COVID-19 infection in pediatric solid organ transplant patients.

BACKGROUND: Adult SOT recipients with COVID-19 have higher mortality rates when compared to general population. There is paucity of data on outcomes in pediatric SOT recipients. METHODS: This is a cross-sectional study investigating the prevalence of COVID-19 infection and outcomes in pediatric SOT (heart, liver, and kidney) recipients. We extracted demographic and clinical characteristics and COVID-19 testing (PCR or [Ab] test) results from medical records. Clinical characteristics were compared between patients who were positive for COVID-19 (PCR or Ab) and those who did not, using Mann-Whitney, Student's t test, or chi-square test. p value <.05 was statistically significant. RESULTS: A total of 108 SOT recipients with a median age of 13.1 (8.4, 17.8) years and median 4.2 (2.7, 7.9) years from transplant were checked for COVID-19 via a PCR or Ab test. A positive PCR was confirmed in 10 patients (9.3%), while 12 patients (11.1%) were positive for COVID-19 Ab. The patients who tested positive in our cohort were 9/50 (18%) heart, 6/68 (8.8%) kidney, and 7/50 (14%) liver transplant recipients. There were no differences in the clinical characteristics between patients with and without COVID-19 infection. All patients were either asymptomatic (50%) or had self-limiting symptoms. No changes were made to the immunosuppressive regimen. Only one patient was hospitalized and none had an oxygen requirement. CONCLUSIONS: In our cohort of pediatric SOT recipients, COVID-19 infection was asymptomatic or mild. This data may aid clinicians in counseling patients and families in this increased-risk population.

Detection of graft fibrosis by vibration-controlled transient elastography in pediatric liver transplant recipients.

Pediatric liver transplant recipients are at risk of developing graft fibrosis which can affect patient survival. VCTE is a non-invasive tool that measures LSM and has been shown to correlate with hepatic fibrosis. The aim of this study was to therefore evaluate the ability of LSM to predict fibrosis in pediatric liver transplant recipients with different graft types. We performed a cross-sectional study evaluating LSM of 28 pediatric liver transplant recipients who underwent a total of 20 liver biopsies within 1 month of LSM. LSM was compared to liver histology as well as graft type: WL or PL. The median LSM of all post-transplant patients was 5.6 kPa (range = 2.7-18.3). There was a statistically significant correlation between LSM and METAVIR fibrosis score (P = .001) and LAF score (P < .001). There was no difference in LSM between graft type (P = .088). The AUROC curve for LSM predicting any significant fibrosis (F ≥ 2) was 0.863. A cutoff value of 7.25 had a sensitivity of 71%, specificity of 100%, NPV of 87%, and PPV of 100% for significant fibrosis. LSM by VCTE is feasible in pediatric liver transplant recipients regardless of graft type. We found a significant correlation between LSM and hepatic fibrosis and established a cutoff value that may help determine which patients warrant further evaluation for graft fibrosis.

Adenomyosis: A Sonographic Diagnosis.

Adenomyosis is a common benign uterine condition and a frequent cause of pelvic pain in premenopausal women. Transvaginal US is now considered the primary imaging modality for the diagnosis of adenomyosis, and thus radiologists should be familiar with its sonographic appearance. US findings can be divided into three categories, which parallel the histology of adenomyosis: (a) ectopic endometrial glands and stroma, (b) muscular hyperplasia/hypertrophy, and (c) increased vascularity. Ectopic endometrial glands manifest as echogenic nodules and striations, radiating from the endometrium into the myometrium. When the glands contain fluid, myometrial cysts and fluid-filled striations may be visible at US. Muscular hyperplasia and hypertrophy cause focal or diffuse myometrial thickening and globular uterine enlargement, often with thin "venetian blind" shadows. The combination of these findings results in a heterogeneous myometrium, with blurring of the endometrial border. Adenomyosis increases uterine vascularity, depicted as a pattern of penetrating vessels at color Doppler US. Other US techniques that are helpful in the diagnosis of adenomyosis include obtaining cine clips and coronal reformatted images, both of which can survey the entire endometrial-myometrial border, and performing saline-infusion sonohysterography, during which ectopic glands frequently fill with either air or fluid. While most cases of adenomyosis develop spontaneously, there are specific inciting causes that include tamoxifen use, postendometrial ablation syndrome, and deep-infiltrating endometriosis. Mimics of adenomyosis include leiomyomas, uterine contractions, neoplasms, and vascular malformations. This article reviews the pathophysiology of adenomyosis and correlates it with the US findings, highlights specific causes of adenomyosis, and describes how to distinguish this common diagnosis from a variety of mimics. Online supplemental material is available for this article. ©RSNA, 2018.

The extensor carpi ulnaris pseudolesion: evaluation with microCT, histology, and MRI.

OBJECTIVE: To determine if magic angle plays a role in apparent central increased signal intensity of the distal extensor carpi ulnaris tendon (ECU) on MRI, to see if histologic findings of tendon degeneration are associated with increased T1 or T2 tendon signal on MR imaging, and to determine the prevalence of the ECU "pseudolesion". MATERIALS AND METHODS: A standard 3 Tesla protocol was utilized to scan ten cadaveric wrists. A 40 mm length of 10 ECU and four extensor carpi radialis brevis (ECRB) tendons were immersion fixed before microCT scanning. Staining with Alcian blue, Masson's trichrome and Safranin O was performed before light microscopy. Fifty clinical wrist MRIs were also reviewed for the presence of increased T1 and/or T2 signal. RESULTS: Central increased T1 and/or T2 signal was observed in 9 of 10 cadaveric ECU tendons, but not in ECRB tendons. MicroCT and histology showed inter-tendinous matrix between the two distal heads of the ECU. Increased mucoid degeneration correlated with increased MRI signal intensity. The tendon fibers were at a maximum of 8.39° to the longitudinal axis on microCT. Clinical MRIs showed increased T1 signal in 6%, increased T2 signal in 8%, increased T1 and T2 signal in 80%, and 6% showing no increased signal. CONCLUSION: Central increased T1 and/or T2 signal in the ECU tendon indicates the presence of normal inter-tendinous ground substance, with increased proteoglycan content (mucoid degeneration) responsible for increased signal intensity. None of the fibers were shown on microCT to approach the magic angle.

Safety and Immunogenicity of Live Viral Vaccines in a Multicenter Cohort of Pediatric Transplant Recipients.

Importance: Live vaccines (measles-mumps-rubella [MMR] and varicella-zoster virus [VZV]) have not been recommended after solid organ transplant due to concern for inciting vaccine strain infection in an immunocompromised host. However, the rates of measles, mumps, and varicella are rising nationally and internationally, leaving susceptible immunocompromised children at risk for life-threating conditions. Objective: To determine the safety and immunogenicity of live vaccines in pediatric liver and kidney transplant recipients. Design, Setting, and Participants: This cohort study included select pediatric liver and kidney transplant recipients who had not completed their primary MMR and VZV vaccine series and/or who displayed nonprotective serum antibody levels at enrollment between January 1, 2002, and February 28, 2023. Eligibility for live vaccine was determined by individual US pediatric solid organ transplant center protocols. Exposures: Exposure was defined as receipt of a posttransplant live vaccine. Transplant recipients received 1 to 3 doses of MMR vaccine and/or 1 to 3 doses of VZV vaccine. Main Outcome and Measure: Safety data were collected following each vaccination, and antibody levels were obtained at 0 to 3 months and 1 year following vaccination. Comparisons were performed using Mann-Whitney U test, and factors associated with development of postvaccination protective antibodies were explored using univariate analysis. Results: The cohort included 281 children (270 [96%] liver, 9 [3%] kidney, 2 [1%] liver-kidney recipients) from 18 centers. The median time from transplant to enrollment was 6.3 years (IQR, 3.4-11.1 years). The median age at first posttransplant vaccine was 8.9 years (IQR, 4.7-13.8 years). A total of 202 of 275 (73%) children were receiving low-level monotherapy immunosuppression at the time of vaccination. The majority of children developed protective antibodies following vaccination (107 of 149 [72%] varicella, 130 of 152 [86%] measles, 100 of 120 [83%] mumps, and 124 of 125 [99%] rubella). One year post vaccination, the majority of children who initially mounted protective antibodies maintained this protection (34 of 44 [77%] varicella, 45 of 49 [92%] measles, 35 of 42 [83%] mumps, 51 of 54 [94%] rubella). Five children developed clinical varicella, all of which resolved within 1 week. There were no cases of measles or rubella and no episodes of graft rejection within 1 month of vaccination. There was no association between antibody response and immunosuppression level at the time of vaccination. Conclusions and Relevance: The findings suggest that live vaccinations may be safe and immunogenic after solid organ transplant in select pediatric recipients and can offer protection against circulating measles, mumps, and varicella.

Functional characterization of transcription factor motifs using cross-species comparison across large evolutionary distances.

We address the problem of finding statistically significant associations between cis-regulatory motifs and functional gene sets, in order to understand the biological roles of transcription factors. We develop a computational framework for this task, whose features include a new statistical score for motif scanning, the use of different scores for predicting targets of different motifs, and new ways to deal with redundancies among significant motif-function associations. This framework is applied to the recently sequenced genome of the jewel wasp, Nasonia vitripennis, making use of the existing knowledge of motifs and gene annotations in another insect genome, that of the fruitfly. The framework uses cross-species comparison to improve the specificity of its predictions, and does so without relying upon non-coding sequence alignment. It is therefore well suited for comparative genomics across large evolutionary divergences, where existing alignment-based methods are not applicable. We also apply the framework to find motifs associated with socially regulated gene sets in the honeybee, Apis mellifera, using comparisons with Nasonia, a solitary species, to identify honeybee-specific associations.

Synthesising Facial Macro- and Micro-Expressions Using Reference Guided Style Transfer.

Long video datasets of facial macro- and micro-expressions remains in strong demand with the current dominance of data-hungry deep learning methods. There are limited methods of generating long videos which contain micro-expressions. Moreover, there is a lack of performance metrics to quantify the generated data. To address the research gaps, we introduce a new approach to generate synthetic long videos and recommend assessment methods to inspect dataset quality. For synthetic long video generation, we use the state-of-the-art generative adversarial network style transfer method-StarGANv2. Using StarGANv2 pre-trained on the CelebA dataset, we transfer the style of a reference image from SAMM long videos (a facial micro- and macro-expression long video dataset) onto a source image of the FFHQ dataset to generate a synthetic dataset (SAMM-SYNTH). We evaluate SAMM-SYNTH by conducting an analysis based on the facial action units detected by OpenFace. For quantitative measurement, our findings show high correlation on two Action Units (AUs), i.e., AU12 and AU6, of the original and synthetic data with a Pearson's correlation of 0.74 and 0.72, respectively. This is further supported by evaluation method proposed by OpenFace on those AUs, which also have high scores of 0.85 and 0.59. Additionally, optical flow is used to visually compare the original facial movements and the transferred facial movements. With this article, we publish our dataset to enable future research and to increase the data pool of micro-expressions research, especially in the spotting task.

Several tetratricopeptide repeat (TPR) motifs of FANCG are required for assembly of the BRCA2/D1-D2-G-X3 complex, FANCD2 monoubiquitylation and phleomycin resistance.

The Fanconi anaemia (FA) FANCG protein is an integral component of the FA nuclear core complex that is required for monoubiquitylation of FANCD2. FANCG is also part of another protein complex termed D1-D2-G-X3 that contains FANCD2 and the homologous recombination repair proteins BRCA2 (FANCD1) and XRCC3. Formation of the D1-D2-G-X3 complex is mediated by serine-7 phosphorylation of FANCG and occurs independently of the FA core complex and FANCD2 monoubiquitylation. FANCG contains seven tetratricopeptide repeat (TPR) motifs that mediate protein-protein interactions and here we show that mutation of several of the TPR motifs at a conserved consensus residue ablates the in vivo binding activity of FANCG. Expression of mutated TPR1, TPR2, TPR5 and TPR6 in Chinese hamster fancg mutant NM3 fails to functionally complement its hypersensitivities to mitomycin C (MMC) and phleomycin and fails to restore FANCD2 monoubiquitylation. Using co-immunoprecipitation analysis, we demonstrate that these TPR-mutated FANCG proteins fail to interact with BRCA2, XRCC3, FANCA or FANCF. The interactions of other proteins in the D1-D2-G-X3 complex are also absent, including the interaction of BRCA2 with both the monoubiquitylated (FANCD2-L) and non-ubiquitylated (FANCD2-S) isoforms of FANCD2. Interestingly, a mutation of TPR7 (R563E), that complements the MMC and phleomycin hypersensitivity of human FA-G EUFA316 cells, fails to complement NM3, despite the mutated FANCG protein co-precipitating with FANCA, BRCA2 and XRCC3. Whilst interaction of TPR7-mutated FANCG with FANCF does appear to be reduced in NM3, FANCD2 is monoubiquitylated suggesting that sub-optimal interactions of FANCG in the core complex and the D1-D2-G-X3 complex are responsible for the observed MMC- and phleomycin-hypersensitivity, rather than a defect in FANCD2 monoubiquitylation. Our data demonstrate that FANCG functions as a mediator of protein-protein interactions and is vital for the assembly of multi-protein complexes including the FA core complex and the D1-D2-G-X3 complex.

Estimation of absolute states of human skeletal muscle via standard B-mode ultrasound imaging and deep convolutional neural networks.

The objective is to test automated in vivo estimation of active and passive skeletal muscle states using ultrasonic imaging. Current technology (electromyography, dynamometry, shear wave imaging) provides no general, non-invasive method for online estimation of skeletal muscle states. Ultrasound (US) allows non-invasive imaging of muscle, yet current computational approaches have never achieved simultaneous extraction or generalization of independently varying active and passive states. We use deep learning to investigate the generalizable content of two-dimensional (2D) US muscle images. US data synchronized with electromyography of the calf muscles, with measures of joint moment/angle, were recorded from 32 healthy participants (seven female; ages: 27.5, 19-65). We extracted a region of interest of medial gastrocnemius and soleus using our prior developed accurate segmentation algorithm. From the segmented images, a deep convolutional neural network was trained to predict three absolute, drift-free components of the neurobiomechanical state (activity, joint angle, joint moment) during experimentally designed, simultaneous independent variation of passive (joint angle) and active (electromyography) inputs. For all 32 held-out participants (16-fold cross-validation) the ankle joint angle, electromyography and joint moment were estimated to accuracy 55 ± 8%, 57 ± 11% and 46 ± 9%, respectively. With 2D US imaging, deep neural networks can encode, in generalizable form, the activity-length-tension state relationship of these muscles. Observation-only, low-power 2D US imaging can provide a new category of technology for non-invasive estimation of neural output, length and tension in skeletal muscle. This proof of principle has value for personalized muscle assessment in pain, injury, neurological conditions, neuropathies, myopathies and ageing.

Objective Analysis of Neck Muscle Boundaries for Cervical Dystonia Using Ultrasound Imaging and Deep Learning.

OBJECTIVE: To provide objective visualization and pattern analysis of neck muscle boundaries to inform and monitor treatment of cervical dystonia. METHODS: We recorded transverse cervical ultrasound (US) images and whole-body motion analysis of sixty-one standing participants (35 cervical dystonia, 26 age matched controls). We manually annotated 3,272 US images sampling posture and the functional range of pitch, yaw, and roll head movements. Using previously validated methods, we used 60-fold cross validation to train, validate and test a deep neural network (U-net) to classify pixels to 13 categories (five paired neck muscles, skin, ligamentum nuchae, vertebra). For all participants for their normal standing posture, we segmented US images and classified condition (Dystonia/Control), sex and age (higher/lower) from segment boundaries. We performed an explanatory, visualization analysis of dystonia muscle-boundaries. RESULTS: For all segments, agreement with manual labels was Dice Coefficient (64 ± 21%) and Hausdorff Distance (5.7 ± 4 mm). For deep muscle layers, boundaries predicted central injection sites with average precision 94 ± 3%. Using leave-one-out cross-validation, a support-vector-machine classified condition, sex, and age from predicted muscle boundaries at accuracy 70.5%, 67.2%, 52.4% respectively, exceeding classification by manual labels. From muscle boundaries, Dystonia clustered optimally into three sub-groups. These sub-groups are visualized and explained by three eigen-patterns which correlate significantly with truncal and head posture. CONCLUSION: Using US, neck muscle shape alone discriminates dystonia from healthy controls. SIGNIFICANCE: Using deep learning, US imaging allows online, automated visualization, and diagnostic analysis of cervical dystonia and segmentation of individual muscles for targeted injection.

Fully automated image-based estimation of postural point-features in children with cerebral palsy using deep learning.

The aim of this study was to provide automated identification of postural point-features required to estimate the location and orientation of the head, multi-segmented trunk and arms from videos of the clinical test 'Segmental Assessment of Trunk Control' (SATCo). Three expert operators manually annotated 13 point-features in every fourth image of 177 short (5-10 s) videos (25 Hz) of 12 children with cerebral palsy (aged: 4.52 ± 2.4 years), participating in SATCo testing. Linear interpolation for the remaining images resulted in 30 825 annotated images. Convolutional neural networks were trained with cross-validation, giving held-out test results for all children. The point-features were estimated with error 4.4 ± 3.8 pixels at approximately 100 images per second. Truncal segment angles (head, neck and six thoraco-lumbar-pelvic segments) were estimated with error 6.4 ± 2.8°, allowing accurate classification (F 1 > 80%) of deviation from a reference posture at thresholds up to 3°, 3° and 2°, respectively. Contact between arm point-features (elbow and wrist) and supporting surface was classified at F 1 = 80.5%. This study demonstrates, for the first time, technical feasibility to automate the identification of (i) a sitting segmental posture including individual trunk segments, (ii) changes away from that posture, and (iii) support from the upper limb, required for the clinical SATCo.

Real-Time Ultrasound Segmentation, Analysis and Visualisation of Deep Cervical Muscle Structure.

Despite widespread availability of ultrasound and a need for personalised muscle diagnosis (neck/back pain-injury, work related disorder, myopathies, neuropathies), robust, online segmentation of muscles within complex groups remains unsolved by existing methods. For example, Cervical Dystonia (CD) is a prevalent neurological condition causing painful spasticity in one or multiple muscles in the cervical muscle system. Clinicians currently have no method for targeting/monitoring treatment of deep muscles. Automated methods of muscle segmentation would enable clinicians to study, target, and monitor the deep cervical muscles via ultrasound. We have developed a method for segmenting five bilateral cervical muscles and the spine via ultrasound alone, in real-time. Magnetic Resonance Imaging (MRI) and ultrasound data were collected from 22 participants (age: 29.0±6.6, male: 12). To acquire ultrasound muscle segment labels, a novel multimodal registration method was developed, involving MRI image annotation, and shape registration to MRI-matched ultrasound images, via approximation of the tissue deformation. We then applied polynomial regression to transform our annotations and textures into a mean space, before using shape statistics to generate a texture-to-shape dictionary. For segmentation, test images were compared to dictionary textures giving an initial segmentation, and then we used a customized Active Shape Model to refine the fit. Using ultrasound alone, on unseen participants, our technique currently segments a single image in [Formula: see text] to over 86% accuracy (Jaccard index). We propose this approach is applicable generally to segment, extrapolate and visualise deep muscle structure, and analyse statistical features online.

Proactive Selective Inhibition Targeted at the Neck Muscles: This Proximal Constraint Facilitates Learning and Regulates Global Control.

While individual muscle function is known, the sensory and motor value of muscles within the whole-body sensorimotor network is complicated. Specifically, the relationship between neck muscle action and distal muscle synergies is unknown. This work demonstrates a causal relationship between regulation of the neck muscles and global motor control. Studying violinists performing unskilled and skilled manual tasks, we provided ultrasound feedback of the neck muscles with instruction to minimize neck muscle change during task performance and observed the indirect effect on whole-body movement. Analysis of ultrasound, kinematic, electromyographic and electrodermal recordings showed that proactive inhibition targeted at neck muscles had an indirect global effect reducing the cost of movement, reducing complex involuntary, task-irrelevant movement patterns and improving balance. This effect was distinct from the effect of gaze alignment which increased physiological cost and reduced laboratory-referenced movement. Neck muscle inhibition imposes a proximal constraint on the global motor plan, forcing a change in highly automated sensorimotor control. The proximal location ensures global influence. The criterion, inhibition of unnecessary action, ensures reduced cost while facilitating task-relevant variation. This mechanism regulates global motor function and facilitates reinforcement learning to change engrained, maladapted sensorimotor control associated with chronic pain, injury and performance limitation.

Diadenosine 5', 5'''-P(1),P(4)-tetraphosphate (Ap4A) is synthesized in response to DNA damage and inhibits the initiation of DNA replication.

The level of intracellular diadenosine 5', 5'''-P(1),P(4)-tetraphosphate (Ap4A) increases several fold in mammalian cells treated with non-cytotoxic doses of interstrand DNA-crosslinking agents such as mitomycin C. It is also increased in cells lacking DNA repair proteins including XRCC1, PARP1, APTX and FANCG, while >50-fold increases (up to around 25 µM) are achieved in repair mutants exposed to mitomycin C. Part of this induced Ap4A is converted into novel derivatives, identified as mono- and di-ADP-ribosylated Ap4A. Gene knockout experiments suggest that DNA ligase III is primarily responsible for the synthesis of damage-induced Ap4A and that PARP1 and PARP2 can both catalyze its ADP-ribosylation. Degradative proteins such as aprataxin may also contribute to the increase. Using a cell-free replication system, Ap4A was found to cause a marked inhibition of the initiation of DNA replicons, while elongation was unaffected. Maximum inhibition of 70-80% was achieved with 20 µM Ap4A. Ap3A, Ap5A, Gp4G and ADP-ribosylated Ap4A were without effect. It is proposed that Ap4A acts as an important inducible ligand in the DNA damage response to prevent the replication of damaged DNA.

Immediate postoperative imaging after uncomplicated endoscopic approach to the anterior skull base: is it necessary?

BACKGROUND: Postoperative imaging is frequently performed to assess for intracranial complications following anterior skull base (ASB) surgery. However, there is little data to suggest that radiologic studies change the management of complications. In this study, the utility of postoperative imaging within 72 hours after uncomplicated ASB surgery was examined. METHODS: A retrospective review was conducted of 143 patients who underwent endoscopic ASB surgery between 2007 and 2013 at Loyola University Medical Center. The main outcomes measures included the ability of head computed tomography (CT) scan to identify postoperative complications and hallmark symptoms associated with complications. RESULTS: Seventy-nine patients underwent postoperative imaging within 72 hours of the initial surgery. The most common finding was pneumocephalus (35/79; 44%). Expanding pneumocephalus requiring surgical intervention developed in 3 cases. Cerebrospinal fluid (CSF) leak was the most common complication, occurring in 22 of 143 (15%) of the patients. Of the 24 patients who developed a postoperative complication, all had clinical signs or symptoms indicative of the need for surgical or medical intervention. The positive predictive value for a head CT scan to detect a complication was 12%, negative predictive value was 92%, and sensitivity and specificity were 63% and 48%, respectively. CONCLUSION: Routine postoperative imaging may be unnecessary after uncomplicated endoscopic ASB surgery because (1) it may not alter patient management; (2) it may not detect the most common complication (CSF leak); and (3) when imaging is positive, the patient has clinical symptoms suggesting a need for intervention.

Preliminary assessment of photoactivated antimicrobial collagen on bioburden in a murine pressure ulcer model.

OBJECTIVE AND BACKGROUND DATA: Overcoming bacterial antibiotic resistance requires alternative strategies. The ability of photoactivated collagen-embedded flavins (PCF) to reduce bioburden in infected pressure ulcers was investigated. DESIGN AND METHODS: Two pressure ulcers were created on the dorsum of female BALB/C mice (n=40, 35 g) maintaining a 5 mm skin bridge between lesions. Ulcers and surrounding skin were covered with Tegaderm™ and inoculated with 0.1 mL of 1 × 10(5) colony-forming units (CFU)/mL methicillin-resistant Staphylococcus aureus (MRSA). Fluid was permitted to reabsorb for 10 min. In experiment 1, one wound from each animal was treated using PCF and photoradiation (PCF+R, n=12) or photoradiation alone (R, n=11). Composite dressing-treated wounds received 1 × 1 cm PCF discs. Overlying Tegaderm was excised, and PCF was placed over the wound and again covered with Tegaderm. Wounds were irradiated at 455 ± 5 nm (350 mW, 1 cm spot diameter, 15 min) using a diode laser 10 min after placement. Controls received no PCF or photoradiation (C, n=12). Animals were euthanized 24 h post-therapy. Quantitative bacterial counts (CFU/g tissue) were determined. In experiment 2, composite dressing-treated wounds were irradiated at 455 ± 5 nm (350 mW, 1 cm spot diameter, 15 min) using a diode laser 10 min after daily PCF placement (0, 1, 2, or 3 treatments, n=8/group). Controls received no treatment. Wounds were cultured daily. Animals were euthanized on day 7 post-infection. Quantitative bacterial counts were determined. RESULTS: PCF+R significantly reduced bacterial counts at 24 h (experiment 1, p<0.0001; experiment 2, p<0.05). The bacterial counts in rats receiving photoradiation alone were no different from those of untreated controls (experiment 1, p=0.24). PCF+R produced a 2-3 log reduction in bacterial counts (experiment 2, p<0.001). Antibacterial effects increased with number of treatments, and persisted for several days post-therapy (p<0.002). CONCLUSIONS: PCF+R inhibited bacterial growth in this model. This effect increased with successive treatments, persisting several days post-therapy. Further studies to optimize this treatment modality are warranted.

Ancient DNA analysis of mid-holocene individuals from the Northwest Coast of North America reveals different evolutionary paths for mitogenomes.

To gain a better understanding of North American population history, complete mitochondrial genomes (mitogenomes) were generated from four ancient and three living individuals of the northern Northwest Coast of North America, specifically the north coast of British Columbia, Canada, current home to the indigenous Tsimshian, Haida, and Nisga'a. The mitogenomes of all individuals were previously unknown and assigned to new sub-haplogroup designations D4h3a7, A2ag and A2ah. The analysis of mitogenomes allows for more detailed analyses of presumed ancestor-descendant relationships than sequencing only the HVSI region of the mitochondrial genome, a more traditional approach in local population studies. The results of this study provide contrasting examples of the evolution of Native American mitogenomes. Those belonging to sub-haplogroups A2ag and A2ah exhibit temporal continuity in this region for 5000 years up until the present day. Of possible associative significance is that archaeologically identified house structures in this region maintain similar characteristics for this same period of time, demonstrating cultural continuity in residence patterns. The individual dated to 6000 years before present (BP) exhibited a mitogenome belonging to sub-haplogroup D4h3a. This sub-haplogroup was earlier identified in the same general area at 10300 years BP on Prince of Wales Island, Alaska, and may have gone extinct, as it has not been observed in any living individuals of the Northwest Coast. The presented case studies demonstrate the different evolutionary paths of mitogenomes over time on the Northwest Coast.