IN VIVO AND IN VITRO EXPRESSION OF SOMATOSTATIN BY HUMAN KIDNEY CELLS. • 2213

Sepsis-associated acute kidney injury (SA-AKI) is a life-threatening syndrome. Lipopolysaccharide (LPS) is a widely used inducer for modeling SA-AKI both in vivo and in vitro. However, due to the innate complexity of the kidney architecture, the mechanisms underlying the pathogenesis of SA-AKI, as well as those involved in LPS-induced kidney injury remain to be clarified. Kidney organoids derived from human pluripotent stem cells (hPSCs) act as a model of multiple types of kidney cells in vitro and eliminate potential confounders in vivo. In the current study, we established LPS-induced kidney injury models both in vivo and in human kidney organoids. Kidney function, pathological changes, and markers of oxidative stress were evaluated with/without the presence of methylprednisolone (MP) treatment both in vivo and in vitro. The extent of LPS-induced oxidative stress and apoptosis in kidney organoids was further investigated in vitro. LPS-induced acute kidney injury in mice, together with pathological changes and increased oxidative stress, as well as enhanced apoptosis in kidney cells were evaluated. These phenomena were ameliorated by MP treatment. Experiments in kidney organoids showed that the LPS-induced apoptotic effects occurred mainly in podocytes and proximal tubular cells. Our experiments demonstrated the efficacy of using kidney organoids as a solid platform to study LPS-induced kidney injury. LPS induced oxidative stress as well as apoptosis in kidney cells independently of changes in perfusion or immune cell infiltration. MP treatment partially alleviated LPS-induced injury by reducing kidney cell oxidative stress and apoptosis.

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The Effect of Vaginal Microbes on in Vivo and in Vitro Expression of Human Papillomavirus 16 E6-E7 Genes

Cancer Detection and Prevention ◽

10.1046/j.1525-1500.1999.00069.x ◽

1999 ◽

Vol 23 (1) ◽

pp. 13-21 ◽

Cited By ~ 4

Author(s):

Patricia J. McNicol ◽

Maria Paraskevas ◽

Fernando B. Guijon

Keyword(s):

Human Papillomavirus ◽

In Vitro Expression ◽

Human Papillomavirus 16

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MST3 is involved in ENaC-mediated hypertension

AJP Renal Physiology ◽

10.1152/ajprenal.00455.2018 ◽

2019 ◽

Vol 317 (1) ◽

pp. F30-F42

Author(s):

Te-Jung Lu ◽

Wei-Chih Kan ◽

Sung-Sen Yang ◽

Si-Tse Jiang ◽

Sheng-Nan Wu ◽

...

Keyword(s):

Kidney Cells ◽

Hypertensive Rats ◽

Madin Darby Canine Kidney ◽

Spontaneously Hypertensive ◽

Hypomorphic Mutation ◽

And Oxidative Stress ◽

Darby Canine Kidney ◽

Canine Kidney

Liddle syndrome is an inherited form of human hypertension caused by increasing epithelial Na+ channel (ENaC) expression. Increased Na+ retention through ENaC with subsequent volume expansion causes hypertension. In addition to ENaC, the Na+-K+-Cl− cotransporter (NKCC) and Na+-Cl− symporter (NCC) are responsible for Na+ reabsorption in the kidneys. Several Na+ transporters are evolutionarily regulated by the Ste20 kinase family. Ste20-related proline/alanine-rich kinase and oxidative stress-responsive kinase-1 phosphorylate downstream NKCC2 and NCC to maintain Na+ and blood pressure (BP) homeostasis. Mammalian Ste20 kinase 3 (MST3) is another member of the Ste20 family. We previously reported that reduced MST3 levels were found in the kidneys in spontaneously hypertensive rats and that MST3 was involved in Na+ regulation. To determine whether MST3 is involved in BP stability through Na+ regulation, we generated a MST3 hypomorphic mutation and designated MST3+/− and MST3−/− mice to examine BP and serum Na+ and K+ concentrations. MST3−/− mice exhibited hypernatremia, hypokalemia, and hypertension. The increased ENaC in the kidney played roles in hypernatremia. The reabsorption of more Na+ promoted more K+ secretion in the kidney and caused hypokalemia. The hypernatremia and hypokalemia in MST3−/− mice were significantly reversed by the ENaC inhibitor amiloride, indicating that MST3−/− mice reabsorbed more Na+ through ENaC. Furthermore, Madin-Darby canine kidney cells stably expressing kinase-dead MST3 displayed elevated ENaC currents. Both the in vivo and in vitro results indicated that MST3 maintained Na+ homeostasis through ENaC regulation. We are the first to report that MST3 maintains BP stability through ENaC regulation.

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671 Comparative Analysis of in Vivo and in Vitro Expression of Kinases in the MAPK Pathway

European Journal of Cancer ◽

10.1016/s0959-8049(12)71316-3 ◽

2012 ◽

Vol 48 ◽

pp. S159

Author(s):

W.M. Dickerson ◽

L.A. Beausang ◽

A. Saab ◽

K. Leong ◽

E.M. Alderman

Keyword(s):

Comparative Analysis ◽

Mapk Pathway ◽

In Vitro Expression

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Synthesis and maturation of lambda receptor in Escherichia coli K-12: in vivo and in vitro expression of gene lamB under lac promoter control.

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.77.3.1491 ◽

1980 ◽

Vol 77 (3) ◽

pp. 1491-1495 ◽

Cited By ~ 16

Author(s):

C. Marchal ◽

D. Perrin ◽

J. Hedgpeth ◽

M. Hofnung

Keyword(s):

Escherichia Coli ◽

In Vitro Expression ◽

K 12 ◽

Lac Promoter

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In vivo and in vitro expression of steroid-converting enzymes in human breast tumours: associations with interleukin-6

British Journal of Cancer ◽

10.1038/sj.bjc.6690749 ◽

1999 ◽

Vol 81 (4) ◽

pp. 690-695 ◽

Cited By ~ 20

Author(s):

V Speirs ◽

D S Walton ◽

M-C Hall ◽

S L Atkin

Keyword(s):

Interleukin 6 ◽

Human Breast ◽

Breast Tumours ◽

In Vitro Expression

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In Vivo Entombment of Bacteria and Fungi during Calcium Oxalate, Brushite and Struvite Urolithiasis

Kidney360 ◽

10.34067/kid.0006942020 ◽

2020 ◽

pp. 10.34067/KID.0006942020

Author(s):

Jessica J. Saw ◽

Mayandi Sivaguru ◽

Elena M. Wilson ◽

Yiran Dong ◽

Robert A. Sanford ◽

...

Keyword(s):

Calcium Oxalate ◽

Stone Formation ◽

Human Kidney ◽

Rrna Gene ◽

Bacterial Cells ◽

Stone Formers ◽

Bacteria And Fungi ◽

Struvite Stones

Background: Human kidney stones form via repeated events of mineral precipitation, partial dissolution and reprecipitation, which are directly analogous to similar processes in other natural and man-made environments where resident microbiomes strongly influence biomineralization. High-resolution microscopy and high-fidelity metagenomic (microscopy-to-omics) analyses, applicable to all forms of biomineralization, have been applied to assemble definitive evidence of in vivo microbiome entombment during urolithiasis. Methods: Stone fragments were collected from a randomly chosen cohort of 20 patients using standard percutaneous nephrolithotomy (PCNL). Fourier transform infrared (FTIR) spectroscopy indicated that 18 of these patients were calcium oxalate (CaOx) stone formers, while one patient each formed brushite and struvite stones. This apportionment is consistent with global stone mineralogy distributions. Stone fragments from 7 of these 20 patients (5 CaOx, 1 brushite and 1 struvite) were thin sectioned and analyzed using brightfield (BF), polarization (POL), confocal, superresolution autofluorescence (SRAF) and Raman techniques. DNA from remaining fragments, grouped according to each of the 20 patients, were analyzed with amplicon sequencing of 16S rRNA gene sequences (V1-V3, V3-V5) and internal transcribed spacer (ITS1, ITS2) regions. Results: Bulk entombed DNA was sequenced from stone fragments in 11 of the 18 CaOx patients, as well as the brushite and struvite patients. These analyses confirmed the presence of an entombed low-diversity community of bacteria and fungi, including Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, and Aspergillus niger. Bacterial cells ~1 µm in diameter were also optically observed to be entombed and well-preserved in amorphous hydroxyapatite spherules and fans of needle-like crystals of brushite and struvite. Conclusions: These results indicate a microbiome is entombed during in vivo CaOx stone formation. Similar processes are implied for brushite and struvite stones. This evidence lays the groundwork for future in vitro and in vivo experimentation to determine how the microbiome may actively and/or passively influence kidney stone biomineralization.

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In Vivo and in Vitro Expression of Foreign Genes in Mouse Testicular Cells

Animal Cell Technology: Basic & Applied Aspects ◽

10.1007/978-94-011-5746-9_56 ◽

1997 ◽

pp. 355-358

Author(s):

T. Muramatsu ◽

O. Shibata ◽

J. Okumura

Keyword(s):

In Vitro Expression ◽

Testicular Cells ◽

Foreign Genes

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3D kidney organoids for bench-to-bedside translation

Journal of Molecular Medicine ◽

10.1007/s00109-020-01983-y ◽

2020 ◽

Author(s):

Navin Gupta✉ ◽

Emre Dilmen ◽

Ryuji Morizane

Keyword(s):

Collecting Duct ◽

Developmental Toxicity ◽

Human Kidney ◽

Great Promise ◽

Duct Systems ◽

Recent Developments ◽

Induced Pluripotent ◽

Kidney Organoids

Abstract The kidneys are essential organs that filter the blood, removing urinary waste while maintaining fluid and electrolyte homeostasis. Current conventional research models such as static cell cultures and animal models are insufficient to grasp the complex human in vivo situation or lack translational value. To accelerate kidney research, novel research tools are required. Recent developments have allowed the directed differentiation of induced pluripotent stem cells to generate kidney organoids. Kidney organoids resemble the human kidney in vitro and can be applied in regenerative medicine and as developmental, toxicity, and disease models. Although current studies have shown great promise, challenges remain including the immaturity, limited reproducibility, and lack of perfusable vascular and collecting duct systems. This review gives an overview of our current understanding of nephrogenesis that enabled the generation of kidney organoids. Next, the potential applications of kidney organoids are discussed followed by future perspectives. This review proposes that advancement in kidney organoid research will be facilitated through our increasing knowledge on nephrogenesis and combining promising techniques such as organ-on-a-chip models.

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