypsophilae and P. agglomerans pv. betae, determine host specificity on gypsophila and beet, respectively. They were previously shown to be DNA-binding proteins imported into host and non-host nuclei and might act as transcriptional activators. Sequence analysis of these effectors did not detect canonical nuclear localization signals (NLSs), but two basic amino acid clusters designated putative NLS1 and NLS2 were detected in their N-terminal and C-terminal regions, respectively. pNIA assay for nuclear import in yeast and bombardment of melon leaves with each of the NLSs fused to a 2xYFP reporter indicated that putative NLS1 and NLS2 were functional in transport of HsvG into the nucleus. A yeast two-hybrid assay showed that HsvB, HsvG, putative NLS1, putative NLS2, HsvG converted into HsvB, or HsvB converted into HsvG by exchanging the repeat domain, all interacted with AtKAP-α and importin-α3 of Arabidopsis thaliana. Deletion analysis of the NLS domains in HsvG suggested that putative NLS1 or NLS2 were required for pathogenicity on gypsophila cuttings and presumably for import of HsvG into the nucleus. This study demonstrates the presence of two functional NLSs in the type III effectors HsvG and HsvB.


Identifying common traits among Australian irrigators using cluster analysis.
In Australia there is a growing awareness that the over-allocation of water entitlements to irrigators needs to be reduced so that environmental flow allocations can be increased. This means that some water will need to be acquired from irrigators and returned to the environment. Most current water reform policies assume that irrigators are solely motivated by profit and will be willing sellers of water, but this might be an untenable approach. Authorities will need to consider new ways of encouraging the participation of irrigators in water reform. The main aim of this research was to identify the non-commercial influences acting on irrigators' behaviour, especially the influence of the values that they hold toward family, land, water, community and lifestyle. The study also aimed to investigate whether it is possible to group irrigators according to these values and then use the groupings to describe how these might affect their willingness to participate in environmental reforms. We clustered the irrigators into three groups with differing orientations; (i) Investors [25%]-profit oriented, (ii) Lifestylers [25%]-lifestyle oriented, (iii) Providers [50%]-family-succession oriented. This research indicates that when designing policy instruments to acquire water for environmental purposes policy-makers should pay more attention to the factors influencing irrigators' decision making, especially non-commercial factors.


A method for evaluating the cost-benefit of different preparedness planning policies against pandemic influenza.
•Our work presents a unifying method to calculate the net-benefit of different preparedness policies against different pandemic influeunza strains. Unlike previous methods, which have focused on evaluating specific strategies against specific pandemics, our method allows assessment of mass immunisation strategies in presence and absence of antiviral drugs for a large range of pandemic influenza strain characteristics and programme features. Overall, the model described here combines two parts to evaluate different preparedness planning policies against pandemic influenza.•The first part is adaptation of an existing transmission model for seasonal influenza to include generalisation across large number of pandemic influenza scenarios.•The second part is development of a tailor-made health economic model devised in collaboration with colleagues at the UK Department of Health and Social Care.


A modified direct lateral approach for neck-preserving total hip arthroplasty: tips and technical notes.
Tissue-sparing surgery for hip replacement aims to minimize muscle damage and conserve the femoral neck through the use of mini-prostheses. We propose a modification of the classical direct lateral access procedure that preserves the gluteus medius. Further advantages during the surgical phase include limited blood loss, visualization of the entire acetabulum, and sparing of the transverse ligament. Precise implantation is facilitated and normal biomechanics are preserved. The gluteus medius is divided longitudinally between the anterior third and posterior two-thirds to provide access to the gluteus minimus, which is detached from the femoral insertion together with a small portion of the vastus lateralis, forming a flap that exposes the underlying articular capsule. When the femoral head is revealed, a decision is made to either continue with its dislocation directly or to resect it and remove it separately to avoid damaging the gluteus medius during dislocation. Upon removal of the femoral head, with the limb flexed and slightly over-rotated, the acetabulum is completely visible. Limb length is maintained through the use of reference stitches on the gluteus minimus tendon and the proximal insertion of the vastus lateralis. In keeping with the minimally invasive philosophy, only pathological tissue is removed (marginal osteophytes, geodes, joint capsule, cartilage to the point of bleeding and pulvinar). We have performed more than 2,000 implants with this procedure since 1990. Advantages and potential critical points are discussed.


Induction of proliferation or hypertrophy of chondrocytes in serum-free culture: the role of insulin-like growth factor-I, insulin, or thyroxine.
In bone forming cartilage in vivo, cells undergo terminal differentiation, whereas most of the cells in normal articular cartilage do not. Chondrocyte hypertrophy can be induced also in vitro by diffusible signals. We have identified growth factors or hormones acting individually on 17-d chick embryo sternal chondrocytes cultured in agarose gels under strictly serum-free conditions. Insulin-like growth factor I or insulin triggered the first steps of chondrocyte maturation, i.e., cell proliferation and increased matrix deposition while the chondrocytic phenotype was maintained. However, cells did not progress to the hypertrophic stage. Proliferation and stimulated collagen production was preceded by a lag period, indicating that synthesis of other components was required before cells became responsive to insulin-like growth factor I or insulin. Very small amounts of FBS exerted effects similar to those of insulin-like growth factor I or insulin. However, FBS could act directly and elicited hypertrophy when constituting greater than 1% of the culture media. Basic FGF has been claimed to be the most potent chondrocyte mitogen, but had negligible effects under serum-free conditions. The same is true for PDGF, a major serum-mitogen. Under the direction of thyroxine, cells did not proliferate but became typical hypertrophic chondrocytes, extensively synthesizing collagen X and alkaline phosphatase.


Antioxidant capacity and meat quality of broilers exposed to different ambient humidity and ammonia concentrations.
To investigate the effect of humidity and ammonia on the antioxidative capacities and meat qualities of broilers, 192 broilers were divided into 2 groups: high (H, 70 ppm) and low (L, 30 ppm) ammonia concentration. These groups were divided into 30% (Treatment humidity, T) and 60% (Control humidity, C) humidity, giving 4 treatments: C+L, C+H, T+L, and T+H. Blood and muscle antioxidative capacities and meat quality were measured. In the H group, body weight (BW), average daily feed intake (ADFI), average daily weight gain (ADG), blood and muscle antioxidative capacities, and postmortem pectoral muscle a* of broilers were significantly decreased (P < 0.05), and pectoral muscle thiobarbituric acid reactive substance (TBARS) contents and drip losses, postmortem pectoral muscle b* (P < 0.05) and L* (P = 0.054), and pectoral muscle shear forces (P = 0.075) increased. In the T condition, BW, ADFI, pectoral muscle superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities, and pectoral muscle L* decreased (P = 0.053), and pectoral muscle shear forces and TBARS contents increased (P < 0.05). In the T+H group, BW, ADFI, ADG, blood antioxidative capacities, pectoral muscle SOD and GSH-Px activities, and postmortem pectoral muscle a* were significantly lower than those of the C+L group, but postmortem pectoral muscle TBARS contents and pectoral muscle drip losses and shear forces significantly increased (P < 0.05). These results revealed that T+H could significantly reduce growth performance, antioxidative capacities, and meat quality of broilers; T intensified these negative effects.


Can dried blood spots be used to accurately measure vitamin D metabolites?
Where conventional blood sampling is challenging, dried blood spots (DBS) provide a practical sample alternative for measuring vitamin D levels. Our study aimed to develop and evaluate a clinical pathology service-based assay suitable for measuring vitamin D in batches of DBS samples collected remote to the testing site. A high throughput liquid chromatography-tandem mass spectrometry (LC-MS/MS) method with derivatisation was developed to measure 25-hydroxyvitamin D metabolites (25OHD3, 25OHD2 and 3-epi-25OHD3) in DBS samples. The assay was validated using paired DBS and plasma samples from 37 healthy adults. The assay reproducibly (<11.5% coefficient of variation) quantified 25OHD3 (range 1-300 nmol/L), 25OHD2 (range 2-300 nmol/L) and 3-epi-25OHD3 (range 1-200 nmol/L) in DBS samples. The 25OHD3 metabolite was detected in all DBS samples, 3-epi-25OHD3 in six plasma (range 2.1-6.3 nmol/L) and paired DBS samples, and 25OHD2 was not detected. Concentrations of 25OHD3 were highly correlated between paired samples: capillary DBS and venous plasma (r = 0.92), venous DBS and venous plasma (r = 0.93), and capillary DBS and venous DBS (r = 0.97). Ordinary least squares regression was used to characterise (β = 0.81) and correct the systematic bias in DBS data (compared to paired plasma). Thereafter, Bland-Altman analysis demonstrated robust agreement between sample-methods. This simple and rapid DBS-based LC-MS/MS assay accurately quantified serum vitamin D metabolites using a paired-sample 'bridging strategy' to correct for the inherent sample-method bias.


Comparison of complications in percutaneous coronary intervention patients mobilized at 3, 4, and 6 hours after femoral arterial sheath removal.
The purpose of this research study was to explore groin complication rates of patients mobilized at 3, 4, and 6 hours after femoral arterial sheath removal following a percutaneous coronary intervention procedure. Participants were recruited from those undergoing coronary angioplasty and coronary stent placement at a large public hospital in Brisbane, Queensland. Participants were randomly allocated to the 3, 4, or 6 hourly mobilization group. After removal of the femoral arterial sheath and again the next day, participants' groins were assessed for evidence of complications including hemorrhage, hematoma formation, and pseudoaneurysm. The results showed that the length of bed rest after arterial sheath removal had no significant effect on bleeding (F 304 = 5.39, P = 0.21) or hematoma formation (F 304 = 0.258, P = 0.612) at the groin puncture site for participants who mobilized at either 3, 4, or 6 hours after percutaneous coronary intervention arterial sheath removal.


The main genetic locus associated with the evolution of gamecocks is centered on ISPD.
Chickens were domesticated >4,000 years ago, probably first for fighting them and only later as a source of food. Fighting chickens, commonly known as gamecocks, continue to be bred throughout the world, but the genetic relationships among geographically diverse gamecocks and with nongame chickens are not known. Here, we sequenced the genomes of 44 geographically diverse gamecocks and of 62 nongame chickens representing a variety of breeds. We combined these sequences with published genomes to generate the most diverse chicken genomes dataset yet assembled, at 307 samples. We found that gamecocks do not form a homogeneous group, yet they share genetic similarities that distinguish them from nongame chickens. Such similarities are likely the result of a common origin before their local diversification into, or mixing with, nongame chickens. Particularly noteworthy is a variant in an intron of ISPD, an extreme outlier present at a frequency of 89% in gamecocks but only 4% in nongame chickens. The ISPD locus has the strongest signal of selection in gamecocks, suggesting it is important for fighting performance. Because ISPD variants that are highly prevalent in gamecocks are still segregating in nongame chickens, selective breeding may help reduce its frequency in farm conditions in which aggression is not a desired trait. Altogether, our work provides genomic resources for agricultural genetics, uncovers a common origin for gamecocks from around the world and what distinguishes them genetically from chickens bred for purposes other than fighting, and points to ISPD as the most important locus related to fighting performance.


Male microchimerism in peripheral blood leukocytes from women with multiple sclerosis.
Fetal microchimerism (F-MC), the persistence of fetal cells in the mother, is frequently encountered following pregnancy. The high prevalence of F-MC in autoimmune disease prompts consideration of the role for immune tolerance and regulation. This study examines the association between F-MC and multiple sclerosis (MS), an autoimmune disorder, of undetermined etiology. 21 out of 51 MS-positive subjects (41%) were classified as positive for F-MC; 4 of 22 (18%) of MS-negative sibling controls, were also positive for MC (p = 0.066). Unanticipated F-MC in controls lead to re-evaluation using 30 female singleton cord blood units (CBUs) as a biological control. Four CBUs were low-level positive. Seventy-three female subjects were assigned to three groups according to disease status and pregnancy history: (1) MS positive (+) women with a history of one male pregnancy before symptom onset (n = 27); (2) MS negative (-) female siblings of MS(+) women with a history of one male pregnancy (n = 22); and (3) MS(+) women that reported never having been pregnant (n = 24). Ten micrograms of genomic DNA obtained from peripheral blood leukocytes of each subject were analyzed for F-MC using allele-specific real-time PCR targeting the SR-Y sequence on the Y-chromosome. MC classification was dichotomous (positive vs. negative) based on PCR results. The association between F-MC and MS warrants further study to define this relationship. F-MC in women self-reporting as nulligravid, supports previous findings that a significant proportion of pregnancies go undetected. This lead to re-validation of a Y-chromosome based assay for F-MC detection.


Applications of isothermal titration calorimetry in pure and applied research from 2016 to 2020.
The last 5 years have seen a series of advances in the application of isothermal titration microcalorimetry (ITC) and interpretation of ITC data. ITC has played an invaluable role in understanding multiprotein complex formation including proteolysis-targeting chimeras (PROTACS), and mitochondrial autophagy receptor Nix interaction with LC3 and GABARAP. It has also helped elucidate complex allosteric communication in protein complexes like trp RNA-binding attenuation protein (TRAP) complex. Advances in kinetics analysis have enabled the calculation of kinetic rate constants from pre-existing ITC data sets. Diverse strategies have also been developed to study enzyme kinetics and enzyme-inhibitor interactions. ITC has also been applied to study small molecule solvent and solute interactions involved in extraction, separation, and purification applications including liquid-liquid separation and extractive distillation. Diverse applications of ITC have been developed from the analysis of protein instability at different temperatures, determination of enzyme kinetics in suspensions of living cells to the adsorption of uremic toxins from aqueous streams.


Functional dyspepsia is associated with a greater symptomatic response to fat but not carbohydrate, increased fasting and postprandial CCK, and diminished PYY.
In patients with functional dyspepsia (FD), symptoms are frequently triggered, or exacerbated, by fatty foods. We hypothesized that in FD patients, a high-fat (high-FAT) meal would induce more symptoms than a high-carbohydrate (high-CHO) meal, associated with an altered secretion of cholecystokinin (CCK), peptide-YY (PYY), and ghrelin and an increased antral size, when compared to healthy subjects (HS). FD symptoms, appetite perceptions, plasma hormones, and antral area were measured in 8 FD patients and 8 HS on three separate days after the ingestion of high-CHO or high-FAT (500 kcal/400 g) meals, or a low-nutrient control (180 kcal/400 g); the energy intake was quantified 60 min later. Nausea (P < 0.01) and pain (P= 0.05) were greater in FD after the high-FAT, when compared to high-CHO and control meals and in HS. Discomfort was greater after all meals in FD when compared to HS (P < 0.05). Fasting CCK and stimulation of CCK by the high-FAT (P < 0.01) meal were greater in FD, while fasting and postprandial PYY were lower (P < 0.001) in FD than in HS, with no differences in fasting, or postprandial, plasma ghrelin between FD and HS. Fasting antral area was greater in FD (P < 0.05), with no differences postprandially between FD and HS. There were no differences in the energy intake between the two groups. In FD patients: (a) a high-FAT meal induces more symptoms than an isocaloric high-CHO meal, and (b) fasting and postprandial plasma CCK concentrations are greater and PYY concentrations are less. Our findings have important implications for the development of diet-based therapies for the treatment of FD.


A prospective study of dairy foods intake and anovulatory infertility.
Dairy foods and lactose may impair fertility by affecting ovulatory function. Yet, few studies have been conducted in humans and their results are inconsistent. We evaluated whether intake of dairy foods was associated with anovulatory infertility and whether this association differed according to fat content. We prospectively followed 18,555 married, premenopausal women without a history of infertility who attempted a pregnancy or became pregnant during an 8-year period. Diet was assessed twice during the study using food-frequency questionnaires. During follow-up, 438 women reported infertility due to an ovulatory disorder. The multivariate-adjusted relative risks (RR) [95% confidence interval (CI); P, trend] of anovulatory infertility comparing women consuming > or = 2 servings per day to women consuming < or = 1 serving per week was 1.85 (1.24-2.77; 0.002) for low-fat dairy foods. The RR (95% CI; P, trend) comparing women consuming > or = 1 serving per day of high-fat dairy foods to those consuming < or = 1 serving per week was 0.73 (0.52-1.01; 0.01). There was an inverse association between dairy fat intake and anovulatory infertility (P, trend = 0.05). Intakes of lactose, calcium, phosphorus and vitamin D were unrelated to anovulatory infertility. High intake of low-fat dairy foods may increase the risk of anovulatory infertility whereas intake of high-fat dairy foods may decrease this risk. Further, lactose (the main carbohydrate in milk and dairy products) may not affect fertility within the usual range of intake levels in humans.


A clinical and pathological study of diffuse type hepatocellular carcinoma.
Six autopsy cases of the rare, diffuse type of hepatocellular carcinoma (HCC), as classified gross anatomically according to the strict definition, have been studied. The prominent clinical feature was the rapid deterioration of the patient's general condition, terminating in hepatic failure. The liver size enlarged quickly, at a perceptible speed, often accompanied by abdominal pain. Diagnosis of this particular type of HCC was difficult, and celiac angiography and scintiscan of the liver were only suggestive when considered together with other laboratory data. Hepatitis B surface antigen was positive in all three patients in whom it was tested. The entire liver was studded with minute, uniformly sized tumor nodules, evenly distributed throughout. Some of them were grossly indistinguishable from cirrhotic nodules. All livers had an underlying cirrhosis which was characterized by relatively small regenerative nodules with thin stromas. Large portal branches at the hilum contained tumor thrombi in all patients, except for one case in which left lobectomy was followed by intraportal dissemination. Histologically, all tumor nodules represented intrahepatic metastases via the portal vein system. Tumor cells were poorly differentiated. These findings suggest that the diffuse type of HCC most frequently, if not always, represents intrahepatic, widespread portal metastases which have occurred within a short period of time.


Frequency and awareness of hepatitis B and C in visitors of Hepatitis Awareness Mela.
To observe the frequency of Hepatitis B and C in a section of society that is presumably healthy and educated and to evaluate the awareness about the modes of transmission of hepatitis B and C, in this segment. A total of 504 individuals visiting the Awareness Mela arranged to observe World Hepatitis Day were screened for Hepatitis B and C. In addition, 106 conveniently selected individuals of age 16 years or more were asked to fill out a questionnaire which enquired on the modes of transmission of the diseases. A total of 504 individuals were screened; 351 males (69.6%) and 153 females (30.4%). Mean age 34.2 +/- 11.7 years; range 11-66 years. Twenty nine (5.75%) were found positive for either hepatitis B or C, mean age being 28.43 years and 92% of the affected being males. Out of the 106 questionnaire takers, 25 were females (23.6%) and 81 males (76.4%). The mean age was 33.2 +/- 11.1 years. Majority (94.7%) believed that used syringes were amongst the main causes of spread of hepatitis B and C while 40.6% did not know about the risks of sharing toothbrushes and towels. Seventy three respondents did not have prior vaccination of Hepatitis B. In addition 2 males and 1 female of study subjects were anti-HCV positive and 1 male HBsAg positive. In the educated class, prevalence of hepatitis B and C appears lower than the general population. They have the knowledge of the mode of spread of these viruses though there is room for further improvement. Many still confuse it with orofaecal transmission. Knowledge on vaccination is also unsatisfactory.


Accelerated degradation of glycogen phosphorylase in denervated and dystrophic mouse skeletal muscle.
Pyridoxal phosphate, the cofactor of glycogen phosphorylase, fulfils the criteria needed of a turnover label for this enzyme. The decay of protein-bound label following administration of [3H]pyridoxine is a good index of the rate of degradation of the enzyme in vivo. This method has been applied to the study of catabolism of the enzyme in normal, denervated and dystrophic mouse skeletal muscle. In both of the pathological conditions the enzyme is degraded more rapidly than normal.


A teratological study of aminoglycoside antibiotic treatment during pregnancy.
The aim of this study was to investigate the teratogenicity of aminoglycoside antibiotics, such as parenteral gentamicin, streptomycin, tobramycin and oral neomycin, during pregnancy. Pair analysis of cases with congenital abnormalities and matched healthy controls was carried out. The setting was the population-based dataset of the Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-96. In total, 38,151 pregnant women who had newborn infants without any defects (control group) and 22,865 pregnant women who had foetuses or newborns with congenital abnormalities were included in the study. 38 (0.16%) and 42 (0.11%) pregnant women in the case and control groups, respectively, were treated with the aminoglycosides studied. A teratogenic potential of gentamicin and neomycin was not indicated by a comparison of the occurrence of aminoglycoside antibiotic treatments in the total control group as referent with the figures of different congenital abnormality groups. In addition, the case-control pair analysis during the second-third months of pregnancy did not show a teratogenic risk of gentamicin and neomycin. The conclusion of this study is that treatment with parenteral gentamicin and oral neomycin during pregnancy presents no detectable teratogenic risk to the foetus, when restricted to structural developmental disturbances.


Absorption, translocation, and detoxification of Cd in two different castor bean (Ricinus communis L.) cultivars.
Cadmium (Cd) is considered to be the most phytotoxic heavy metal pollutant. The selection of castor bean cultivars with Cd tolerance and the exploration of the physiological mechanisms involved in Cd tolerance are critical steps for improving phytoremediation performance. In this study, a hydroponic experiment was used to investigate variations in Cd transportation, chelation, and subcellular distribution in two different castor bean cultivars, namely JX-22 and ZB-9. Both cultivars had high tolerance index scores, indicating that both cultivars were tolerant to Cd. The findings of the present study indicate that Cd is significantly more mobile in JX-22 than in ZB-9 during xylem and phloem transportation, resulting in the accumulation of Cd in the shoots of JX-22 was 7.67 times that in ZB-9. Subcellular distribution assessment verified that more Cd was bound to the biologically detoxified metal fractions than the metal sensitive fractions in JX-22. The contents of the non-protein thiol pool and glutathione in the leaves were higher in JX-22 than ZB-9 when exposed to Cd. These results indicate that JX-22 has a greater ability to accumulate Cd, and well-coordinated physiological changes in JX-22 afford greater Cd tolerance in comparison to ZB-9 under Cd exposure, indicating that JX-22 is suitable for use in the remediation of Cd-contaminated soils.


A mechanistic classification of closed, indirect fractures and dislocations of the lower cervical spine.
Closed, indirect fractures and dislocations of the lower cervical spine occur in families or groups within which there is a spectrum of anatomic damage to a cervical motion segment. This study of 165 cases demonstrates the various spectra of injury, called phylogenies, and develops a classification based on the mechanism of injury. The common groups are compressive flexion, vertical compression, distractive flexion, compressive extension, distractive extension, and lateral flexion. The probability of an associated neurologic lesion relates directly to the type and severity of cervical spine injury. With use of the classification, it is possible to formulate a rational treatment plan for injuries to the cervical spine.


Disparities in Beef Tapeworm Identification Rates in the Abattoirs of Gauteng Province, South Africa: A Descriptive Epidemiologic Study.
Bovine Taenia saginata cysticercus infections (also called bovine cysticercosis or beef measles) is usually diagnosed in cattle only during post-mortem meat inspection. The aim of this study was to investigate the identification rates of these infections in and to identify predictors/determinants of variations in the identification rates in abattoirs in Gauteng province, South Africa. Retrospective data for over 1.4 million cattle carcasses inspected in 26 abattoirs between January 2010 and December 2013 were used for the study. The identification rates (proportion of bovine Taenia saginata cysticercus positive carcasses) were computed and generalized estimating equations used to identify predictors/determinants of identification rates. The overall identification rate was 0.70% (95% CI: 0.45, 0.95). Significantly (p< 0.05) lower rates were reported during summer (0.55%) than other seasons. Some geographic areas reported significantly (p<0.05) higher rates than others. The identification rates in high throughput abattoirs was significantly (p<0.05) higher (RR: 9.4; 95% CI: 4.7-19.1) than in low throughput abattoirs. Similarly, the identification rates among animals from feedlots were significantly (p<0.05) higher (RR: 1.6; 95% CI: 1.7-3.5) than those from non-feedlot sources. No significant (p>0.05) association was identified between identification rates and either the number of meat inspectors per abattoir or the provider of inspection services. Although no significant association was found between identification rates and provider of inspection services, follow-up studies will need to be done to specifically investigate the potential conflict of interest arising from the fact that abattoir owners hire meat inspection services directly. Capture of abattoir surveillance data needs to include farm address and for each case to be reported separately. Finally, information on the type of identified cysts (alive or calcified) need to be collected to help better estimate risk to consumers. This study provides useful baseline data to guide future studies, surveillance and control efforts.


Cellular and molecular mechanisms of the bone marrow sparing effects of the glucose chloroethylnitrosourea chlorozotocin.
1,(2-Chloroethyl)-3-cyclohexyl-1-nitrosourea (CCNU) and other chloroethylnitrosourea anticancer agents in clinical use produce severe and cumulative bone marrow toxicity. Chlorozotocin, a glucose analogue, has demonstrated reduced hematologic toxicity while retaining full antitumor activity. The biochemical-pharmacologic properties of chlorozotocin and CCNU were compared in human bone marrow. After a 2-hr incubation with a 0.1-mM drug concentration, total cellular uptake of chlorozotocin in whole marrow was 2.47 +/- 0.80 pmole/10(4) cells and was not significantly different compared to the uptake of 1.94 +/- 0.53 pmole/10(4) cells with CCNU. The quantitative alkylation of bone marrow DNA by chlorozotocin, 22.8 +/- 1.2 pmole/mg DNA, was equivalent to that produced by CCNU, 22.9 +/- 0.5 pmole/mg DNA. Bone marrow was separated into 14 fractions by centrifugal elutriation. CCNU uptake was found to be greater than that of chlorozotocin in 3 fractions that were primarily composed of lymphocytes, monocytes, and normoblasts. Chlorozotocin uptake was greater than CCNU in 6 fractions that contained primarily mature and immature myeloid cells as well as the highest CFU-GM activity. The two drugs produced a comparable degree of DNA strand breakage and DNA-protein cross-linking as measured by alkaline elution of pooled fractions of elutriated bone marrow. DNA interstrand crosslinking was not found with either drug. The most significant finding of this study is the differences in the site of drug alkylation by chlorozotocin and CCNU in bone marrow chromatin. Endonuclease digestions with MCN, DNase I, and DNase II showed nonrandom alkylation of specific regions of chromatin by the two drugs: CCNU demonstrated a preferential binding to the transcriptionally active regions of chromatin, whereas chlorozotocin predominantly alkylated the transcriptionally inactive regions. These data suggest that the lethal damage of nitrosourea alkylation in human bone marrow is principally expressed in transcriptionally active regions of chromatin.


[Reaction of the protein-synthesizing apparatus of the major stomach cells in rats to thyroxine administration].
Electron microscopy and cytochemical analysis have shown that protein-synthesizing apparatus of major stomach cells was characterized by the most pronounced changes observed in cell structures. Comparison between ultrastructural and biochemical analysis demonstrating leucine and orotic acid incorporation into the total protein and RNA, respectively, has revealed the correlation between the structure of nucleus, nucleolia and biosynthesizing activity of secretory cells. It was established that, while in parietal cells the enzymes at the organella level affected mainly mitochondria, in major cells protein-synthesizing apparatus was involved.


Ear drops for the removal of ear wax.
Problems attributed to the accumulation of wax (cerumen) are one of the most common reasons for people to present to their general practitioners with ear trouble (Sharp 1990). Treatment for this condition often involves use of a wax softening agent (cerumenolytic) in order to disperse the cerumen and reduce the need for syringing, or to facilitate syringing should it prove necessary, but there is no consensus on the effectiveness of the wide variety of cerumenolytics in use. To assess the effectiveness of ear drops (cerumenolytics) for the removal of symptomatic ear wax. We searched the Cochrane ENT Group Register, the Cochrane Central Register of Controlled Trials (CENTRAL, Issue 1, 2003), and MEDLINE and EMBASE up to March 2003. Reference lists of all trials were also manually searched. We identified all randomised controlled trials (with or without blinding) in which a cerumenolytic was evaluated in comparison with either no treatment, a placebo, or other cerumenolytics in participants with hard or impacted ear wax, and in which the proportion of participants with sufficient clearance of the external canal to make further mechanical clearance unnecessary (primary outcome measure) was stated or calculable. The full text articles of all the retrieved trials of possible relevance were reviewed by the two reviewers and the inclusion criteria applied independently. Any differences in opinion about which studies to include in the review were resolved by discussion. Trials were graded for methodological quality using the Cochrane approach. Data extraction was performed in a standardised manner by one reviewer and rechecked by the other reviewer, and where necessary investigators were contacted to obtain missing information. Meta-analysis was neither possible nor considered appropriate because of the heterogeneity of the treatments, treatment amounts and durations, trial procedures, and scoring systems. A narrative overview of the results is therefore presented. Eight trials satisfied the inclusion criteria, the majority of which were of poor quality. In all, 587 participants received one of nine different cerumenolytics. One trial compared active treatments with no treatment, two trials compared active treatments with water or a saline 'placebo', and all eight trials placed two or more active treatments in head-to-head comparisons. Seven trials included syringing as a secondary treatment where necessary.Overall, results were inconclusive. One trial found a significant difference between one of three active agents (Cerumol) in comparison to no treatment, but no statistically significant difference was found between these three agents (sodium bicarbonate ear drops; Cerumol; sterile water). In two trials no statistical difference was found between the effectiveness of either sodium bicarbonate ear drops, Cerumol, Cerumenex or Colace versus a sterile water or saline 'placebo'. Three trials (from the same source) found statistically significant differences in favour of the same active agent (Exterol) in comparison to glycerol and Cerumol. Three trials found no statistically significant difference between two or more cerumenolytics (Otocerol versus Cerumol; Audax versus Earex; sodium bicarbonate ear drops versus Cerumol). Two trials comparing the same two cerumenolytics (Cerumenex versus Colace) also failed to show any significant benefit of one over the other. No serious adverse effects were reported from any of the interventions. Trials to date have been heterogeneous and of poor quality, making it difficult to offer any definitive recommendations on the effectiveness of cerumenolytics for the removal of symptomatic ear wax. Future trials should be of high methodological quality, have large sample sizes, and compare both oil-based and water-based solvents with placebo and/or no treatment.


[Radiogenic nephropathy].
Patient-individual dosimetric analyses are a useful tool in external beam radiotherapy (EBR) to protect patients from side effects such as radiogenic nephropathy. At this point in time, individual dosimetry is not used as a standard in patient treated with radiolabelled antibody fragments or polypeptides. The reasons are a number of problems, which make patient dosimetry more challenging than in EBR. While in EBR, the dose is distributed evenly in the organ and the organ volume can exactly be determined, in internal radiotherapy the tracer is not evenly distributed within the organ leading to a non-uniform dose distribution. In addition, the dose rate of the most commonly used radionuclides is lower than in EBR and the range of their radiation differ, so that the radiobiological effects are differing considerably in comparison to EBR. More complex models have to be used for clinical kidney dosimetry in internal radiotherapy. In this paper, we give a concise overview of the reasons for accumulation of radiotracers in the kidney, the most recent developments in kidney dosimetry, and approaches to reduce the kidney uptake of radiotracers in order to avoid radiogenic nephropathy.


Proteasome- and ethanol-dependent regulation of HCV-infection pathogenesis.
This paper reviews the role of the catabolism of HCV and signaling proteins in HCV protection and the involvement of ethanol in HCV-proteasome interactions. HCV specifically infects hepatocytes, and intracellularly expressed HCV proteins generate oxidative stress, which is further exacerbated by heavy drinking. The proteasome is the principal proteolytic system in cells, and its activity is sensitive to the level of cellular oxidative stress. Not only host proteins, but some HCV proteins are degraded by the proteasome, which, in turn, controls HCV propagation and is crucial for the elimination of the virus. Ubiquitylation of HCV proteins usually leads to the prevention of HCV propagation, while accumulation of undegraded viral proteins in the nuclear compartment exacerbates infection pathogenesis. Proteasome activity also regulates both innate and adaptive immunity in HCV-infected cells. In addition, the proteasome/immunoproteasome is activated by interferons, which also induce "early" and "late" interferon-sensitive genes (ISGs) with anti-viral properties. Cleaving viral proteins to peptides in professional immune antigen presenting cells and infected ("target") hepatocytes that express the MHC class I-antigenic peptide complex, the proteasome regulates the clearance of infected hepatocytes by the immune system. Alcohol exposure prevents peptide cleavage by generating metabolites that impair proteasome activity, thereby providing escape mechanisms that interfere with efficient viral clearance to promote the persistence of HCV-infection.


Optimal photon energies for IUdR K-edge radiosensitization with filtered x-ray and radioisotope sources.
The purpose of this work is to determine the most physically effective radiation energy for K-edge absorption of x- or gamma-rays by iododeoxyuridine (IUdR) on Chinese hamster ovary (CHO) cells. Brachytherapy sources (Sm-145, I-125, Yb-169 and Am-241) and x-ray beams (30 kVp, 100 kVp and 100 kVp with gold, gadolinium, lead or tungsten filtration) were investigated for their preferential absorption qualities by IUdR sensitized DNA. The 30 kVp, 100 kVp and 100 kVp with tungsten filtration were then used to irradiate CHO cells, with or without IUdR incorporation (i.e. 10(-5) M of IUdR for 3 days). Radiation absorption calculations were performed to determine the increase in energy absorption in DNA with and without IUdR incorporated. In order to measure the in vitro biological effects of K-edge absorption, cell survival experiments were performed. The radiation physics calculations yielded an iodine dose enhancement ratio (DER) of 1.4+/-0.15. 1.8+/-0.15 and 2.7+/-0.15 for the 30 kVp, 100 kVp and tungsten filtered 100 kVp respectively, for 18% IUdR replacement of thymidine in DNA. The corresponding cell sensitization enhancement ratios (SER), determined from the cell survival assay, were determined to be 1.24+/-0.2, 1.8+/-0.2 and 2.3+/-0.3 for the 30 kVp, 100 kVp and tungsten filtered 100 kVp respectively, for cells with 18+/-2% IUdR incorporation. These SER values are in reasonable agreement with the DER values of 1.4, 1.8 and 2.7. From these radiation calculations and radiobiology experiments we confirm that using x-radiation energies above the K-edge of iodine (33.2 keV) can have a significant effect on cell survival. This effect is due mainly to the increase in the local dose to the DNA for IUdR-sensitized cells compared with the normal DNA which lacks the iodine contrast agent. Our results support the clinical application of IUdR and low-energy brachytherapy, perhaps using new technologies such as the x-ray needle or new isotopes such as Yb-169.


Analysis of CATMA transcriptome data identifies hundreds of novel functional genes and improves gene models in the Arabidopsis genome.
Since the finishing of the sequencing of the Arabidopsis thaliana genome, the Arabidopsis community and the annotator centers have been working on the improvement of gene annotation at the structural and functional levels. In this context, we have used the large CATMA resource on the Arabidopsis transcriptome to search for genes missed by different annotation processes. Probes on the CATMA microarrays are specific gene sequence tags (GSTs) based on the CDS models predicted by the Eugene software. Among the 24 576 CATMA v2 GSTs, 677 are in regions considered as intergenic by the TAIR annotation. We analyzed the cognate transcriptome data in the CATMA resource and carried out data-mining to characterize novel genes and improve gene models. The statistical analysis of the results of more than 500 hybridized samples distributed among 12 organs provides an experimental validation for 465 novel genes. The hybridization evidence was confirmed by RT-PCR approaches for 88% of the 465 novel genes. Comparisons with the current annotation show that these novel genes often encode small proteins, with an average size of 137 aa. Our approach has also led to the improvement of pre-existing gene models through both the extension of 16 CDS and the identification of 13 gene models erroneously constituted of two merged CDS. This work is a noticeable step forward in the improvement of the Arabidopsis genome annotation. We increased the number of Arabidopsis validated genes by 465 novel transcribed genes to which we associated several functional annotations such as expression profiles, sequence conservation in plants, cognate transcripts and protein motifs.


Association of a single nucleotide polymorphism near the interleukin-28B gene with response to hepatitis C therapy in HIV/hepatitis C virus-coinfected patients.
Given that peginterferon-ribavirin treatment is poorly tolerated, there is interest in the identification of predictors of response, particularly in HIV/hepatitis C virus (HCV)-coinfected patients that respond less than HCV-monoinfected individuals. A single nucleotide polymorphism (SNP) near the IL28B gene (rs12979860) has been shown to predict treatment response in HCV-monoinfected patients carrying genotype 1. Information is lacking for HIV/HCV-coinfected individuals and/or other HCV genotypes. From 650 HIV/HCV-coinfected patients, we identified those who had completed a course of peginterferon-ribavirin therapy with a validated outcome and available repository DNA. The rs12979860 SNP was examined in a blinded fashion. A total of 164 patients were included in the final IL28B genotyping analysis, 90 (55%) of whom achieved sustained virological response (SVR). HCV genotype distribution was as follows: HCV-1 58%, HCV-3 31% and HCV-4 11%. Overall, the SVR rate was higher in patients with CC than in those CT/TT genotypes: 56 of 75 (75%) versus 34 of 89 (38%) (P < 0.0001). The effect of the SNP was seen in HCV genotypes 1 and 4 but not in HCV genotype 3 carriers. In the multivariable analysis (odds ratio; 95% confidence interval; P value), the rs12979860 CC genotype was a strong predictor of SVR (3.7; 1.6-8.5; 0.002), independent of HCV genotype 3 (8.0; 3.1-21.0; <0.001), serum HCV-RNA less than 600,000 IU/ml (11.9; 3.8-37.4; <0.001) and lack of advanced liver fibrosis (3.5; 1.4-8.9; 0.009). The rs12979860 SNP located near the IL28B gene is associated with HCV treatment response in HIV-infected patients with chronic hepatitis C due to genotypes 1 or 4. Thus, IL28B genotyping should be considered as part of the treatment decision algorithm in this difficult-to-treat population.


Factors That Help and Hinder the Implementation of Digital Depression Prevention Programs: School-Based Cross-sectional Study.
Digital prevention programs that are delivered in a school environment can inoculate young people against depression. However, little is known about the school-based factors that help and hinder the implementation of these programs. Staff members are integral for supporting mental health programs in schools and are likely to have a wealth of expertise and knowledge about the factors that affect implementation. The primary objective of this study was to explore the barriers and facilitators to implementing a digital depression prevention program in Australian secondary schools with teachers, counselors, and principals. The secondary objective was to explore variations in these factors across different school contexts, including the school type (government or nongovernment), location (capital city, regional/or rural areas), and socioeconomic status (SES) (low, medium, high). This quantitative cross-sectional survey study assessed the barriers and facilitators to implementing a hypothetical digital prevention program in Australian schools. The survey was taken by 97 teachers (average age 38.3 years), 93 counselors (average age 39.5 years), and 11 principals (average age 50.9 years) across Australia between November 2017 and July 2018. A range of barriers and facilitators relating to logistics and resources, staff support, and program factors were endorsed by the surveyed staff. Consistent with prior research, common barriers included a lack of time and resources (ie, staff and rooms). These barriers were particularly evident in government, rural/regional, and low socioeconomic schools. Other barriers were specific to digital delivery, including privacy issues and a lack of clarity around staff roles and responsibilities. Facilitators included upskilling staff through training, embedding the program into the curriculum, and other program factors including universal delivery, screening of students' mental health, and clear referral pathways. Knowledge about the program efficacy was also perceived as important by a large proportion of the respondents. The digital depression prevention program was perceived as suitable for use within different schools in Australia, although certain factors need to be considered to enable effective implementation. Logistics and resources, support, and program factors were identified as particularly important for school-based implementation. To maximize the effectiveness in delivering digital programs, implementation may need to be tailored to the staff roles and school types.


Polycyclic aromatic hydrocarbons, tobacco smoke, and epigenetic remodeling in asthma.
Environmental determinants including aerosolized pollutants such as polycyclic aromatic hydrocarbons (PAHs) and tobacco smoke have been associated with exacerbation and increased incidence of asthma. The influence of aerosolized pollutants on the development of immune dysfunction in asthmatics has been suggested to be mediated through epigenetic remodeling. Genome accessibility and transcription are regulated primarily through DNA methylation, histone modification, and microRNA transcript silencing. Epigenetic remodeling has been shown in studies to be associated with Th2 polarization and associated cytokine and chemokine regulation in the development of asthma. This review will present evidence for the contribution of the aerosolized pollutants PAH and environmental tobacco smoke to epigenetic remodeling in asthma.


[Characteristics of regional lymph nodes in breast cancer (quantitative histochemical study)].
The changes in axillary lymph nodes in mammary gland carcinoma of different histological types, metastasizing and nonmetastasizing, as well as after radiation therapy and in fibroadenomatosis were studied. The study was carried out on cryostate sections by histological and histochemical methods. Signs of activation of lymph nodes were clearly seen only in solid carcinoma, not always manifested in adenocarcinomas and scirrhous carcinomas, and undetectable in fibroadenomatosis. The quantitative determination of enzymes and nucleic acids showed differences in their activity between fibroadenomatosis and carcinomas. Proliferation processes dominated significantly over lymphocyte differentiation in carcinoma, increasing even more in metastasizing tumors. Pre-operative irradiation did not inhibit metabolism or proliferative activity of the cells.


Genetic Predictors of Benign Course of Ulcerative Colitis-A North American Inflammatory Bowel Disease Genetics Consortium Study.
A subset of patients with ulcerative colitis (UC) have a benign course and an overall favorable prognosis. Early identification of these low-risk patients may allow for a less aggressive therapeutic approach and possible reduction of therapy-associated risks. The aim of this project was to identify the genetic predictors of benign UC phenotype. UC patients were selected from the National Institute of Diabetes and Digestive and Kidney Diseases Inflammatory Bowel Disease Genetics Consortium. Benign phenotype was defined as no need for immunomodulatory or biological therapy, hospitalizations, or colectomy. The association between benign UC phenotype and known loci linked to the risk of inflammatory bowel disease (IBD) was evaluated. The results for 156 index single-nucleotide polymorphisms (SNPs) from the known IBD loci were extracted for the main analysis. The association of the benign phenotype to a genetic burden score was also evaluated. None of the index SNPs from the IBD loci reached the predefined threshold of 1 × 10. In the exploratory analysis of the remaining Immunochip SNPs and imputed major histocompatibility complex data, 5 distinct suggestive association signals are identified (rs1697950, rs2523639, rs17836409, rs11742854, and rs75001121). No SNPs from IBD susceptibility loci were found to be associated (at our predefined threshold of 1 × 10) with a benign UC disease course. The rs11742570 variant on chromosome 5 was the one with the greatest association to benign disease although the association did not reach the predefined significant threshold. Given the modest power of our study, the findings suggested on the exploratory analysis merit extension to larger discovery cohorts.


Recombination and the origin of sequence diversity in the DRB MHC class II locus in chamois (Rupicapra spp.).
We examined the evolutionary processes contributing to genetic diversity at the major histocompatibility complex (MHC) class II DRB locus in chamois (Rupicapra spp., subfamily Caprinae). We characterised the pattern of intragenic recombination (or homologous gene conversion) and quantified the amount of recombination in the genealogical history of the two chamois species, Pyrenean chamois (Rupicapra pyrenaica) and Alpine chamois (Rupicapra rupicapra). We found evidence for intragenic recombination, and the estimated amount of population recombination suggests that recombination has been a significant process in generating DRB allelic diversity in the genealogical history of the genus Rupicapra. Moreover, positive selection appears to act on the same peptide-binding residues in both analysed chamois species, but not in identical intensity. Recombination coupled with positive selection drives the rapid evolution at the peptide-binding sites in the MHC class II DRB gene. Many chamois MHC class II DRB alleles are thus much younger than previously assumed.


CRISPR/Cas9-based genome editing in mice uncovers 13 testis- or epididymis-enriched genes individually dispensable for male reproduction†.
Developing a safe and effective male contraceptive remains a challenge in the field of medical science. Molecules that selectively target the male reproductive tract and whose targets are indispensable for male reproductive function serve among the best candidates for a novel non-hormonal male contraceptive method. To determine the function of these genes in vivo, mutant mice carrying disrupted testis- or epididymis-enriched genes were generated by zygote microinjection or electroporation of the CRISPR/Cas9 components. Male fecundity was determined by consecutively pairing knockout males with wild-type females and comparing the fecundity of wild-type controls. Phenotypic analyses of testis appearance and weight, testis and epididymis histology, and sperm movement were further carried out to examine any potential spermatogenic or sperm maturation defect in mutant males. In this study, we uncovered 13 testis- or epididymis-enriched evolutionarily conserved genes that are individually dispensable for male fertility in mice. Owing to their dispensable nature, it is not feasible to use these targets for the development of a male contraceptive.


Etiologies and Clinical Outcomes of Patients With Secondary Hemophagocytic Lymphohistiocytosis at a Tertiary PICU.
To assess the etiologies and outcomes of patients with secondary hemophagocytic lymphohistiocytosis in the PICU. Prospective observational cohort study. A single PICU at a pediatric tertiary hospital in Hanoi, Vietnam. Pediatric patients meeting the criteria for secondary hemophagocytic lymphohistiocytosis. None. Between June 2017 and May 2018, 25 consecutive patients with a mean (SD) age of 23.3 months (21.6 mo) were included. Collected variables included etiologies of hemophagocytic lymphohistiocytosis and clinical and laboratory findings at admission. The Pediatric Index of Mortality 2 score at admission was calculated. Outcomes were death and multiple organ dysfunction. The severity of multiple organ dysfunction was assessed by the Pediatric Logistic Organ Dysfunction 2 score. The mean (SD) Pediatric Index of Mortality 2 predicted mortality rate was 5.6% (7.6%). Cytomegalovirus and Epstein-Barr virus coinfections (60%) were the most common suspected etiology of hemophagocytic lymphohistiocytosis. Other etiologies included Epstein-Barr virus sole infections (20%), cytomegalovirus sole infections (16%), and one unknown cause (4%). Multiple organ dysfunction (excluding hematologic failure) was found in 22 patients (88%) with death occurring in 14 patients (56%). The mean (SD) Pediatric Logistic Organ Dysfunction 2 predicted mortality rate among patients with multiple organ dysfunction was 11.9% (11.2%). Despite having lower Pediatric Index of Mortality 2 predicted mortality rates at admission, Epstein-Barr virus-cytomegalovirus coinfection cases with multiple organ dysfunction had slightly greater Pediatric Logistic Organ Dysfunction 2 predicted mortality rates than Epstein-Barr virus sole infection cases with multiple organ dysfunction: 12.2% (10.5%) versus 11.3% (11.0%). However, these rates were lower than cytomegalovirus sole infection cases with multiple organ dysfunction (14.4% [16.3%]). Area under the curve values for Pediatric Index of Mortality 2 and Pediatric Logistic Organ Dysfunction 2 were 0.74 (95% CI, 0.52-0.95) and 0.78 (95% CI, 0.52-1.00), respectively, suggesting that both scales were fair to good at predicting mortality. Viral infections, particularly Epstein-Barr virus-cytomegalovirus coinfections, were a common cause of secondary hemophagocytic lymphohistiocytosis. The implication of these coinfections on the clinical course of hemophagocytic lymphohistiocytosis needs to be delineated.


Testicular Cancer-Surgical Treatment.
Testicular Germ Cell Tumor (GCT) is the most common solid tumor in men between the ages of 20-44. Men diagnosed with GCT have excellent survival rates due to advances in the multimodal treatment paradigm of chemotherapy, radiation therapy, and surgery. When considering the adequate treatment, several variables should be investigated and known to select the proper procedure. Therefore, when considering Testicular Intra-Epithelial Neoplasia, organ-sparring treatment, such as radiotherapy or organ-sparring surgery should be considered, reaching a cure rate of 98%. However, when the case is of a seminoma or a non-seminoma, orchiectomy is usually the chosen procedure, reaching an oncological cure rate of 80-85%, when there is no metastasis. Retroperitoneal Lymph Node Dissection (RPLND) is generally considered as a treatment option for non-seminomas, when lymph nodes are compromised. There are three different RPLND techniques: open, laparoscopic, and robotic. The open approach is as effective as the other two in its oncological efficiency. Although, when considering both laparoscopic and robotic approach, hospital stays are significantly reduced, better cosmetic results, and less complications when compared to the open approach. Both laparoscopic and robotic approaches require extensive experience and have a steep learning curve, while also providing similar outcome, however, recent studies have been pointing out a slight increase of advantages on the robotic approach. Therefore, further studies are necessary to assert the robotic approach superiority. Also, it is noteworthy that new technologies are on the rise, improving the laparoscopic approach, requiring further studies after their uses are consolidated.


A multicentre review comparing long term outcomes of endoscopic vein harvesting versus open vein harvesting for coronary artery bypass surgery.
Utilisation of the Endoscopic Vein Harvesting (EVH) technique has been increasing for coronary artery bypass grafting (CABG) for the last two decades. Some surgeons remain concerned about the long-term patency of the long saphenous vein harvested endoscopically compared to traditional Open Vein Harvesting (OVH). The aim of this study was to perform a retrospective analysis of the outcomes between EVH and OVH from three UK centres with 10 years follow-up. 27,024 patients underwent CABG with long saphenous vein harvested by EVH (n=13,794) or OVH (n=13,230) in three UK centres between 2007 and 2019. Propensity modelling was used to calculate the Inverse Probability of Treatment Weights (IPTW). The primary endpoint was mortality from all causes and secondary endpoints were length of hospital stay, postoperative complications, and incidence of repeat coronary re-vascularisation for symptomatic patients. IPTW was used to balance the two intervention groups for baseline and preoperative co-morbidities. Median follow-up time was 4.54 years for EVH and 6.00 years for OVH. Death from any cause occurred in 13.8% of the EVH group versus 20.8% in the OVH group over the follow-up period. The hazard ratio of death (EVH to OVH) was 0.823 (95% CI: 0.767, 0.884). Length of hospital stay was similar between the groups (p=0.86). Post-operative pulmonary complications were more common in EVH vs OVH (14.7% vs. 12.8%, p<0.001), but repeat coronary re-vascularisation was similar between the groups. This large retrospective multicentre analysis indicates that EVH has a lower risk of mortality compared with OVH during the follow-up period of the study. The observed benefits of EVH may outweigh the risks but should be considered on a case-by-case basis. We hope this review gives confidence to other cardiac centres that offering an EVH approach to conduit harvesting does not affect long term patient outcomes.


Structure and mechanical properties of the ribosomal L1 stalk three-way junction.
The L1 stalk is a key mobile element of the large ribosomal subunit which interacts with tRNA during translocation. Here, we investigate the structure and mechanical properties of the rRNA H76/H75/H79 three-way junction at the base of the L1 stalk from four different prokaryotic organisms. We propose a coarse-grained elastic model and parameterize it using large-scale atomistic molecular dynamics simulations. Global properties of the junction are well described by a model in which the H76 helix is represented by a straight, isotropically flexible elastic rod, while the junction core is represented by an isotropically flexible spherical hinge. Both the core and the helix contribute substantially to the overall H76 bending fluctuations. The presence of wobble pairs in H76 does not induce any increased flexibility or anisotropy to the helix. The half-closed conformation of the L1 stalk seems to be accessible by thermal fluctuations of the junction itself, without any long-range allosteric effects. Bending fluctuations of H76 with a bulge introduced in it suggest a rationale for the precise position of the bulge in eukaryotes. Our elastic model can be generalized to other RNA junctions found in biological systems or in nanotechnology.


[CYP2D6 and CYP2C19 genotyping: effectivity in psychiatric practice]
International guidelines recommend in patients with an in- or decreased CYP2D6 and CYP2C19 metabolism to adjust the dose of medication metabolized by these enzymes. This is in purpose to increase effectiveness and to lower the risk of side-effects of this medication. However, it is still unclear if dose adjustment based on genotype results in better clinical outcomes. To provide an update regarding CYP2D6 and CYP2C19 genotyping in psychiatry in relation to ethnic diversity.To provide an update regarding CYP2D6 and CYP2C19 genotyping in psychiatry in relation to ethnic diversity. We conducted a comprehensive meta-analysis to the prevalence of non-normal metabolizers as the equivalent of the sum-prevalence of poor, intermediate and ultrarapid metabolizer CYP2D6 and CYP2C19 predicted phenotypes. For the prevalence and effectiveness study, a total of 166 Antilleans living in the Netherlands and 269 psychiatric patients (on the island Curaçao) were genotyped for CYP2D6 and CYP2C19. Of the psychiatric patients, 45 non-normal CYP2D6 metabolizers using medication metabolized by CYP2D6, were included for dose adjustment and were matched with 41 normal metabolizers. All 45 patients were using antipsychotic medication for a minimum of two years. Four months after dose adjustment they were reassessed. The mean total probability estimates of having a non-normal predicted phenotype worldwide were 36% and 62% for CYP2D6 and CYP2C19, respectively. There was a large interethnic variability (min-max 2.7-61.2% (CYP2D6) and minmax 31.7-80.1% (CYP2C19)). No significant difference was found in the phenotypes of psychiatric patients, Dutch Caribbean subjects from the general population, and European populations. There were no beneficial effects of dose adjustments to phenotype in the non-normal CYP2D6 metabolizers. More than 75% of the world population has a non-normal CYP2D6 and/or CYP2C19 phenotype. Dose adjustment to the CYP2D6 phenotype according to international guidelines in patients on long-term antipsychotic treatment showed no beneficial effect. Further research to CYP genotyping in psychiatry is warranted.


Overexpression of PD-L1 significantly associates with tumor aggressiveness and postoperative recurrence in human hepatocellular carcinoma.
The aberrant expression of programmed cell death 1 ligands 1 and 2 (PD-Ls) on tumor cells dampens antitumor immunity, resulting in tumor immune evasion. In this study, we investigated the expression of PD-Ls in human hepatocellular carcinoma (HCC) to define their prognostic significance after curative surgery. Immunohistochemistry was used to investigate PD-Ls expression as well as granzyme B+ cytotoxic and FoxP3+ regulatory T cell infiltration on tissue microarrays containing 240 randomly selected HCC patients who underwent surgery. The results were further verified in an independent cohort of 125 HCC patients. PD-Ls expression on HCC cell lines was detected by Western blot assay. Patients with higher expression of PD-L1 had a significantly poorer prognosis than patients with lower expression. Although patients with higher expression of PD-L2 also had a poorer survival, the difference in recurrence was not statistically significant. Multivariate analysis identified tumor expression of PD-L1 as an independent predictor for postoperative recurrence. No correlation was found between PD-Ls expression and granzyme B+ lymphocyte infiltration, whereas a significant positive correlation was detected between PD-Ls expression and FoxP3+ lymphocyte infiltration. In addition, tumor-infiltrating cytotoxic and regulatory T cells were also independent prognosticators for both survival and recurrence. The prognostic value of PD-L1 expression was validated in the independent data set. Our data suggest for the first time that PD-L1 status may be a new predictor of recurrence for HCC patients and provide the rationale for developing a novel therapy of targeting the PD-L1/PD-1 pathway against this fatal malignancy.


A multistable movement display: evidence for two separate motion systems in human vision.
Two competing sensations of apparent movement were produced by the rapid alternation of two multielement stimulus frames. Either sensation could be made dominant by, appropriate manipulations of the stimulus display. The results suggest that there are two systems capable of generating movement signals in man. One system depends on preliminary processing of form, and the second system does not.


Pre-B cells in mouse bone marrow: immunofluorescence stathmokinetic studies of the proliferation of cytoplasmic mu-chain-bearing cells in normal mice.
By using a technique that combines metaphase arrest with immunofluorescence labeling, the proliferation of specifically identified pre-B cells in mouse bone marrow has been analyzed under physiological conditions in vivo. Pre-B cells bearing cytoplasmic mu-chains and no surface mu-chains constituted 12% of marrow nucleated cells, or 27 X 10(5) cells/femur, whereas surface mu-bearing B lymphocytes totaled 33 X 10(5) cells/femur. Pre-B cells measured 7 to 14 micron in diameter, the small number seen in metaphase (1 to 2%) being large cells (greater than 10 microns). After vincristine injection, the metaphase incidence (Imet) of pre-B cells increased with cell size; a broad-dose range of vincristine gave similar Imet values. Mitoses were arrested for 4 hr with no apparent cell death. Linear regression analysis of the increase in Imet of pre-B cells 2 to 4 hr after vincristine revealed a rate of entry into mitosis of 6.3%/hr, relative to all pre-B cells (average compartment turnover time, 16 hr), and 15.3%/hr for the large proliferating pre-B cell subset. This represented a pre-B cell production of 1.3 X 10(5) cells/femoral shaft/hr or 0.5 X 10(8) cells/whole bone marrow organ/day, emphasizing the magnitude of B lymphocyte genesis in normal bone marrow. Combined with reported renewal rates for small pre-B cells and small B lymphocytes, these values form a kinetic model of B lymphocyte development. The results reveal an apparent overproduction of large pre-B cells, consistent with a speculative post-mitotic loss of some immature primary B lymphocytes.


Danggui Buxue Decoction exerts its therapeutic effect on rheumatoid arthritis through the inhibition of Wnt/β-catenin signaling pathway.
Danggui Buxue Decoction (DBD) is a traditional Chinese medicine prescription, which has the functions of benefiting Qi, generating blood and regulating the immune system. At present, various clinical reports suggest that DBD has some efficacy in Rheumatoid arthritis (RA), but its mechanism of action is still unclear. Thus, the present study explored mechanism of this preparation on RA. The effect of DBD was evaluated by tumor necrosis factor (TNF)-α-induced Human fibroblast-like synoviocyte of rheumatoid arthritis (HFLS-RA) cell model and collagen-induced arthritis (CIA) rat model, respectively. Inflammatory factors including TNF-ɑ, IL-1β, IL-6 and IL-10 in the culture supernatants or rat serum were measured using ELISA. The related indexes including fur luster, mental state and activity of rat and the symptoms including swelling and deformation of toes and ankles were also measured. In vitro results showed that DBD cannot only inhibit the proliferation of HFLS-RA cells but also reduce the levels of pro-inflammatory factors while increasing the level of anti-inflammatory factors. Similar results were obtained from in vivo experiments. Rats receiving DBD showed a decrease in the severity of rheumatoid arthritis in rat models. Moreover, the protein levels of c-myc and β-catenin decreased significantly, while the protein level of SFRP4 increased, which indicated that DBD might inhibit the inflammatory reaction by regulating Wnt/β-catenin signaling pathway, thus alleviating the symptoms of RA. Our findings not only provide insights for understanding the molecular mechanism of DBD in treating RA, but also provide the theoretical basis for further clinical prevention and treatment.


Wind turbines and health: An examination of a proposed case definition.
Renewable energy demands have increased the need for new wind farms. In turn, concerns have been raised about potential adverse health effects on nearby residents. A case definition has been proposed to diagnose "Adverse Health Effects in the Environs of Industrial Wind Turbines" (AHE/IWT); initially in 2011 and then with an update in 2014. The authors invited commentary and in turn, we assessed its scientific merits by quantitatively evaluating its proposed application. We used binomial coefficients to quantitatively assess the potential of obtaining a diagnosis of AHE/IWT. We also reviewed the methodology and process of the development of the case definition by contrasting it with guidelines on case definition criteria of the USA Institute of Medicine. The case definition allows at least 3,264 and up to 400,000 possibilities for meeting second- and third-order criteria, once the limited first-order criteria are met. IOM guidelines for clinical case definitions were not followed. The case definition has virtually no specificity and lacks scientific support from peer-reviewed literature. If applied as proposed, its application will lead to substantial potential for false-positive assessments and missed diagnoses. Virtually any new illness that develops or any prevalent illness that worsens after the installation of wind turbines within 10 km of a residence could be considered AHE/IWT if the patient feels better away from home. The use of this case definition in the absence of a thorough medical evaluation with appropriate diagnostic studies poses risks to patients in that treatable disorders would be overlooked. The case definition has significant potential to mislead patients and its use cannot be recommended for application in any health-care or decision-making setting.


Epithelial displacement after stereotactic 11-gauge directional vacuum-assisted breast biopsy.
Displaced epithelial fragments at percutaneous biopsy of ductal carcinoma in situ (DCIS) may mimic stromal invasion. This study was undertaken to determine the frequency of epithelial displacement in DCIS lesions of patients who underwent stereotactic 11-gauge directional vacuum-assisted breast biopsy. We retrospectively reviewed 28 consecutive DCIS lesions in patients who underwent stereotactic 11-gauge directional vacuum-assisted breast biopsy followed by surgery. Surgical specimens were examined for histologic evidence of epithelial displacement, consisting of fragments of epithelium in artifactual spaces in breast parenchyma or in lymphovascular channels, accompanied by hemorrhage, fat necrosis, inflammation, hemosiderin-laden macrophages, or granulation tissue. The median number of specimens obtained per lesion was 14 (range, seven to 45). The median interval from stereotactic biopsy to surgery was 27 days (range, 10-59 days). Surgery revealed DCIS in 19 (68%) of 28 lesions, DCIS and infiltrating carcinoma in four lesions (14%), and no residual carcinoma in five lesions (18%). Reactive changes at the biopsy site were identified in all cases. Displacement of benign epithelium into granulation tissue at the stereotactic biopsy site was identified in two cases (7%). We found no evidence of displacement of malignant epithelium. Epithelial displacement is uncommon after stereotactic 11-gauge directional vacuum-assisted biopsy of the breast. We observed displacement of benign epithelium in two (7%) of 28 DCIS lesions and no displacement of malignant epithelium.


Sarcoidosis with optic nerve and retinal involvement.
A patient with the typical funduscopic changes of sarcoidosis died following an accident. The histopathological findings in one eye were correlated with the ophthalmoscopic observations made several days prior to death. Gross and histopathological examination revealed that the characteristic whitish-yellow focal retinal exudates, the perivascular exudates with an appearance like candle wax drippings, the preretinal and intravitreal nodules, and the localized tumefactions on the inner surface of the retina and optic nerve head noted ophthalmoscopically were a result of the nodular proliferation of epithelioid cells. These cells appeared to be derived primarily from blood vessels in the retina and optic nerve head. Although subpigment epithelial granulomas were found, there were no granulomas present within the choroid or elsewhere within the uvea.


Role of CXC group chemokines in lung cancer development and progression.
Clinical and translational research on lung cancer patients undergoing surgical treatment can provide valuable scientific data and unique opportunity to study tumor microenvironment. CXC chemokines, which are members of a big family of cytokines, are undoubtedly involved in tumor growth regulation and metastasizing pathways. For better understanding of CXC chemokine involvement in the process of carcinogenesis we have studied the cohort of early stage non-small cell lung cancer patients undergoing surgery with curative intent. Our aim was to assess CXC chemokine ligand (CXCL) levels in patient blood samples representing systemic circulation and tumor microenvironment; assess CXC chemokine receptor (CXCR) expression in tumor tissue; and measure tumor infiltrating immune cell subpopulations. A total of 54 patients with NSCLC had radical lung resection were enrolled in a single center prospective study and were followed-up annually for up to six years. During surgical procedure peripheral and tumor draining blood samples were taken. CXCL1, CXCL4, CXCL5, CXCL6, CXCL7, CXCL8, CXCL9, CXCL10, CXCL11 and CXCL12 levels were determined by ELISA, and chemokine concentration gradient was calculated. Tumor infiltrating immune cells (T helper cells, T cytotoxic cells, macrophages, B cells, plasma cells) and expression of CXCR1, CXCR2, CXCR3 and CXCR4 in tumor tissue were assessed by immunohistochemistry. Statistically significant decrease in chemokine concentration was found for CXCL4 (P=0.002) and CXCL5 (P=0.011), and statistically significant concentration increase was found for CXCL7 (P=0.001) in total cohort. We have found statistically significant CXC chemokine concentration change for majority of chemokines-CXCL1 (P=0.002), CXCL4 (P=0.001), CXCL5 (P=0.013), CXCL7 (P=0.036), CXCL8 (P=0.026), CXCL9 (P=0.034) and CXCL10 (P=0.032) in a group of patients who had good clinical result after surgery with no evidence of relapse, on the other hand patients with cancer recurrence including local and systemic cancer spread did not show any change of chemokine concentration in blood except for CXCL1 (P=0.041). We have also found that chemokine levels and gradients correlate with CXC receptor expression and number of tumor infiltrating immune cell subpopulations. Assessment of tumor microcirculation is useful for evaluation of different types of circulating biomarkers and application of our method can be very wide, integrating thoracic surgeons into translational cancer research.


Trans-cellular migration: cell-cell contacts get intimate.
Trans-cellular migration, the movement of one cell directly through another, seems an unlikely, counterintuitive, and even bizarre process. Trans-cellular migration has been reported for nearly half a century in leukocyte transendothelial migration in vivo, but is not well enough accepted to widely feature in textbook accounts of diapedesis. Recently, the first in vitro and additional in vivo observations of trans-cellular diapedesis have been reported. Mechanisms by which this occurs are just beginning to be elucidated and point to podosome-like protrusive activities in leukocytes and specific fusogenic functions in endothelial cells. Emerging evidence for a quantitatively significant contribution of trans-cellular migration to leukocyte trafficking in increasingly diverse settings suggests that this phenomenon represents an important and physiologic cell biological process.


Dugong (Dugong dugon) vocalization patterns recorded by automatic underwater sound monitoring systems.
To quantitatively examine the diurnal, or tidal, effects on dugong behavior, we employed passive acoustic observation techniques to monitor the animals. Automatic underwater sound monitoring systems for dugongs (AUSOMS-D) were deployed on the sea floor at depths of about 5 m south of Talibong Island, Thailand. The AUSOMS-D recorded underwater sound in stereo at a sampling frequency of 44.1 kHz for more than 116 consecutive hours. Dugong calls were automatically detected by newly developed software with a detection rate of 36.1% and a false alarm rate of 2.9%. In total, 3453 calls were detected during the 164 h of recording. The autocorrelation of the call rate indicated an attendance cycle of about 24 or 25 h, and the most frequent vocalizations were observed from 0300 to 0500 h. The calculated bearings of the sound sources, i.e., dugongs, were used as an indicator to track the relative numbers of dugongs during the monitoring periods.


Assessing the Accuracy of Citizen Scientist Reported Measurements for Agrichemical Contaminants.
Citizen science is a research tool capable of addressing major environmental challenges, including contamination of water resources by agrichemicals, such as nutrients and pesticides. The objectives of this study were (1) to identify the proportion of accurate observations by citizen scientists using rapid assessment water quality tools, and (2) to characterize how a user's prior experience with water quality tools was associated with the accuracy of citizen scientists. To achieve these objectives, we conducted group testing with over 136 citizen scientists and compared their results from water quality testing of water samples to results obtained using laboratory analytical methods. Following brief training, we observed that accuracy of reported results varies based on the user's experience level where experienced and expert users shared consistent and reliable measurements. Where erroneous measures were reported, citizen scientists tend to overestimate contaminant concentrations when using colorimetric water quality tools. Additionally, we identified differences in accuracy related to the types of water quality assessment tools used by citizen scientists from each experience group. This study demonstrates the importance of evaluating participant background experience in designing citizen science campaigns.


Ethnicity, infant-feeding practices, and childhood adiposity.
There has been professional concern that the type of milk used for infant-feeding may lead to adiposity. Studies of the relationship between infant milk-feeding and adiposity, however, have led to inconsistent results. This study investigated the relationship of infant-feeding practices to three indicators of adiposity: body weight, body mass index (BMI) and sum of seven skinfolds. The sample includes children at 3 or 4 years of age, in three ethnic groups. Multivariate techniques assessed the relationship among practices of infant-feeding with three indicators of adiposity, while considering potential confounding variables. Although a weak bivariate relationship was detected between the duration of breastfeeding and body weight, none of the measures of infant-feeding were related to the three indicators of adiposity. Black-American girls had smaller skinfolds than Anglo- or Mexican-American girls, with no ethnic group differences among boys. Concerns about adiposity due to methods of infant-feeding can be allayed, at least among 3- or 4-year-old children.


Detrital shadows: estuarine food web connectivity depends on fluvial influence and consumer feeding mode.
We measured the influence of landscape setting on estuarine food web connectivity in five macrotidal Pacific Northwest estuaries across a gradient of freshwater influence. We used stable isotopes (δ13 C, δ15 N, δ34 S) in combination with a Bayesian mixing model to trace primary producer contributions to suspension- and deposit-feeding bivalve consumers (Mytilus trossulus and Macoma nasuta) transplanted into three estuarine vegetation zones: emergent marsh, mudflat, and eelgrass. Eelgrass includes both Japanese eelgrass (Zostera japonica) and native eelgrass (Zostera marina). Fluvial discharge and consumer feeding mode strongly influenced the strength and spatial scale of observed food web linkages, while season played a secondary role. Mussels displayed strong cross-ecosystem connectivity in all estuaries, with decreasing marine influence in the more fluvial estuaries. Mussel diets indicated homogenization of detrital sources within the water column of each estuary. In contrast, the diets of benthic deposit-feeding clams indicated stronger compartmentalization in food web connectivity, especially in the largest river delta where clam diets were trophically disconnected from marsh sources of detritus. This suggests detritus deposition is patchy across space, and less homogenous than the suspended detritus pool. In addition to fluvial setting, other estuary-specific environmental drivers, such as marsh area or particle transport speed, influenced the degree of food web linkages across space and time, often accounting for unexpected patterns in food web connectivity. Transformations of the estuarine landscape that alter river hydrology or availability of detritus sources can thus potentially disrupt natural food web connectivity at the landscape scale, especially for sedentary organisms, which cannot track their food sources through space.


Imitating actions on objects in early-onset and regressive autism: effects and implications of task characteristics on performance.
This study was designed to examine the nature of object imitation performance in early autism. We hypothesized that imitation would be relatively preserved when behaviors on objects resulted in salient instrumental effects. We designed tasks in which, in one condition, the motor action resulted in a salient, meaningful effect on an object, whereas in the other condition, the same action resulted in a less salient effect because of differing object characteristics. The motor aspects of the tasks did not vary across conditions. Four participant groups of 2- to 5-year-olds were examined: 17 children with early-onset autism, 24 children with regressive onset autism, 22 children with developmental delays, and 22 children with typical development. Groups were matched on nonverbal skills, and differences in verbal development were examined as a moderator of imitative ability. Results revealed an interaction of group by condition, with the combined autism group failing more tasks than the combined comparison groups, and failing more tasks in the less salient condition than in the more salient condition, as hypothesized. Analyses of autism subgroups revealed these effects were primarily because of the regression onset group. Accuracy of motor performance was examined by analyzing errors. Among children passing imitative acts, there were no group differences and no condition effects in the number, type, or pattern of performance errors. Among children passing the tasks, the group with autism did not demonstrate more emulation errors (imitating the goal but not the means) than other groups. There was no evidence that either motor or attentional aspects of the tasks contributed to the poorer imitative performance of the children with autism.


Loss of EZH2-like or SU(VAR)3-9-like proteins causes simultaneous perturbations in H3K27 and H3K9 tri-methylation and associated developmental defects in the fungus Podospora anserina.
Selective gene silencing is key to development. It is generally accepted that H3K27me3-enriched heterochromatin maintains transcriptional repression established during early development and regulates cell fate. Conversely, H3K9me3-enriched heterochromatin prevents differentiation but constitutes protection against transposable elements. We exploited the fungus Podospora anserina, a valuable alternative to higher eukaryote models, to question the biological relevance and functional interplay of these two distinct heterochromatin conformations. We established genome-wide patterns of H3K27me3 and H3K9me3 modifications, and found these marks mutually exclusive within gene-rich regions but not within repeats. We generated the corresponding histone methyltransferase null mutants and showed an interdependence of H3K9me3 and H3K27me3 marks. Indeed, removal of the PaKmt6 EZH2-like enzyme resulted not only in loss of H3K27me3 but also in significant H3K9me3 reduction. Similarly, removal of PaKmt1 SU(VAR)3-9-like enzyme caused loss of H3K9me3 and substantial decrease of H3K27me3. Removal of the H3K9me binding protein PaHP1 provided further support to the notion that each type of heterochromatin requires the presence of the other. We also established that P. anserina developmental programs require H3K27me3-mediated silencing, since loss of the PaKmt6 EZH2-like enzyme caused severe defects in most aspects of the life cycle including growth, differentiation processes and sexual reproduction, whereas loss of the PaKmt1 SU(VAR)3-9-like enzyme resulted only in marginal defects, similar to loss of PaHP1. Our findings support a conserved function of the PRC2 complex in fungal development. However, we uncovered an intriguing evolutionary fluidity in the repressive histone deposition machinery, which challenges canonical definitions of constitutive and facultative heterochromatin.


Facial scars after a road accident--combined treatment with pulsed dye laser and Q-switched Nd:YAG laser.
We report the case of a woman who presented with several facial scars following a road accident. Treatment was carried out using combined laser treatment with pulsed dye laser (PDL) and Q-switched neodymium:yttrium-aluminum-garnet laser (QS Nd:YAG laser). No side effects or complications from treatment were noted or reported. The patient had very good cosmetic results with this combined technique. A variety of facial scars - erythematous, pigmented, atrophic, and hypertrophic - may occur as a result of trauma, surgery, burns, and skin disease. Surgery with other adjunctive methods including radiotherapy, intralesional steroids, and pressure therapy have shown variable results. Laser treatment has been attempted for scar revision since the 1980s. The PDL is the optimal treatment for reducing scar bulk and symptoms. It also decreases erythema and telangiectasia associated with scars, normalizes the skin surface texture, and improves the scar pliability. The QS Nd:YAG laser (1064 nm) is highly effective for traumatic tattoo removal, resulting in complete clearance in the majority of cases.


Frontiers in epidermal barrier homeostasis--an approach to mathematical modelling of epidermal calcium dynamics.
Intact epidermal barrier function is crucial for survival and is associated with the presence of gradients of both calcium ion concentration and electric potential. Although many molecules, including ion channels and pumps, are known to contribute to maintenance of these gradients, the mechanisms involved in epidermal calcium ion dynamics have not been clarified. We have established that a variety of neurotransmitters and their receptors, originally found in the brain, are expressed in keratinocytes and are also associated with barrier homeostasis. Moreover, keratinocytes and neurons show some similarities of electrochemical behaviour. As mathematical modelling and computer simulation have been employed to understand electrochemical phenomena in brain science, we considered that a similar approach might be applicable to describe the dynamics of epidermal electrochemical phenomena associated with barrier homeostasis. Such methodology would also be potentially useful to address a number of difficult problems in clinical dermatology, such as ageing and itching. Although this work is at a very early stage, in this essay, we discuss the background to our approach and we present some preliminary results of simulation of barrier recovery.


HIV impairs TNF-alpha mediated macrophage apoptotic response to Mycobacterium tuberculosis.
The factors that contribute to the exceptionally high incidence of Mycobacterium tuberculosis (MTb) disease in HIV(+) persons are poorly understood. Macrophage apoptosis represents a critical innate host cell response to control MTb infection and limit disease. In the current study, virulent live or irradiated MTb (iMTbRv) induced apoptosis of differentiated human U937 macrophages in vitro, in part dependent on TNF-alpha. In contrast, apoptosis of differentiated HIV(+) human U1 macrophages (HIV(+) U937 subclone) was markedly reduced in response to iMTbRv and associated with significantly reduced TNF-alpha release, whereas apoptosis and TNF-alpha release were intact to TLR-independent stimuli. Furthermore, reduced macrophage apoptosis and TNF-alpha release were independent of MTb phagocytosis. Whereas surface expression of macrophage TLR2 and TLR4 was preserved, IL-1 receptor associated kinase-1 phosphorylation and NF-kappaB nuclear translocation were reduced in HIV(+) U1 macrophages in response to iMTbRv. These findings were confirmed using clinically relevant human alveolar macrophages (AM) from healthy persons and asymptomatic HIV(+) persons at clinical risk for MTb infection. Furthermore, in vitro HIV infection of AM from healthy persons reduced both TNF-alpha release and AM apoptosis in response to iMTbRv. These data identify an intrinsic specific defect in a critical macrophage cellular response to MTb that may contribute to disease pathogenesis in HIV(+) persons.


Antimicrobial prophylaxis in patients with immune thrombocytopenia treated with rituximab: a retrospective multicenter analysis.
The primary aim of this study was to describe the use of primary anti-infective prophylaxis (AP) in common clinical practice in patients affected by immune thrombocytopenia (ITP) and treated with RTX. Population studied consisted of patients affected by ITP (age ≥ 18 years) who had received at least one dose of RTX from January 2008 to June 2018. Five Italian haematology centres participated in the current study. Data were retrospectively collected: demographic data (age, gender), concomitant comorbidities and previous therapies for ITP, characteristics of AP, the occurrence of infections and their management. The ITP cohort consisted of 67 patients sub-grouped into two categories according to the administration of AP: (1) treated with AP (N= 34; 51%) and (2) not treated with AP (N=33, 49%). AP consisted of combined trimethoprim/sulfamethoxazole (TMP/SMX) and acyclovir (AC) in half of patients. TPM/SMX as a single agent was adopted in 32% patients and one patient received only AC. Overall, infections were experienced in 15% of patients during follow-up with a similar proportion in the 2 groups (treated and not treated) of patients (14.7% vs 15%). Clinical course of infections was however, less severe in patients treated with AP, where all infections were grade 2 and did not require hospitalization. In neither group of patients was reported Pneumocystis pneumonia. In conclusion, despite the absence of clear evidence, our analysis shows that AP in patients with ITP receiving RTX is frequently adopted, even if in the absence of well-defined criteria. Prophylaxis administration is quite consistent within the same haematological Center; thus, it seems related to clinicians' experience.


Cerebral metastases from epithelial ovarian carcinoma.
We describe four patients with epithelial ovarian carcinoma and cerebral metastases. Cerebral metastases are still an infrequent occurrence in this malignancy but, as treatment for ovarian cancer produces more long-term survivors, such uncommon patterns of spread may become more frequent. Part of a future successful cure for this disorder may require the recognition of the central nervous system as a potential sanctuary site for metastatic disease.


Site and land-use associations of soil bacteria and fungi define core and indicative taxa.
Soil microbial diversity has major influences on ecosystem functions and services. However, due to its complexity and uneven distribution of abundant and rare taxa, quantification of soil microbial diversity remains challenging and thereby impeding its integration into long-term monitoring programs. Using metabarcoding, we analyzed soil bacterial and fungal communities at 30 long-term soil monitoring sites from the three land-use types arable land, permanent grassland, and forest with a yearly sampling between snowmelt and first fertilization over five years. Unlike soil microbial biomass and alpha-diversity, microbial community compositions and structures were site- and land-use-specific with CAP reclassification success rates of 100%. The temporally stable site core communities included 38.5% of bacterial and 33.1% of fungal OTUs covering 95.9% and 93.2% of relative abundances. We characterized bacterial and fungal core communities and their land-use associations at the family-level. In general, fungal families revealed stronger land-use associations as compared to bacteria. This is likely due to a stronger vegetation effect on fungal core taxa, while bacterial core taxa were stronger related to soil properties. The assessment of core communities can be used to form cultivation-independent reference lists of microbial taxa, which may facilitate the development of microbial indicators for soil quality and the use of soil microbiota for long-term soil biomonitoring.