Bovine viral diarrhoea viruses from New Zealand belong predominantly to the BVDV-1a genotype.

AIM: To determine which genotypes of bovine viral diarrhoea virus (BVDV) circulate among cattle in New Zealand. METHODS: Samples comprised BVDV-1-positive sera sourced from submissions to veterinary diagnostic laboratories in 2019 (n = 25), 2020 (n = 59) and 2022 (n = 74) from both beef and dairy herds, as well as archival BVDV-1 isolates (n = 5). Fragments of the 5' untranslated region (5' UTR) and glycoprotein E2 coding sequence of the BVDV genome were amplified and sequenced. The sequences were aligned to each other and to international BVDV-1 sequences to determine their similarities and phylogenetic relationships. The 5' UTR sequences were also used to create genetic haplotype networks to determine if they were correlated with selected traits (location, type of farm, and year of collection). RESULTS: The 5' UTR sequences from New Zealand BVDV were closely related to each other, with pairwise identities between 89% and 100%. All clustered together and were designated as BVDV-1a (n = 144) or BVDV-1c (n = 5). There was no evidence of a correlation between the 5' UTR sequence and the geographical origin within the country, year of collection or the type of farm. Partial E2 sequences from New Zealand BVDV (n = 76) showed 74-100% identity to each other and clustered in two main groups. The subtype assignment based on the E2 sequence was the same as based on the 5' UTR analysis. This is the first comprehensive analysis of genomic variability of contemporary New Zealand BVDV based on the analysis of the non-coding (5' UTR) and coding (E2) sequences. CONCLUSIONS AND CLINICAL RELEVANCE: Knowledge of the diversity of the viruses circulating in the country is a prerequisite for the development of effective control strategies, including a selection of suitable vaccines. The data presented suggest that New Zealand BVDV are relatively homogeneous, which should facilitate eradication efforts including selection or development of the most suitable vaccines.

Magnetic-Competition-Induced Colossal Magnetoresistance in n-Type HgCr_{2}Se_{4} under High Pressure.

The n-type HgCr_{2}Se_{4} exhibits a sharp semiconductor-to-metal transition (SMT) in resistivity accompanying the ferromagnetic order at T_{C}=106 K. Here, we investigate the effects of pressure and magnetic field on the concomitant SMT and ferromagnetic order by measuring resistivity, dc and ac magnetic susceptibility, as well as single-crystal neutron diffraction under various pressures up to 8 GPa and magnetic fields up to 8 T. Our results demonstrate that the ferromagnetic metallic ground state of n-type HgCr_{2}Se_{4} is destabilized and gradually replaced by an antiferromagnetic, most likely a spiral magnetic, and insulating ground state upon the application of high pressure. On the other hand, the application of external magnetic fields can restore the ferromagnetic metallic state again at high pressures, resulting in a colossal magnetoresistance (CMR) as high as ∼ 3×10^{11}% under 5 T and 2 K at 4 GPa. The present study demonstrates that n-type HgCr_{2}Se_{4} is located at a peculiar critical point where the balance of competition between ferromagnetic and antiferromagnetic interactions can be easily tipped by external stimuli, providing a new platform for achieving CMR in a single-valent system.

Development of cubic anvil type high pressure apparatus for neutron diffraction.

High-pressure neutron diffraction (HPND) experiments in extended pressure and temperature ranges can provide invaluable information for understanding many pressure-induced emergent phenomena, such as unusual phase transitions and quantum critical behavior involving spin, orbital, charge and structural degrees of freedom, in strongly correlated materials. Many apparatuses for different purposes of HPND experiments have been developed in several laboratories. Recently, a clamp-type cubic anvil high pressure cell that can generate pressure over 7 GPa at 3 K was developed for low-temperature HPND measurements. In this paper, characteristics of the clamp-type cubic anvil high pressure cell are presented and its performances are demonstrated by measuring magnetic neutron scattering under pressure on MnP single crystal samples.

Frustrated magnetic interactions in an S = 3/2 bilayer honeycomb lattice compound Bi3Mn4O12(NO3).

An inelastic neutron scattering study has been performed in an S = 3/2 bilayer honeycomb lattice compound Bi3Mn4O12(NO3) at ambient and high magnetic fields. Relatively broad and monotonically dispersive magnetic excitations were observed at ambient field, where no long-range magnetic order exists. In the magnetic-field-induced long-range ordered state at 10 T, the magnetic dispersions become slightly more intense, albeit still broad as in the disordered state, and two excitation gaps, probably originating from an easy-plane magnetic anisotropy and intrabilayer interactions, develop. Analyzing the magnetic dispersions using the linear spin-wave theory, we estimated the intraplane and intrabilayer magnetic interactions, which are almost consistent with those determined by ab initio density functional theory calculations [M. Alaei et al., Phys. Rev. B 96, 140404(R) (2017)], except the third and fourth neighbor intrabilayer interactions. Most importantly, as predicted by the theory, there is no significant frustration in the honeycomb plane but frustrating intrabilayer interactions probably give rise to the disordered ground state.

Orbital Glass State of the Nearly Metallic Spinel Cobalt Vanadate.

Strain, magnetization, dielectric relaxation, and unpolarized and polarized neutron diffraction measurements were performed to study the magnetic and structural properties of spinel Co_{1-x}V_{2+x}O_{4}. The strain measurement indicates that, upon cooling, ΔL/L in the order of ∼10^{-4} starts increasing below T_{C}, becomes maximum at T_{max}, and then decreases and changes its sign at T^{*}. Neutron measurements indicate that a collinear ferrimagnetic order develops below T_{C} and upon further cooling noncollinear ferrimagnetic ordering occurs below T_{max}. At low temperatures, the dielectric constant exhibits a frequency dependence, indicating slow dynamics. These results indicate the existence of an orbital glassy state at low temperatures in this nearly metallic frustrated magnet.

Factors that determine body composition of female systemic lupus erythematosus (SLE) patients in Sri Lanka: a comparative study using dual-energy x-ray absorptiometry.

Studies on body composition and its determinants among SLE patients are limited. Estimation of body composition, analysis of determinants and associations of different body compartments are important in planning long-term care of these patients. The aim of the study was to identify the changes in body composition among SLE patients and assess the effect of corticosteroid use, patient and disease-related variables on body composition. We compared lean mass, fat mass, bone mineral density (BMD), and bone mineral content (BMC) determined by dual-energy x-ray absorptiometry technology, in a group of premenopausal women with SLE (n = 27) and an age-matched healthy group of women (n = 27). The median (IQR) duration of SLE was 3 (2-5) years while median (IQR) duration and dose of prednisolone therapy were 108 (88 - 172) weeks and 9730 (6160-15360) mg, respectively. No significant difference was observed in body mass index (BMI) or total fat mass between the two groups. SLE patients, however, had significantly lower lean mass (p < 0.001), BMD (p < 0.001) and BMC (p < 0.005) than healthy controls. Among cases, compared with lean mass, total body fat content showed stronger associations with total body BMD (r = 0.49, p < 0.01) and total body BMC (r = 0.63, p < 0.01). When a stepwise regression model was fitted, lean mass among controls and total fat mass among cases emerged as the best predictors of BMC/BMD. No significant correlations were found between the disease duration or cumulative glucocorticosteroid dose and total body BMD, total body BMC, lean mass or total fat content in SLE patients.

The orphan tyrosine kinase receptor, ROR2, mediates Wnt5A signaling in metastatic melanoma.

Tyrosine kinase receptors represent targets of great interest for cancer therapy. Here we show, for the first time, the importance of the orphan tyrosine kinase receptor, ROR2, in melanoma progression. Using melanoma tissue microarrays, we show that ROR2 is expressed predominantly in metastatic melanoma. As ROR2 has been shown to specifically interact with the non-canonical Wnt ligand, Wnt5A, this corroborates our earlier data implicating Wnt5A as a mediator of melanoma metastasis. We show here that increases in Wnt5A cause increases in ROR2 expression, as well as the PKC-dependent, clathrin-mediated internalization of ROR2. WNT5A knockdown by siRNA decreases ROR2 expression, but silencing of ROR2 has no effect on WNT5A levels. ROR2 knockdown does, however, result in a decrease in signaling downstream of Wnt5A. Using in vitro and in vivo metastasis assays, we show that ROR2 is necessary for the Wnt5A-mediated metastasis of melanoma cells. These data imply that ROR2 may represent a novel target for melanoma therapy.

Claudin-1 overexpression in melanoma is regulated by PKC and contributes to melanoma cell motility.

Serial analysis of gene expression followed by pathway analysis implicated the tight junction protein claudin-1 (CLDN1) in melanoma progression. Tight junction proteins regulate the paracellular transport of molecules, but staining of a tissue microarray revealed that claudin-1 was overexpressed in melanoma, and aberrantly expressed in the cytoplasm of malignant cells, suggesting a role other than transport. Indeed, melanoma cells in culture demonstrate no tight junction function. It has been shown that protein kinase C (PKC) can affect expression of claudin-1 in rat choroid plexus cells, and we observed a correlation between levels of activated PKC and claudin expression in our melanoma cells. To determine if PKC could affect the expression of CLDN1 in human melanoma, cells lacking endogenous claudin-1 were treated with 200 nM phorbol myristic acid (PMA). PKC activation by PMA caused an increase in CLDN1 transcription in 30 min, and an increase in claudin-1 protein by 12 h. Inhibition of PKC signaling in cells with high claudin-1 expression resulted in decreased claudin-1 expression. CLDN1 appears to contribute to melanoma cell invasion, as transient transfection of melanoma cells with CLDN1 increased metalloproteinase 2 (MMP-2) secretion and activation, and subsequently, motility of melanoma cells as demonstrated by wound-healing assays. Conversely, knockdown of CLDN1 by siRNA resulted in the inhibition of motility, as well as decreases in MMP-2 secretion and activation. These data implicate claudin-1 in melanoma progression.

Genome-wide analysis of gene expression in adult Anopheles gambiae.

With their genome sequenced, Anopheles gambiae mosquitoes now serve as a powerful tool for basic research in comparative, evolutionary and developmental biology. The knowledge generated by these studies is expected to reveal molecular targets for novel vector control and pathogen transmission blocking strategies. Comparisons of gene-expression profiles between adult male and nonblood-fed female Anopheles gambiae mosquitoes revealed that roughly 22% of the genes showed sex-dependent regulation. Blood-fed females switch the majority of their metabolism to blood digestion and egg formation within 3 h after the meal is ingested, in detriment to other activities such as flight and response to environment stimuli. Changes in gene expression are most evident during the first, second and third days after a blood meal, when as many as 50% of all genes showed significant variation in transcript accumulation. After laying the first cluster of eggs (between 72 and 96 h after the blood meal), mosquitoes return to a nongonotrophic stage, similar but not identical to that of 3-day-old nonblood-fed females. Ageing and/or the nutritional state of mosquitoes at 15 days after a blood meal is reflected by the down-regulation of approximately 5% of all genes. A full description of the large number of genes regulated at each analysed time point and each biochemical pathway or biological processes in which they are involved is not possible within the scope of this contribution. Therefore, we present descriptions of groups of genes displaying major differences in transcript accumulation during the adult mosquito life. However, a publicly available searchable database (http://www.angagepuci.bio.uci.edu/) has been made available so that detailed analyses of specific groups of genes based on their descriptions, functions or levels of gene expression variation can be performed by interested investigators according to their needs.

Relative lack of clinical disease among household contacts of tuberculosis patients compared to leprosy households.

The incidence of clinical tuberculosis and clinical leprosy among household members of tuberculosis and leprosy patients in Sri Lanka was studied. The study period was approximately 20 years (January 1981 to December 2001) and the total number of patients and contacts were 325 and 968 for tuberculosis and 726 and 3066 for leprosy, respectively. While none of the tuberculosis patient households had more than 1 patient nor any contacts who developed clinical disease during the observation period, 20% (148/726) of the leprosy patients had more than 1 patient in the family and 0.9% (13/1403) of their contacts who were followed-up developed clinical leprosy during the observation period. Although the tuberculosis patient household contacts did not develop clinical disease, in 79% (88/112) of contacts who were tested by Western blot analysis, there was serologic evidence of Mycobacterium tuberculosis infection. These data show that in populations of comparable socio-economic, environmental and geographic locations, tuberculosis and leprosy show very different transmission patterns. In general, in tuberculosis household contacts, in spite of exposure, infection did not proceed to clinical disease. In contrast, a significant number of leprosy household contacts developed clinical leprosy. These findings have implications in the design and implementation of control programmes for these two diseases.

Synchronization of coupled rotators: Josephson junction ladders and the locally coupled Kuramoto model.

We show that the resistively shunted junction (RSJ) equations describing a ladder array of overdamped, critical-current disordered Josephson junctions that are current biased along the rungs of the ladder can be mapped onto a Kuramoto model with nearest neighbor, sinusoidal couplings. This result is obtained by an averaging method, in which the fast dynamics of the RSJ equations are integrated out, leaving the dynamics which describe the time scale over which neighboring junctions along the rungs of the ladder phase and frequency synchronize. We quantify the degree of frequency synchronization of the rung junctions by calculating the standard deviation of their time-averaged voltages, sigma(omega), and the phase synchronization is quantified by calculating the time average of the modulus of the Kuramoto order parameter, <|r|>. We test the results of our averaging process by comparing the values of sigma(omega) and <|r|> for the original RSJ equations and our averaged equations. We find excellent agreement for dc bias currents of I(B)/ greater, similar 3, where is the average critical current of the rung junctions, and critical current disorders of up to 10%. We also study the effects of thermal noise on the synchronization properties of the overdamped ladder. Finally, we find that including the effects of junction capacitance can lead to a discontinuous synchronization transition as the strength of the coupling between neighboring junctions is smoothly varied.

Neurological manifestations of snake bite in Sri Lanka.

BACKGROUND AND AIMS: Snake bite is an important cause of mortality and morbidity in certain parts of Sri Lanka. This study was designed to determine the offending snakes, neurological manifestations, disease course, and outcome in neurotoxic envenomation. METHODS AND MATERIAL: Fifty six consecutive patients admitted with neurological manifestations following snake bite were studied prospectively. Data were obtained regarding the offending snakes, neurological symptoms, time taken for onset of symptoms, neurological signs, and time taken for recovery. RESULTS: The offending snake was Russell's viper in 27(48.2%), common and Sri Lankan krait in 19(33.9%), cobra in 3(5.4%), and unidentified in 7(12.5%). Ptosis was the commonest neurological manifestation seen in 48(85.7%) followed by ophthalmoplegia (75%), limb weakness (26.8%), respiratory failure (17.9%), palatal weakness (10.7%), neck muscle weakness (7.1%), and delayed sensory neuropathy (1.8%). Neurological symptoms were experienced usually within 6 hours after the bite. Following administration of antivenom, the signs of recovery became evident within a few hours to several days. The duration for complete recovery ranged from four hours to two weeks. CONCLUSIONS: Complete recovery of neuromuscular weakness was observed in all patients except for one who died with intracerebral haemorrhage shortly after admission.

In Wuchereria bancrofti filariasis, asymptomatic microfilaraemia does not progress to amicrofilaraemic lymphatic disease.

BACKGROUND: In lymphatic filariasis due to Wuchereria bancrofti infections, the relationship between the natural course of infection and development of clinical disease remains controversial. The two hypotheses that are widely considered are (1) microfilaraemia represents an early stage of infection which progresses to amicrofilaraemic clinical disease and (2) microfilaraemia and clinical disease are two sequentially unrelated independent entities of the filarial infection and disease. Aim To determine whether microfilaraemic individuals develop lymphatic disease. METHODS: The study was conducted in Sri Lanka during the period 1982 to 1998. There were two components, firstly a cross-sectional study and then a longitudinal study. Microfilaraemia was determined by microscopic examination of night blood films. Microfilaraemia associated anti-filarial antibodies were determined by ELISA. Clinical examinations were performed to determine if the test subjects had evidence of acute and chronic lymphoedema. RESULTS: Two major observations were made. First, the incidence and development of adenolymphangitis and lymphoedema in microfilaraemic individuals were very rare and these subjects maintained asymptomatic microfilaraemic status for very long periods of time. Second, in contrast to microfilaraemic subjects, the incidence and development of lymphangitis and lymphoedema were significantly higher in amicrofilaraemic anti-filarial antibody-positive subjects. CONCLUSION: Microfilaraemia does not represent a precondition to development of clinical disease (except male genital involvement).

Mycobacterium tuberculosis in the United Arab Emirates: evidence of local transmission with unique strains.

A retrospective epidemiological analysis was performed of Mycobacterium tuberculosis infections in the Al Ain Medical District, Al Ain, United Arab Emirates (U.A.E.) during the period 1995-2000. The mean incidence for the study period was 7.1%, more than 3 times that reported for the period 1983-1992 (2.1%). For the years 1997 through 2000, the highest incidences (approximately 5-7% of tested) were from health care facilities that cater exclusively for citizens and long-term residents of the U.A.E. Corresponding rates for the immigrant visa applicants (non-citizens) were lower and showed a dramatic decrease from approximately 18% in 1995 to approximately 2% in 2000. Most importantly, the number of multidrug-resistant cases showed an increase from 1.4% during the period August 1997-December 1998 to 8.5% during the period January 1999-July 2000. Analysis of 7 different isolates by restriction fragment length polymorphism (RFLP) showed RFLP patterns that did not match > 4000 individual patterns from 32 countries, suggesting the possible presence of M. tuberculosis strains unique to the U.A.E. Our data demonstrate local transmission of M. tuberculosis in the Al Ain Medical Region of the U.A.E.

Upregulation of a raf kinase and a DP-1 family transcription factor in epidermal growth factor (EGF) stimulated filarial parasites.

Previously we have shown that in the filarial parasite Brugia malayi, stimulation with murine epidermal growth factor (EGF) upregulated the expression of the nuclear GTPase, Ran. In this paper we provide further evidence that filarial parasites possess the ability to respond to mammalian EGF. Stimulation of B. malayi microfilariae with EGF increased transcription of a Raf kinase, increased the physical interaction between Ran and at least eight unidentified proteins, abolished the association of a putative EGF receptor with the nuclear GTPase Ran and enhanced phosphorylation of native microfilarial proteins. In the cattle filarial parasite Setaria digitata, stimulation of adult worms with EGF was probably responsible for up-regulation of a DP-1 family transcription factor. These data suggest that filarial parasites possess the ability to respond to mammalian EGF and that mammalian growth factors may regulate developmental maturation of filarial parasites.

Evaluation of PCR-based methods for the diagnosis of infection in bancroftian filariasis.

The value of the polymerase chain reaction (PCR) in the diagnosis of Wuchereria bancrofti infection was evaluated in comparison to microscopical examination of night blood smears, Nuclepore filtration, serology and ultrasonography. No correlation was found between PCR-based deoxyribonucleic acid (DNA) probing and serology. We did not find any evidence of free filarial DNA in either blood plasma or chylocoele fluid. We conclude that the 2 PCR-based techniques evaluated are not more sensitive than Nuclepore filtration for detection of W. bancrofti microfilaraemia, need at least 1 intact microfilaria in the volume of blood used for DNA extraction, and were much inferior to ultrasonography for detection of amicrofilaraemic adult worm carriers.

Evaluation of recombinant chitinase and SXP1 antigens as antimicrofilarial vaccines.

Prior studies indicate that a microfilarial stage-specific chitinase is a possible candidate antigen for a transmission-blocking vaccine against Brugian filariasis. The antigen is a functional enzyme that progressively appears as microfilariae mature and become able to infect and develop in a susceptible mosquito vector. It is recognized by a monoclonal antibody that reduces microfilaremia in infected animals and by a subset of sera from infected persons who remain amicrofilaremic. Immunization of jirds with recombinant chitinase induced partial protection against microfilaremia resulting from subsequent infection with Brugia malayi, but did not reduce adult worm burdens. Vaccination was much less effective when administered during the prepatent stage of infection and was ineffective when given to microfilaremic jirds. The protective epitope appears to be located close to the carboxy terminus of the chitinase molecule. Immunization of jirds with SXP1, an antigen present in multiple worm stages, also reduced microfilaremia and, in some experiments, adult worm burdens, but hyperimmunization with a recombinant filarial myosin was not protective. These observations indicate that the relative timing of immunization and infection is an important factor in the efficacy of antimicrofilarial vaccines.

The 1996 Lindberg Award. Calcium antagonists alter cell shape and induce procollagenase synthesis in keloid and normal human dermal fibroblasts.

Fibroblast cytomorphology is tightly coupled to phenotypic expression, particularly as it relates to extracellular matrix protein synthesis and degradation. We have observed that calcium antagonists, such as verapamil and trifluoperazine, depolymerize actin filaments and alter fibroblast cell shape from bipolar to spherical. Characteristically, the depolymerization of actin filaments, which mediates the cell shape change, turns on procollagenase gene expression in normal human skin fibroblasts. We have found the same effects of calcium antagonists on cell shape, cytoskeletal components, and induction of procollagenase in the keloid fibroblasts of three cell lines, CB792, CW792, and WT949. Rounded cells were seen in 74.8% of verapamil-treated and 86.7% of trifluoperazine-treated cells, whereas only 1.1% of the control cells were spherical. The percentage of cells that synthesized collagenase in the control, verapamil-treated, and trifluoperazine-treated groups was 3.8%, 42.8%, and 53.4%, respectively. Approximately 60% of rounded cells exhibited increased collagenase synthesis when the cells were treated with a calcium antagonist. These results indicate considerable heterogeneity in the phenotypic response to morphologic change. The amount of procollagenase synthesized in a cell was estimated by the fluorescence intensity of the fluorescein-labeled antibody. The normalized fluorescence intensity of procollagenase in the control cells was about 2 to 2.6 times that of background. In contrast, the normalized fluorescence intensity of procollagenase in the calcium antagonist-treated cells was about 2.4 to 12 times that of background. This high intensity level indicates an increase in procollagenase production in the calcium antagonist-treated cells. Calcium green dye used to study cytosolic calcium revealed that after cells were treated with verapamil, the cytosolic calcium ion concentration first increased and then decreased. The change of cytosolic calcium ion concentration may be related to the depolymerization of actin filaments and the alteration of cell shape.

Rapid regulation of neurite outgrowth and retraction by phospholipase A2-derived arachidonic acid and its metabolites.

Arachidonic acid and lipoxygenase metabolites have been proposed to act as retrograde synaptic messengers and as early mediators of neuronal injury, but few studies have analyzed their roles in controlling neurite behavior within a time window of minutes to hours. Phospholipase A2 inhibitors (BPB, ONO-RS-082, quinacrine and AACOCF3) and the lipoxygenase inhibitor AA861 delayed the initial outgrowth of NG108-15 cell neurites on laminin. Inhibitors of diacylglycerol lipase (RHC 80267), cyclooxygenase (indomethacin) and free radicals (N-acetyl cysteine and vitamin E) did not produce similar effects. Phospholipase A2 and lipoxygenase inhibitors also prevented acute neurite retraction in response to lysophosphatidic acid and eight other agents tested, and decreased F-actin staining at cell margins. Conversely, exogenous arachidonic acid (1 microM) enhanced the responses of neurites in outgrowth and retraction assays. Phospholipase A2 and lipoxygenase pathways appear to have a general role in maintaining the ability of neurites to respond rapidly to external stimuli, possibly via regulating the ability of the cytoskeleton to remodel.

Neuroglian-mediated cell adhesion induces assembly of the membrane skeleton at cell contact sites.

The protein ankyrin links integral membrane proteins to the spectrin-based membrane skeleton. Ankyrin is often concentrated within restricted membrane domains of polarized epithelia and neurons, but the mechanisms responsible for membrane targeting and its segregation within a continuous lipid bilayer remain unexplained. We provide evidence that neuroglian, a cell adhesion molecule related to L1 and neurofascin, can transmit positional information directly to ankyrin and thereby polarize its distribution in Drosophila S2 tissue culture cells. Ankyrin was not normally associated with the plasma membrane of these cells. Upon expression of an inducible neuroglian minigene, however, cells aggregated into large clusters and ankyrin became concentrated at sites of cell-cell contact. Spectrin was also recruited to sites of cell contact in response to neuroglian expression. The accumulation of ankyrin at cell contacts required the presence of the cytoplasmic domain of neuroglian since a glycosyl phosphatidylinositol-linked form of neuroglian failed to recruit ankyrin to sites of cell-cell contact. Double-labeling experiments revealed that, whereas ankyrin was strictly associated with sites of cell-cell contact, neuroglian was more broadly distributed over the cell surface. A direct interaction between neuroglian and ankyrin was demonstrated using yeast two-hybrid analysis. Thus, neuroglian appears to be activated by extracellular adhesion so that ankyrin and the membrane skeleton selectively associate with sites of cell contact and not with other regions of the plasma membrane.