Production of functional proteins: balance of shear stress and gravity

The present invention provides a method for production of functional proteins including hormones by renal cells in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-a-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel

Production of functional proteins: balance of shear stress and gravity

A method for the production of functional proteins including hormones by renal cells in a three dimensional culturing process responsive to shear stress uses a rotating wall vessel. Natural mixture of renal cells expresses the enzyme 1-.alpha.-hydroxylase which can be used to generate the active form of vitamin D: 1,25-diOH vitamin D.sub.3. The fibroblast cultures and co-culture of renal cortical cells express the gene for erythropoietin and secrete erythropoietin into the culture supernatant. Other shear stress response genes are also modulated by shear stress, such as toxin receptors megalin and cubulin (gp280). Also provided is a method of treating an in-need individual with the functional proteins produced in a three dimensional co-culture process responsive to shear stress using a rotating wall vessel

Correcting errors in synthetic DNA through consensus shuffling

Although efficient methods exist to assemble synthetic oligonucleotides into genes and genomes, these suffer from the presence of 1–3 random errors/kb of DNA. Here, we introduce a new method termed consensus shuffling and demonstrate its use to significantly reduce random errors in synthetic DNA. In this method, errors are revealed as mismatches by re-hybridization of the population. The DNA is fragmented, and mismatched fragments are removed upon binding to an immobilized mismatch binding protein (MutS). PCR assembly of the remaining fragments yields a new population of full-length sequences enriched for the consensus sequence of the input population. We show that two iterations of consensus shuffling improved a population of synthetic green fluorescent protein (GFPuv) clones from ∼60 to >90% fluorescent, and decreased errors 3.5- to 4.3-fold to final values of ∼1 error per 3500 bp. In addition, two iterations of consensus shuffling corrected a population of GFPuv clones where all members were non-functional, to a population where 82% of clones were fluorescent. Consensus shuffling should facilitate the rapid and accurate synthesis of long DNA sequences

Association of physical function with predialysis blood pressure in patients on hemodialysis

BACKGROUND: New information from various clinical settings suggests that tight blood pressure control may not reduce mortality and may be associated with more side effects. METHODS: We performed cross-sectional multivariable ordered logistic regression to examine the association between predialysis blood pressure and the short physical performance battery (SPPB) in a cohort of 749 prevalent hemodialysis patients in the San Francisco and Atlanta areas recruited from July 2009 to August 2011 to study the relationship between systolic blood pressure and objective measures of physical function. Mean blood pressure for three hemodialysis sessions was analyzed in the following categories: <110 mmHg, 110-129 mmHg (reference), 130-159 mmHg, and ≥160 mmHg. SPPB includes three components: timed repeated chair stands, timed 15-ft walk, and balance tests. SPPB was categorized into ordinal groups (≤6, 7-9, 10-12) based on prior literature. RESULTS: Patients with blood pressure 130-159 mmHg had lower odds (OR 0.57, 95% CI 0.35-0.93) of scoring in a lower SPPB category than those whose blood pressure was between 110 and 129 mmHg, while those with blood pressure ≥160 mmHg had 0.56 times odds (95% CI 0.33-0.94) of scoring in a lower category when compared with blood pressure 110-129 mmHg. When individual components were examined, blood pressure was significantly associated with chair stand (130-159 mmHg: OR 0.59, 95% CI 0.38-0.92) and gait speed (≥160 mmHg: OR 0.59, 95% CI 0.35-0.98). Blood pressure ≥160 mmHg was not associated with substantially higher SPPB score compared with 130-159 mmHg. CONCLUSIONS: Patients with systolic blood pressure at or above 130 mmHg had better physical performance than patients with lower blood pressure in the normotensive range. The risk-benefit tradeoff of aggressive blood pressure control, particularly in low-functioning patients, should be reexamined

Wisconsin 1936

Manuscript map ; Map used in Kaysen's 1937 book entitled: "The railroads of Wisconsin 1827-1937." Map listed in: Manuscript and annotated maps in the American Geographical Society Library: a cartobibliography, 2010.James P. Kaysen was a civil engineer, Wisconsin railroad historian and author of The Railroads of Wisconsin, 1827-1937 (Boston: Railway and Locomotive Historical Society, 1937). The Kaysen maps in the AGS Library include several manuscript maps (entries #95-99) in addition to a number of annotated U.S. Geological Survey topographic quadrangle sheets (Appendices A and B). The maps were given to the AGS Library by Mrs. James P. Kaysen in the mid-1990s. This collection complements the James P. Kaysen Papers, 1930-1979, held by the Wisconsin Historical Society Archives in Madison.Grayscale1:633,60

Kickapoo Hills, Oak Creek, Wisconsin 1930

"University of Wisconsin, 623 W. State St., Milwaukee, Wis." Map listed in: Manuscript and annotated maps in the American Geographical Society Library: a cartobibliography, 2010. James P. Kaysen was a civil engineer, Wisconsin railroad historian and author of The Railroads of Wisconsin, 1827-1937 (Boston: Railway and Locomotive Historical Society, 1937). The Kaysen maps in the AGS Library include several manuscript maps (entries #95-99) in addition to a number of annotated U.S. Geological Survey topographic quadrangle sheets (Appendices A and B). The maps were given to the AGS Library by Mrs. James P. Kaysen in the mid-1990s. This collection complements the James P. Kaysen Papers, 1930-1979, held by the Wisconsin Historical Society Archives in Madison.Grayscale;1:1,20

Lake Spur, Wisconsin 1938

Blueline print. Original was a manuscript map. "Feb. 6, 1938 J.P.K."Color1:15,84

Milwaukee, Wisconsin 1928

Manuscript map.; Includes notes.; "July 1928."; James P. Kaysen was a civil engineer, Wisconsin railroad historian and author of The Railroads of Wisconsin, 1827-1937 (Boston: Railway and Locomotive Historical Society, 1937). The Kaysen maps in the AGS Library include several manuscript maps (entries #95-99) in addition to a number of annotated U.S. Geological Survey topographic quadrangle sheets (Appendices A and B). The maps were given to the AGS Library by Mrs. James P. Kaysen in the mid-1990s. This collection complements the James P. Kaysen Papers, 1930-1979, held by the Wisconsin Historical Society Archives in Madison.; Map listed in: Manuscript and annotated maps in the American Geographical Society Library: a cartobibliography, 2010.;Grayscale;Scale approximately 1:50,000. 1 1/4 in. = 1 mile