Emerging Genomic Trends on Rabies Virus in Davao Region, Philippines, 2018-2021.

Rabies, caused by the rabies virus (RABV), remains a significant public health issue in the Philippines despite efforts to control it. To eliminate rabies by 2030, effective surveillance strategies are crucial. In this study, we examined RABV evolution and phylodynamics in the Davao Region using genome sequences from Davao City and nearby provinces. We adapted the RABV ARTIC Protocol for Oxford Nanopore High-Throughput Sequencing to optimize workflow efficiency under limited resources. Comparing new virus samples collected from June 2019 to June 2021 (n = 38) with baseline samples from June 2018 to May 2019 (n = 49), new sub-clades were observed in the phylogenetic tree, suggesting divergence from older variants that were previously undetected. Most of the new viruses belonged to the Asian SEA4_A1.1.1 lineage, but new (SEA4_B1 and SEA4_B1.1) and emerging (SEA4_B1.1_E1) lineages that have never been reported in the Philippines were also identified. The baseline study reported phylogeographic clustering of RABV isolates from the same areas. However, this pattern was disrupted in the current biosurveillance, with variants detected in areas outside the original cluster. Furthermore, our findings revealed significant transmission routes between Davao City and neighboring provinces, contrasting with the predominantly intra-city transmission observed in the baseline study. These results underscore the need for ongoing and timely genomic surveillance to monitor genetic diversity changes and the emergence of novel strains, as well as to track alterations in transmission pathways. Implementing cost-effective next-generation sequencing workflows will facilitate the integration of genomic surveillance into rabies control programs, particularly in resource-limited settings. Collaborations between different sectors can empower local laboratories and experts in genomic technologies and analysis.

Periodontal bacteria DNA findings in human cardiac tissue - Is there a link of periodontitis to heart valve disease?

BACKGROUND: The aim of the study was to detect periodontal pathogens DNA in atrial and myocardial tissue, and to investigate periodontal status and their connection to cardiac tissue inflammation. METHODS: In 30 patients, biopsy samples were taken from the atrium (A) and the ventricle myocardium (M) during aortic valve surgery. The dental examination included the dental and periodontal status (PS) and a collection of a microbiological sample. The detection of 11 periodontal pathogens DNA in oral and heart samples was carried out using PCR. The heart samples were prepared for detecting the LPS-binding protein (LBP), and for inflammation scoring on immunohistochemistry (IHC), comprising macrophages (CD68), LPS-binding protein receptor (CD14), and LBP (big42). RESULTS: 28 (93%) patients showed moderate to severe periodontitis. The periodontal pathogens in the oral samples of all patients revealed a similar distribution (3-93%). To a lesser extent and with a different distribution, these bacteria DNA were also detected in atrium and myocardium (3-27%). The LBP was detected in higher amount in atrium (0.22±0.16) versus myocardium (0.13±0.13, p=0.001). IHC showed a higher inflammation score in atrial than myocardial tissue as well as for CD14, CD68 and for LBP. Additional, periodontal findings showed a significant correlation to CD14 and CD68. CONCLUSION: The results provide evidence of the occurrence of oral bacteria DNA at the cardiac tissue, with a different impact on atrial and myocardial tissue inflammation. Influence of periodontal findings was identified, but their relevance is not yet distinct. Therefore further clinical investigations with long term implication are warranted.

[Post-denervation renal artery stenosis - a matter of concern?]. / Les sténoses artérielles rénales post-dénervation - une problématique préoccupante ?

Renal denervation, an invasive technique indicated in resistant hypertension patients insufficiently controlled by antihypertensive drugs, has a good safety profile. However, an increasing number of post-denervation renal artery stenosis cases has recently been reported. We describe the case of a 49-year-old woman with resistant hypertension who was referred to our university hypertension center for renal sympathetic denervation. Her daily treatment included six antihypertensive drugs. CT angiography prior to denervation showed no renal artery stenosis or vessel wall lesions. A standard renal denervation procedure using the St Jude protocol was performed. After an initial improvement in blood pressure profile, she presented with a blood pressure impairment at 3 months after renal denervation leading to the diagnosis of a severe right renal artery stenosis.

Cytocompatibility of brushite and monetite cell culture scaffolds made by three-dimensional powder printing.

This study investigated the cytocompatibility of low-temperature direct 3-D printed calcium phosphate scaffolds in vitro. The fabrication of the scaffolds was performed with a commercial 3-D powder printing system. Diluted phosphoric acid was printed into tricalcium phosphate powder, leading to the formation of dicalcium phosphate dihydrate (brushite). Hydrothermal conversion of the brushite matrices led to the formation of dicalcium phosphate anhydrous (monetite). The biocompatibility was investigated using the osteoblastic cell line MC3T3-E1. Cell viability and the expression of alkaline phosphatase served as parameters. The culture medium was analyzed for pH value, concentration of free calcium and phosphate ions and osteocalcin. Both types of scaffolds showed a considerable increase of cell proliferation and viability; the monetite matrices were a little inferior compared with the brushite ones. The activity of alkaline phosphatase showed a similar pattern. Optical and electron microscopy revealed an obvious cell growth on the surface of both materials. Analysis of the culture medium showed minor alterations of pH value within the physiological range. The concentrations of free calcium and phosphate ions were obviously different among brushite and monetite cultures, due to their different solubility. The content of osteocalcin of the culture medium was reduced by the printed scaffolds due to adsorption. We conclude that the powder printed brushite and monetite matrices have a suitable biocompatibility for their use as cell culture scaffolds. Both materials enable osteoblastic cells in vitro to proliferate and differentiate due to the expression of typical osteoblastic markers.

Niche-specificity and the variable fraction of the Pectobacterium pan-genome.

We compare genome sequences of three closely related soft-rot pathogens that vary in host range and geographical distribution to identify genetic differences that could account for lifestyle differences. The isolates compared, Pectobacterium atrosepticum SCRI1043, P. carotovorum WPP14, and P. brasiliensis 1692, represent diverse lineages of the genus. P. carotovorum and P. brasiliensis genome contigs, generated by 454 pyrosequencing ordered by reference to the previously published complete circular chromosome of P. atrosepticum genome and each other, account for 96% of the predicted genome size. Orthologous proteins encoded by P. carotovorum and P. brasiliensis are approximately 95% identical to each other and 92% identical to P. atrosepticum. Multiple alignment using Mauve identified a core genome of 3.9 Mb conserved among these Pectobacterium spp. Each core genome is interrupted at many points by species-specific insertions or deletions (indels) that account for approximately 0.9 to 1.1 Mb. We demonstrate that the presence of a hrpK-like type III secretion system-dependent effector protein in P. carotovorum and P. brasiliensis and its absence from P. atrosepticum is insufficient to explain variability in their response to infection in a plant. Additional genes that vary among these species include those encoding peptide toxin production, enzyme production, secretion proteins, and antibiotic production, as well as differences in more general aspects of gene regulation and metabolism that may be relevant to pathogenicity.

[Why screening for a renal artery stenosis?]. / Pourquoi dépister une sténose artérielle rénale?

Diagnosis of renal artery stenosis (RAS) should be discussed in numerous clinical situations including refractory high blood pressure (HBP), HBP in a polyvascular patient, degradation of renal function following renin angiotensin inhibitor or flash pulmonary edema. Ultrasound-doppler coupled with gadolinium-enhanced MR or CT angiography has proven adequate for most patients with RAS. Digital subtraction angiography should be limited to revascularisation procedures. Functional testing are not sensitive or specific enough because the degree of renin activation differs widely among patients with RAS. Renal percutaneous angioplasty induces a light to moderate decrease in blood pressure, has no effect on renal function but allows to reduce the number of anti-hypertensive drugs. Stenting completed angioplasty is worthwhile in most patients with atherosclerotic RAS. ACE inhibitors decrease mortality and increase renal function in patients with RAS.

Aerosol optical depth, aerosol composition and air pollution during summer and winter conditions in Budapest.

The dependence of aerosol optical depth (AOD) on air particulate concentrations in the mixing layer height (MLH) was studied in Budapest in July 2003 and January 2004. During the campaigns gaseous (CO, SO(2), NO(x), O(3)), solid components (PM(2.5), PM(10)), as well as ionic species (ammonium, sulfate and nitrate) were measured at several urban and suburban sites. Additional data were collected from the Budapest air quality monitoring network. AOD was measured by a ground-based sun photometer. The mixing layer height and other common meteorological parameters were recorded. A linear relationship was found between the AOD and the columnar aerosol burden; the best linear fit (R(2)=0.96) was obtained for the secondary sulfate aerosol due to its mostly homogeneous spatial distribution and its optically active size range. The linear relationship is less pronounced for the PM(2.5) and PM(10) fractions since local emissions are very heterogeneous in time and space. The results indicate the importance of the mixing layer height in determining pollutant concentrations. During the winter campaign, when the boundary layer decreases to levels in between the altitudes of the sampling stations, measured concentrations showed significant differences due to different local sources and long-range transport. In the MLH time series unexpected nocturnal peaks were observed. The nocturnal increase of the MLH coincided with decreasing concentrations of all pollutants except for ozone; the ozone concentration increase indicates nocturnal vertical mixing between different air layers.

Role of pre-infarction angina and inflammatory status in the extent of microvascular obstruction detected by MRI in myocardial infarction patients treated by PCI.

BACKGROUND/OBJECTIVES: The extent of microvascular obstruction (MVO) during myocardial infarction referred to as the "no-reflow phenomenon", may determine myocardial damage. Our study aimed to investigate the incidence and the influencing factors of MVO in patients with ST-elevation myocardial infarction (STEMI) treated by primary percutaneous intervention (PCI). PATIENTS, METHODS: Using contrast-enhanced MRI, microvascular obstruction was defined as early hypoenhancement. Contrast defects were scored from 0 (no hypoenhancement) to 3 (strong hypoenhancement). 50 patients (56+/-11 years) with STEMI underwent PCI. Contrast-enhanced MRI (6+/-2 days after STEMI) and biochemical parameters were evaluated. RESULTS: Microvascular obstruction (score 1 to 3) was observed in 90% of the patients and major microvascular obstruction (score 2-3) in 54%. In univariate analysis, leukocytes and CRP levels were associated with MVO, whereas pre-infarction angina and prior medication by aspirin or calcium channel antagonist appeared protective. Microvascular obstruction intensity positively correlated with baseline inflammation status assessed by C-reactive protein and leukocytes (rho=0.43 and rho=0.44; p=0.003), the peak of CK (rho=0.56; p=0.01) or Troponin I (rho=0.59; p=0.01) and negatively correlated with LVEF (rho=-0.44; p=0.002). Multivariate analysis identified the absence of pre-infarction angina as the only independent predictor for microvascular obstruction (odds ratio, 8.35, 95% confidence interval 1.27-54.71; p=0.027). CONCLUSION: MRI-detected microvascular obstruction has a high incidence in patients with STEMI treated by primary PCI and determines post-MI LVEF even in patients with post PCI TIMI 3 flow score. Pre-infarction angina appears to be an independent determinant of the extent of MVO detected by MRI.

[Seasonal fluctuations and influence of nutrition on macular pigment density]. / Jahreszeitliche Schwankungen und Einfluss der Ernährung auf die makuläre Pigmentdichte.

PURPOSE: We have previously reported on measuring macular pigment density (MPD) with a scanning laser ophthalmoscope (HRA, Heidelberg Engineering, Heidelberg, Germany). This study war undertaken to evaluate the variation of MPD over a period of 1 year in healthy subjects. METHOD: We used autofluorescence images recorded with a HRA to evaluate MPD with a 2 degrees circle centered on the fovea. Healthy subjects were included in the study and MPD measurements were repeated every 2 months over a period of 1 year. RESULTS: We included a total of 30 healthy subjects aged 19-34 years (mean: 23+/-2 years). Mean MPD at time point 1 was 0.215+/-0.056 density units (DU), at time point 2 0.235+/-0.051 DU, at time point 3 0.218+/-0.055 DU, at time point 4 0.228+/-0.057 DU, at time point 5 0.225+/-0.053 DU, and at time point 6 0.203+/-0.050 DU. The statistical analysis revealed no significant variation of MPD over the follow-up period of 1 year. CONCLUSION: This study demonstrates that MPD shows no variation over a period of 1 year in healthy subjects.

Automated oral hygiene self-care devices: making evidence-based choices to improve client outcomes.

Dental hygienists have historically been the primary promoters of health, prevention, and wellness in dental practice. With the large number of automated self-care devices available, many clients expect their dental hygienists to be experts and advisors on such purchases. It is vitally important for dental hygienists to understand and evaluate the available clinical research findings, and along with their clients, apply them in a collaborative decision making process. The purpose of this course is for dental hygienists to learn the scientific and practical implications of evidence-based care in the selection of automated oral hygiene self-care devices. Measurable clinical outcomes in plaque removal, reduction in gingivitis, bleeding, probing depth, periodontal pathogens, and the role of inflammatory mediators will be discussed. Automated self-care products currently available will be identified. Successful completion of the course objectives will be assessed by a post-test, to be completed by the learner after reading the article in its entirety. Two hours of continuing education course credit will be awarded following successful completion of the post test.

Epiretinal membranes after extracapsular cataract surgery(1).

PURPOSE: To determine whether uneventful extracapsular cataract extraction (ECCE) with posterior chamber intraocular lens (PC IOL) implantation induces epiretinal membranes (ERMs). SETTING: Private practice, with statistical evaluation at a university department in Germany. METHODS: This prospective study comprised 296 consecutive patients (332 eyes) who had uneventful ECCE with PC IOL implantation for senile cataract. Eyes were examined within 2 weeks preoperatively and 2 weeks and 6, 9, and 12 months postoperatively. The macula was examined with a 78.0 diopter Volk lens. The main outcome measure was the presence of ERM. The membranes were classified as present when cellophane macular reflex (CMR) or macular pucker (MP) was observed. RESULTS: The median age of the patients was 78 years (range 50 to 97 years). At baseline, ERM was present in 49 of 332 eyes (14.8%), 40 (12.1%) with CMR and 9 (2.7%) with MP. Six months postoperatively, ERM was present in 84 of 332 eyes (25.3%), 72 (21.7%) with CMR and 12 (3.6%) with MP. The difference between the baseline and 6 month incidence was significant (P <.001). All new cases of ERM were of the CMR type. One year after surgery, ERM was present in 54 of 198 eyes (27.3%), 47 (23.7%) with CMR and 7 (3.5%) with MP. The ERMs rarely influenced visual acuity; however, 6 of 12 eyes (50.0%) with MP and 13 of 72 (18.1%) with CMR had metamorphopsia. CONCLUSIONS: The prevalence of ERM increased by 71.4% during the first 6 months after uneventful ECCE with PC IOL implantation. Thereafter, the prevalence remained about the same. Newly formed ERMs were probably induced by the uneventful surgery. Although new-onset membranes usually do not preclude good visual acuity, they can cause metamorphopsia postoperatively.

Determination of cutin-bound residues of chlorothalonil by immunoassay.

An indirect competitive enzyme-linked immunosorbent assay (ELISA) was used to determine photochemically cutin-bound residues of chlorothalonil in enzymatically isolated tomato and apple fruit cuticles. The samples were spiked, irradiated, exhaustively extracted, and depolymerized with boron trifluoride complex resulting in a soluble depolymerisate. With this procedure, the ELISA could be calibrated with free target molecules for the quantification of originally bound chlorothalonil residues. In fruit cuticles that were irradiated for 8 h by simulated sunlight, 0.030 and 0.068 mg/g photoinduced cutin-bound residues of wax-free cuticles (calculated as chlorothalonil) were determined for tomatoes and apples, respectively. For the used antibody mAb chl. 4/11, cross-reactivities with derivatives of chlorothalonil simulating different types of cuticle-bound residues are given and discussed.

Myocardial flow reserve parametric map, assessed by first-pass MRI compartmental analysis at the chronic stage of infarction.

Regional myocardial flow and flow reserve (MFR) were assessed by compartmental analysis of Gd-enhanced MRI first-pass data in 7 patients with atypical chest pain, and in 15 patients with previous transmural myocardial infarction. The FE product (Flow x Extraction coefficient), derived from the modified Kety equation, was measured in regions corresponding to the Tetrofosmine-SPECT fixed defect and in remote normal regions. The FE product at rest and hyperemic FE product were similar in healed revascularized tissues (70.5 +/- 15.6 and 112.5 +/- 19.5 ml/mn/100 g, respectively) and in normal myocardium (76.2 +/- 18.3 and 142.2 +/- 33.0, respectively). In contrast, the FE index (48.8 +/- 15.2 and 60.7 +/- 18.0, respectively, P < 0.01 versus the two previous groups) and the MFR (1.27 +/- 0.20 vs. 1.91 +/- 0.29 in normal regions) were reduced in healed fibrotic tissues when the infarct-related artery remained occluded. Myocardial flow reserve maps allowed correct identification of regions corresponding to an occluded infarct-related artery.

[Cartography of absolute myocardial perfusion with magnetic resonance imaging. Methods and results]. / Cartographie de la perfusion myocardique absolue avec imagerie par résonance magnétique. Méthodes et résultats.

Ultra-rapid dynamic MRI (one image per heart beat) can follow the progression of the intra-myocardial signal during the first passage of diffusable gadolinium chelates injected as a bolus through a peripheral vein. A quantitative evaluation of myocardial perfusion is possible using a compartmental model of analysis. Absolute myocardial flow can be measured at rest and during hyperaemia induced by dipyridamole. It is possible to associate functional mapping, corresponding to parametric images of the flow indices, to the global evaluation. The ratio between the values obtained during hyperaemia and under basal conditions correspond to the myocardial reserve. The principles, results and limitations of this method are discussed in the light of published results, underlining the advantages of absolute flow measurement and of the differences between the results of MRI and myocardial scintigraphy.

New approaches in the management of choroidal neovascular membrane in age-related macular degeneration.

Age-related macular degeneration (AMD) is a leading cause of blindness in the elderly population. The prevalence is reported to be 1.2-1.4% in several population-based epidemiological studies. Currently 25-30 million people worldwide are blind due to AMD. With the aging world population it is bound to increase significantly, and could become a significant public health problem in next two decades, with serious socio-economic implications. Several strategies are today available to treat the wet form of AMD, which is responsible for significant visual loss. These were until recently confined to laser photocoagulation, and subretinal surgery, but today two other modalities, namely, radiation and photodynamic therapy, are available. These treatment modalities however, are aimed at preservation of vision only, and not at reversing the process of the disease. Further research on antiangiogenic drugs and gene therapy could significantly help AMD patients.

Differentiation of chromatin during DNA elimination in Euplotes crassus.

In Euplotes crassus, most of the micronuclear genome is eliminated during formation of a transcriptionally active macronucleus. To understand how this is mediated throughout the genome, we have examined the chromatin structure of the macronucleus-destined sequences and Tec transposons, which are dispersed in 15,000 copies in the micronuclear genome and completely eliminated during formation of the macronuclear genome. Whereas the macronucleus-destined sequences show a typical pattern of nucleosomal repeats in micrococcal nuclease digests, the Tec element chromatin structure digests to a nucleosome-like repeat pattern that is not typical: the minimum digestion products are approximately 300-600 base pairs, or "subnucleosomal," in size. In addition, the excised, circular forms of the Tec elements are exceedingly resistant to nucleases. Nevertheless, an underlying nucleosomal structure of the Tec elements can be demonstrated from the size differences between repeats in partial micrococcal nuclease digests and by trypsin treatment of nuclei, which results in mononucleosome-sized products. Characterization of the most micrococcal nuclease-resistant DNA indicates that micronuclear telomeres are organized into a chromatin structure with digestion properties identical to those of the Tec elements in the developing macronucleus. Thus, these major repetitive sequence components of the micronuclear genome differ in their chromatin structure from the macronuclear-destined sequences during DNA elimination. The potential role of developmental stage-specific histone variants in this chromatin differentiation is discussed.

[Perioperative prevalence of epiretinal membranes in eyes with senile cataract]. / Perioperative Prävalenz von epiretinalen Membranen bei Augen mit seniler Katarakt.

BACKGROUND: Results from epidemiologic studies suggest that cataract extraction is a risk-factor for the formation of epiretinal membranes (EM). PATIENTS AND METHODS: 250 consecutive eyes which underwent uneventful phakoemulsification with implantation of a posterior chamber intraocular lens were examined within two weeks prior and an two weeks after surgery for the presence of epiretinal membranes. Cellophane macular reflex (CMR) was distinguished from macular pucker (MP). RESULTS: We found an increase in the prevalence of epiretinal membranes from preoperatively 12/250 (4.8%) to post-operatively 33/250 (13.2%). The prevalence of CMR increased from 8/250 (3.2%) to 27/250 (10.8%). The prevalence of MP remained almost the same 4/250 (1.6%) prior and 6/250 (2.4%) after surgery. CONCLUSIONS: Increased prevalence of epiretinal membranes following cataract extraction observed in epidemiological studies can be explained by easier detection of such membranes in pseudophakic eyes where the observer gets a clear view of the fundus. Therefore it is questionable whether uneventful extracapsular cataract extraction can induce the formation of epiretinal membranes.

Characterization of the Euplotes crassus macronuclear rDNA and its potential as a DNA transformation vehicle.

We have cloned the macronuclear linear DNA molecule carrying the ribosomal RNA genes from the ciliated protozoan Euplotes crassus. DNA sequence analysis was carried out to locate coding regions and to determine whether sequences that have been mutated to confer antibiotic resistance are conserved in the E. crassus genes. The beginning and end of the primary transcript were mapped. In order to determine whether conserved sequences that might serve as replication origins were present, the 5' and 3' non-coding sequences from E. crassus were compared to the corresponding sequences from the macronuclear linear rDNA molecules from the following euplotid species: Euplotes vannus, Euplotes minuta, Euplotes raikovii and Euplotes rariseta. A DNA transformation construct was made by generating a putative anisomycin resistant mutation along with a mutation generating a restriction site polymorphism. Microinjection of the construct into the developing macronucleus of mated cells resulted in exconjugant cell lines with increased resistance to anisomycin. The injected rDNA with the restriction site polymorphism is detectable in the anisomycin resistant cells and appears to represent a minor fraction of the rDNA.

Differential expression of linker histone variants in Euplotes crassus.

Two genes have been cloned from the ciliate Euplotes crassus that encode proteins with sequence similarity to the linker histones from a variety of organisms. One gene, H1-1, is present on a 1.3-kb macronuclear DNA molecule and encodes a 16.2- kDa protein. The second gene, H1-2, is present on a 0.7-kb DNA molecule and encodes an 18.8-kDa protein. Both H1-1 and H1-2 are expressed in vegetative cells, but the two genes exhibit very different patterns of expression during macronuclear development. H1-1 transcripts accumulate during conjugation and during the final rounds of DNA amplification. H1-2 transcripts accumulate after the onset of polytene chromosome formation and remain high throughout the remainder of macronuclear development. H1-1 is the major perchloric-acid-soluble protein from macronuclei. The pattern of gene expression and the macronuclear location of the H1-1 protein indicate that H1-1 is the predominant linker histone in vegetative macronuclei.