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The Reynolds number in the renal tubule is extremely low, consistent with laminar flow. Consequently, luminal flow can be described by the Hagen-Poiseuille laminar flow equation. This equation calculates the volumetric flow rate from the axial pressure gradient and flow resistance, which is dependent on the length and diameter of each renal tubule segment. Our goal was to calculate the pressure drop along each segment of the renal tubule and to determine the points of highest resistance. When the Hagen-Poiseuille equation was used for rat superficial nephrons based on known tubule flow rates, lengths, and diameters, it was found that the maximum pressure drop occurred in two segments: the thin descending limbs of Henle and the inner medullary collecting ducts. The high resistance in the thin descending limbs is due to their small diameters. The steep pressure drop observed in the inner medullary collecting ducts is due to the convergent structure of the tubules, which channels flow into fewer and fewer tubules toward the papillary tip. For short-looped nephrons, the calculated glomerular capsular pressure matched measured values, even with the high collecting duct flow rates seen in water diuresis, provided that tubule compliance was taken into account. In long-looped nephrons, the greater length of thin limb segments is likely compensated for by a larger luminal diameter. Simulation of the effect of proximal diuretics, namely acetazolamide or type 2 sodium-glucose transporter inhibitors, predicts a substantial back pressure in Bowman’s capsule, which may contribute to observed decreases in glomerular filtration rate.

Current clinical methods to treat articular cartilage lesions provide temporary relief of the symptoms but fail to permanently restore the damaged tissue. Tissue engineering, using mesenchymal stem cells (MSCs) combined with scaffolds and bioactive factors, is viewed as a promising method for repairing cartilage injuries. However, current tissue engineered constructs display inferior mechanical properties compared to native articular cartilage, which could be attributed to the lack of structural organization of the extracellular matrix (ECM) of these engineered constructs in comparison to the highly oriented structure of articular cartilage ECM. We previously showed that we can guide MSCs undergoing chondrogenesis to align using microscale guidance channels on the surface of a two-dimensional (2-D) collagen scaffold, which resulted in the deposition of aligned ECM within the channels and enhanced mechanical properties of the constructs. In this study, we developed a technique to roll 2-D collagen scaffolds containing MSCs within guidance channels in order to produce a large-scale, three-dimensional (3-D) tissue engineered cartilage constructs with enhanced mechanical properties compared to current constructs. After rolling the MSC-scaffold constructs into a 3-D cylindrical structure, the constructs were cultured for 21 days under chondrogenic culture conditions. The microstructure architecture and mechanical properties of the constructs were evaluated using imaging and compressive testing. Histology and immunohistochemistry of the constructs showed extensive glycosaminoglycan (GAG) and collagen type II deposition. Second harmonic generation imaging and Picrosirius red staining indicated alignment of neo-collagen fibers within the guidance channels of the constructs. Mechanical testing indicated that constructs containing the guidance channels displayed enhanced compressive properties compared to control constructs without these channels. In conclusion, using a novel roll-up method, we have developed large scale MSC based tissue-engineered cartilage that shows microscale structural organization and enhanced compressive properties compared to current tissue engineered constructs. Statement of significance Tissue engineered cartilage constructs made with human mesenchymal stem cells (hMSCs), scaffolds and bioactive factors are a promising solution to treat cartilage defects. A major disadvantage of these constructs is their inferior mechanical properties compared to the native tissue, which is likely due to the lack of structural organization of the extracellular matrix of the engineered constructs. In this study, we developed three-dimensional (3-D) cartilage constructs from rectangular scaffold sheets containing hMSCs in micro-guidance channels and characterized their mechanical properties and metabolic requirements. The work led to a novel roll-up method to embed 2-D microscale structures in 3-D constructs. Further, micro-guidance channels incorporated within the 3-D cartilage constructs led to the production of aligned cell-produced matrix and enhanced mechanical function.

Previous studies have demonstrated that both the V2-receptor agonist, 1-desamino-8-D-arginine vasopressin (dDAVP), and the V1a-receptor agonist, [Phe2, Orn8]vasotocin (PO-VT), increase intracellular calcium concentration ([Ca2+]i) in the rat inner medullary collecting duct (IMCD). The present studies were done to clarify the receptor subtype(s) responsible for calcium mobilization. Measurements of [Ca2+]i, using fura 2 in microdissected IMCD segments, confirmed that arginine vasopressin (AVP), dDAVP, and PO-VT stimulate an increase in [Ca2+]i and that the response to all three agents could be blocked by the specific V2-receptor antagonist, [d(CH2)5(1),D-Ile2, Ile4, Arg8]vasopressin (II-VP). These results would suggest that all three agents acted through the V2 receptor. Furthermore, we showed that PO-VT increased cAMP production in IMCD suspensions and water permeability in isolated perfused tubules. These responses were also blocked by II-VP, indicating that PO-VT is also a V2 agonist in the IMCD. Finally, we utilized the quantitative reverse transcription-polymerase chain reaction technique of Wiesner (Nucleic Acids Res. 20: 5863-5864, 1992) to evaluate V1a and V2 mRNA levels in rat collecting duct. In terminal IMCD, we estimated > 30 copies/cell for V2 receptor mRNA but less than 1 copy/cell of V1a receptor mRNA, thus there is littler or no V1a mRNA expression in the terminal IMCD. These results suggest that calcium mobilization in response to vasopressin analogues is associated with the V2 receptor and that the V2 receptor is linked to both adenylyl cyclase and calcium mobilization in the rat IMCD.

The urine concentrating effect of the mammalian renal inner medulla has been attributed to countercurrent multiplication of a transepithelial osmotic difference arising from passive absorption of NaCl from thin ascending limbs of long loops of Henle. This study assesses, both mathematically and experimentally, whether the permeability criteria for effective function of this passive hypothesis are consistent with transport properties measured in long loops of Henle of chinchilla. Mathematical simulations incorporating loop of Henle transepithelial permeabilities idealized for the passive hypothesis generated a steep inner medullary osmotic gradient, confirming the fundamental feasibility of the passive hypothesis. However, when permeabilities measured in chinchilla were used, no inner medullary gradient was generated. A key parameter in the apparent failure of the passive hypothesis is the long-loop descending limb (LDL) urea permeability, which must be small to prevent significant transepithelial urea flux into inner medullary LDL. Consequently, experiments in isolated perfused thin LDL were conducted to determine whether the urea permeability may be lower under conditions more nearly resembling those in the inner medulla. LDL segments were dissected from 30–70% of the distance along the inner medullary axis of the chinchilla kidney. The factors tested were NaCl concentration (125–400 mM in perfusate and bath), urea concentration (5–500 mM in perfusate and bath), calcium concentration (2–8 mM in perfusate and bath), and protamine concentration (300 micrograms/ml in perfusate). None of these factors significantly altered the measured urea permeability, which exceeded 20 x 10(-5) cm/s for all conditions. Simulation results show that this moderately high urea permeability in LDL is an order of magnitude too high for effective operation of the passive countercurrent multiplier.

The vasopressin-regulated urea carrier and the vasopressin-regulated water channel are distinct transporters present in the apical membrane of the inner medullary collecting duct (IMCD) cells. To assess whether these transporters may be activated by common mechanisms, we investigated the time course of increase of urea and water permeability in response to vasopressin in isolated perfused terminal IMCD segments. The permeability responses were determined through the use of a specially designed continuous-flow fluorometer for rapid analysis of collected tubule fluid samples. The time courses of activation of the two transporters by vasopressin were virtually identical. Both urea and water permeability displayed a rapid initial increase for the first 10 min followed by a slower secondary response lasting at least 30 additional min. The lag periods between vasopressin addition and the initial rise in permeability were the same for urea (34.2 +/- 8.8 s) and water (34.8 +/- 8.9 s) transport activation. Furthermore, the initial rate of permeability increase (normalized by the total increase) was not significantly different for the two transport processes. The lag periods for the increase in urea permeability in response to 8-bromoadenosine 3',5'-cyclic monophosphate and vasopressin were not significantly different. The results are consistent with the view that the rate-limiting step in vasopressin-induced activation is the same for both the urea carrier and water channel and may lie at a step beyond generation of adenosine 3',5'-cyclic monophosphate.

Only a few studies have concentrated on elements such as thallium (TI). Uranium (U) has been studied as a radionuclide of concern in food and the environment. Foodstuffs contain 10-100 ng U{center_dot}{sup -1} with vegetables and cereals contributing most heavily to the daily intake of ca 1.5 ug U. Between 10-30% of ingested U is absorbed, with most being stored in bone. Rainbow trout (onchorynchus mykiss) and longnose sucker (Catostomus catostomus) from a lake with naturally high radioactivity contained < 5 ng U{center_dot}g{sup -1} in the flesh. Trout bone contained 40 ng U{center_dot}g{sup -1}. Higher tissue U concentrations occurred in fish from areas receiving U mining wastes. Bioconcentration factors for bone and flesh were estimated to be low, 118 and 14.7, respectively. This paper describes the Inductively coupled plasma-mass spectrometry (ICP-MS) determination of Tl and U in digestive gland tissue from lobsters captured in the vicinity of Belledune Harbor, New Brunswick, Canada. The harbor is the site of a lead smelter, a fertilizer plant, and a coal-fired power station (the latter due to enter production in late 1993) and thus has the potential of adding significant amounts of Tl to the local marine environment. The accumulation of Tl from watermore » by marine shellfish is low, at least for bivalves, and the accumulated Tl is eliminated in a number of days when the animals are transferred to clean water. Bioconcentration factors for U in finfish ranged from 0.4-17 for larger species. However, because of the high concentrations of various trace elements in lobster digestive gland, its desirability as a foodstuff, and its relatively large size (approximately 20% of the edible tissue yield), we have investigated Tl and U concentrations and {sup 235U}/{sup 238U} ratios in it. 15 refs., 1 fig., 3 tabs.« less

Acute pulmonary edema can be induced by phorbol myristate acetate (PMA). Oxygen radicals released from the neutrophils have been considered to play an important role in the pathogenesis of PMA-induced pulmonary edema. In the present experiment, we studied the effect of dimethylthiourea (DMTU) on PMA-induced pulmonary injuries in isolated perfused lungs of rats. DMTU is a potent scavenger of the hydroxyl radical and hydrogen peroxide. PMA infusion into the isolated lung increased pulmonary arterial pressure (delta PAP) by 37.8 +/- 3.9 mmHg. The lung weight gain (LWG) and lavage albumin concentration (LAC) amounted to 6.2 +/- 1.2 g and 102.0 +/- 22.9 mg/dl, respectively. DMTU (100 mM) pretreatment significantly reduced the PAP increase (delta PAP = 4.6 +/- 0.8 mmHg, p less than 0.001), LWG (0.3 +/- 0.1 g, p less than 0.01) and LAC (25.3 +/- 1.7 mg/dl, p less than 0.01). Additional in vitro experiments demonstrated that DMTU depressed the chemiluminescence released from neutrophils activated by PMA (17.9 +/- 2.6 mV.min to 2.6 +/- 0.5 mV.min, p less than 0.01). The results suggest that DMTU, a scavenger of toxic radicals, decreases the lung edema through both attenuation of pulmonary hypertension and protection of vascular permeability from PMA injury.

In 1980 cadmium (Cd) concentrations in digestive gland from American Lobster (Homarus americanus), captured at Belledune Harbour, New Brunswick, Canada were found to range 47.6-372 mg/kg wet weight. Since the digestive gland is commonly eaten the harbor fishery was closed. The adjacent area was decreed a controlled fishery zone. The cadmium source was a lead smelter on the harbor shore. The company installed an aqueous effluent treatment plant and improved handling procedures within the plant area. Aqueous Cd discharges were subsequently reduced by more than 95%. Cadmium levels in lobsters have been monitored annually since 1980 by sampling in early spring, immediately after the lobsters emerged from their overwintering locations. The results from the 1981-1985 monitoring program were used to model the system, analyze the data and predict the years in which certain changes to the management plan might be implemented.

A method was developed for measuring volume flow rate in free-flowing proximal tubules. Multiple injections of a solution containing fluorescein isothiocyanate-dextran (mol wt 20,000) were made into the tubular fluid stream while fluorescence was detected by videomicroscopy. Progression of the dye bolus between two points along the tubule was measured to determine fluid velocity. Tubule diameter was measured by video image shearing and flow rate was calculated from the product of velocity and cross-sectional area. The technique was validated by simultaneous collections in the same tubule just downstream. There was good correlation between the results obtained with the two methods (r = 0.89). The estimated average injection volume was 0.0165 +/- 0.00016 nl. An increase in tubular volume flow caused by repetitive injections during micropuncture collections was too small to be detected. This new method should be useful in experimental situations that require uninterrupted delivery of fluid to the macula densa, or when better temporal resolution is required than can be obtained with the conventional method.

The ability of the kidney to increase sodium and water excretion in response to increases in perfusion pressure has been recognized for more than 50 years. Because glomerular filtration rate is tightly autoregulated, pressure natriuresis occurs as the result of decreased tubular sodium reabsorption rather than increased filtered load. Micropuncture and microperfusion data support the contention that acute changes in arterial pressure can alter proximal tubule reabsorption; however, studies have failed to show a consistent association between changes in sodium excretion and peritubular, interstitial, or tubular pressures. Thus, the specific intrarenal mechanism for the change in tubular reabsorption in response to an acute change in arterial pressure does not appear to be related to the peritubular physical factors at the level of outer cortical nephrons. The possible roles of angiotensin and prostaglandins as humoral mediators of pressure natriuresis are considered in this report. Although angiotensin II is a powerful modulator of the slope of the pressure natriuresis relationship, the responsiveness of sodium excretion to arterial pressure is actually enhanced by angiotensin-converting enzyme inhibitors. These data suggest that angiotensin does not mediate the basic phenomenon. Recent experiments indicate that intrarenal prostaglandins also modulate the magnitude of the pressure natriuresis relationship, but these hormones do not appear to be essential for its basic manifestation.