Skin punch biopsy

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Persistence of Borrelia burgdorferi in experimentally infected dogs after antibiotic treatment

Persistence of Borrelia burgdorferi in experimentally infected dogs after antibiotic treatment

PCR. Skin punch biopsy and postmortem tissues were tested for borrelia DNA with specific primer pairs targeting the chro- mosomal 23S rRNA gene or the OspA gene on plasmids. The results are presented in Table 2 and Fig. 1. PCR was positive for skin punch biopsy samples from all dogs with the exception of dog A95-2/5 before antibiotic treatment was initiated. Within 2 weeks of initiation of the antibiotic treatment, skin punch biopsy samples from two of six doxycycline- and two of five amoxicillin-treated dogs became PCR negative while bi- opsy samples from untreated infected control dogs remained PCR positive. However, between 51 and 134 days after treat- ment was completed, skin punch biopsy samples from all in- fected and antibiotic-treated dogs showed positive PCRs again (Table 2; Fig. 1). In addition, with the exception of dog A95- 2/5, which remained uninfected throughout the experiment, multiple tissues from three of four doxycycline-treated, two of three amoxicillin-treated, and two of two untreated infected dogs had positive PCRs (Table 2). Tissues from two uninfected SPF dogs, as well as from dog A95-2/5, that served as negative controls were PCR negative.
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Culture Confirmed Reinfection of a Person with Different Strains of Borrelia burgdorferi Sensu Stricto

Culture Confirmed Reinfection of a Person with Different Strains of Borrelia burgdorferi Sensu Stricto

In recent years, the utility of serum-based diagnostic testing for Lyme disease has improved substantially; however, recovery by culture of the bacterium from skin biopsies of suspected patients is still the only definitive laboratory test. Reinfection of patients has been assumed to occur but as yet has not been documented by serial isolates from the same person. We present a case of culture-confirmed reinfection of a patient in Menominee County, Michigan. Borrelia burgdorferi was isolated from the skin punch biopsy specimens during each episode of erythema migrans (EM) and was subjected to molecular strain typing, genetic analysis of two outer surface protein genes, protein profile analysis, and serum antibody response testing. Results show that these isolates are distinct strains of the bacterium and that the two episodes of EM were caused by independent infections. This report describes the documented, culture-confirmed reinfection of a human by two different strains of B. burgdorferi.
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PCR Based Quantification of Borrelia burgdorferi Organisms in Canine Tissues over a 500 Day Postinfection Period

PCR Based Quantification of Borrelia burgdorferi Organisms in Canine Tissues over a 500 Day Postinfection Period

Isolation of B. burgdorferi from tissue samples. Skin punch biopsy specimens (diameter, 4 mm) were collected sterilely under local anesthesia at 4-week intervals. Additional skin biopsy samples were collected 14 days after the initi- ation of the antibiotic therapy. Twenty-five different tissues were collected from each dog at necropsy, with frequent changes of instruments to avoid cross- contamination. Tissues collected at necropsy included skin from the left and right sides, synovial membranes from six joints (shoulder, elbow, and knee), muscle and fascia from front limbs (musculus triceps, fascia antebrachii), and hind limbs (musculus adductor, fascia lata), superficial cervical, axillary, and popliteal lymph nodes, pericardium, peritoneum, and meninges. Skin samples were ground in 0.2 ml of modified Barbour-Stoenner-Kelly medium containing kanamycin and ri- fampin (BSK-II plus KR) with a sterile pellet pestle and were placed into 6.5 ml of medium as described previously (1). Tissues retrieved postmortem were sus- pended in 3 ml of medium and were treated in a tissue homogenizer (Stomacher; Teckmar, Cincinnati, Ohio). The suspension was placed into 27 ml of prewarmed medium. The medium was incubated at 34°C for 5 weeks and was examined at 1, 3, and 5 weeks by dark-field microscopy for the presence of live spirochetes.
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Nonerythropoietic Tissue Protective Compounds Are Highly Effective Facilitators of Wound Healing

Nonerythropoietic Tissue Protective Compounds Are Highly Effective Facilitators of Wound Healing

Erythropoietin (EPO) is a type I cytokine that utilizes different receptor isoforms either to maintain hematopoiesis or protect against injuries that arise from widely diverse etiologies. EPO also facilitates healing by reducing inflammation and mobilizing en- dothelial progenitor cells to participate in restorative neoangiogenesis, but it is unclear which EPO receptor isoform is responsible for healing and whether this receptor use varies according to the type of wound. In the present studies carried out in the rat, we have utilized receptor-selective derivatives of EPO to determine which receptor type operates in (i) a nonischemic wound (skin punch biopsy), (ii) a permanently ischemic wound (raised musculocutaneous flap), (iii) an intermittent ischemic reperfusion wound (pressure or decubitus ulcer), or (iv) wounds complicated by infection (cecal ligation and perforation). Using these mod- els, we demonstrate that nonerythropoietic tissue protective compounds administered immediately following injury limit wound size and accelerate eschar closure independent of wound type. Moreover, in a model of peritonitis-induced adhesions, daily ad- ministration of the nonerythropoietic derivative carbamyl-EPO (10 μ g/kg-bw) was associated with significantly lower serum TNF α concentration, illness scores, increased survival, as well as decreased adhesion formation. These results confirm that wound heal- ing is mediated by the tissue protective receptor isoform and argue that nonerythropoietic tissue protective molecules constitute promising new therapeutics for treatment of a wide variety of surgical wounds.
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Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

Increased CXC Ligand 10 Levels and Gene Expression in Type 1 Leprosy Reactions

was calculated. Statistical analysis of data. For measurements of serum CXCL10, statistical significance between groups at a single time point was tested using the Mann- Whitney 2-tailed test. The SAS statistics package GLM procedure (14) was used to analyze data as a repeated-measures design in a split-plot arrangement of treatments. This design allows more powerful tests with a smaller sample size because of the better control of error in blocking on subjects, with each subject serving as his/her own control, so that intersubject variation is excluded from the residual error (11). When overall analysis of variance indicated significance, post hoc pairwise comparisons were conducted with Tukey’s test for main effects and the t test of least-squares means for interaction effects. Spearman correlation coefficients were calculated. The Wilcoxon matched-pairs signed-rank test was used to assess the statistical significance of cell counts in paired biopsy specimens in immunostaining studies. All comparisons were considered significant at a P value of ⬍0.05.
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Facit 4046 4047 Punch Reader Operator Guide Edition 1 Aug86 pdf

Facit 4046 4047 Punch Reader Operator Guide Edition 1 Aug86 pdf

Punch/Readeraddress Service Request Punch Service Request Reader Punch mode, IEEE/Punch/Reader Parity Transparent Punch Transparent copy Transparent reading Time out Control Code Process[r]

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Topical Application of Tat-Rac1 Promotes Cutaneous Wound Healing in Normal and Diabetic Mice

Topical Application of Tat-Rac1 Promotes Cutaneous Wound Healing in Normal and Diabetic Mice

therapeutic gel, is approved by the Food and Drug Administration (FDA) to treat diabetic wounds. These facts highlight a sense of urgency for scientific discovery-based therapeutic interventions. Various studies have identified Rac1, a small GTPase, as playing an important role in wound healing. Deletion or inhibition of Rac1 in either skin epidermis or oral epithelia resulted in delayed wound healing in the skin or the oral mucosa [4, 5]. Similarly, inducible deletion of the Rac1 gene in fibroblasts resulted in delayed wound closure with reduced collagen production and myofibroblast formation [6]. Con- versely, transgenic mice expressing a constitutively active Rac1 in smooth muscle cells accelerate cutaneous wound repair through promoting angiogenesis [7]. In addition to its role in normal wound healing, a study reported that ganglioside GM3 depletion in diabetic mouse wounds improved healing by requiring IGF-1 receptor-mediated Rac1 activation to restore keratinocyte motility [8, 9]. Additionally, ultrasonic treatment–mediated promo- tion of wound healing in mouse models of diabetic and aging skin wounds involves activation of the calcium/CamKinaseII/Tiam1/Rac1 pathway that promotes fibroblast migration[10]. These data suggest that impaired wound healing lacks sufficient Rac1 activity and that activation of endogenous Rac1 should be explored as a therapeutic strategy to improve wound healing. In the present study, we sought to determine if Rac1 protein can be utilized pharmacologically to promote wound healing. Because Rac1 is an intracellular protein, it has to be delivered to the inside of cells to exert its functions. To this end, we employed a protein transduction strategy to deliver Rac1 into wounded cells. We chose a protein transduction peptide derived from the Tat protein of the HIV virus, and fused it to human Rac1 to create a Tat-Rac1 protein. Our study provides proof of principle for using Tat-Rac1 as a pharmacological biologic to promote cutaneous wound healing.
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Solvent effects in permeation assessed in vivo by skin surface biopsy

Solvent effects in permeation assessed in vivo by skin surface biopsy

Classic physical penetration enhancement strategies include iontophoresis [3], electroporation [4], phono- phoresis [5-7] or supersaturated solutions [8,9], but recently novel approaches have been developed such as compressed gas propulsion [10] and the use of microfab- ricated microneedles to pierce the stratum corneum [11]. Several chemical substances have been shown to possess the ability to enhance permeation across the skin, and are therefore commonly included in transdermal systems. These include low molecular weight alcohols [12], alkyl methanol sulphoxides [13], non ionic surfactants [14] (polysorbates, polyethoxylated alkyl ethers and esters and poloxamers), oleic acid in synergy with propylene glycol [15,16] and azone [17,18]. In spite of the wide use of chemical enhancers, there are still few guidelines for their rational choice and the exact mechanisms of action in the skin have not yet been fully established. In this context, the skin surface biopsy (SSB) technique can be regarded as an innovative strategy in the study of transcutaneous pen- etration. This simple and versatile approach to the study of the stratum corneum was introduced by Goldschmidt and Kligman in the 1960's [19] and was later developed by Marks and Dawber [20]. If consecutive biopsies are per- formed in the same area it becomes possible to follow the permeation of a compound through the stratum corneum [21,22]. SSB alone does not provide any data regarding the amount of chemical permeated through the epidermis and dermis into systemic circulation, but it can be com- bined with the determination of blood and urine concen- tration in pharmacokinetic studies. Corneosurfametry is also a technique that relies on SSB to assess the impact on the stratum corneum of surfactants used in cosmetics [23].
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The retinal pathology of ovine ceroid lipofuscinosis : a thesis presented in fulfilment of the requirement for the degree of Master of Veterinary Science at Massey University

The retinal pathology of ovine ceroid lipofuscinosis : a thesis presented in fulfilment of the requirement for the degree of Master of Veterinary Science at Massey University

Skin biopsies were taken from the axillary region because of the abundance of apocrine sweat glands found in this area. The selected area of skin was disinfected and small pieces, approximately 0.5 cm in diameter were removed using sharp scissors. The fresh biopsies were attached to microtome chucks, frozen, and 8 � sections were cut and mounted in glycerol for fluorescence microscopy. Some skins were sectioned at 6, 8 and 10 )JlTI because the degree of fluorescence could have been related to section thickness. The sections were examined on a Reichert I mrnunopan microscope , fitted with a quartz halogen lamp, using excitor filter 30, 8x1 F ITC3 and barrier filter 18 x 3 OG 515 GG9 giving a wave length of 500 nm . In some cases a portion
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Angioimmunoblastic T Cell Lymphoma Mimicking Chronic Urticaria

Angioimmunoblastic T Cell Lymphoma Mimicking Chronic Urticaria

He had been diagnosed and suffering from “recurrent idiopathic angioedema and urticaria” for the past 2 years. The first episode involved right ankle edema, which progressed over the course of three weeks to the entire right body and face. The patient was admitted and treated with intravenous steroids and discharged on oral prednisone. Extensive work- up failed to find a trigger, and amlodipine was discontinued “as a precaution.” Over the next 2 years he developed multiple episodes of angioedema associated with urticaria- like lesions (erythematous, pruritic rash) lasting at least one week and sometimes longer, which resolved with steroids and antihistamines. Work-up at the time revealed monoclonal gammopathy of undetermined significance (MGUS) and normal bone marrow biopsy, skin biopsy, and flow cytometry.
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Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes

Comparison of LPS-stimulated release of cytokines in punch versus transwell tissue culture systems of human gestational membranes

Immunohistochemical labeling of IL-8 was performed on paraffin-embedded sections to identify cellular ori- gins of this cytokine within the amnion, chorion laeve and decidua. IL-8 was selected for evaluation due to its significant secretion difference between punch cultures and both compartments of the transwell cultures. Fig- ure 2 shows representative immunohistological sections of control and LPS-treated membranes from punch and transwell culture systems. Unstimulated punch and transwell membranes showed limited IL-8 positive label- ing localized intracellularly within the cytoplasm of his- tiocytes. After stimulation with LPS, marked increases of intracellular IL-8 were visualized in the chorion leave and decidua; less marked increases were observed in the amniotic mesoderm.
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Small Fiber Neuropathy in Children: Two Case Reports Illustrating the Importance of Recognition

Small Fiber Neuropathy in Children: Two Case Reports Illustrating the Importance of Recognition

Small fiber neuropathy (SFN) is a debilitating condition that often leads to pain and autonomic dysfunction. In the last few decades, SFN has been gaining more attention, particularly in adults. However, literature about SFN in children remains limited. The present article reports the cases of 2 adolescent girls diagnosed with SFN. The first patient (14 years of age) complained about painful itch and tingling in her legs, as well as dysautonomia symptoms for years. She also reported a red/purple-type discoloration of her legs aggravated by warmth and standing, compatible with erythromelalgia. The diagnosis of SFN was confirmed by a reduced intraepidermal nerve fiber density (IENFD) in skin biopsy sample. No underlying conditions were found. Symptomatic neuropathic pain treatment was started with moderate effect. The second patient (16 years of age) developed painful sensations in both feet and hands 6 weeks after an ICU admission for diabetic ketoacidosis, which included dysautonomia symptoms. She also exhibited some signs of erythromelalgia. The patient was diagnosed with predominant SFN (abnormal IENFD and quantitative sensory testing) as well as minor large nerve fiber involvement. Treatment with duloxetine, combined with a rehabilitation program, resulted in a marked improvement in her daily functioning. Although the SFN diagnosis in these 2 cases could be established according to the definition of SFN used in adults, additional diagnostic tools are needed that may be more appropriate for children. Additional information about the course of SFN in children may result in better treatment options.
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Probiotic supplements containing Lactobacillus reuteri does not affect the levels of matrix metalloproteinases and interferons in oral wound healing

Probiotic supplements containing Lactobacillus reuteri does not affect the levels of matrix metalloproteinases and interferons in oral wound healing

The study had a randomized, placebo-controlled, dou- ble-blind cross-over design. The protocol was approved by The Research Ethics Committee of the Capital Region of Denmark (H-16025546) and the study was registered at Clintrials.gov (NCT03210779). During a 1-week run-in period, the participants were assigned to let an active or placebo lozenge slowly dissolve in their mouth, two times per day. After 8 days, a biopsy was taken under local anesthesia containing adrenalin with aid of a standardized punch (4  mm diameter) in the free buccal oral mucosa in the upper left or right premolar region. After compression with cotton gauges for 30 min, the created wound was left uncovered and no sutures were used. The subjects were thoroughly instructed to continue with their assigned lozenges for another 8 days. In addition, they were asked to topically apply one drop of oil (probiotic or placebo) directly on the wound, once daily for 8 days with aid of a plas- tic micro-brush. No food or nutritional restrictions were given, and all subjects were asked to maintain their normal oral hygiene routines during the course of the study. After a 4-week washout period, all the
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Congenital Pilar and Smooth Muscle Nevus

Congenital Pilar and Smooth Muscle Nevus

Hair follicles asso- ciated with intersecting bundles of smooth muscle in the reticular dermis were seen in a punch biopsy specimen (Fig 2).. The lesion showed no gross changes 2#{189} y[r]

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Ability of sodium copper chlorophyllin complex to repair photoaged skin by stimulation of biomarkers in human extracellular matrix

Ability of sodium copper chlorophyllin complex to repair photoaged skin by stimulation of biomarkers in human extracellular matrix

This clinical study demonstrates a statistically significant improvement in repair of photoaged skin by topical treatment with a test formulation containing a liposomal dispersion of 0.05% sodium copper chlorophyllin, based on changes in key epidermal and dermal biomarkers. The degree of repair seen with the sodium copper chlorophyllin complex and the positive control (tretinoin cream, United States Pharmacopeia 0.025%), based on immunohistological analysis, was not sta- tistically different. The results support and validate recently published in vitro antihyaluronidase activity for sodium copper chlorophyllin complex. Topical retinoid and sodium copper chlorophyllin complex for repair of photoaged skin should be evaluated further in this biopsy model to quantify whether there is an additive or synergistic response with this unique combination of topical agents.
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Sensitivity of PCR Targeting the IS2404 Insertion Sequence of Mycobacterium ulcerans in an Assay Using Punch Biopsy Specimens for Diagnosis of Buruli Ulcer

Sensitivity of PCR Targeting the IS2404 Insertion Sequence of Mycobacterium ulcerans in an Assay Using Punch Biopsy Specimens for Diagnosis of Buruli Ulcer

Punch biopsy specimens from Mycobacterium ulcerans disease lesions were used to compare the sensitivities and specificities of direct smear, culture, PCR, and histopathology in making a diagnosis of M. ulcerans disease in a field setting. PCR for the insertion element IS2404 was modified to include uracil-N-glycosylase and deoxyuridine triphosphate instead of deoxythymidine triphosphate to reduce the risk of cross contamination. The “gold standard” for confirmation of clinically diagnosed Buruli ulcer was a definite histological diagnosis, a positive culture for M. ulcerans, or a smear positive for acid-fast bacilli (AFB), together with a possible histological diagnosis. For 70 clinically diagnosed cases of M. ulcerans disease, the modified PCR was 98% sensitive and gave a rapid result. The sensitivities of microscopy, culture, and histology were 42%, 49%, and 82%, respectively. The use of a 4-mm punch biopsy specimen was preferred to a 6-mm punch biopsy specimen since the wound was less likely to bleed and to need stitching. Given adequate technical expertise and the use of controls, the PCR was viable in a teaching hospital setting in Ghana; and in routine practice, we would recommend the use of Ziehl-Neelsen staining of biopsy specimens to detect AFB, followed by PCR, in AFB- negative cases only, in order to minimize costs. Histology and culture remain important as quality control tests, particularly in studies of treatment efficacy.
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Granulomatous/sarcoid-like lesions associated with checkpoint inhibitors: a marker of therapy response in a subset of melanoma patients

Granulomatous/sarcoid-like lesions associated with checkpoint inhibitors: a marker of therapy response in a subset of melanoma patients

Six months after completion of ipilimumab therapy, surveillance studies revealed interval progression of disease with new bilateral pulmonary nodules, medi- astinal and hilar lymphadenopathy, a liver lesion, and an adrenal nodule, suggestive of metastatic melanoma. Pembrolizumab therapy (2 mg/kg) was subsequently initiated, and computed tomography (CT) scans 3 months after initiation of pembrolizumab revealed radiographic improvement and decreased size of pul- monary, liver, and adrenal lesions. The patient toler- ated 22 cycles of pembrolizumab with minimal adverse reactions, and CT scans obtained 15 months after initiation of therapy revealed stable disease with- out new lesions. Restaging positron emission tomog- raphy/CT (PET/CT) scans obtained 18 months after initiation of pembrolizumab therapy revealed enlar- ging gastric, retroperitoneal, and paratracheal lymph nodes, and after 20 months of pembrolizumab ther- apy (27 cycles), the patient presented with erythema and swelling of the forearms. Physical examination re- vealed symmetrical, firm subcutaneous nodules involv- ing bilateral dorsal hands, forearms, and elbows (Fig. 1a-b). Ultrasound examination revealed a 4.3 × 1 × 2.3 cm soft tissue nodule (Fig. 1c), and needle core and skin punch biopsies showed a collection of epithelioid histiocytes forming non-caseating granu- lomata (Fig. 1d-e). Fite, Gomori methenamine silver, and gram stains and tissue cultures were negative for microorganisms. Melanoma was not identified. PET/ CT performed at 23 months of pembrolizumab ther- apy (30 cycles), revealed prominent fluorodeoxyglu- cose (FDG) avidity of mediastinal / bilateral hilar LAD and subcutaneous nodules (Fig. 1f). In view of the clinical, radiographic, and histologic findings, these features were consistent with a cutaneous granulomatous/sarcoid-like lesions associated with CPI involving the skin and pulmonary lymph nodes. Given the patient’s excellent response to pembrolizu- mab therapy and the development of these therapy related granulomatous/sarcoid-like lesions, his therapy was subsequently discontinued.
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MAST CELL LEUKEMIA PRESENTING AS URTICARIA PIGMENTOSA

MAST CELL LEUKEMIA PRESENTING AS URTICARIA PIGMENTOSA

Special Giemsa staining of the previous skin biopsy taken at 9 months of age revealed the true character of the infiltrative cells to be tissue mast cells. Repeat skin and lymph node bio[r]

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Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

Exosomes from acellular Wharton’s jelly of the human umbilical cord promotes skin wound healing

During skin healing, there is a dynamic interplay be- tween cells in granulation tissue and extracellular matri- ces. While dermal fibroblasts contribute to the secretion of extracellular matrix components [56], there is a feed- back loop where components of extracellular matrices affect fibroblasts and activate some of the signaling path- ways important for wound healing such as wnt/β-catenin signaling [57]. The abundance of α2M in exosomes, to- gether with the promising effect of exosomes on wound healing in vitro and in vivo, raises the possibility that exosomes enhance wound healing, at least partly, through the paracrine effects of α2M. We exogenously added α2M to cells after conducting a scratch assay to determine its effect on cell migration. Using different doses of α2M [58] it was revealed that the high dose of 1000 ng/ml had the greatest effect on cell migration into the scratch zone while the lower doses did not affect cell migration. It is likely that exosomes containing an abun- dant amount of α2M in their cargo are engulfed by cells in the wound area which enhances cell migration. Our viability assay with 100 ng/ml α2M exogenously added to fibroblasts showed that α2M had the greatest increase in viability compared with whole exosomes and controls. For keratinocytes, similarly, 100 ng/ml showed signifi- cant improvement in keratinocyte viability. These data suggest that α2M increases two important characteristics of cells during healing (migration and viability) and this might be the underlying mechanism behind the positive effect of exosomes on wound healing that we herein re- port. It is not clear whether the inhibition of proteinases, and hence an increase in growth factors, is affecting the cellular characteristics in favor of wound healing or if α2M directly affects the cells.
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PDP 1 DDT Listing Feb63 pdf

PDP 1 DDT Listing Feb63 pdf

/title punch and punch format setup... dap fa jmp Iss.[r]

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