Ljungan virus present in intrauterine fetal death diagnosed by both immunohistochemistry and PCR

OBJECTIVES: Following up on prior evidence from animal and human studies of Ljungan virus (LV) in intrauterine fetal death (IUFD), we examine additional cases of IUFD using two standard assays of viral detection: immunohistochemistry (IHC) and real time RT‐PCR. MATERIALS AND METHODS: Frozen and formalin‐fixed specimens from IUFD cases were tested for the presence of LV using real time RT‐PCR and IHC, respectively. Formalin‐fixed organs from terminated pregnancies diagnosed as trisomy 21 were used as controls in the IHC assay. RESULTS: Presence of LV was demonstrated in all five IUFD cases by IHC and further confirmed in three of these cases by real time RT‐PCR. Only one of 18 trisomy 21 controls was LV positive by IHC. CONCLUSION: The presence of LV in IUFD patients has been confirmed by two different assays. Birth Defects Research (Part A), 2009. © 2009 Wiley‐Liss, Inc.     

Intrauterine death, fetal malformation, and delayed pregnancy in Ljungan virus-infected mice

BACKGROUND: A picornavirus (Ljunganvirus [LV]) has recently been associated with disease during pregnancy in its natural rodent reservoir and in humans. A study of laboratory mice infected under controlled conditions was therefore undertaken. METHODS: CD-1 female mice were infected gestational day two and subjected to varying regimes of stress. RESULTS: LV infection in combination with stress resulted in uterine resorptions, malformations, and neonatal death. A short delay in time to first pregnancy and births was observed in pairs infected in utero. CONCLUSIONS: LV is found in different species of native animals in both Europe and the United States and human epidemiological evidence connects LV and human reproduction, while the observations here indicate that LV is responsible for reproductive problems in a laboratory mouse model. The current findings suggest that the hypothesis that LV also causes disease in pregnant women and their offspring deserves further study.     

Impact of Acute Changes of Left Ventricular Contractility on the Transvalvular Impedance: Validation Study by Pressure-Volume Loop Analysis in Healthy Pigs

Background

The real-time and continuous assessment of left ventricular (LV) myocardial contractility through an implanted device is a clinically relevant goal. Transvalvular impedance (TVI) is an impedentiometric signal detected in the right cardiac chambers that changes during stroke volume fluctuations in patients. However, the relationship between TVI signals and LV contractility has not been proven. We investigated whether TVI signals predict changes of LV inotropic state during clinically relevant loading and inotropic conditions in swine normal heart.

Methods

The assessment of RVTVI signals was performed in anesthetized adult healthy anesthetized pigs (n = 6) instrumented for measurement of aortic and LV pressure, dP/dt_max and LV volumes. Myocardial contractility was assessed with the slope (Ees) of the LV end systolic pressure-volume relationship. Effective arterial elastance (Ea) and stroke work (SW) were determined from the LV pressure-volume loops. Pigs were studied at rest (baseline), after transient mechanical preload reduction and afterload increase, after 10-min of low dose dobutamine infusion (LDDS, 10 ug/kg/min, i.v), and esmolol administration (ESMO, bolus of 500 µg and continuous infusion of 100 µg·kg−1·min−1).

Results

We detected a significant relationship between ESTVI and dP/dtmax during LDDS and ESMO administration. In addition, the fluctuations of ESTVI were significantly related to changes of the Ees during afterload increase, LDDS and ESMO infusion.

Conclusions

ESTVI signal detected in right cardiac chamber is significantly affected by acute changes in cardiac mechanical activity and is able to predict acute changes of LV inotropic state in normal heart.     

Correlated recovery of five lizard populations following eradication of invasive mammals

Many conservation decisions rely on the assumption that multiple populations will respond similarly to management. However, few attempts have been made to evaluate indicators of population trends (i.e. population indicator species). Eradication of introduced mammals from offshore islands is a commonly used management technique for conservation of native taxa in New Zealand. Pacific rats and rabbits were eradicated from Korapuki Island in 1986/1987 enabling population recovery of native species that had been suppressed by predation or competition. However, the degree to which species’ responses were correlated has not been evaluated. We investigated correlations among lizard population trends on Korapuki Island as a test of the population indicator species concept. Our dataset consisted of captures of the five resident lizard species (three skinks, two geckos) from biannual pitfall trapping over a 10 year period (1986–1995) immediately following rodent eradication. We used a Bayesian modelling approach to examine correlations in population trends (based on mean annual counts) between species. Population trends were positively correlated for all species pairs (substantively for 90 % of pairs) and we detected no negative correlations. Systematic searches for single lizard species may indicate correlated recovery of lizard populations following rodent eradication and provide a cost-effective alternative to traditional ‘whole community’ monitoring. Our findings support evidence-based use of the population indicator species concept in cases where a shared ecological driver can be identified.     

Placental hCG immunohistochemistry and serum free-Beta-hCG at 11–13 weeks’ gestation in intrauterine fetal demise

Intrauterine fetal demise (IUFD) is a continuing problem that can result in severe psychosocial trauma for expecting parents. Our aim was to analyze placental human chorionic gonadotropin (hCG) expression at the third trimester and free-Beta-hCG levels measured at 11–13 weeks in cases of IUFD that occurred after 34 weeks’ gestation, alongside a parallel analysis of a set of controls. In this retrospective study we present immunohistochemical data of a tissue microarray that included the following: 12 placentas where IUFD occurred (24 samples); 28 control placentas from first and early second trimester (56 samples); and 30 control placentas at term of pregnancy (60 samples). We used immunohistochemistry to analyze the expression of hCG. Data are also presented from 3,240 first trimester trisomies screening tests, of which 21 pregnancies resulted in IUFD (15 after 22 weeks’ gestation and 6 after 34 weeks). All pregnancies took place between 2001 and 2010. For each case, our analysis took account of pregnancy-related data that we gathered from the relevant clinical files. Small for gestational age (SGA) was defined as neonatal weight <10th centile. Our results show that full-term placentas displayed a decreased immunohistochemical expression of hCG in comparison with those at the first trimester (p < 0.05). Moreover, low hCG expression in placentas at the third trimester was shown to be an independent risk factor for IUFD after 34 weeks’ gestation (under multivariate analysis with p < 0.05). When we reviewed first trimester screening results, free-Beta-HCG was found to be lower for the group of IUFD after 34 weeks’ gestation than in the group of live births (p < 0.05). This difference was heavily weighted by non small for gestational age (non-SGA) associated cases of IUFD: these presented a free-Beta-hCG MoM log of ?0.27 (±0.09) in contrast to just ?0.01 (±0.03) in SGA-associated IUFD (p < 0.05). Our results show that low hCG is an independent risk factor for IUFD after 34 weeks’ gestation, and that levels of the hormone are significantly lower in non-SGA associated cases of IUFD.     

Diabetes related cardiomyopathy time dependent echocardiographic evaluation in an experimental rat model

Type I diabetes is associated with a unique form of cardiomyopathy in the absence of atherosclerosis. The mechanisms involved in this phenomenon are not defined, but in humans this is associated with initial diastolic dysfunction followed by altered contractile performance. A relevant animal model would provide opportunities for mechanistic studies and experimental therapeutics, but none have been previously established for this unique form of cardiac pathophysiology, particularly with respect to clinically relevant and time-dependent diastolic and systolic assessments. Here we tested the hypothesis that the streptozotocin rat model mimics human phenomena with respect to time-dependent diastolic and systolic performance deficits, and investigated a role for cardiac hypertrophy and/or fibrosis. Streptozotocin was dosed 65 mg/kg i.p. and cardiac performance was assessed longitudinally for 56 days using noninvasive echocardiographic techniques. Significant hyperglycemia was detected within 3 days and remained elevated throughout the study (p<0.05). Significant reductions in HR and diastolic performance (transmitral flow velocities and slopes) were observed within 3 days relative to age matched controls, and these reductions progressed throughout the 56 day study. In contrast, statistically significant systolic dysfunction (LV fractional shortening, cardiac output) and LV dilation were detected only after 35 days. Increases in LV size and/or extent of fibrosis were not observed at any time. These results demonstrate the value of echocardiographic methods for time-dependent diastolic and systolic assessments in rodent models. Furthermore, diastolic dysfunction precedes contractile abnormalities in the streptozotocin model, similar to events that occur in humans.     

Spatial variation in abundance of the junin virus hosts in endemic and nonendemic Argentine haemorrhagic fever zones

Abstract Argentine haemorrhagic fever (AHF) is caused by Junin (JUN) virus, which is hosted by the drylands vesper mouse (Calomys musculinus). In this work we monitored population abundance of C. musculinus and rodent assemblages for 3 years in and outside the AHF endemic zones (northern Buenos Aires, southern Córdoba and Santa Fe Provinces, Argentina). The study area was divided into endemic and nonendemic zones. In the endemic zone epidemic sites were recognized, characterized by recent emergence and maintenance of AHF cases, and also historical sites, characterized by decreased incidence or disappearance of AHF human cases. In the nonendemic zone AHF has never been recognized. Although differences were statistically significant only during some periods, population abundance of C. musculinus was usually lower in the nonendemic sites. The pattern and magnitude of seasonal fluctuations in C. musculinus populations were also distinct in the nonendemic sites as compared to endemic sites. The relative abundance of C. musculinus in rodent assemblage was lower in nonendemic sites than in the endemic sites. The lower population densities and dampened seasonal dynamics may be at least partly responsible for the absence of AHF cases in the nonendemic zone. It is suggested that the balance between intra and interspecific interactions might be the cause of the pattern of incidence and prevalence of pathogens in the host species.     

Ungulate bocaparvovirus 4 and rodent bocavirus are different genotypes of the same species of virus

Bocaviruses are associated with many human infectious diseases, such as respiratory tract infections, gastroenteritis, and hepatitis. Rats are known to be reservoirs of bocaviruses, including rodent bocavirus and rat bocavirus. Recently, ungulate bocaparvovirus 4, a known porcine bocavirus, has also been found in rats. Thus, investigating bocaviruses in rats is important for determining the origin of the viruses and preventing and controlling their transmission. To the best of our knowledge, no study to date has investigated bocaviruses in the livers of rats. In this report, a total of 624 rats were trapped in southern China between 2014 and 2017. Liver and serum samples from rats were tested for the prevalence of bocaviruses using PCR. Sequences related to ungulate bocaparvovirus 4 and rodent bocavirus were detected in both liver and serum samples. Interestingly, the prevalence of ungulate bocaparvovirus 4 (reference strain:KJ622366.1) was higher than that of rodent bocavirus (reference strain:KY927868.1) in both liver (2.24% and 0.64%, respectively) and serum samples (2.19% and 0.44%, respectively). The NS1 regions of ungulate bocaparvovirus 4 and rodent bocavirus related sequences displayed over 84% and 88% identity at the nucleic acid and amino acid levels, respectively. Furthermore, these sequences had similar genomic structure, genomic features, and codon usage bias, and shared a common ancestor. These viruses also displayed greater adaptability to rats than pigs. Our results suggested that ungulate bocaparvovirus 4 and rodent bocavirus may originate from rats and may be different genotypes of the same bocavirus species.     

Human and rodent bone marrow mesenchymal stem cells that express primitive stem cell markers can be directly enriched by using the CD49a molecule

Bone marrow (BM) from human and rodent species contains a population of multipotential cells referred to as mesenchymal stem cells (MSCs). Currently, MSCs are isolated indirectly by using a culture step and then the generation of fibroblast colony-forming units (CFU-fs). Unprocessed or native BM MSCs have not yet been fully characterised. We have previously developed a direct enrichment method for the isolation of MSCs from human BM by using the CD49a protein (alpha1-integrin subunit). As the CD49a gene is highly conserved in mammals, we have evaluated whether this direct enrichment can be employed for BM cells from rodent strains (rat and mouse). We have also studied the native phenotype by using both immunodetection and immunomagnetic methods and have compared MSCs from mouse, rat and human BM. As is the case for human BM, we have demonstrated that all rodent multipotential CFU-fs are contained within the CD49a-positive cell population. However, in the mouse, the number of CFU-fs is strain-dependent. Interestingly, all rat and mouse Sca-1-positive cells are concentrated within the CD49a-positive fraction and also contain all CFU-fs. In human, the colonies have been detected in the CD49a/CD133 double-positive population. Thus, the CD49a protein is a conserved marker that permits the direct enrichment of BM MSCs from various mammalian species; these cells have been phenotyped as true BM stem cells.     

川西平原农田啮齿动物群落动态：年间变动和季节变动

为了研究农田啮齿动物群落的动态规律,用标志重捕的方法给出了川西平原农田的啮齿动物群落的物种数、结合种群密度、生物量、物种多样性的Shannon指数与Simpson指数等5个变量的时间序列资料,并分析了这些时间序列的年间变动和季节变动的特点。结果表明：（1）5个变量总是处于不断的变化之中,特别是年间交替升降十分明显;（2）群落的全部8个物种从未同时出现过,同一个月最多有5个物种,最少只有1个物种;（3）5个变量季节变动的幅度均较大并各具有1至3个不等的明显的峰值,而年间变动的幅度则相对较小;（4）5个变量的最大值均出现在夏季,最小值则出现在春季、夏季或冬季;（5）5个变量的季节性均不强;（6）优势种大足鼠（Rattus nitidus)的种群密度分别与群落结合种群密度、生物量的年间变动和季节变动均具有相似的变动规律;此外,优势种大足鼠的种群密度、生物量分别占群落结合种群密度、生物量的比例均较高,而且这两个比例季节变动的幅度都较大而年间变动则较为稳定。     

Essential dynamics of reversible peptide folding: memory-free conformational dynamics governed by internal hydrogen bonds

A principal component analysis has been applied on equilibrium simulations of a beta-heptapeptide that shows reversible folding in a methanol solution. The analysis shows that the configurational space contains only three dense sub-states. These states of relatively low free energy correspond to the "native" left-handed helix, a partly helical intermediate, and a hairpin-like structure. The collection of unfolded conformations form a relatively diffuse cloud with little substructure. Internal hydrogen-bonding energies were found to correlate well with the degree of folding. The native helical structure folds from the N terminus; the transition from the major folding intermediate to the native helical structure involves the formation of the two most C-terminal backbone hydrogen bonds. A four-state Markov model was found to describe transition frequencies between the conformational states within error limits, indicating that memory-effects are negligible beyond the nanosecond time-scale. The dominant native state fluctuations were found to be very similar to unfolding motions, suggesting that unfolding pathways can be inferred from fluctuations in the native state. The low-dimensional essential subspace, describing 69% of the collective atomic fluctuations, was found to converge at time-scales of the order of one nanosecond at all temperatures investigated, whereas folding/unfolding takes place at significantly longer time-scales, even above the melting temperature.     

Part II: beneficial effects of the peroxynitrite decomposition catalyst FP15 in murine models of arthritis and colitis

BACKGROUND: Peroxynitrite is a reactive oxidant species produced from nitric oxide and superoxide, which has been indirectly implicated in the pathogenesis of many inflammatory conditions including arthritis and colitis. Here, using a novel peroxynitrite decomposition catalyst, FP15, we directly investigate the role of peroxynitrite in the pathogenesis of arthritis and colitis in rodent models. METHODS: Arthritis was induced in mice by intradermal collagen injection; incidence and severity of arthritis was monitored using a macroscopic scoring system. At the end of the experiment paws were taken for determination of neutrophil infiltration (myeloperoxidase [MPO] activity), oxidative stress (malondialdehyde [MDA] level), and cytokine/chemokine levels. Colitis was induced in mice by 5% dextran sodium sulfate (DSS) in their drinking water. Colitis symptoms were assessed 10 days later, the parameters determined included body weight, rectal bleeding, colon length, colonic MPO and MDA levels, and colon histologic damage. RESULTS: Treatment with FP15 significantly reduced the inflammation and oxidative stress in arthritis and colitis. FP15 reduced both the incidence and severity of arthritis in mice and this was associated with reduced paw MPO and MDA levels. Similarly, in colitis, FP15 reduced colon damage, and this was associated with reduced colon neutrophil infiltration and oxidative stress. CONCLUSIONS:The protective effect of FP15 suggests that peroxynitrite plays a significant pathogenetic role in arthritis and colitis in the currently employed rodent models. Further work is needed to determine whether neutralization of peroxynitrite also represents a promising strategy to treat human inflammatory diseases such as arthritis and colitis.     

Isolation and characterization of two vitellins from eggs of the spider Polybetes pythagoricus (Araneae: Sparassidae)

Despite vitellins being essential yolk proteins, their presence in spiders remains almost unknown. Two vitellins from the spider Polybetes pythagoricus, named LV1 and LV2, were isolated and their size, shape, lipids, fatty acids, proteins and carbohydrates moieties were determined. LV1 has a density similar to that of HDL with 49.3% lipids, and LV2 has a density similar to that of VHDL with 9.7% lipids. The major neutral lipid present in both vitellins was found to be esterified cholesterol, 16% for LV1 and 24% for LV2. The major fatty acid was 18:1n-9 in LV1 and LV2. Results from native PAGE showed a lipoprotein of 550 kDa for LV1 and three lipoproteins of 571, 400 and 257 kDa for LV2. SDS-PAGE evidenced two major apolipoproteins of 64 and 25 kDa in LV1. The three lipoproteins of LV2 were electroeluted and analyzed by SDS-PAGE, showing different proportions of the same apolipoproteins (181, 67 and 60 kDa). LVs were analyzed by spectrophotometry, immunochemical and electron microscopy, showing that the respiratory pigment hemocyanin was not present as apolipoprotein. This fact evidenced that these LVs were not related to hemolymphatic lipoproteins.     

Insulin assembly damps conformational fluctuations: Raman analysis of amide I linewidths in native states and fibrils

The crystal structure of insulin has been investigated in a variety of dimeric and hexameric assemblies. Interest in dynamics has been stimulated by conformational variability among crystal forms and evidence suggesting that the functional monomer undergoes a conformational change on receptor binding. Here, we employ Raman spectroscopy and Raman microscopy to investigate well-defined oligomeric species: monomeric and dimeric analogs in solution, native T(6) and R(6) hexamers in solution and corresponding polycrystalline samples. Remarkably, linewidths of Raman bands associated with the polypeptide backbone (amide I) exhibit progressive narrowing with successive self-assembly. Whereas dimerization damps fluctuations at an intermolecular beta-sheet, deconvolution of the amide I band indicates that formation of hexamers stabilizes both helical and non-helical elements. Although the structure of a monomer in solution resembles a crystallographic protomer, its encagement in a native assembly damps main-chain fluctuations. Further narrowing of a beta-sheet-specific amide I band is observed on reorganization of insulin in a cross-beta fibril. Enhanced flexibility of the native insulin monomer is in accord with molecular dynamics simulations. Such conformational fluctuations may initiate formation of an amyloidogenic nucleus and enable induced fit on receptor binding.     

Spicing up restoration: can chili peppers improve restoration seeding by reducing seed predation?

Seed predation by rodents presents a significant barrier to native plant recruitment and can impede restoration seeding efforts. In nature, some plants contain secondary defense compounds that deter seed predators. If these natural defense compounds can be applied to unprotected seeds to inhibit rodent granivores, this approach could improve restoration seeding. Capsaicin is the active ingredient in chili pepper (Capsicum spp.) seeds that creates the burning sensation associated with human consumption of hot peppers. This compound has a similar effect on other mammals and is believed to have evolved as a deterrent to rodent seed predators. We used seed‐coating techniques to attach powder ground from Bhut Jolokia (Capsicum chinense) peppers to native plant seeds and evaluated the efficacy of these seed coatings for deterring rodent seed predation and enhancing native plant recruitment using laboratory and field experiments. Laboratory feeding trials demonstrated that native deer mice (Peromyscus maniculatus) consumed far fewer pepper‐coated seeds compared to untreated control seeds. Field seed‐addition experiments consistently demonstrated that rodent seed predation reduced native plant recruitment over the 4‐year study. Coating techniques used in the first 3 years were not persistent enough to reduce rodent seed predation effects on plant recruitment. However, a more persistent coating applied in conjunction with late‐winter sowing negated rodent seed predation effects on recruitment in year 4. Our results demonstrate that coating seeds with natural plant defense compounds may provide an effective, economical way to improve the efficacy of plant restoration by deterring seed predation by ubiquitous rodent granivores.     

Survey of neonatal screening for primary hypothyroidism in England,Wales, and Northern Ireland 1982-4

National screening for congenital hypothyroidism was established in the United Kingdom in 1982. During 1982-4, 488 infants with primary congenital hypothyroidism were detected by the 25 regional screening laboratories in England, Wales, and Northern Ireland. In addition, one infant had signs of cretinism at birth and was investigated before the screening test was done and four infants were known to have been missed by the screening programme; among these four infants the initial thyroid stimulating hormone concentrations were normal in two with inherited defects of synthesis of thyroxine, not measured in one, and false negative in one. The overall incidence of primary hypothyroidism was 1:3937 births (boys 1:6640, girls 1:2756). The incidence seemed to be reduced in infants born to black mothers (two cases only) and increased in those born to Asian mothers (61 cases). Congenital anomalies other than those of the thyroid gland were reported in 36 children (7%), and 15 (3%) died from various causes before the age of 4. Infants who were considered to show unequivocal evidence of hypothyroidism started treatment at a median age of 17 days (5th and 95th centiles 10 and 42 days) compared with a median age of 14 days (5th and 95th centiles 9 and 21 days) for infants with classic phenylketonuria also detected by national screening.     

Noninvasive Doppler-derived myocardial performance index in rats with myocardial infarction: validation and correlation by conductance catheter

The rodent model of myocardial infarction (MI) is extensively used in heart failure studies. However, long-term follow-up of echocardiographic left ventricular (LV) function parameters such as the myocardial performance index (MPI) and its ratio with the fractional shortening (LVFS/MPI) has not been validated in conjunction with invasive indexes, such as those derived from the conductance catheter (CC). Sprague-Dawley rats with left anterior descending coronary artery ligation (MI group, n = 9) were compared with a sham-operated control group (n = 10) without MI. Transthoracic echocardiography (TTE) was performed every 2 wk over an 8-wk period, after which classic TTE parameters, especially MPI and LVFS/MPI, were compared with invasive indexes obtained by using a CC. Serial TTE data showed significant alterations in the majority of the noninvasive functional and structural parameters (classic and novel) studied in the presence of MI. Both MPI and LVFS/MPI significantly (P < 0.05 for all reported values) correlated with body weight (r = -0.58 and 0.76 for MPI and LVFS/MPI, respectively), preload recruitable stroke work (r = -0.61 and 0.63), LV end-diastolic pressure (LVEDP) (r = 0.82 and -0.80), end-diastolic volume (r = 0.61 and -0.58), and end-systolic volume (r = 0.46 and -0.48). Forward stepwise linear regression analysis revealed that, of all variables tested, LVEDP was the only independent determinant of MPI (r = 0.84) and LVFS/MPI (r = 0.83). We conclude that MPI and LVFS/MPI correlate strongly and better than the classic noninvasive TTE parameters with established, invasively assessed indexes of contractility, preload, and volumetry. These findings support the use of these two new noninvasive indexes for long-term analysis of the post-MI LV remodeling.