Ventriculocisternal perfusions were carried out on chronically prepared, unanesthetized goats. Measurements were made of steady-state rates at which inulin, fructose, creatinine, urea, K, Na, and labeled water (TOH) were removed from perfusion fluid at various hydrostatic and osmotic pressures. The ventricular system is virtually impermeable to inulin. Inulin is removed from subarachnoid spaces by bulk absorption at rates which vary linearly with hydrostatic pressure. Net absorption ceases at –15 cm H2O. Rate of net formation of cerebrospinal fluid (CSF) is equal to inulin clearance plus the difference between outflow and inflow rates; normally it averages 0.16 cm3 min–1 and is essentially independent of hydrostatic pressures in the range –10 to +30 cm H2O. Net rate of formation is linearly related to total osmotic pressure differences between plasma and CSF. The coefficient of osmotic flow is greater than that measured from diffusion of TOH, as in other porous membranes. Passive permeability characteristics resemble those of the vasopressin-treated toad bladder.

Respiratory responses to inhaled CO2 were measured in unanesthetized goats during repeated perfusions of the ventriculocisternal system through chronically implanted cannulas. [HCO3–] and pH were measured in carotid loop blood and cisternal outflow. Average steady-state alveolar ventilation increased fourfold when cerebrospinal fluid (CSF)-[HCO3–] was reduced from 30 to 15 mm/liter at constant, normal CO2 pressure or threefold when CSF pH changed from 7.32 to 7.21 at constant, normal CSF-[HCO3–]. Sensitivity was two- to sevenfold greater than reported for anesthetized animals. At constant CSF pH the ventilatory response to inhaled CO2 was 60% of the isobicarbonate response. Pco2 in cisternal outflow was shown to approximate that in cerebral tissue. HCO3– flux was measured as a function of CSF-[HCO3–] and concentration profiles between CSF and capillary blood were considered. Alveolar ventilation is a single linear function of [H+] in tissue fluid located two-thirds to three-fourths of the distance along the functional concentration gradient of HCO3– between CSF and blood at all values of Pco2 and CSF-[HCO3–] which we investigated.

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Type 1 diabetes mellitus (T1DM) is caused by immune destruction of insulin-producing pancreatic β-cells. Commonly used insulin injection therapy does not provide a dynamic blood glucose control to prevent long-term systemic T1DM-associated damages. Donor shortage and the limited long-term success of islet transplants have stimulated the development of novel therapies for T1DM. Gene therapy-based glucose-regulated hepatic insulin production is a promising strategy to treat T1DM. We have developed gene constructs which cause glucose-concentration–dependent human insulin production in liver cells. A novel set of human insulin expression constructs containing a combination of elements to improve gene transcription, mRNA processing, and translation efficiency were generated as minicircle DNA preparations that lack bacterial and viral DNA. Hepatocytes transduced with the new constructs, ex vivo, produced large amounts of glucose-inducible human insulin. In vivo, insulin minicircle DNA (TA1m) treated streptozotocin (STZ)-diabetic rats demonstrated euglycemia when fasted or fed, ad libitum. Weight loss due to uncontrolled hyperglycemia was reversed in insulin gene treated diabetic rats to normal rate of weight gain, lasting ∼1 month. Intraperitoneal glucose tolerance test (IPGT) demonstrated in vivo glucose-responsive changes in insulin levels to correct hyperglycemia within 45 minutes. A single TA1m treatment raised serum albumin levels in diabetic rats to normal and significantly reduced hypertriglyceridemia and hypercholesterolemia. Elevated serum levels of aspartate transaminase, alanine aminotransferase, and alkaline phosphatase were restored to normal or greatly reduced in treated rats, indicating normalization of liver function. Non-viral insulin minicircle DNA-based TA1m mediated glucose-dependent insulin production in liver may represent a safe and promising approach to treat T1DM.

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