A second generation leishmanization vaccine with a markerless attenuated leishmania major strain using CRISPR gene editing
expression; cells; growth; immunity; immunization; protection; immunogenicity; balb/c mice; centrin deleted parasites; cutaneous leishmaniasis
Leishmaniasis is a neglected tropical disease caused by Leishmania protozoa transmitted by infected sand flies. Vaccination through leishmanization with live Leishmania major has been used successfully but is no longer practiced because it resulted in occasional skin lesions. A second generation leishmanization is described here using a CRISPR genome edited L. major strain (LmCen(-/-)). Notably, LmCen(-/-) is a genetically engineered centrin gene knock-out mutant strain that is antibiotic resistant marker free and does not have detectable off-target mutations. Mice immunized with LmCen(-/-) have no visible lesions following challenge with L. major-infected sand flies, while non-immunized animals develop large and progressive lesions with a 2-log fold higher parasite burden. LmCen(-/-) immunization results in protection and an immune response comparable to leishmanization. LmCen(-/-) is safe since it is unable to cause disease in immunocompromised mice, induces robust host protection against vector sand fly challenge and because it is marker free, can be advanced to human vaccine trials. Here, the authors engineer an attenuated knock-out Leishmania (LmCen(-/-)) vaccine that is safe in immunocompromised mice and induces an immune response and protection similar to leishmanization with wild-type Leishmania. Since LmCen(-/-) is antibiotic resistant marker free, it is a candidate for clinical development.
Zhang W; Karmakar S; Gannavaram S; Dey R; Lypaczewski P; Ismail N; Siddiqui A; Simonyan V; Oliveira F; Coutinho-Abreu I; DeSouza-Vieira T; Meneses C; Oristian J; Serafim TD; Musa Abu; Nakamura R; Saljoughian N; Volpedo G; Satoskar M; Satoskar S; Dagur PK; McCoy JP; Kamhawi S; Valenzuela JG; Hamano S; Satoskar AR; Matlashewski G; Nakhasi HL
Nature Communications
2020
2020-07-10
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
journalArticle
<a href="http://doi.org/10.1038/s41467-020-17154-z" target="_blank" rel="noreferrer noopener">10.1038/s41467-020-17154-z</a>
Intramuscular antagonism of the G-protein coupled estrogen receptor 1 partially affects dimorphic characteristics of the syrinx, but is ineffective within the neural song circuit of zebra finches
androgen receptor; behavior; brain; control nuclei; Drug delivery; estradiol; Estrogen receptor; expression; GPER1 antagonist; gpr30; growth; Sexual dimorphism; sexual-differentiation; Songbird; Syrinx; system
Within the zebra finch song system, robust sex differences exist that enable singing behavior in males, but not females. Estradiol is a potent contributor to this process, but how and through which receptor(s) it acts is not clear. Historically, pharmacological manipulations of nuclear estrogen receptors have yielded conflicting results possibly due to method of drug delivery. More recently, the membrane bound G-protein coupled estrogen receptor 1 (GPER1) has also been identified as a potential candidate, but its function has not been fully described. To further investigate the role of GPER1, and the importance of the route of drug administration, a specific antagonist (G-15) was intramuscularly administered to zebra finches for 25 days, starting on the day of hatching. G-15 significantly decreased muscle fiber sizes of ventralis and dorsalis in the syrinx of males only. Dimorphic characteristics of the neural song system were unaffected by this manipulation in either sex. These results contrast with a study in which G-15 was intracranially delivered. In males, select song nuclei were decreased in volume, and in females, syrinx muscle fiber size was increased. Together, these results support the hypothesis that estrogens acting through GPER1 influence dimorphic development of the song system, and that method of drug administration is important in this species.
Tehrani Mahtab Attarhaie; Veney Sean L
General and Comparative Endocrinology
2020
2020-07-01
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
journalArticle
<a href="http://doi.org/10.1016/j.ygcen.2020.113492" target="_blank" rel="noreferrer noopener">10.1016/j.ygcen.2020.113492</a>
Experiments With Osteoblasts Cultured Under Varying Orientations With Respect To The Gravity Vector
attachment; averaged gravity; Biotechnology & Applied Microbiology; bone; Cell Biology; cell-culture; clinostat; gene-expression; growth; growth rate; in-vitro; inversion; microgravity; osteoblasts; proliferation; space; substrate
Substrate attachment is crucial for normal growth and differentiation of many cell types. To better understand the role of gravity in osteoblast attachment and growth in vitro, 17-day-old embryonic chick calvarial osteoblasts were subjected to directional variations with respect to gravity. Osteoblasts, grown in MEM or DME supplemented with 10% FBS and attached to type I collagen-coated coverslips, were loaded into cylindrical containers completely filled with medium and oriented so that cells were either atop or beneath, or coverslips continuously rotated (similar to2 rpm) in a clinostat, thereby continuously changing their orientation with respect to gravity. Cells in these three conditions were collected daily for up to 6 days, and cell viability, two osteoblast functions, and proliferation were assessed. Data suggest the number and function of attached osteoblasts is unaltered by inversion or clino-rotation in initially confluent cultures. In sparsely plated cultures, however, osteoblast viability was significantly decreased (similar to50%) in inverted and rotated cultures during the first 3 days of sampling, but from days 4 - 6 no significant difference was found in viable cell number for the three conditions. Decreases in viable cell number within the first days of the experiments could result from death followed by detachment, detachment followed by death, differences in proliferation rate, or lag-phase duration. To help distinguish among these, BrdU labeling for 2 or 24 hr was used to assess cell proliferation rate. Log-phase growth rates were calculated and were unchanged among the three conditions tested. These results point to an increase in lag-phase duration in inverted and rotated cultures. In summary, changing the cell-substrate attachment direction with respect to gravity causes an immediate response in the form of diminished viable osteoblast number in sparse, early cultures, but the effect disappears after 3 - 4 days and does not occur in mature, confluent cultures.
Kacena M A; Todd P; Gerstenfeld L C; Landis W J
Cytotechnology
2002
2002
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1023/a:1023936503105" target="_blank" rel="noreferrer noopener">10.1023/a:1023936503105</a>
Comparison Of Different Chondrocytes For Use In Tissue Engineering Of Cartilage Model Structures
articular-cartilage; auricular cartilage; Cell Biology; construct; growth; in-vitro; regeneration; scaffold; shape; stem-cells; vivo
This study compares bovine chondrocytes harvested from four different animal locations-nasoseptal, articular, costal, and auricular-for tissue-engineered cartilage modeling. While the work serves as a preliminary investigation for fabricating a human ear model, the results are important to tissue-engineered cartilage in general. Chondrocytes were cultured and examined to determine relative cell proliferation rates, type II collagen and aggrecan gene expression, and extracellular matrix production. Respective chondrocytes were then seeded onto biodegradable poly(L-lactide-epsilon-caprolactone) disc-shaped scaffolds. Cell-copolymer constructs were cultured and subsequently implanted in the subcutaneous space of athymic mice for up to 20 weeks. Neocartilage development in harvested constructs was assessed by molecular and histological means. Cell culture followed over periods of up to 4 weeks showed chondrocyte proliferation from the tissue sources varied, as did levels of type II collagen and aggrecan gene expression. For both genes, highest expression was found for costal chondrocytes, followed by nasoseptal, articular, and auricular cells. Retrieval of 20-week discs from mice revealed changes in construct dimensions with different chondrocytes. Greatest disc diameter was found for scaffolds seeded with auricular chondrocytes, followed by those with costal, nasoseptal, and articular cells. Greatest disc thickness was measured for scaffolds containing costal chondrocytes, followed by those with nasoseptal, auricular, and articular cells. Retrieved copolymer alone was smallest in diameter and thickness. Only auricular scaffolds developed elastic fibers after 20 weeks of implantation. Type II collagen and aggrecan were detected with differing expression levels on quantitative RT-PCR of discs implanted for 20 weeks. These data demonstrate that bovine chondrocytes obtained from different cartilaginous sites in an animal may elicit distinct responses during their respective development of a tissue-engineered neocartilage. Thus, each chondrocyte type establishes or maintains its particular developmental characteristics, and this observation is critical in the design and elaboration of any tissue-engineered cartilage model.
Isogai N; Kusuhara H; Ikada Y; Ohtani H; Jacquet R; Hillyer J; Lowder E; Landis W J
Tissue Engineering
2006
2006-04
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1089/ten.2006.12.691" target="_blank" rel="noreferrer noopener">10.1089/ten.2006.12.691</a>
Hesperornis Escapes Plesiosaur Attack
birds; Geology; growth; Hesperornis; ichthyosaur; Late Cretaceous; Mosasaur; Paleontology; Pathology; Pierre Shale; Plesiosaur; poultry
Martin L D; Rothschild B M; Burnham D A
Cretaceous Research
2016
2016-08
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1016/j.cretres.2016.02.005" target="_blank" rel="noreferrer noopener">10.1016/j.cretres.2016.02.005</a>
Lipid Deposits And Lipo-mucosomes In Human Cholecystitis And Epithelial Metaplasia In Chronic Cholecystitis
biliary-tract tumors; carcinoma; cholecystitis; cholelithiasis; epithelium metaplasia; expression; fine-structure; gallbladder; gallstones; growth; lipo-mucosomes; Microscopy; Pathology; precursor lesions; sex steroid treatment; syrian-hamster gallbladder; ultrastructure
Gilloteaux J; Tomasello L M; Elgison D A
Ultrastructural Pathology
2003
2003-09
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1080/01913120390239962" target="_blank" rel="noreferrer noopener">10.1080/01913120390239962</a>
Low Power Millimeter Waves Modify Phospholipid Model Membranes Structure
growth; inhibition; microwaves; Oncology
Filippelli L; Beneduci A; Chidichimo G; Westerman P W; Malmer M; Servello A
Anticancer Research
2004
2004-09
Journal Article or Conference Abstract Publication
n/a
THE USE OF CULTURED EPITHELIAL AUTOGRAFTS IN THE WOUND CARE OF SEVERELY BURNED PATIENTS
skin; Pediatrics; Surgery; cells; growth; burns; clinical-application; coverage; epithelial autografts; human epidermal-keratinocytes; skin replacement
McAree K G; Klein R L; Boeckman C R
Journal of Pediatric Surgery
1993
1993-02
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1016/s0022-3468(05)80266-0" target="_blank" rel="noreferrer noopener">10.1016/s0022-3468(05)80266-0</a>
Oncologic mathematics - Evolution of a new specialty
Surgery; nonlinear dynamics; growth; tumors; breast-cancer; chaos; biology; prognostic factors; models; complexity; fractals
Hypothesis: Mathematical methods and their derivatives have practical applications to oncology. They can be used to describe fundamental aspects of tumor behavior, such as loss of genetic stability, tumor growth, immunologic identity, genesis of diversity, and methods of prognosticating cancer. Data Sources: Descriptive models and published literature in the fields of oncology and applied mathematics. Data Synthesis: Cancer does not conform to simple mathematical principles. Its irregular mode of carcinogenesis, erratic tumor growth, variable response to tumoricidal agents, and poorly understood metastatic patterns constitute highly variable clinical behavior. Defining this process requires an accurate understanding of the interactions between tumor cells and host tissues and ultimately determines prognosis. Applying time-tested and evolving mathematical methods to oncology may provide new tools with inherent advantages for the description of tumor behavior, selection of therapeutic modes, prediction of metastatic patterns, and providing an inclusive basis for prognostication. We term this combined field of research "oncologic mathematics." As surgeons, we have the unique opportunity to be active participants and assume leadership in research that affects selection of the optimal anticancer treatment for our patients. Mathematicians describe equations that define tumor growth and behavior, whereas surgeons actively deal with biological processes. Oncologic mathematics applies these principles to clinical settings. Conclusion: Experimentally testable, oncologic mathematics may provide a framework to determine clinical outcome on a patient-specific basis and increase the growing awareness that mathematical models help simplify seemingly complex and random tumor behavior.
Chandawarkar R Y; Guyton D P
Archives of Surgery
2002
2002-12
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.1001/archsurg.137.12.1428" target="_blank" rel="noreferrer noopener">10.1001/archsurg.137.12.1428</a>
EVOLUTIONARY HISTORY OF THE ROBUST AUSTRALOPITHECINES - GRINE,FE
Anthropology; boisei; brain; east-africa; Evolutionary Biology; growth; hominids; Homo; Kenya; miocene; neogene
Ward S C
Journal of Human Evolution
1991
1991-12
Journal Article
<a href="http://doi.org/10.1016/0047-2484(91)90097-f" target="_blank" rel="noreferrer noopener">10.1016/0047-2484(91)90097-f</a>
The antiviral xanthate compound D609 inhibits herpes simplex virus type 1 replication and protein phosphorylation
antiviral drug; cells; culture; D609; gene; growth; HSV-1; identification; interruption; kinase; Pharmacology & Pharmacy; phosphorylation; protein; protein-kinase; pseudorabies virus; purification; us3; US3 PK; Virology
The mechanism of antiviral action of tricyclodecan-9-yl-xanthogenate (D609) was investigated in vitro. D609 inhibited herpes simplex virus type 1 (HSV-1) replication without apparent cytotoxicity. It reduced phosphorylation of virus-infected cell polypeptides and inhibited the HSV-1 encoded protein kinase (US3 PK) and, to a lesser extent, cellular protein kinase C in vitro. Virus production was reduced by D609 at concentrations greater than 3.8 mu M, With complete inhibition at 75.2 mu M at an MOI of 1 PFU/cell or less. Addition of D609 could be delayed until 7 h post-infection and still inhibit virus replication. Phosphorylation of infected cell viral polypeptides of 34 (similar molecular weight to the substrate of the viral US3 protein kinase) and 69 kDa was inhibited at 18.4 mu M Treatment of infected or uninfected cells with 37.6 mu M D609 reduced protein phosphorylation to background levels. A concentration of 1.9 mu M D609 in vitro inhibited the viral US3-encoded PK, which had been purified from infected cell lysates by affinity chromatography and identified by specific antibody. Purified cellular protein kinase C was inhibited at 75.2 mu M D609 whereas other cellular kinases including casein kinase 1 and cAMP dependent kinase were not inhibited at concentrations as high as 188 mu M D609. Collectively these data indicate that the mechanism of antiviral action of D609 is by inhibition of protein kinases and protein phosphorylation affecting a late step in HSV replication. (C) 1997 Elsevier Science B.V.
Walro D G; Rosenthal K S
Antiviral Research
1997
1997-09
Journal Article
<a href="http://doi.org/10.1016/s0166-3542(97)00040-5" target="_blank" rel="noreferrer noopener">10.1016/s0166-3542(97)00040-5</a>
Synergistic antitumor activity of vitamins C and K-3 on human urologic tumor cell lines
2-methyl-1; 4-naphthoquinone; bladder; cancer-chemotherapy; carcinoma cells; growth; isolated hepatocytes; l-ascorbic-acid; menadione; metabolism; mitomycin-c; oxidative stress; Pharmacology & Pharmacy; prostate; Research & Experimental Medicine; urologic neoplasms; vitamin C
A micro-tetrazolium assay was employed to evaluate vitamin C (VC), vitamin K-3 (VK3) and vitamin C/vitamin K-3 combinations (VC/VK3) for their antitumor activity against eight human urologic tumor cell lines. While the individual vitamins exhibited antitumor activity at high concentrations, co-administration of the vitamin in a VC : VK3 ratio of 100 : 1 potentiated antitumor activity 4- to 61-fold even when exposure times were as short as 1 hour. Administration of exogenous catalase destroyed the antitumor activity of the vitamins and suggested that catalase destroyed the antitumor activity of the vitamins and suggested that hydrogen peroxide and perhaps other reactive oxygen species were involved in the antitumor mechanism of these vitamins. Electron micrographs taken in a previous study demonstrated that vitamin treatment damaged mitochondria and may have impaired ATP synthesis. Analysis of cellular ATP and thiol levels as well as DNA and protein synthesis during the first five hours following a one hour VC/VK3 treatment, revealed: a transient increase in ATP production, a substantial decrease in DNA synthesis, an increase in protein synthesis and a decrease in thiol levels. These results suggested that redox cycling of the vitamin combination increased oxidative stress until it surpassed the reducing ability of the cellular thiols and cellular or genetic damage ensured.
Venugopal M; Jamison J M; Gilloteaux J; Koch J A; Summers M; Giammar D; Sowick C; Summers J L
Life Sciences
1996
1996-09
Journal Article
<a href="http://doi.org/10.1016/0024-3205(96)00466-3" target="_blank" rel="noreferrer noopener">10.1016/0024-3205(96)00466-3</a>
Age estimation in bowhead whales using tympanic bulla histology and baleen isotopes
Arctic; Balaena; Balaena mysticetus; bone; carbon; corpora; GLGs; growth; LAGs; layers; Marine & Freshwater Biology; minke whales; stable isotopes; tympanic bulla; Zoology
Tympanic bullae and baleen plates from bowhead whales of the Western Arctic population were examined. Growth layer groups (GLGs) in the involucrum of the tympanic bone were used to estimate age of the whales, and compared to stable isotope signatures along transects of baleen plates and the involucrum. The involucrum of the tympanic bone consists of three regions that form in utero, during nursing in the first year, and during the first decades of life, respectively. Life history events, such as annual migration, are recorded in the bowhead tympanic bulla. It is likely that bone growth in the bowhead tympanic occurs during periods of high food intake, while slow or arrested growth occurs during periods of low food intake. Comparisons between numbers of GLGs in the tympanic, number of isotopic oscillations in a baleen plate, length of the baleen plate, and total whale length show correlation coefficients as high as 0.97. The tympanic GLG method is particularly useful for estimating the age of whales up to 20 yr old.
Sensor J D; George J C; Clementz M T; Lovano D M; Waugh D A; Givens G H; Suydam R; Stimmelmayr R; Thewissen J G M
Marine Mammal Science
2018
2018-04
Journal Article
<a href="http://doi.org/10.1111/mms.12476" target="_blank" rel="noreferrer noopener">10.1111/mms.12476</a>
Tail Growth Tracks the Ontogeny of Prehensile Tail Use in Capuchin Monkeys (Cebus albifrons and C. apella)
alouatta-palliata; Anthropology; bending strength; Evolutionary Biology; growth; macaca-mulatta; musculoskeletal system; ontogeny; platyrrhine caudal vertebrae; positional behavior; postnatal; segment distribution; squirrel-monkeys; tails; tissue composition; world monkeys
Physical anthropologists have devoted considerable attention to the structure and function of the primate prehensile tail. Nevertheless, previous morphological studies have concentrated solely on adults, despite behavioral evidence that among many primate taxa, including capuchin monkeys, infants and juveniles use their prehensile tails during a greater number and greater variety of positional behaviors than do adults. In this study, we track caudal vertebral growth in a mixed longitudinal sample of white-fronted and brown capuchin monkeys (Cebus albifrons and Cebus apella). We hypothesized that young capuchins would have relatively robust caudal vertebrae, affording them greater tail strength for more frequent tail-suspension behaviors. Our results supported this hypothesis. Caudal vertebral bending strength (measured as polar section modulus at midshaft) scaled to body mass with negative allometry, while craniocaudal length scaled to body mass with positive allometry, indicating that infant and juvenile capuchin monkeys are characterized by particularly strong caudal vertebrae for their body size. These findings complement previous results showing that long bone strength similarly scales with negative ontogenetic allometry in capuchin monkeys and add to a growing body of literature documenting the synergy between postcranial growth and the changing locomotor demands of maturing animals. Although expanded morphometric data on tail growth and behavioral data on locomotor development are required, the results of this study suggest that the adult capuchin prehensile-tail phenotype may be attributable, at least in part, to selection on juvenile performance, a possibility that deserves further attention. Am J Phys Anthropol 146:465-473, 2011. (C) 2011 Wiley-Liss, Inc.
Russo G A; Young J W
American Journal of Physical Anthropology
2011
2011-11
Journal Article
<a href="http://doi.org/10.1002/ajpa.21617" target="_blank" rel="noreferrer noopener">10.1002/ajpa.21617</a>
Ontogeny of intrinsic digit proportions in laboratory rats (Rattus norvegicus): a test of the grasping theory of primate hand and foot growth.
allometry; development; grip; growth; life history; phalangeal index
Young Jesse W; Hyde Alexander; German Rebecca
Biological Journal of the Linnean Society
2019
2019-07
<a href="https://doi.org/10.1093/biolinnean/blz066">https://doi.org/10.1093/biolinnean/blz066</a>
Using the Sauvegrain method to predict peak height velocity in boys and girls.
*Growth; Adolescence; Adolescent; Age Determination by Skeleton – Methods; Age Determination by Skeleton/*methods; Body Height – Physiology; Body Height/*physiology; Child; Elbow – Radiography; Elbow/*diagnostic imaging; Female; Follow-Up Studies; Growth; Human; Humans; Male; Observer Bias; Observer Variation; Prospective Studies; Reproducibility of Results; Reproduction; Sex Characteristics
BACKGROUND: Correlating peak height velocity (PHV) with assessments of skeletal maturity has important implications in the treatment of scoliosis and other pediatric orthopaedic disorders. This study aims to compare the appearance of the elbow to the PHV in both boys and girls. METHODS: We selected 20 children who participated in the Brush Inquiry, a comprehensive study of the development of healthy children. The PHV was identified for each subject. Three observers used the Sauvegrain method to score the elbow maturity of these subjects at 5 visits (PHV -2 years, PHV -1 year, PHV, PHV +1 year, PHV +2 years). Reliability was tested with intraclass correlation coefficients, and maturity scores were compared with the PHV timing. RESULTS: An interrater reliability score of r = 0.915 and an intrarater reliability score of r = 0.909 indicate that this method can be reliably and consistently applied to differentiate elbow x-rays of varying skeletal maturities in children. The mean total scores of boys and girls seem to be equal at the 5 visits. There were no total scores of 26 or higher for boys or girls at PHV. CONCLUSIONS: The Sauvegrain score in adolescent boys and girls is reliable, and a score of 26 or higher indicates that the child has passed PHV. There is a strong trend for the mean total score of boys to equal that of girls at each stage relative to the PHV. LEVEL OF EVIDENCE: Diagnostic Level III. See Instructions to Authors for a complete description of levels of evidence.
Hans Sarah D; Sanders James O; Cooperman Daniel R
Journal of pediatric orthopedics
2008
2008-12
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1097/BPO.0b013e31818ee3c4" target="_blank" rel="noreferrer noopener">10.1097/BPO.0b013e31818ee3c4</a>