78 EFFICACY OF USING IN VITRO-PRODUCED CATTLE EMBRYOS TO DEVELOP AN EQUINE EMBRYO CRYOPRESERVATION SYSTEM

2009 ◽  
Vol 21 (1) ◽  
pp. 139
Author(s):  
C. M. Syverson ◽  
A. M. Paprocki ◽  
R. W. Koppang ◽  
J. R. Dobrinsky
Keyword(s):  
The United States ◽  
Blastocyst Stage ◽  
Embryo Cryopreservation ◽  
State University ◽  
Pump System ◽  
In Vitro System ◽  
Colorado State University ◽  
Fetal Bovine

Unfortunately, there is no reliable in vitro system available to provide a routine and economical source of equine embryos for research and commercial use. We explore the use of standard in vitro-produced (IVP) cattle embryos for the development of a direct transfer (dt) equine embryo cryopreservation system. Ovaries were collected from mature females at an abattoir and transported to our laboratory. Cumulus–oocyte complexes (COC) were aspirated from 2- to 6-mm follicles with an 18-gauge needle fixed to a vacuum pump system. Only COC surrounded by 2 or more layers of compact cumulus investment and containing oocytes of equal size were placed into a commercial TCM-199-based IVM system (Minitube of America Inc., Verona, WI). After 22 h of IVM, mature COC were placed into standard IVF. Prospective embryos were cultured for 120 h in CR-1/BSA (Minitube of America Inc.), and then supplemented with 10% fetal bovine serum (FBS) and cultured for an additional 48 h. Only excellent to good morula-blastocyst stage IVP cattle embryos were used in this study. As a control, the Colorado State University equine embryo dt-vitrification system (CSU; Carnevale EM Vet. Clin. North Am. Equine Pract 22, 831–841) was used on IVP cattle blastocysts. Two different Minitube EquiPRO-based dt-media with either 10% FBS or Minitube BSA-V (Fraction-V; Minitube of America Inc.) were tested on IVP cattle embryos. Minitube BSA-V is highly defined and internationally compliant BSA approved for use in raw form or in culture medium in the United States and European Union. Of 103 IVP cattle embryos cryopreserved by the CSU method, 45 (43.7%) embryos were viable after 24 h of culture. Of 121 IVP cattle embryos cryopreserved with Minitube EquiPRO + 10% FBS, 52 (42.9%) embryos were viable after 24 h of culture. Of 90 IVP cattle embryos cryopreserved with Minitube EquiPRO + BSA-V, 40 (44.4%) embryos were viable after 24 h of culture. Minitube EquiPRO + BSA-V was used with in vivo-produced equine morulae (no capsule). A total of 9 mares were flushed on Day 6.5, and 8 excellent to good morulae were recovered. All 8 embryos were dt-vitrified in the Minitube EquiPRO + BSA-V system. All 8 embryos were later warmed and transferred to synchronous recipient mares. On Day 14 of presumptive pregnancy, 7 (87.5%) mares were confirmed pregnant by real-time ultrasound examination. Not all presumptive foals were needed, so 4 recipient mares were randomly selected and intentionally aborted. Three mares were selected to carry presumptive foals to term. All 3 mares produced live and healthy foals at the term of gestation. Standard IVP cattle embryos can serve as a relatively inexpensive model for the development and testing of equine embryo cryopreservation systems.

Bone Formation by Human Oseoblasts on Implant Materials

1998 ◽  
Vol 4 (S2) ◽  
pp. 938-939
Author(s):  
G. Gronowicz ◽  
M. Ahmad
Keyword(s):  
In Vitro System ◽  
Implant Materials ◽  
Sequence Of Events ◽  
Ecm Proteins ◽  
Transmission Electron ◽  
Fluorescence Light ◽  
Primary Human Osteoblasts ◽  
Fetal Bovine

We have developed an in vitro system to study the process by which osteoblasts attach and secrete extracellular matrix (ECM) proteins that subsequently mineralize in response to implant materials. In vivo mineralization is characterized by the sequential expression by osteoblasts of specific ECM proteins that are capable of calcifying in an orderly manner. A similar sequence of events is shown to occur in this in vitro system. The extent and pattern of mineralization was visualized by fluorescence light microscopy and transmission electron microscopy (TEM), and was dependent on the substrate to which osteoblasts adhered.Materials and MethodsDisks (22 mm diameter) composed of Tivanium (Tiv) (Ti6AI4V, ASTM F-136) and Zimaloy (Zim)(CoCrMo, ASTM F-75), prepared identically to clinical orthopaedic implants, were supplied by Zimmer, Inc. Glass disks were used as controls. Osteoblast-like cells (Saos-2), derived from a human osteosarcoma, or primary human osteoblasts from a 41 year-old male patient were added to the disks in a-MEM medium with 10% fetal bovine serum,

Repurposing N,N-Dimethylacetamide (DMA), a Pharmaceutical Excipient, as a Prototype Novel Anti-inflammatory Agent for the Prevention and/or Treatment of Preterm Birth

2018 ◽  
Vol 24 (9) ◽  
pp. 989-992 ◽  
Author(s):  
Samir Gorasiya ◽  
Juliet Mushi ◽  
Ryan Pekson ◽  
Sabesan Yoganathan ◽  
Sandra E. Reznik
Keyword(s):  
Preterm Birth ◽  
Ex Vivo ◽  
The United States ◽  
Occurrence Rate ◽  
Pharmaceutical Excipient ◽  
Ongoing Work ◽  
Exciting Line ◽  
Pregnant Mice

Background: Preterm birth (PTB), or birth that occurs before 37 weeks of gestation, accounts for the majority of perinatal morbidity and mortality. As of 2016, PTB has an occurrence rate of 9.6% in the United States and accounts for up to 18 percent of births worldwide. Inflammation has been identified as the most common cause of PTB, but effective pharmacotherapy has yet to be developed to prevent inflammation driven PTB. Our group has discovered that N,N-dimethylacetamide (DMA), a readily available solvent commonly used as a pharmaceutical excipient, rescues lipopolysaccharide (LPS)-induced timed pregnant mice from PTB. Methods: We have used in vivo, ex vivo and in vitro approaches to investigate this compound further. Results: Interestingly, we found that DMA suppresses cytokine secretion by inhibiting nuclear factor-kappa B (NF-κB). In ongoing work in this exciting line of investigation, we are currently investigating structural analogs of DMA, some of them novel, to optimize this approach focused on the inflammation associated with PTB. Conclusion: Successful development of pharmacotherapy for the prevention of PTB rests upon the pursuit of multiple strategies to solve this important clinical challenge.

scholarly journals Photosensitizing Medications and Skin Cancer: A Comprehensive Review

Cancers ◽  
2021 ◽  
Vol 13 (10) ◽  
pp. 2344
Author(s):  
Elisabeth A. George ◽  
Navya Baranwal ◽  
Jae H. Kang ◽  
Abrar A. Qureshi ◽  
Aaron M. Drucker ◽  
...  
Keyword(s):  
Skin Cancer ◽  
Cancer Risk ◽  
Cell Carcinoma ◽  
The United States ◽  
In Vivo Studies ◽  
Cancer Risk Factors ◽  
Skin Cancer Risk ◽  
Number Of Patients

(1) The incidence of skin cancer is increasing in the United States (US) despite scientific advances in our understanding of skin cancer risk factors and treatments. In vitro and in vivo studies have provided evidence that suggests that certain photosensitizing medications (PSMs) increase skin cancer risk. This review summarizes current epidemiological evidence on the association between common PSMs and skin cancer. (2) A comprehensive literature search was conducted to identify meta-analyses, observational studies and clinical trials that report on skin cancer events in PSM users. The associated risks of keratinocyte carcinoma (squamous cell carcinoma and basal cell carcinoma) and melanoma are summarized, for each PSM. (3) There are extensive reports on antihypertensives and statins relative to other PSMs, with positive and null findings, respectively. Fewer studies have explored amiodarone, metformin, antimicrobials and vemurafenib. No studies report on the individual skin cancer risks in glyburide, naproxen, piroxicam, chlorpromazine, thioridazine and nalidixic acid users. (4) The research gaps in understanding the relationship between PSMs and skin cancer outlined in this review should be prioritized because the US population is aging. Thus the number of patients prescribed PSMs is likely to continue to rise.

scholarly journals Effects of Hyperglycemia on Vascular Smooth Muscle Ca2+Signaling

2017 ◽  
Vol 2017 ◽  
pp. 1-16 ◽  
Author(s):  
Nahed El-Najjar ◽  
Rashmi P. Kulkarni ◽  
Nancy Nader ◽  
Rawad Hodeify ◽  
Khaled Machaca
Keyword(s):  
Insulin Resistance ◽  
Smooth Muscle ◽  
Sarcoplasmic Reticulum ◽  
Vascular Smooth Muscle ◽  
Complex Disease ◽  
Prolonged Exposure ◽  
In Vitro System ◽  
A7r5 Cells

Diabetes is a complex disease that is characterized with hyperglycemia, dyslipidemia, and insulin resistance. These pathologies are associated with significant cardiovascular implications that affect both the macro- and microvasculature. It is therefore important to understand the effects of various pathologies associated with diabetes on the vasculature. Here we directly test the effects of hyperglycemia on vascular smooth muscle (VSM) Ca2+signaling in an isolated in vitro system using the A7r5 rat aortic cell line as a model. We find that prolonged exposure of A7r5 cells to hyperglycemia (weeks) is associated with changes to Ca2+signaling, including most prominently an inhibition of the passive ER Ca2+leak and the sarcoplasmic reticulum Ca2+-ATPase (SERCA). To translate these findings to the in vivo condition, we used primary VSM cells from normal and diabetic subjects and find that only the inhibition of the ER Ca2+leaks replicates in cells from diabetic donors. These results show that prolonged hyperglycemia in isolation alters the Ca2+signaling machinery in VSM cells. However, these alterations are not readily translatable to the whole organism situation where alterations to the Ca2+signaling machinery are different.

scholarly journals Expression and Characterization of the Flocculin Flo11/Muc1, a Saccharomyces cerevisiae Mannoprotein with Homotypic Properties of Adhesion

2007 ◽  
Vol 6 (12) ◽  
pp. 2214-2221 ◽  
Author(s):  
Lois M. Douglas ◽  
Li Li ◽  
Yang Yang ◽  
A. M. Dranginis
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Biofilm Formation ◽  
In Vitro System ◽  
Glycosylphosphatidylinositol Anchor ◽  
Fungal Adhesion ◽  
Very High ◽  
Molecular Weight Form

ABSTRACT The Flo11/Muc1 flocculin has diverse phenotypic effects. Saccharomyces cerevisiae cells of strain background Σ1278b require Flo11p to form pseudohyphae, invade agar, adhere to plastic, and develop biofilms, but they do not flocculate. We show that S. cerevisiae var. diastaticus strains, on the other hand, exhibit Flo11-dependent flocculation and biofilm formation but do not invade agar or form pseudohyphae. In order to study the nature of the Flo11p proteins produced by these two types of strains, we examined secreted Flo11p, encoded by a plasmid-borne gene, in which the glycosylphosphatidylinositol anchor sequences had been replaced by a histidine tag. A protein of approximately 196 kDa was secreted from both strains, which upon purification and concentration, aggregated into a form with a very high molecular mass. When secreted Flo11p was covalently attached to microscopic beads, it conferred the ability to specifically bind to S. cerevisiae var. diastaticus cells, which flocculate, but not to Σ1278b cells, which do not flocculate. This was true for the 196-kDa form as well as the high-molecular-weight form of Flo11p, regardless of the strain source. The coated beads bound to S. cerevisiae var. diastaticus cells expressing FLO11 and failed to bind to cells with a deletion of FLO11, demonstrating a homotypic adhesive mechanism. Flo11p was shown to be a mannoprotein. Bead-to-cell adhesion was inhibited by mannose, which also inhibits Flo11-dependent flocculation in vivo, further suggesting that this in vitro system is a useful model for the study of fungal adhesion.

How Does the Inclusion of Twins Conceived via Fertility Treatments Influence the Results of Twin Studies?

2012 ◽  
Vol 15 (6) ◽  
pp. 746-752 ◽  
Author(s):  
S. Alexandra Burt ◽  
Kelly L. Klump
Keyword(s):  
Twin Studies ◽  
The United States ◽  
Fertility Treatment ◽  
State University ◽  
Behavioral Differences ◽  
Vitro Fertilization ◽  
Fertility Treatments ◽  
Control And Prevention ◽  
Twin Families

Rates of twinning have risen dramatically over the last 30 years, from 1 in 53 births in 1980 to 1 in 30 births in 2009 (Martin et al. (January 2012). Three decades of twin births in the United States, 1980–2009. Atlanta, GA: Centers for Disease Control and Prevention, National Center for Health Statistics). This increase is largely attributable to increases in the use of fertility treatments (i.e., ovulation induction and in vitro fertilization) combined with delays in parenthood. Although this increase means that more twins are available for recruitment into twin studies, it also has potential consequences for the heritability estimates obtained in these studies. This study sought to evaluate this possibility, making use of the ongoing Michigan Twins Project (N = 7,261 families with twins aged 3–17 years), an arm of the Michigan State University Twin Registry. Results revealed that, on average, twins conceived via fertility treatments had lower rates of behavior problems than those conceived naturally, although these behavioral differences could be explained largely by demographic and socio-economic differences across the two types of twin families. Twin similarity did not meaningfully differ across fertility treatment status. We thus conclude that estimates of genetic and environmental influences obtained from twin studies over the last 10–15 years are more or less unaffected by the inclusion of twins conceived via fertility treatments in their samples.

scholarly journals Activation of yeast polymerase II transcription by herpesvirus VP16 and GAL4 derivatives in vitro.

1989 ◽  
Vol 9 (11) ◽  
pp. 4746-4749 ◽  
Author(s):  
D I Chasman ◽  
J Leatherwood ◽  
M Carey ◽  
M Ptashne ◽  
R D Kornberg
Keyword(s):  
Rna Polymerase Ii ◽  
Fusion Proteins ◽  
In Vitro System ◽  
Transcription System ◽  
Natural Mechanism ◽  
Transcription In Vitro ◽  
Rna Polymerase Ii Transcription ◽  
Fold Stimulation

Fusion proteins known to activate transcription in vivo were tested for the ability to stimulate transcription in vitro in a recently developed Saccharomyces cerevisiae RNA polymerase II transcription system. One fusion protein, whose activation domain was derived from the herpesvirus transcriptional activator VP16, gave more than 100-fold stimulation in the in vitro system. The order of effects of the various proteins was the same for transcription in vitro and in vivo, suggesting that the natural mechanism of activation is preserved in vitro.

Accurate transcription of a plant mitochondrial gene in vitro

1991 ◽  
Vol 11 (4) ◽  
pp. 2035-2039
Author(s):  
P J Hanic-Joyce ◽  
M W Gray
Keyword(s):  
Mitochondrial Gene ◽  
Plant Mitochondria ◽  
In Vitro Transcription ◽  
Dna Templates ◽  
In Vitro System ◽  
Transcription System ◽  
Translation Initiation Codon ◽  
Mitochondrial Extract

To investigate transcriptional mechanisms in plant mitochondria, we have developed an accurate and efficient in vitro transcription system consisting of a partially purified wheat mitochondrial extract programmed with cloned DNA templates containing the promoter for the wheat mitochondrial cytochrome oxidase subunit II gene (coxII). Using this system, we localize the coxII promoter to a 372-bp region spanning positions -56 to -427 relative to the coxII translation initiation codon. We show that in vitro transcription of coxII is initiated at position -170, precisely the same site at which transcription is initiated in vivo. Transcription begins within the sequence GTATAGTAAGTA (the initiating nucleotide is underlined), which is similar to the consensus yeast mitochondrial promoter motif, (A/T)TATAAGTA. This is the first in vitro system that faithfully reproduces in vivo transcription of a plant mitochondrial gene.

Severe Hemolytic Reaction Due to Anti-JK3

1999 ◽  
Vol 123 (10) ◽  
pp. 949-951
Author(s):  
Carol S. Marshall ◽  
Denis Dwyre ◽  
Robin Eckert ◽  
Liisa Russell
Keyword(s):  
The United States ◽  
Cell Phenotype ◽  
Prior History ◽  
Ethnic Variability ◽  
Hemolytic Transfusion Reaction ◽  
History Of ◽  
Rare Phenotype ◽  
Antibody Screen

Abstract A 35-year-old gravida 3, para 3 Filipino woman with a negative antibody screen, no prior history of transfusion, and no hemolytic disease of the newborn in her children suffered a massive postpartum hemorrhage requiring transfusion. A severe hemolytic transfusion reaction occurred 5 days after delivery. Subsequently, a panagglutinin on a routine antibody identification panel was identified as anti-Jk3. The patient's red blood cell phenotype was Jk(a−b−) and all of her children were Jk(a−b+), yet the antibody that formed reacted with equal strength against all Jka- or Jkb-positive cells. The rare Jk(a−b−) phenotype is more common in Polynesians. Anti-Jk3, like other Kidd system antibodies, is difficult to detect because in vivo production may be absent between provocative episodes and because these antibodies often show weak in vitro reactions. The increasing numbers of Pacific Islanders in the United States could result in more frequent encounters with this rare phenotype. Increased awareness of ethnic variability in blood phenotypes and of the capricious nature of Kidd antibodies can help pathologists and technologists deal more effectively with these cases.

Virulence and Genetic Characterization of Six Baculovirus Strains Isolated From Different Populations of Spodoptera Frugiperda (Lepidoptera: Noctuidae)

2021 ◽  
Author(s):  
Ingrid Zanella-Saenz ◽  
Elisabeth A. Herniou ◽  
Jorge E. Ibarra ◽  
Ma.Cristina Del Rincón-Castro ◽  
Ilse Alejandra Huerta-Arredondo
Keyword(s):  
Biological Control ◽  
Cell Cultures ◽  
Spodoptera Frugiperda ◽  
Fall Armyworm ◽  
The United States ◽  
Infection Mechanisms ◽  
Different Populations

Abstract Fall armyworm (FAW), Spodoptera frugiperda (Smith, 1797), is a polyphagous, voracious, and economically important agricultural pest. Biological control of FAW is a strategy that must be further explored. This study evaluated six baculovirus strains isolated from infected FAW larvae from Mexico, Argentina, Honduras, and the United States. Five alphabaculoviruses (SfNPV-An2, SfNPV-Arg, SfNPV-Fx, SfNPV-Ho and SfNPV-Sin) and one betabaculovirus (SfGV-RV), were tested against FAW larvae, showing a wide diversity of virulence levels among strains when their estimated LC50s were compared, being SfNPVArg, SfNPV-Ho and SfNPV-Fx more virulent than SfNPV-An 2 , SfNPV-Sin and SfGV-RV. To determine any virulence difference in vitro studies of these isolates, Sf9 cell cultures were used. Interestingly, only ODVs from four of the test SfNPV strains showed infectivity on Sf9 cell cultures, and some differences in virulence were observed. Genomic restriction analyses and partial sequences of lef-8, lef-9 , and polh/granulin genes showed little variability among alphabaculoviruses, both, among them and with previously reported sequences. However, sequences from SfGV-RV were closer to previously reported sequences from the SfGVVG008 strain than the SfGV-Arg and SfGV-VG014 strains. The great difference in the in vivo virulence was not correlated with great similarity among the isolates. The characterization of these six baculoviruses isolates offers the basis for exploring their potential as biological control agents against S. frugiperda, as well the initial studies on their specific infection mechanisms, evolution, and ecology.

Sign in / Sign up

Export Citation Format

Share Document