Matrix metalloproteinase 13-deficient mice are resistant to osteoarthritic cartilage erosion but not chondrocyte hypertrophy or osteophyte development.
To investigate the role of matrix metalloproteinase 13 (MMP-13; collagenase 3) in osteoarthritis (OA). OA was surgically induced in the knees of MMP-13-knockout mice and wild-type mice, and mice were compared. Histologic scoring of femoral and tibial cartilage aggrecan loss (0-3 scale), erosion (0-7 scale), and chondrocyte hypertrophy (0-1 scale), as well as osteophyte size (0-3 scale) and maturity (0-3 scale) was performed. Serial sections were stained for type X collagen and the MMP-generated aggrecan neoepitope DIPEN. Following surgery, aggrecan loss and cartilage erosion were more severe in the tibia than femur (P<0.01) and tibial cartilage erosion increased with time (P<0.05) in wild-type mice. Cartilaginous osteophytes were present at 4 weeks and underwent ossification, with size and maturity increasing by 8 weeks (P<0.01). There was no difference between genotypes in aggrecan loss or cartilage erosion at 4 weeks. There was less tibial cartilage erosion in knockout mice than in wild-type mice at 8 weeks (P<0.02). Cartilaginous osteophytes were larger in knockout mice at 4 weeks (P<0.01), but by 8 weeks osteophyte maturity and size were no different from those in wild-type mice. Articular chondrocyte hypertrophy with positive type X collagen and DIPEN staining occurred in both wild-type and knockout mouse joints. Our findings indicate that structural cartilage damage in a mouse model of OA is dependent on MMP-13 activity. Chondrocyte hypertrophy is not regulated by MMP-13 activity in this model and does not in itself lead to cartilage erosion. MMP-13 deficiency can inhibit cartilage erosion in the presence of aggrecan depletion, supporting the potential for therapeutic intervention in established OA with MMP-13 inhibitors.


Induction of apoptosis reduces the severity of caerulein-induced pancreatitis in mice.
The recent observation that the severity of pancreatitis is inversely related to the extent of apoptosis in five experimental models of the disease has suggested the possibility that apoptosis might protect against pancreatic injury in pancreatitis. This hypothesis was tested by inducing pancreatitis in mice during a phase of extensive apoptosis. Mice were fed a raw soya diet for five weeks to stimulate pancreatic growth and then switched to a regular chow diet for 27 hrs to permit involution of the hypertrophied gland. That involution is characterized by extensive apoptosis of acinar cells. Pancreatitis was induced, in either control mice or mice undergoing pancreatic involution, by repeated intraperitoneal administration of a supramaximally stimulating dose of caerulein (50 microg/kg given each hr for 12 hrs). The magnitude of hyperamylasemia, degree of inflammation, and extent of necrosis were reduced in the mice receiving caerulein during pancreatic involution. We conclude that induction of apoptosis may protect against acinar cell injury and reduce the severity of pancreatitis.


Ostreid herpesvirus 1 latent infection and reactivation in adult Pacific oysters, Crassostrea gigas.
Ostreid herpesvirus 1 (OsHV-1) is one of the most economically important pathogens of Pacific oysters. Understanding the pathogenesis of this virus is critical to developing tools to control outbreaks on shellfish farms. OsHV-1 is genetically related to vertebrate herpesviruses, which have a lytic and a latent stage, with the latent stage capable of being reactivated to the lytic stage. Here, OsHV-1 latency in Pacific oysters was investigated in experimentally and naturally infected oysters. Lytic infection in one-year-old oysters injected with the Tomales Bay strain of OsHV-1 was detectable between 1 and 4 days post-injection (dpi) but was not detectable after 5 dpi. The injected oysters shed 1 × 102 to 1 × 104 DNA copies/ml into the water during the 4-day acute phase. Lytic shedding was not detectable in two-year-old oysters injected similarly with the same strain of OsHV-1; however, the OsHV-1 genome was detectable by qPCR in the adductor muscle, gill, mantle, and hemocytes within the first 3 dpi, after which it became undetectable. No OsHV-1 was detectable in the adductor muscle, gill, or mantle from experimentally infected oysters on days 15 and 21 post-injection or from oysters sampled 9 months after surviving an OsHV-1 mortality event; however, OsHV-1 DNA could be detected in hemocytes of both experimentally infected oysters at 21 dpi and naturally infected oysters using nested PCR. In addition, lytic viral gene transcription was detectable in hemocytes of experimentally infected oysters between 1 and 21 dpi and in hemocytes of naturally infected oysters. Furthermore, OsHV-1 reactivation from latency was induced in experimentally infected oysters at 21 dpi and in naturally infected oysters 12 months after an OsHV-1 outbreak.


Anxiety levels in people who stutter: a randomized population study.
The question of whether people who stutter are generally more anxious than people who do not stutter has not yet been resolved. One major methodological barrier to determining whether differences exist has been the type of stuttering sample used. Studies investigating anxiety levels of those who stutter have mostly assessed people referred to stuttering therapy clinics, which is arguably a biased sample. To date, no studies have been published that have measured the anxiety levels of people who stutter in the community using random selection procedures. Such a sample is more likely to be representative of the population of people who stutter. The present study involved a random selection and telephone interview of people in 4,689 households. The telephone respondent was given a description of stuttering and asked if any person living in their household stuttered. If yes, a number of corroborative questions were asked, and permission was requested to tape the speech of the person believed to stutter over the telephone. A definite case of stuttering was based on (a) a positive detection of stuttering from the tape and (b) at least one of the corroborative questions supporting the diagnosis. A total of 87 people were identified as definite cases of stuttering across all ages, and 63 participants who were 15 years or older completed a trait anxiety questionnaire over the telephone. Mean trait anxiety levels were significantly higher than levels generally found in society, though differences were not large. Implications of these results are discussed.


Transforming growth factor-beta 1 (TGF-beta1) promotes IL-2 mRNA expression through the up-regulation of NF-kappaB, AP-1 and NF-AT in EL4 cells.
Transforming growth factor beta1 (TGF-beta1) has been previously shown to modulate interleukin 2 (IL-2) secretion by activated T-cells. In the present studies, we determined that TGF-beta1 induced IL-2 mRNA expression in the murine T-cell line EL4, in the absence of other stimuli. IL-2 mRNA expression was significantly induced by TGF-beta1 (0.1-1 ng/ml) over a relatively narrow concentration range, which led to the induction of IL-2 secretion. Under identical condition, we examined the effect of TGF-beta1 on the activity of nuclear factor AT (NF-AT), nuclear factor kappaB (NF-kappaB), activator protein-1 (AP-1) and octamer, all of which contribute to the regulation of IL-2 gene expression. Electrophoretic mobility shift assays showed that TGF-beta1 markedly increased NF-AT, NF-kappaB and AP-1 binding to their respective cognate DNA binding sites, whereas octamer binding remained constant, as compared with untreated cells. Employing a reporter gene expression system with p(NF-kappaB)3-CAT, p(NF-AT)3-CAT and p(AP-1)3-CAT, TGF-beta1 treatment of transfected EL4 cells induced a dose-related increase in chloramphenicol acetyltransferase activity that correlated well with the DNA binding profile found in the electrophoretic mobility shift assay studies. These results show that TGF-beta1, in the absence of any additional stimuli, up-regulates the activity of key transcription factors involved in IL-2 gene expression, including NF-AT, NF-kappaB and AP-1, to help promote IL-2 mRNA expression by EL4 cells.


Immunomodulation of hepatic ischemic injury via increased Bcl-X(L) and decreased Bcl-X(S).
Although classic ischemia-reperfusion injury is mediated by reactive oxygen intermediaries, increasing evidence implicates a role for immune-mediated apoptosis during ischemic injury in transplantation. Herein, we report the effects of polyclonal rabbit anti-thymocyte globulin (rATG) on mediators of hepatic apoptosis during cold storage. Three-month-old male Lewis rats were placed under halothane anesthesia and the portal vein cannulated. University of Wisconsin (UW) solution (35 ml) with (n = 5) and without (n = 5) 20 mg/kg anti-rat rATG was infused before hepatectomy. The liver was stored in UW solution +/- rATG (143 ng/ml) at 4 degrees C for various times up to 24 h. Specimens were terminal deoxyuridine nick end labeling-stained for apoptosis. Tissue lysates were analyzed by Western blotting and densitometry. Compared to UW alone, significantly fewer apoptotic cells were present in UW + rATG perfused and stored livers. There were early and sustained significant increases in Bcl-X(L) and decreases in Bcl-X(S) with rATG. There was an initial, but not sustained, significant decrease in Bax with rATG. Moreover, there was a significant one-third decrease in caspase-9 production with rATG at 0, 6, 12, and 18 h. Decreased proapoptotic Bcl-X(S) and increased antiapoptotic Bcl-X(L), as well as decreased downstream proapoptotic caspase-9 expression, during liver ischemia after treatment with rATG, all favor cell survival. Because apoptotic ischemic injury results in allograft dysfunction, preservation strategies that ameliorate such immunological effects may improve organ function.


Impact of biogas digesters on cookhouse volatile organic compound exposure for rural Kenyan farmwomen.
Women living on rural Kenyan smallholder dairy farms burn wood as biofuel in family cookhouses. Unventilated biofuel combustion produces harmful levels of respirable particles and volatile organic compound (VOC) emissions in indoor environments. Biogas digesters, which can generate high methane-content biogas from livestock manure composting were recently installed on 31 farms. The study objectives were to compare VOC exposure profiles for women cooking on farms with and without biogas digesters, and to compare seasonal variations in VOC exposures for those women cooking with biogas. Participants (n=31 biogas farms, n=31 referent farms) wore passive thermal desorption VOC sampling tubes and recorded cookhouse fuel use on time activity sheets for 7 days. Women using biogas spent significantly less time (mean=509 min/week) exposed to cookhouse wood smoke compared with the referent group (mean=1122 min/week) (P<0.01). Total VOC exposure did not differ between farm groups (P=0.14), though concentrations of trans-1,3-dichloropropene, bromoform, and 1,4-dichlorobenzene in biogas cookhouses were significantly lower than in referent cookhouses, even after Bonferroni correction. The composition of VOC species was also significantly different, reflecting the different fuel sources. Biogas digester technologies have great potential for reducing exposure to wood smoke VOCs in low-income countries.


A role for PHANTASTICA in medio-lateral regulation of adaxial domain development in tomato and tobacco leaves.
Diverse leaf forms in nature can be categorized into two groups: simple and compound. A simple leaf has a single blade unit, whilst a compound leaf is dissected into leaflets. For both simple and compound leaves, a MYB domain transcription factor PHANTASTICA (PHAN) plays an important role in establishing the adaxial domain in the leaf. Absence of PHAN in arabidopsis and antirrhinum leaves supresses blade development, and in tomato suppresses leaflet development. However, in the rachis and petiole regions of tomato leaves where PHAN and the adaxial domain coexist, it has been unclear why leaf blade and leaflets are not formed. We hypothesized that PHAN regulates the medio-lateral extent of the adaxial domain, thereby determining compound leaf architecture. To test this hypothesis, we generated and analysed transgenic tomato plants expressing tomato PHAN (SlPHAN) under the Cauliflower mosaic virus (CaMV) 35S promoter in both sense and antisense orientations, and tobacco plants that over-express tomato SlPHAN. Modulations in SlPHAN resulted in a variety of leaf morphologies such as simple, ternate and compound in either a peltate or non-peltate arrangement. Measurements of the extent of the adaxial domain along the wild-type tomato leaf axis showed that the adaxial domain is narrowed in the rachis and petiole in comparison with regions where laminar tissue arises. In antiSlPHAN transgenic leaves, no blade or leaflet was formed where the adaxial domain was medio-laterally narrowed, and KNOX gene expression was correlatively upregulated. CaMV35S::SlPHAN expression led to widening of the adaxial domain and ectopic blade outgrowth in the rachis of tomato and in the petiole of tobacco. Taken together, these results suggest that SlPHAN plays a role in medio-lateral extension of the adaxial domain and contributes to final leaf morphology in tomato. This study provides a novel insight into leaf architecture in tomato and highlights how changes in the expression domain of a master regulator gene such as SlPHAN can be translated into diverse final leaf morphologies.


Characterization of Japanese quail yellow as a genomic deletion upstream of the avian homolog of the mammalian ASIP (agouti) gene.
ASIP is an important pigmentation gene responsible for dorsoventral and hair-cycle-specific melanin-based color patterning in mammals. We report some of the first evidence that the avian ASIP gene has a role in pigmentation. We have characterized the genetic basis of the homozygous lethal Japanese quail yellow mutation as a >90-kb deletion upstream of ASIP. This deletion encompasses almost the entire coding sequence of two upstream loci, RALY and EIF2B, and places ASIP expression under control of the RALY promoter, leading to the presence of a novel transcript. ASIP mRNA expression was upregulated in many tissues in yellow compared to wild type but was not universal, and consistent differences were not observed among skins of yellow and wild-type quail. In a microarray analysis on developing feather buds, the locus with the largest downregulation in yellow quail was SLC24A5, implying that it is regulated by ASIP. Finally, we document the presence of ventral skin-specific isoforms of ASIP mRNA in both wild-type quails and chickens. Overall, there are remarkable similarities between yellow in quail and lethal yellow in mouse, which involve a deletion in a similar genomic position. The presence of ventral-specific ASIP expression in birds shows that this feature is conserved across vertebrates.


[Which transplantation strategies in primary hyperoxaluria type 1?].
The following main conclusions concerning the transplantation strategies to be adopted in primary hyperoxaluria type 1 (PH1) were drawn from the data collected from 22 patients who received combined liver-kidney grafts and 2 patients who received isolated liver grafts in Europe from June 1984 to March 1990. In end-stage renal failure due to PH1 liver-kidney transplantation yields better results than conventional renal transplantation. An isolated liver graft should be planned in patients with GFR between 25 and 60 ml/min/1.73 m2 whereas a combined liver-kidney graft is to be recommended as soon as the GFR falls below 25 ml/min/1.73 m2. Such patients should not be maintained on dialysis for more than a few months since they would unavoidably develop oxalosis with the risk of disabling lesions in the skeleton and cardiovascular system. Besides, oxalosis would be regularly followed by long-standing hyperoxaluria, with the risk of damage to the kidney graft, despite the correction of the enzyme deficit brought up by the liver graft.


Role of toxic oxygen products from phagocytic cells in tissue injury.
The generation of toxic oxygen products from activated phagocytic cells represents an important pathogenic mechanism in tissue damage associated with inflammatory reactions that are characterized by the involvement of neutrophils or other phagocytic cells. The production of these toxic products from activated phagocytic cells is related to receptor activation on the surface of the cells. Most emphasis has been placed on studies in the lung where the production of these toxic metabolites by phagocytic cells, either in the vasculature or in the airway compartment of the lungs, has been demonstrated to result in endothelial cell injury or damage and destruction of alveolar lining cells. In some cases these reactions are progressive, resulting in parenchymal collapse and fibrosis. Naturally occurring protective factors have been demonstrated both within cells and in the plasma and interstitial fluid compartments. Although there is increasing evidence that in the shock syndrome complement activation products and toxic oxygen metabolites from activated leukocytes play important roles, the most direct evidence for related mechanisms comes in detailed studies of immune complex induced injury where a role for both O-2 and H2O2 has been clearly demonstrated. These facts indicate that toxic oxygen metabolites from activated phagocytic cells play important roles in a variety of pathological situations, and have provided new insights into intervention therapy.


Development of one-step TaqMan® real-time reverse transcription-PCR and conventional reverse transcription-PCR assays for the detection of equine rhinitis A and B viruses.
Equine rhinitis viruses A and B (ERAV and ERBV) are common equine respiratory viruses belonging to the family Picornaviridae. Sero-surveillance studies have shown that these two viral infections are prevalent in many countries. Currently, the diagnosis of ERAV and ERBV infections in horses is mainly based on virus isolation (VI). However, the sensitivity of VI testing varies between laboratories due to inefficient viral growth in cell culture and lack of cytopathic effect. Therefore, the objective of this study was to develop molecular diagnostic assays (real-time RT-PCR [rRT-PCR] and conventional RT-PCR [cRT-PCR] assays) to detect and distinguish ERAV from ERBV without the inherent problems traditionally associated with laboratory diagnosis of these infections. Three rRT-PCR assays targeting the 5'-UTR of ERAV and ERBV were developed. One assay was specific for ERAV, with the two remaining assays specific for ERBV. Additionally, six cRT-PCR assays targeting the 5'-UTR and 3D polymerase regions of ERAV and ERBV were developed. Both rRT-PCR and cRT-PCR assays were evaluated using RNA extracted from 21 archived tissue culture fluid (TCF) samples previously confirmed to be positive for ERAV (n = 11) or ERBV (n = 10) with mono-specific rabbit antisera. The ERAV rRT-PCR and cRT-PCR assays could only detect ERAV isolates and not ERBV isolates. Similarly, the ERBV rRT-PCR and cRT-PCR assays could only detect ERBV isolates and not ERAV isolates. None of the rRT-PCR or cRT-PCR assays cross-reacted with any of the other common equine respiratory viruses. With the exception of one cRT-PCR assay, the detection limit of all of these assays was 1 plaque forming unit per ml (pfu/ml). The newly developed rRT-PCR and cRT-PCR assays provide improved diagnostic capability for the detection and differentiation of ERAV and ERBV. However, a larger number of clinical specimens will need to be tested before each assay is adequately validated for the detection of ERAV and/or ERBV in suspect cases of either viral infection.


Vasopressin release in response to nausea-producing agents and cholecystokinin in monkeys.
Administration of lithium chloride and copper sulfate to adult monkeys caused marked elevations in plasma vasopressin (AVP) levels without significant increases in plasma oxytocin (OT) levels. Emesis was produced in five of the seven animals given these agents, in support of nausea as the main stimulus to AVP release. A similar pattern of AVP release without OT release was found after administration of cholecystokinin (CCK). Although most monkeys vomited in response to 10 micrograms/kg of CCK, a significant increase in plasma AVP levels also was produced with a dose of 1 microgram/kg, which did not produce emesis in any animal. These findings are in marked contrast with previous results in rats, which indicated that lithium chloride, copper sulfate, and CCK each stimulated OT rather than AVP release. Despite this interspecies difference, the significant neurohypophysial hormone secretion in response to both nausea-producing agents and CCK suggests that AVP secretion in monkeys, similar to OT secretion in rats, might reflect activation of central pathways mediating nausea and/or inhibition of food intake, even when overt illness is not produced.


Identification of a short basic peptide motif able to drive copy-number dependent nuclear accumulation of a linked protein.
The repetitive [RTRG](6) peptide was fortuitously identified as a potent nuclear localization signal when linked to the green fluorescent reporter protein. Replacing the arginines by lysines, or the threonines by glycines, both resulted in a decreased nuclear targeting ability of the peptide within this context. By contrast, the sequence [RT](12) proved able to drive nuclear accumulation of the linked protein as efficiently as the starting peptide. Remarkably, [RTRG](n) peptides where n=2 to 6 showed a gradual, copy-number dependent, increase in their ability to target the green fluorescent protein to the cell nucleus. As a consequence, the nuclear to cytoplasmic concentration ratio of the linked protein within the cell could be adjusted to different values depending on the number of repeats used in the fusion. Our observation may open the way to the use of [RTRG](n) repeats of given lengths (n=2 to 6) for fixing the nuclear-cytoplasmic partition of shuttling protein domains in the course of their functional study.


Insights into the chemical logic and enzymatic machinery of NRPS assembly lines.
Appreciation that some cyclic peptide antibiotics such as gramicidin S and tyrocidine were nonribosomally synthesized has been known for 50 years. The past two decades of research including advances in bacterial genetics, genomics, protein biochemistry and mass spectrometry have codified the principles of assembly line enzymology for hundreds of nonribosomal peptides and in parallel for thousands of polyketides. The advances in understanding the strategies used for chain initiation, elongation and termination from these assembly lines have revitalized natural product biosynthetic communities.


Update on the cost-effectiveness of screening for colorectal neoplasia.
Colorectal cancer is an ideal target for population screening because it is a prevalent disease with an identifiable precursor lesion that, when treated, favorably alters the natural history of the disease. Several strategies for screening have illustrated efficacy, including fecal occult blood testing, sigmoidoscopy and colonoscopy. Cost-effectiveness analyses have been performed to determine whether the resources required to implement screening are justified by potential gains. The U.S. Preventive Services Task Force recently commissioned a study on the cost-effectiveness of colorectal cancer screening, which revealed that screening was cost-effective compared to no screening. However, it could not be confirmed which strategy would save the most life-years, nor which was most cost-effective. Since publication of this review, several additional cost-effectiveness analyses have been performed. These studies confirm that screening average risk patients at age 50 by a variety of available strategies is likely to be reasonable by current standards for resource utilization, and that either colonoscopy every 10 years (or once at age 65) or the combination of annual fecal occult blood testing with sigmoidoscopy every 5 years are viable alternatives. Additional economic analyses have examined the use of aspirin chemoprophylaxis to prevent colorectal cancer either alone or as an adjunct to screening strategies. These studies reaffirm the cost-effectiveness of colorectal cancer screening, but illustrate that aspirin chemoprophylaxis is unlikely to be associated with gains for which society would be willing to pay. At present, the decision to choose one colorectal screening strategy over another is based on availability of screening modalities, patient and provider preferences, and associated adherence to screening recommendations. Assessment of preference and development of interventions to increase adherence to screening should be a focus of research in the future.


The influence in vivo of murine colony-stimulating factor-1 on myeloid progenitor cells in mice recovering from sublethal dosages of cyclophosphamide.
Pure murine colony-stimulating factor-1 (CSF-1) was assessed for its effects in vivo in mice pretreated seven days earlier with a sublethal dosage of cyclophosphamide. The multipotential (CFU-GEMM), erythroid (BFU-E), and granulocyte-macrophage (CFU-GM) progenitor cells in these mice were in a slowly cycling or noncycling state. Intravenous administration of 20,000 units of CSF-1 to these mice stimulated the hematopoietic progenitors into a rapidly cycling state in the marrow and spleen within three hours. Significant increases in absolute numbers of marrow and spleen CFU-GM and spleen BFU-E and CFU-GEMM were also detected. No endotoxin was detected in the CSF-1 preparation by Limulus lysate assay, and treatment of CSF-1 at 100 degrees C for 20 to 30 minutes completely inactivated the in vitro and in vivo stimulating effects. The effects of CSF-1 were not mimicked by the in vivo administration of 0.1 to 10 ng Escherichia coli lipopolysaccharide. These results suggest that the effects of CSF-1 in vivo were not due to contaminating endotoxin or to a nonspecific protein effect. CSF-1 did not enhance colony formation by BFU-E or stimulate colony formation by CFU-GEMM in vitro, thus suggesting that at least some of the effects of CSF-1 noted in vivo are probably indirect and mediated by accessory cells.


Can Innovative Trial Designs in Orphan Diseases Drive Advancement of Treatments for Common Neurological Diseases?
Global regulatory agencies have transformed their approach to approvals in their processes for formal review of the safety and efficacy of new drugs. Opportunities for innovation have expanded because of the coronavirus disease 2019 (COVID-19) pandemic. Several regulatory-led initiatives have progressed rapidly during the past year, including patient-focused drug development, model-informed drug development, real-world evidence, and complex innovative trial designs. Collectively, these initiatives have accelerated the rate of approvals. Despite demands to focus on urgent needs imposed by the COVID-19 pandemic, the number of new drug approvals over the past year, particularly for rare diseases, has outpaced expectations. Advancing therapeutics for nervous system disorders requires adaptive strategies that align with rapid developments in the field. Three relentlessly progressive diseases, amyotrophic lateral sclerosis, Duchenne muscular dystrophy, and Parkinson's disease are in urgent need of new treatments. Herein, we propose new regulatory initiatives, including innovative trial designs and patient-focused drug development that accelerate clinical trial conduct while meeting critical regulatory requirements for therapeutic approval.


Advantages of technetium pyrophosphate scintigraphy over plasma enzyme analysis in estimation of anterior myocardial infarct size.
Infarct size was estimated by cumulative creatine kinase MB isoenzyme (CKMB-r) release and by technetium 99m stannous pyrophosphate (TcPYP) scintigraphy in 27 patients with acute anterior myocardial infarction. In eight patients, scintigraphy showed a central area of reduced tracer uptake surrounded by a peripheral rim of increased TcPYP accumulation ("doughnut" pattern). This appearance occurred only in large infarcts and the maximal scintigraphic area (51.3 +/- 2.8 cm2, mean +/- SEM) in this group was significantly greater than that in the remainder (28.1 +/- 2.5 cm2). Correlation between CKMB-r and maximal scintigraphic infarct area was moderate in the whole group. Exclusion of patients, however, with "doughnut" scintigrams in which correlation was very poor, resulted in substantial improvement in the remainder. It is suggested that in the central regions of large "doughnut" infarcts, reduced blood flow hinders the efflux of CKMB from the centre causing an underestimate of infarct size. Pyrophosphate scintigraphy appears to be more accurate than CKMB release in measuring the size of these large anterior infarcts.


Reproducibility of assessing fecal microbiota in chronic constipation.
While limited data suggest that the fecal microbiota in healthy people is stable over time, the intraindividual variability of the fecal microbiota in constipated patients is unknown. This study evaluated the intraindividual reproducibility of fecal microbiota analyzed with 16S rRNA gene sequencing in two stool samples collected without and after a laxative, respectively, in 25 healthy people and 25 constipated women. Participants completed a food record for 3 d before the stool collection. Colonic transit was measured with scintigraphy. The constipated patients were older (48±15 vs 39±10 y, P=.02) than healthy participants but had a similar BMI. The total daily caloric intake was less (P=.005) in constipated (1265±350 kcal) than healthy participants (1597±402 kcal). Fourteen patients but only two controls (P<.005), had delayed colonic transit. For most measures of alpha (eg, Observed OTU number, Shannon index) and beta diversity (eg, Bray-Curtis dissimilarity, UniFrac, phyla level abundance), the ICCs between two stool samples were high, indicating moderate or strong agreement, and similar in healthy people and constipated patients. The ICC for the weighted UniFrac distance, which is weighted by abundance, was lower than its unweighted counterpart, indicating that the unweighted measure is more robust and reproducible. The intraindividual reproducibility of fecal microbiota in constipated patients is high and comparable to healthy participants. For most purposes, evaluating the fecal microbiota in a single stool sample should generally suffice in adequately powered studies of healthy and constipated patients.


Electoral reform and public policy outcomes in Thailand: the politics of the 30-Baht health scheme.
How do changes in electoral rules affect the nature of public policy outcomes? The current evidence supporting institutional theories that answer this question stems almost entirely from quantitative cross-country studies, the data of which contain very little within-unit variation. Indeed, while there are many country-level accounts of how changes in electoral rules affect such phenomena as the number of parties or voter turnout, there are few studies of how electoral reform affects public policy outcomes. This article contributes to this latter endeavor by providing a detailed analysis of electoral reform and the public policy process in Thailand through an examination of the 1997 electoral reforms. Specifically, the author examines four aspects of policy-making: policy formulation, policy platforms, policy content, and policy outcomes. The article finds that candidates in the pre-1997 era campaigned on broad, generic platforms; parties had no independent means of technical policy expertise; the government targeted health resources to narrow geographic areas; and health was underprovided in Thai society. Conversely, candidates in the post-1997 era relied more on a strong, detailed national health policy; parties created mechanisms to formulate health policy independently; the government allocated health resources broadly to the entire nation through the introduction of a universal health care system, and health outcomes improved. The author attributes these changes in the policy process to the 1997 electoral reform, which increased both constituency breadth (the proportion of the population to which politicians were accountable) and majoritarianism.


Long-acting insulin analogs: a review of "real-world" effectiveness in patients with type 2 diabetes.
A systematic review was conducted to evaluate the effectiveness of initiating insulin treatment with insulin glargine or insulin detemir for type 2 diabetes in a routine clinical practice setting. Medline, EMBASE and Cochrane literature databases were searched to identify published "real world" reports. Studies were included in the review if they reported the following; insulin dose, change in HbA1c, change in body weight and hypoglycemic events. In routine practice, decreases inHbA1c associated with insulin glargine and insulin detemir were variable and ranged between approximately -0.3% to -1.5% depending on prior treatment, but switching to insulin analogs was associated with less weight gain than previously reported. Compared with data reported in published trials, hypoglycemic event rates associated with basal analog insulin use in clinical practice were lower in patients initiating insulin and comparable in patients using basal-bolus regimens. Most patients were treated with low doses of insulin analog administered as once daily injections. In routine clinical practice most patients stopped concomitant use of sulfonylureas when initiating insulin analogs and it is likely that this together with the lower dose of insulin influenced outcomes. Nevertheless, despite initiating insulin therapy, it was also apparent that for many patients in routine clinical practice attainment of glycemic goals remains elusive.


Can primary care team-based transition to insulin improve outcomes in adults with type 2 diabetes: the stepping up to insulin cluster randomized controlled trial protocol.
Type 2 diabetes (T2D) brings significant human and healthcare costs. Its progressive nature means achieving normoglycaemia is increasingly difficult, yet critical to avoiding long term vascular complications. Nearly one-half of people with T2D have glycaemic levels out of target. Insulin is effective in achieving glycaemic targets, yet initiation of insulin is often delayed, particularly in primary care. Given limited access to specialist resources and the size of the diabetes epidemic, primary care is where insulin initiation must become part of routine practice. This would also support integrated holistic care for people with diabetes. Our Stepping Up Program is based on a general practitioner (GP) and practice nurse (PN) model of care supported appropriately by endocrinologists and credentialed diabetes educator-registered nurses. Pilot work suggests the model facilitates integration of the technical work of insulin initiation within ongoing generalist care. This protocol is for a cluster randomized controlled trial to examine the effectiveness of the Stepping Up Program to enhance the role of the GP-PN team in initiating insulin and improving glycaemic outcomes for people with T2D. 224 patients between the ages of 18 and 80 years with T2D, on two or more oral hypoglycaemic agents and with an HbA1c ≥7.5% in the last six months will be recruited from 74 general practices. The unit of randomization is the practice.Primary outcome is change in glycated haemoglobin HbA1c (measured as a continuous variable). We hypothesize that the intervention arm will achieve an absolute HbA1c mean difference of 0.5% lower than control group at 12 months follow up. Secondary outcomes include the number of participants who successfully transfer to insulin and the proportion who achieve HbA1c measurement of <7.0%. We will also collect data on patient psychosocial outcomes and healthcare utilization and costs. The study is a pragmatic translational study with important potential implications for people with T2D, healthcare professionals and funders of healthcare though making better use of scarce healthcare resources, improving timely access to therapy that can improve disease outcomes. Australian and New Zealand Clinical Trials Registry ACTRN12612001028897.


Serum levels of hepatitis C virus core antigen as a marker of infection and response to therapy.
Hepatitis C virus (HCV) core antigen is a recently developed marker of hepatitis C infection. We compared the predictive power of HCV core antigen with reverse transcription polymerase chain reaction (RT-PCR) and branched DNA assay for HCV-RNA as markers of infection and response to interferon therapy. Four hundred and forty-four sera from 111 patients (65 men, 52 yr) with chronic hepatitis C, receiving ribavirin together with standard interferon (n = 61) or pegylated interferon (n = 50) were retrospectively investigated. Pretreatment, RT-PCR, branched DNA (median 621,887 IU/ml), and HCV core antigen (median 57 pg/ml) gave positive results in 100%, 99%, and 94% of the sera; the correlation between HCV core antigen and branched DNA was 0.75. The median HCV RNA level among the 7 of 111 (6%) patients that had a negative core Ag result was 15,016 IU/ml. Pretreatment levels of HCV core antigen were significantly lower in the 41 patients with a sustained virological response than in the 39 relapsers and 31 nonresponders (17 pg/ml, 114 pg/ml, 58 pg/ml; p-value 0.005). Independently of treatment schedule, wk 12 more than 2 log(10) reduction of viremia or a negative result for HCV core antigen had 100% negative predictive value (NPV) for a response to therapy compared to 94% for negative RT-PCR. The positive predictive value (PPV) of HCV core antigen and branched DNA was only 47% and 48%. In conclusion, the HCV core antigen is a less sensitive test of HCV viremia than HCV-RNA assays and is competitive with the bDNA assay as an early predictor of a nonresponse.


Co-design of a Smartphone App for People Living With Dementia by Applying Agile, Iterative Co-design Principles: Development and Usability Study.
The benefits of involving those with lived experience in the design and development of health technology are well recognized, and the reporting of co-design best practices has increased over the past decade. However, it is important to recognize that the methods and protocols behind patient and public involvement and co-design vary depending on the patient population accessed. This is especially important when considering individuals living with cognitive impairments, such as dementia, who are likely to have needs and experiences unique to their cognitive capabilities. We worked alongside individuals living with dementia and their care partners to co-design a mobile health app. This app aimed to address a gap in our knowledge of how cognition fluctuates over short, microlongitudinal timescales. The app requires users to interact with built-in memory tests multiple times per day, meaning that co-designing a platform that is easy to use, accessible, and appealing is particularly important. Here, we discuss our use of Agile methodology to enable those living with dementia and their care partners to be actively involved in the co-design of a mobile health app. The aim of this study is to explore the benefits of co-design in the development of smartphone apps. Here, we share our co-design methodology and reflections on how this benefited the completed product. Our app was developed using Agile methodology, which allowed for patient and care partner input to be incorporated iteratively throughout the design and development process. Our co-design approach comprised 3 core elements, aligned with the values of patient co-design and adapted to meaningfully involve those living with cognitive impairments: end-user representation at research and software development meetings via a patient proxy; equal decision-making power for all stakeholders based on their expertise; and continuous user consultation, user-testing, and feedback. This co-design approach resulted in multiple patient and care partner-led software alterations, which, without consultation, would not have been anticipated by the research team. This included 13 software design alterations, renaming of the product, and removal of a cognitive test deemed to be too challenging for the target demographic. We found patient and care partner input to be critical throughout the development process for early identification of design and usability issues and for identifying solutions not previously considered by our research team. As issues addressed in early co-design workshops did not reoccur subsequently, we believe this process made our product more user-friendly and acceptable, and we will formally test this assumption through future pilot-testing.


A genetically tagged Psb27 protein allows purification of two consecutive photosystem II (PSII) assembly intermediates in Synechocystis 6803, a cyanobacterium.
Photosystem II (PSII) is a large membrane bound molecular machine that catalyzes light-driven oxygen evolution from water. PSII constantly undergoes assembly and disassembly because of the unavoidable damage that results from its normal photochemistry. Thus, under physiological conditions, in addition to the active PSII complexes, there are always PSII subpopulations incompetent of oxygen evolution, but are in the process of undergoing elaborate biogenesis and repair. These transient complexes are difficult to characterize because of their low abundance, structural heterogeneity, and thermodynamic instability. In this study, we show that a genetically tagged Psb27 protein allows for the biochemical purification of two monomeric PSII assembly intermediates, one with an unprocessed form of D1 (His27ΔctpAPSII) and a second one with a mature form of D1 (His27PSII). Both forms were capable of light-induced charge separation, but unable to photooxidize water, largely because of the absence of a functional tetramanganese cluster. Unexpectedly, there was a significant amount of the extrinsic lumenal PsbO protein in the His27PSII, but not in the His27ΔctpAPSII complex. In contrast, two other lumenal proteins, PsbU and PsbV, were absent in both of these PSII intermediate complexes. Additionally, the only cytoplasmic extrinsic protein, Psb28 was detected in His27PSII complex. Based on these data, we have presented a refined model of PSII biogenesis, illustrating an important role of Psb27 as a gate-keeper during the complex assembly process of the oxygen-evolving centers in PSII.


The PEP Project - Synergistic Community Based Action in Prevention, Early Detection and Palliative Care, to Impact the Cancer Burden in India.
With unprecedented surge in the incidence and prevalence of cancer in India, it has become imperative to strengthen the workforce for all the domains of cancer care. A large proportion of the activity required for prevention as well as for palliation, lie outside of tertiary institutions, in the community. Palliative care (PC) as a field is expanding exponentially across the country and the service providers often engage and work actively within the local community in their region. This article describes the scope for reducing the cancer burden in the community, through capacity building of community based PC healthcare functionaries in the domains of Prevention, and Early detection of common cancers along with Palliative care - the PEP domains. It suggests aligning and enhancing the workforce already active within the community for PC, for screening, and if feasible, for early detection of common cancers. The article describes possibilities of initiating PEP activities and offers a set of screening questionnaire that may be used when engaged with family/ community setting. The aim is to integrate the activities done, to detect the need for palliative care in a family / community, with that required to detect need for evaluation of most common cancers- oral, breast and cervix. The PEP concept may be adapted to different levels, based on the team presence in the communities, degree of engagement, and availability of trainers and healthcare personnel.


Site-specific cancer incidence and mortality after cerebral angiography with radioactive thorotrast.
Few opportunities exist to evaluate the carcinogenic effects of long-term internal exposure to alpha-particle-emitting radionuclides. Patients injected with Thorotrast (thorium-232) during radiographic procedures, beginning in the 1930s, provide one such valuable opportunity. We evaluated site-specific cancer incidence and mortality among an international cohort of 3,042 patients injected during cerebral angiography with either Thorotrast (n = 1,650) or a nonradioactive agent (n = 1,392) and who survived 2 or more years. Standardized incidence ratios (SIR) for Thorotrast and comparison patients (Denmark and Sweden) were estimated and relative risks (RR), adjusted for population, age and sex, were generated with multivariate statistical modeling. For U.S. patients, comparable procedures were used to estimate standardized mortality ratios (SMR) and RR, representing the first evaluation of long-term, site-specific cancer mortality in this group. Compared with nonexposed patients, significantly increased risks in Thorotrast patients were observed for all incident cancers combined (RR = 3.4, 95% CI 2.9-4.1, n = 480, Denmark and Sweden) and for cancer mortality (RR = 4.0, 95% CI 2.5-6.7, n = 114, U.S.). Approximately 335 incident cancers were above expectation, with large excesses seen for cancers of the liver, bile ducts and gallbladder (55% or 185 excess cancers) and leukemias other than CLL (8% or 26 excess cancers). The RR of all incident cancers increased with time since angiography (P < 0.001) and was threefold at 40 or more years; significant excesses (SIR = 4.0) persisted for 50 years. Increasing cumulative dose of radiation was associated with an increasing risk of all incident cancers taken together and with cancers of the liver, gallbladder, and peritoneum and other digestive sites; similar findings were observed for U.S. cancer mortality. A marginally significant dose response was observed for the incidence of pancreas cancer (P = 0.05) but not for lung cancer. Our study confirms the relationship between Thorotrast and increased cancer incidence at sites of Thorotrast deposition and suggests a possible association with pancreas cancer. After injection with >20 ml Thorotrast, the cumulative excess risk of cancer incidence remained elevated for up to 50 years and approached 97%. Caution is needed in interpreting the excess risks observed for site-specific cancers, however, because of the potential bias associated with the selection of cohort participants, noncomparability with respect to the internal or external comparison groups, and confounding by indication. Nonetheless, the substantial risks associated with liver cancer and leukemia indicate that unique and prolonged exposure to alpha-particle-emitting Thorotrast increased carcinogenic risks.


Impact of a counter-rotating planetary rotation system on thin-film thickness and uniformity.
Planetary rotation systems incorporating forward- and counter-rotating planets are used as a means of increasing coating-system capacity for large oblong substrates. Comparisons of planetary motion for the two types of rotating systems are presented based on point tracking for multiple revolutions as well as comparisons of quantitative thickness and uniformity. Counter-rotation system geometry is shown to result in differences in thin-film thickness relative to standard planetary rotation for precision optical coatings. This systematic error in thin-film thickness will reduce deposition yields for sensitive coating designs.


Identification of distinctive interferon gene signatures in different types of myositis.
Activation of the type 1 interferon (IFN1) pathway is a prominent feature of dermatomyositis (DM) muscle and may play a role in the pathogenesis of this disease. However, the relevance of the IFN1 pathway in patients with other types of myositis such as the antisynthetase syndrome (AS), immune-mediated necrotizing myopathy (IMNM), and inclusion body myositis (IBM) is largely unknown. Moreover, the activation of the type 2 interferon (IFN2) pathway has not been comprehensively explored in myositis. In this cross-sectional study, our objective was to determine whether IFN1 and IFN2 pathways are differentially activated in different types of myositis by performing RNA sequencing on muscle biopsy samples from 119 patients with DM, IMNM, AS, or IBM and on 20 normal muscle biopsies. The expression of IFN1- and IFN2-inducible genes was compared between the different groups. The expression of IFN1-inducible genes was high in DM, moderate in AS, and low in IMNM and IBM. In contrast, the expression of IFN2-inducible genes was high in DM, IBM, and AS but low in IMNM. The expression of IFN-inducible genes correlated with the expression of genes associated with inflammation and muscle regeneration. Of note, ISG15 expression levels alone performed as well as composite scores relying on multiple genes to monitor activation of the IFN1 pathway in myositis muscle biopsies. IFN1 and IFN2 pathways are differentially activated in different forms of myositis. This observation may have therapeutic implications because immunosuppressive medications may preferentially target each of these pathways.


Endoscopic parathyroidectomy: report of an initial experience.
Preoperative localization of parathyroid lesions and intraoperative quick parathyroid hormone (PTH) assay have been proposed to minimize the extent of operation in primary hyperparathyroidism. To this purpose, endoscopic procedures have been introduced recently. During a period of 13 months, 39 of 65 consecutive patients with primary hyperparathyroidism were selected for endoscopic parathyroidectomy on the basis of the following criteria: preoperative echographic diagnosis of a single adenoma, absence of nodular goiter, and no prior neck operations. Unilateral neck exploration and excision of the adenoma was performed through a gasless procedure combined with intraoperative PTH measurements. Mean follow-up after the operation was 7 months (range 1 to 13 months). Thirty-nine parathyroid adenomas were removed; the mean diameter was 21 mm (range 5 to 30 mm). The mean operative time was 65 minutes (range 30 to 180 minutes). In all cases PTH concentration decreased significantly. Patients who underwent endoscopic parathyroidectomy had less postoperative pain compared with patients who underwent conventional hemithyroidectomy. At follow-up, serum calcium and PTH levels were normal in all cases. Endoscopic parathyroidectomy proved to be a feasible surgical procedure that can be performed in an acceptable operative time with an excellent cosmetic result. The gasless approach avoided any emphysema.


The background occurrence of selected clinical conditions prior to the start of an extensive national vaccination program in Japan.
The COVID-19 pandemic caused by SARS-CoV-2 has now affected tens of millions of people globally. It is the hope that vaccines against SARS-CoV-2 will deliver a comprehensive solution to this global pandemic; however, this will require extensive national vaccination programs. Ultimately, clinical conditions and even sudden unexplained death will occur around the time of vaccination, thus a distinction needs to be made between events that are causally related to the vaccine or temporally related to vaccination. This study aimed to estimate the background occurrence of 43 clinical conditions in the Japanese population. A retrospective cohort study was conducted from 2013 to 2019 using data from two large healthcare claims databases (MDV and JMDC) in Japan. The estimated number of new cases and incidence were calculated based on the actual number of new cases identified in the databases. The PubMed and Ichushi-web databases, as well as grey literature such as guidelines and government statistics, were also searched to identify any publications related to incidence of these conditions in Japan. The estimates of the number of total cases and incidence were similar for the MDV and JMDC databases for some diseases. In addition, some estimates were similar to those in the scientific literature. For example, from the MDV and JMDC databases, estimates of incidence of confirmed Bell's palsy in 2019 were 41.7 and 47.9 cases per 100,000 population per year, respectively. These estimates were of the same order from the scientific publication. Determining whether clinical conditions occurring around the time of vaccination are causally or only temporally related to vaccination will be critical for public health decision makers as well as for the general public. Comparison of background occurrence at the population level may provide some additional objective evidence for the evaluation of temporality or causality.


Increased intra-mitochondrial lipofuscin aggregates with spherical dense body formation in mitochondrial myopathy.
Lipofuscin aggregation may result from incomplete degradation of damaged mitochondria by autophagy-lysosome pathway, and intra-mitochondrial lipofuscin aggregation may exacerbate mitochondrial abnormalities in mitochondrial myopathy (MM) and mitochondrial disease. We examined vastus lateralis muscle biopsies from 24 patients with pathologically diagnosed MM and clinically diagnosed chronic progressive external ophthalmoplegia, in comparison to the biopsies from 3 other groups:10 patients with inclusion body myositis (IBM), 11 younger adults, and 10 older subjects with no to minimal myopathic changes. Lipofuscin aggregation in muscle fibres was assessed on autofluorescence microscopy, some histochemical stains, and electron microscopy (EM). EM analyses demonstrated intra-mitochondrial lipofuscin aggregates, spherical dense bodies (SDBs), and paracrystalline inclusions (PCIs) which were semi-quantitatively assessed. Intra-mitochondrial lipofuscin aggregates showed no significant differences between groups of MM patients and older subjects or IBM patients, but significant differences between groups of younger adults and others with associated age-related changes. Intra-mitochondrial SDBs were significantly more in MM patients than in older subjects, IBM patients, and younger adults. There was a significant positive correlation between intra-mitochondrial lipofuscin aggregates and SDBs. These findings suggest that intra-mitochondrial formation of lipofuscin SDBs is more in MM and contributing to the pathophysiology of mitochondrial disease.


Staying connected when coming apart: The psychological correlates of contact and sex with an ex-partner.
Although marital separation is an inherently social experience, most research on adults' psychological adjustment following a romantic separation focuses on intrapersonal characteristics, or individual differences (e.g., attachment style, personality, longing) that condition risk for poor psychological outcomes. We know little about how these individual differences interact with interpersonal processes, such as contact between ex-partners. In the current study, we sought to understand how adults' continued attachment to (and longing for) an ex-partner, and both nonsexual and sexual contact with an ex-partner (CWE and SWE, respectively), are related to adults' post-separation psychological adjustment among 137 (n = 50 men) adults reporting recent marital separations. Data revealed that (1) less separation acceptance was associated with poorer psychological adjustment; (2) among people having CWE, those reporting less acceptance reported significantly poorer adjustment relative to those reporting more acceptance; (3) among people reporting more acceptance, those having CWE reported significantly better adjustment relative to those not having CWE; (4) among people not having SWE, those reporting less acceptance reported significantly poorer adjustment relative to those reporting more acceptance; and (5) among people reporting less acceptance, those having SWE reported significantly better adjustment relative to those not having SWE. We discuss the findings in terms of adult attachment, pair-bonding, and the loss of coregulatory processes following marital separation.


Effects of intravenous S-9780, an angiotensin-converting enzyme inhibitor, in normotensive subjects.
S-9780 is the active diacid metabolite of the new angiotensin-converting enzyme (ACE) inhibitor perindopril. In a double-blind, randomised, crossover study, the effects of 1, 2, and 4 mg of S-9780 administered intravenously (i.v.) were compared with placebo in eight normotensive subjects. All active doses caused immediate, maximal, and similar inhibition of plasma ACE with 40% inhibition persisting after 48 h. Plasma renin activity was elevated 4 and 8 h after dosing, but no effect on plasma aldosterone, adrenaline or noradrenaline levels was detected. Diastolic blood pressure was lowered by 4 mg of S-9780 until 24 h after dosing. Heart rate did not change. The pharmacokinetics of S-9780 fitted a three-compartment model with a terminal half-life (t1/2) of 31 h. Inhibition of plasma ACE was closely related to observed drug concentration, with 1.8 +/- 0.9 ng/ml (mean +/- S.D.) producing 50% inhibition of the enzyme. S-9780 caused predictable effects on the cardiovascular and renin angiotensin systems.


Acetabular liner fixation by cement.
Many situations in revision THA require the exchange of a PE liner in the setting of a well-fixed cementless acetabular shell. Unfortunately, a replacement liner is not always available, the locking mechanism of the metal shell may be damaged or incompatible with the desired liner, or the shell is malpositioned. Revision of a well-fixed cementless acetabular shell has been associated with considerable morbidity. This raises several questions: can a new PE liner be fixed in the existing shell using bone cement, and if so, which techniques can improve the end result, and in which patients should they be used? Biomechanical testing of cemented PE liners has shown initial fixation strengths that exceed conventional locking mechanisms. It is not known during what period this initial fixation will fail, but clinical reports with followup of as many as 6 years have shown survival in approximately 90% of cases. These studies have shown the importance of proper patient selection, accurate sizing of the PE liner, careful preparation of the substrate of the liner and the shell, and good cement technique. The potential advantages of this technique are less surgical morbidity, more rapid surgery and patient recovery, the ability to incorporate antibiotics in the cement, and more liner options.


Cofilin phosphorylation by protein kinase testicular protein kinase 1 and its role in integrin-mediated actin reorganization and focal adhesion formation.
Testicular protein kinase 1 (TESK1) is a serine/threonine kinase with a structure composed of a kinase domain related to those of LIM-kinases and a unique C-terminal proline-rich domain. Like LIM-kinases, TESK1 phosphorylated cofilin specifically at Ser-3, both in vitro and in vivo. When expressed in HeLa cells, TESK1 stimulated the formation of actin stress fibers and focal adhesions. In contrast to LIM-kinases, the kinase activity of TESK1 was not enhanced by Rho-associated kinase (ROCK) or p21-activated kinase, indicating that TESK1 is not their downstream effector. Both the kinase activity of TESK1 and the level of cofilin phosphorylation increased by plating cells on fibronectin. Y-27632, a specific inhibitor of ROCK, inhibited LIM-kinase-induced cofilin phosphorylation but did not affect fibronectin-induced or TESK1-induced cofilin phosphorylation in HeLa cells. Expression of a kinase-negative TESK1 suppressed cofilin phosphorylation and formation of stress fibers and focal adhesions induced in cells plated on fibronectin. These results suggest that TESK1 functions downstream of integrins and plays a key role in integrin-mediated actin reorganization, presumably through phosphorylating and inactivating cofilin. We propose that TESK1 and LIM-kinases commonly phosphorylate cofilin but are regulated in different ways and play distinct roles in actin reorganization in living cells.


Relationship between faecal microbiota and plasma metabolome in rats fed NK603 and MON810 GM maize from the GMO90+ study.
Safety concerns arising from the consumption of foods derived from genetically modified (GM) crops remains a controversial subject. We report here a faecal microbiota compositional analysis in Wistar rats from the GMO90 + study, which fed glyphosate-tolerant NK603 (+/- Roundup application) and Bt toxin MON810 GM maize for 6 months in comparison to their closest non-GM isogenic lines. We first integrated the faecal microbiota compositional data with results from plasma metabolomics to understand which bacterial species can influence host metabolism. Coriobacteriaceae and Acetatifactor significantly predicted plasma metabolic profile in males, while Bifidobacterium and Ruminococcus were able to predict female plasma metabolites. We then investigated the differences in fecal microbiota composition between group of rats fed MON810 or NK603 GM maize in comparison to their isogenic lines. Bacterial community richness was not altered by the test diets. There were no statistically significant differences in taxa abundance in the rat faecal microbiota that we could attribute to the consumption of either MON810 or NK603. We show that the consumption of the widely cultivated GM maize varieties NK603 and MON810 even up to 33% of the total diet had no effect on the status of the faecal microbiota compared to non-GM near isogenic lines.


An Initial examination of fear of negative and positive evaluation in youth.
Fear of negative evaluation (FNE) and fear of positive evaluation (FPE) are both core features of social anxiety. The majority of research with these constructs has been done with older adolescents and adults, with only one previous study examining FNE and FPE in childhood. However, this previous work relied exclusively on parent-report of youth FNE and FPE. Here, we examined the factor structure of FNE and FPE using youth self-reports. Moreover, we examined the associations with dimensions of internalizing and externalizing problems. We found that two-factor structure of FNE and FPE was a marginal fit to the data. Exploratory models identified three items that showed significant cross-loadings on non-target factors. Overall, we found that FNE was associated with dimensions of internalizing problems reported by youth and their mothers. FPE was associated with internalizing problems reported by youth, but not parents. Associations between FNE and clinical outcomes were stronger than those for FPE. This study demonstrates promise of FNE and FPE in youth and highlights important directions for future research.


Distribution of ganglionic sympathetic neurons supplying the subcutaneous, perirenal and mesentery fat tissue depots in the pig.
Previous morphological studies revealed that the adipose tissue is innervated by adrenergic nerve fibers. Furthermore, physiological studies showed that the metabolism of adipose tissue is controlled by the adrenergic component of the nervous system. However, nothing is known on the sources of innervation of different fat tissue depots. Therefore, we decided to study the distribution of ganglionic sympathetic neurons innervating adipose tissue in the pig by means of a retrograde tracing method. We used 9 male and 9 female pigs of approximately 50 kg body weight. The retrograde tracer, Fast Blue (FB), was injected into the subcutaneous, perirenal and mesentery fat tissue depots. Results of the present study showed that numerous centers of the sympathetic nervous system innervate adipose tissue in the pig. FB+ neurons projecting to the subcutaneous fat tissue were placed in the thoraco-lumbar region of the sympathetic chain ganglia (SChG). However, neurons supplying perirenal and mesentery fat tissue depots were found in both the SChG and prevertebral ganglia (PVG). We conclude that different adipose tissue depots (subcutaneous, perirenal and mesentery) have different sources of innervation and that there is no significant difference in the distribution of neurons innervating adipose tissue in male and female pigs.


Posterior intravaginal slingplasty (infracoccygeal sacropexy) for severe posthysterectomy vaginal vault prolapse--a preliminary report on efficacy and safety.
The infracoccygeal sacropexy, or posterior intravaginal slingplasty, was first reported by Petros as a minimally invasive procedure for the treatment of vault prolapse. This report is a prospective observational study that confirms the efficacy and safety of this new procedure. In this operation the IVS Tunneller (Tyco Healthcare, USA) is used to insert an 8 mm polypropylene tape between the perineum and the vaginal vault. The resulting artificial neoligament reinforces the atrophied uterosacral ligaments. The rectovaginal fascia and perineal body are then repaired by a technique that includes using a bridge of vaginal epithelium to strengthen the central vaginal epithelium at the point of maximum weakness. The symptomatic cure rates for prolapse were 91%, urgency 79%, nocturia 82% and pelvic pain 78%. All patients were discharged home within 24 hours. There were minimal surgical complications and no transfusions were required. The technique is safe, minimally invasive, has a short learning curve, and the skills needed are those of any competent pelvic surgeon.


Spatial tuning of neurons in the inferior colliculus of the big brown bat: effects of sound level, stimulus type and multiple sound sources.
We examined factors that affect spatial receptive fields of single units in the central nucleus of the inferior colliculus of Eptesicus fuscus. Pure tones, frequency- or amplitude-modulated sounds, or noise bursts were presented in the free-field, and responses were recorded extracellularly. For 58 neurons that were tested over a 30 dB range of sound levels, 7 (12%) exhibited a change of less than 10 degrees in the center point and medical border of their receptive field. For 28 neurons that were tested with more than one stimulus type, 5 (18%) exhibited a change of less than 10 degrees in the center point and medial border of their receptive field. The azimuthal response ranges of 19 neurons were measured in the presence of a continuous broadband noise presented from a second loudspeaker set at different fixed azimuthal positions. For 3 neurons driven by a contralateral stimulus only, the effect of the noise was simple masking. For 11 neurons driven by sound at either side, 8 were unaffected by the noise and 1 showed a simple masking effect. For the remaining 2, as well as for 5 neurons that were excited by contralateral sound and inhibited by ipsilateral sound, the peak of the azimuthal response range shifted toward the direction of the noise.


Distribution of epidermal growth-factor receptors in normal and neoplastic mammary tissues.
Epidermal growth factor (EGF) is considered to be mitogenic for proliferation of mammary glands in animals. The action of EGF is mediated by specific EGF receptors (EFG-R). In the present study, we investigated distribution of EGF receptors during various physiological stages of mammary glands, N-methyl-N-nitrosourea (MNU)-induced mammary tumors in rats and human breast cancer samples. EGF receptor concentrations were determined by Scatchard analyses in the membrane fraction of the tissues. Results showed increased EGF receptor levels in the structurally differentiated mammary tissues from pregnant rats; whereas lower concentrations were observed in the functionally differentiated glands from lactating rats. EGF receptors were absent in the majority of the tumors induced by MNU. The loss of EGF receptor was not observed during the first 20 days post carcinogen treatment, but appeared to be correlated with the onset of the tumor. Consistent with the literature, the majority of the steroid receptor positive human breast cancer samples were EGF receptor negative, whereas steroid receptor negative samples contained EGF receptors. These results suggest that the loss of EGF receptors in ovarian hormone dependent mammary tumors does not occur gradually during carcinogenesis but appears to be a characteristic of hormone dependent mammary tumor cells.


Anti-histone antibodies in idiopathic and drug-induced lupus recognize distinct intrahistone regions.
We have identified regions within core histones that are antigenic for autoantibodies in systemic lupus erythematosus (SLE) and drug-induced lupus. An immunoblotting technique was used to determine the reactivity of lupus antibodies for intact histones and for trypsin-resistant histone fragments that lack the amino- and carboxyl-terminal amino acids that are normally exposed in native nucleosomes. In SLE, the predominant anti-histone response was restricted to epitopes in the trypsin-sensitive regions. Of 20 SLE sera that had strong antibody activity for multiple intact histones, 17 showed minimal activity with any of the corresponding trypsin-resistant fragments. A markedly different pattern of reactivity was present in sera of patients with procainamide (Pr)-induced lupus in which antibodies to H2A, H2B, and the H2A-H2B complex had strong fragment activity. Interestingly, recognition of trypsin-resistant fragments was also noted in a small number of SLE sera that contained antibodies to the H2A-H2B complex. In contrast to both SLE and Pr-induced lupus, antibodies induced by hydralazine (Hy) reacted primarily with H3 and H4. Furthermore, these antibodies bound equally well to the corresponding trypsin-resistant regions that are thought to be relatively unexposed in native nucleosomes. Thus, the specificities of anti-histone antibodies in SLE, Pr-induced lupus, and Hy-induced lupus are markedly different, but in each disease reactivity appears to be restricted to a limited number of histone determinants. The data raise the possibility that autoantigen in the form of native nucleosomes may be recognized in SLE and possibly in Pr-induced lupus. In contrast, the propensity of Hy to induce autoantibodies to determinants usually not recognized in SLE or Pr-induced lupus may suggest a different immunogenic stimulus in this disease.


A sequence-specific conformational epitope on U1 RNA is recognized by a unique autoantibody.
An autoantibody from a patient with lupus-overlap syndrome was found to bind a specific region of U1 RNA. By using RNA sequence analysis, immunoprecipitation, and competition experiments with in vitro synthesized fragments of U1 RNA, a region of 40 nucleotides representing a stem-loop secondary structure was found to be an immunoreactive domain. This antibody recognized a conformational epitope because neither the RNA stem nor the RNA loop alone was immunoprecipitable. Antisense U1 RNA, U1 DNA, and other small RNAs were not reactive with the antibody. While the origins of nucleic acid-binding antibodies are unknown, this RNA-specific autoantibody probably originated by direct presentation to the immune system or as an anti-idiotype against a more common U1 small nuclear ribonucleoprotein-specific autoantibody. Thus, these findings have implications for the mechanisms of autoimmune recognition and provide an immunological approach to probing RNA structure and protein-RNA interactions.


DNA adducts and PM(10) exposure in traffic-exposed workers and urban residents from the EPIC-Florence City study.
Air pollution and particulate matter in urban areas have been associated with increased mortality from cardiovascular and respiratory diseases and increased cancer risk. Carcinogenic effects of particulate matter have been related to the contents of specific compounds, such as polycyclic aromatic hydrocarbons. The latter may form bulky DNA adducts, that may be considered as candidate markers of cancer risk. We have recently shown that traffic-exposed workers and the general population in Florence have divergent levels of DNA adducts, possibly related to different levels of exposure to genotoxic agents from vehicle emissions. In the current study, in a series of 214 Florence City healthy adults enrolled in a prospective study in the period 1993-1998 (152 residents / 62 traffic-exposed workers), we investigated the correlation between individual levels of DNA bulky adducts and PM(10) exposure scores based on daily environmental measures provided by the local Environmental Protection Agency for the whole study period, by two types of urban monitoring stations (high- and low-traffic). We found that PM(10) cumulative scores from high-traffic stations over the last 1-2 weeks prior to blood drawing significantly correlated (r=0.58, p=0.02) with DNA adduct levels among non-smoking traffic-exposed workers (but not among residents with no occupational exposure to vehicle emissions). A multivariate regression analysis adjusted for possible confounders confirmed these findings. PM(10) scores from low-traffic stations did not show any correlation. These results show that DNA adduct levels in non-smoking workers reflect the average levels of exposure to PM(10) in high-traffic urban areas experienced over a time period of 1-2 weeks. Since DNA adduct levels have been found predictive of lung cancer risk, our findings provide clues relevant to the reduction of genotoxic damage and possibly cancer risk among traffic-exposed urban workers.


Growth factors modulate gonadotropin receptor induction in granulosa cell cultures.
Epidermal growth factor and fibroblast growth factor inhibited follicle-stimulating hormone-dependent induction of luteinizing hormone receptor in cultured ovarian granulosa cells of the rat. In contrast, platelet-derived growth factor potentiated the induction of luteinizing hormone receptor by follicle-stimulating hormone. These results indicate that growth factors, well known for their effects on mitosis and DNA synthesis in cultured mammalian cells, are also able to modulate hormone-dependent differentiation in an endocrine target cell.


Chronic nicotine exposure has dissociable behavioural effects on control and beta2-/- mice.
Nicotine exerts beneficial effects on various neurological and psychiatric pathologies, yet its effects on cognitive performance remain unclear. Mice lacking the beta2 subunit of the nicotinic receptor (beta2-/-) show characteristic deficits in executive functions and are suggested as reliable animal models for some specific endophenotypes of human pathologies, notably ADHD. We use beta2-/- and their controls to investigate the consequences of chronic nicotine exposure on cognitive behaviour. We show that in control mice, this treatment elicits somewhat slight effects, particularly affecting nocturnal activity and self-grooming. By contrast, in beta2-/- mice, chronic nicotine treatment had restorative effects on exploratory behaviour in the open-field and affected rearing, but did not modify motor functions. We confirmed that beta2-/- mice exhibit impaired exploratory and social behaviour, and further demonstrated their nocturnal hyperactivity. These data support the proposal that beta2-/- mice represent a relevant model for cognitive disorders in humans and that nicotine administered chronically at low dose may relieve some of these.


Investigating the relationship between worker demographics and nature of injury on Federal Department of Defense workers' compensation injury rates and costs from 2000 to 2008.
This is the first study of workers' compensation injuries and costs in Department of Defense workers that examined whether any demographic factors including age, sex, occupation, and nature of injury altered the risks or costs of an injury or illness over time. Department of Defense Workers' Compensation claims for period 2000 to 2008 were analyzed (n = 142,115) using Defense Portal Analysis and Defense Manpower Data Center to calculate injury rates and costs. Regression analysis was done using SPSS to examine the change in the risk of injury or illness over time from 2000 to 2008. The age group of 30 to 34 years had the lowest costs per claim and highest claims rate, 332 per 10,000. The age group of 65 to 70 years had the lowest claims rate of 188 per 10,000 but the highest costs per claim. Claims cost increased $69 for each 5-year group, and older workers had a threefold increase in costs per claim. Younger workers get hurt more often, but older workers tend to have more expensive claims.


Stress inversions to forecast magma pathways and eruptive vent location.
When a batch of magma reaches Earth's surface, it forms a vent from which volcanic products are erupted. At many volcanoes, successive batches may open vents far away from previous ones, resulting in scattered, sometimes seemingly random spatial distributions. This exposes vast areas to volcanic hazards and makes forecasting difficult. Here, we show that magma pathways and thus future vent locations may be forecast by combining the physics of magma transport with a Monte Carlo inversion scheme for the volcano stress history. We validate our approach on a densely populated active volcanic field, Campi Flegrei (Italy), where we forecast future vents on an onshore semiannular belt located between 2.3 and 4.2 km from the caldera center. Our approach offers a mechanical explanation for the vent migration over time at Campi Flegrei and at many calderas worldwide and may be applicable to volcanoes of any type.


Unraveling the intricate connection between dietary factors and the success in long-term space missions.
In recent decades of spaceflight, inadequate caloric intake has posed significant nutritional challenges, contributing to muscle degradation, weakened immune and cardiovascular systems during and after space missions. This challenge becomes more acute on longer exploration missions, where transporting all food for the entire mission becomes a logistical challenge. This places immense pressure on the food system, requiring energy-dense, varied, stable, and palatable food options. Prolonged storage can lead to nutrient degradation, reducing their bioavailability and bioaccessibility to astronauts. Research is essential not only to improve the quality and stability of space food but also to enhance nutrient bioavailability, thereby reducing weight and volume of food. Muscle and bone loss represent major risks during extended spaceflight, prompting extensive efforts to find exercise countermeasures. However, increased exercise requires additional energy intake, and finding the optimal balance between energy needs and the preservation of muscle and bone mass is challenging. Currently, there is no reliable way to measure total energy expenditure and activity-related energy expenditures in real-time. Systematic research is necessary to develop onboard technology for accurate energy expenditure and body composition monitoring. This research should aim to establish an optimal exercise regimen that balances energy requirements while maintaining astronaut strength and minimizing food transport. In summary, this overview outlines key actions needed for future exploration missions to maintain body mass and physical strength of space travellers. It addresses the requirements for food processing and preservation, considerations for space food formulation and production, and the essential measures to be implemented.


Tumor-Derived Thymic Stromal Lymphopoietin Expands Bone Marrow B-cell Precursors in Circulation to Support Metastasis.
Immature B cells in the bone marrow emigrate into the spleen during adult lymphopoiesis. Here, we report that emigration is shifted to earlier B-cell stages in mice with orthotopic breast cancer, spontaneous ovarian cancer, and possibly in human breast carcinoma. Using mouse and human bone marrow aspirates and mouse models challenged with highly metastatic 4T1 breast cancer cells, we demonstrated that this was the result of secretion of thymic stromal lymphopoietin (TSLP) by cancer cells. First, TSLP downregulated surface expression of bone marrow (BM) retention receptors CXCR4 and VLA4 in B-cell precursors, increasing their motility and, presumably, emigration. Then, TSLP supported peripheral survival and proliferation of BM B-cell precursors such as pre-B-like cells. 4T1 cancer cells used the increased pool of circulating pre-B-like cells to generate metastasis-supporting regulatory B cells. As such, the loss of TSLP expression in cancer cells alone or TSLPR deficiency in B cells blocked both accumulation of pre-B-like cells in circulation and cancer metastasis, implying that the pre-B cell-TSLP axis can be an attractive therapeutic target. SIGNIFICANCE: Cancer cells induce premature emigration of B-cell precursors from the bone marrow to generate regulatory B cells.


PhytoPipe: a phytosanitary pipeline for plant pathogen detection and diagnosis using RNA-seq data.
Detection of exotic plant pathogens and preventing their entry and establishment are critical for the protection of agricultural systems while securing the global trading of agricultural commodities. High-throughput sequencing (HTS) has been applied successfully for plant pathogen discovery, leading to its current application in routine pathogen detection. However, the analysis of massive amounts of HTS data has become one of the major challenges for the use of HTS more broadly as a rapid diagnostics tool. Several bioinformatics pipelines have been developed to handle HTS data with a focus on plant virus and viroid detection. However, there is a need for an integrative tool that can simultaneously detect a wider range of other plant pathogens in HTS data, such as bacteria (including phytoplasmas), fungi, and oomycetes, and this tool should also be capable of generating a comprehensive report on the phytosanitary status of the diagnosed specimen. We have developed an open-source bioinformatics pipeline called PhytoPipe (Phytosanitary Pipeline) to provide the plant pathology diagnostician community with a user-friendly tool that integrates analysis and visualization of HTS RNA-seq data. PhytoPipe includes quality control of reads, read classification, assembly-based annotation, and reference-based mapping. The final product of the analysis is a comprehensive report for easy interpretation of not only viruses and viroids but also bacteria (including phytoplasma), fungi, and oomycetes. PhytoPipe is implemented in Snakemake workflow with Python 3 and bash scripts in a Linux environment. The source code for PhytoPipe is freely available and distributed under a BSD-3 license. PhytoPipe provides an integrative bioinformatics pipeline that can be used for the analysis of HTS RNA-seq data. PhytoPipe is easily installed on a Linux or Mac system and can be conveniently used with a Docker image, which includes all dependent packages and software related to analyses. It is publicly available on GitHub at https://github.com/healthyPlant/PhytoPipe and on Docker Hub at https://hub.docker.com/r/healthyplant/phytopipe .


Evaluation for antioxidant and renoprotective activity of olmesartan using nephrectomy rats.
Angiotensin II type 1 receptor blockers, which inhibit the rennin-angiotensin system, are used in the treatment of hypertension. In addition to their ability to lower blood pressure, these compounds have also been reported to protect organs, such as kidney and heart. Although the mechanisms of these protective effects are not fully understood, it is generally thought that their antioxidant effects likely play a role. The aim of the present study was to characterize the relationship between the antioxidant activity of olmesartan and its pharmacological actions such as renoprotective or blood pressure lowering effects, using 5/6 nephrectomy rats. In 5/6 nephrectomy rats, the potential antioxidant power, the ratio of oxidized to unoxidized albumin, as a marker of protein oxidation in blood, both systolic and diastolic blood pressure, plasma creatinine concentration, and amounts of protein excreted into the urine were significantly higher than the corresponding values for sham operated rats. However, olmesartan significantly suppressed these parameters within 8 weeks after oral administration in 5/6 nephrectomy rats. The oxidized albumin ratio was significantly decreased 4 weeks after the administration of olmesartan and these lower levels were maintained at 8 weeks. Furthermore, olmesartan improved radical scavenging activity of isolated albumin from rat plasma. Interestingly, a good correlation was found between the oxidized albumin ratios and renal function, whereas no correlation was found in the case of blood pressure. Based on those findings, we conclude that the antioxidant properties of olmesartan may be related to its renoprotective action rather than an antihypertensive effect.


Molecular testing in lung cancer: the time is now.
In the past few years, we have witnessed a revolution in the molecular understanding of non-small cell lung cancer. Major progress has also been made in the clinic, with the introduction of EGFR-targeted and anti-angiogenic therapies. These advances have led to the development of a multitude of commercially available prognostic and predictive biomarkers. In particular, EGFR mutation and EML4/ALK testing have reached clinical validation and are incorporated into current treatment paradigms. This overview will present the scientific background of the biology of the relevant biomarkers and the studies conducted for their clinical validation. The technical challenges and shortcomings of these assays are also discussed. Furthermore, ongoing biomarker-driven clinical studies and the appropriate clinical use of available tests will be reviewed to assist the clinician with the proper incorporation of molecular testing into the routine care of patients with non-small cell lung cancer.


Determinants of susceptibility and resistance to feline leukemia virus infection. I. Role of macrophages.
The role of autochthonous peritoneal feline macrophages (M theta) in the age-related resistance of cats to feline leukemia virus (FeLV) was investigated by a study of the functional properties and FeLV susceptibility of M theta from kittens and adult cats and the effect of hydrocortisone (HC) and silica on M theta-FeLV interactions. Although the phagocytic functions of isolated M theta from kittens and adults were equivalent, the mean FeLV susceptibility of M theta from kittens was five times that of M theta from adult cats, thus establishing a direct correlation between the age-related susceptibility of cats and M theta from cats to FeLV. M theta of viremic cats were found to be infected with FeLV in vivo; virus titers were slightly higher than those obtained after in vitro infection of M theta, M theta from cats that had experienced regressive FeLV infection were not significantly more resistant to FeLV infection in vitro than were M theta from naive adult specific-pathogen-free cats. HC, which has been shown to enhance the in vivo FeLV susceptibility of cats, also enhanced the permissiveness of M theta from cats to FeLV in vitro (600-fold for M theta from adult cats and 200-fold for M theta) from kittens. M theta permissiveness to FeLV was highly sensitive to HC and occurred in M theta infected in vivo or in vitro. In parallel with the effect of HC on the natural resistance of cats to FeLV, administration of silica before virus inoculation also markedly enhanced the FeLV susceptibility of adult cats. Silica was toxic for isolated M theta but not for lymphocytes in vitro, and silica produced monocytopenia and neutrophilia, delayed skin allograft rejection, and augmented feline oncovirus-associated cell membrane antigen antibody responses in vivo. These experiments indicate that M theta were linked to the natural resistance of cats to FeLV and that the temporary elimination of M theta functions (e.g., by silica) and/or the conversion of the M theta-FeLV relationship from a nonpermissive to a permissive state (e.g., by corticosteroids) resulted in failure of early virus containment, in persistent virus amplification in hemolymphatic tissues, and in subsequent FeLV-related proliferative or antiproliferative disease.


Non-invasive myocardial iron assessment in thalassaemic patients. T2 relaxometry and magnetization transfer ratio measurements.
To compare T2 relaxometry and magnetization transfer ratio (MTR) measurements of myocardial tissue in normal volunteers and thalassaemic patients for assessment of the myocardial iron levels. All examinations were done on a 1 T MR system using a multi-echo spin-echo sequence with 8 echoes for T2 measurements and a gradient echo sequence for MTR measurements. Diastolic cardiac triggering was used in both sequences. Ten patients and 10 normal subjects were included in the study. T2 and MTR measurements were correlated with serum ferritin levels. Regression analysis between T2 and MTR measurements and ferritin demonstrated a reversed linear relationship, (r=-0.932, p<0.05) and (r= -0.824, p<0.05), respectively. Mean T2 relaxation time and mean MTR of the normal subjects (57.95+/-4.9 ms and 43.70+/-3.3%) was significantly higher than that of the thalassaemic patients (38.8+/-6.2 ms and 26.40+/-6.1%) (p<0.01), respectively. MTR measurements can be used to complement T2 measurements for non-invasive myocardial iron assessment.


Risk factors for arm lymphedema following breast cancer diagnosis in Black women and White women.
Purpose Lymphedema of the arm is a potential complication of breast cancer therapy. This study examines pre-disposing factors that may operate in conjunction with treatment-related factors in the development of arm lymphedema in a large cohort of White and Black breast cancer survivors. Methods 494 women (271 White and 223 Black) with in situ to Stage III-A primary breast cancer completed a baseline interview within 18 months of diagnosis. Information on lymphedema was collected during a follow-up interview, conducted on average 50 months after diagnosis. Self-reported data were used to classify women with or without lymphedema. Multivariable logistic regression models were developed to identify risk factors for arm lymphedema. Results Arm lymphedema was associated with younger age at diagnosis (odds ratio, OR per year of age = 0.96; 95% confidence interval, CI = 0.93-0.99), positive history of hypertension (OR = 2.31; 95% CI = 1.38-3.88), obesity (OR for body mass index, BMI> or =30 = 2.48; 95% CI = 1.05-5.84) and having had surgery where 10 or more lymph nodes were excised (OR = 2.16; 95% CI = 1.12-4.17). While Black women had higher prevalence of arm lymphedema than White women (28% vs. 21%), race was not associated with lymphedema risk in models adjusted for multiple factors (adjusted OR = 1.01; 95% CI = 0.63-1.63). Conclusion Risk of arm lymphedema did not differ significantly for Black and White women. Risk factors identified in this study offer opportunities for interventions (weight loss, control of blood pressure, use of sentinel node biopsy where possible) for reducing incidence of lymphedema or controlling the symptoms associated with this condition.


Removing contaminants from databases of draft genomes.
Metagenomic sequencing of patient samples is a very promising method for the diagnosis of human infections. Sequencing has the ability to capture all the DNA or RNA from pathogenic organisms in a human sample. However, complete and accurate characterization of the sequence, including identification of any pathogens, depends on the availability and quality of genomes for comparison. Thousands of genomes are now available, and as these numbers grow, the power of metagenomic sequencing for diagnosis should increase. However, recent studies have exposed the presence of contamination in published genomes, which when used for diagnosis increases the risk of falsely identifying the wrong pathogen. To address this problem, we have developed a bioinformatics system for eliminating contamination as well as low-complexity genomic sequences in the draft genomes of eukaryotic pathogens. We applied this software to identify and remove human, bacterial, archaeal, and viral sequences present in a comprehensive database of all sequenced eukaryotic pathogen genomes. We also removed low-complexity genomic sequences, another source of false positives. Using this pipeline, we have produced a database of "clean" eukaryotic pathogen genomes for use with bioinformatics classification and analysis tools. We demonstrate that when attempting to find eukaryotic pathogens in metagenomic samples, the new database provides better sensitivity than one using the original genomes while offering a dramatic reduction in false positives.


The adipose tissue stromal vascular fraction secretome enhances the proliferation but inhibits the differentiation of myoblasts.
Adipose tissue is an excellent source for isolation of stem cells for treating various clinical conditions including injuries to the neuromuscular system. Many previous studies have focused on differentiating these adipose stem cells (ASCs) towards a Schwann cell-like phenotype (dASCs), which can enhance axon regeneration and reduce muscle atrophy. However, the stromal vascular fraction (SVF), from which the ASCs are derived, also exerts broad regenerative potential and might provide a faster route to clinical translation of the cell therapies for treatment of neuromuscular disorders. The aim of this study was to establish the effects of SVF cells on the proliferation and differentiation of myoblasts using indirect co-culture experiments. A Growth Factor PCR Array was used to compare the secretomes of SVF and dASCs, and the downstream signaling pathways were investigated. SVF cells, unlike culture-expanded dASCs, expressed and secreted hepatocyte growth factor (HGF) at concentrations sufficient to enhance the proliferation of myoblasts. Pharmacological inhibitor studies revealed that the signal is mediated via ERK1/2 phosphorylation and that the effect is significantly reduced by the addition of 100 pM Norleual, a specific HGF inhibitor. When myoblasts were differentiated into multinucleated myotubes, the SVF cells reduced the expression levels of fast-type myosin heavy chain (MyHC2) suggesting an inhibition of the differentiation process. In summary, this study shows the importance of HGF as a mediator of the SVF effects on myoblasts and provides further evidence for the importance of the secretome in cell therapy and regenerative medicine applications.


[Repair]
Transplantation of fetal neural cells represents an attractive replacement strategy for the treatment of certain neurodegenerative diseases. This is the case with Parkinson's disease, which results from a selective loss of dopaminergic neurons of the substantia nigra. Experimentation with animal models has demonstrated the feasibility of this approach. Grafting studies in patients have shown that intrastriatal implantation of solid grafts or cells obtained from human fetal mesencephalon usually results in a clinical benefit in patients. Despite continuous methodological progress, transplantation requires both conceptual and technical improvements. Current research aims at preventing the extensive death of donor dopaminergic neurons during the grafting procedure. However, the possibility of new sources of cells is currently being investigated. These include xenogeneic porcine neurons, or human cells programmed to produce dopamine or neurotrophic factors. A promising approach is based on the use of pluripotent stem cells derived from the brain, the bone marrow or early embryos. It is hoped that it will be possible to tightly control their proliferation and differentiation into dopaminergic neurons. Hence, it seems possible that transplantation will be widely used in the clinic in the future.


Association between frailty and patient outcomes after cancer surgery: a population-based cohort study.
Frailty is associated with poor postoperative outcomes, but existing data do not describe frailty's interaction with tumour characteristics at the time of cancer surgery. Our objective was to estimate the association between frailty and long-term survival, and to explore any interaction with tumour stage and grade. This was a population-based cohort study conducted using linked provincial health administrative data in Ontario, Canada (2009-20). Using a cancer registry, we identified adults having elective cancer surgery. Frailty was measured using a validated index (range 0-1; higher score=greater frailty). Associations between frailty and long-term postoperative survival (primary outcome) were estimated using proportional hazards regression. Secondary outcomes were length of stay, discharge destination, days alive at home, and healthcare costs. We identified and included 52 012 patients. Mean frailty score was 0.13 (standard deviation 0.07). During follow-up, 19 378 (37.3%) patients died. After adjustment for risk factors, each 10% increase in frailty was associated with a 1.60-fold relative decrease in survival (95% confidence interval: 1.56-1.64). The frailty-survival association was strongest for patients with lower stage and grade cancers. Increased frailty was associated with longer hospital stays (3 days), fewer days alive and at home (42 days yr-1), more frequent discharge to a nursing facility (2.38-fold), and increased healthcare costs ($6048). Patient frailty is associated with decreased long-term survival after cancer surgery. The association is stronger for early-stage and -grade cancers, which would otherwise have a better survival prognosis.