Tubular Length Research Articles

Novel sorbitan monostearate organogels

Sorbitan monostearate, a hydrophobic nonionic surfactant, gels a number of organic solvents such as hexadecane, isopropyl myristate, and a range of vegetable oils. Gelation is achieved by dissolving/dispersing the organogelator in hot solvent to produce an organic solution/dispersion, which, on cooling sets to the gel state. Cooling the solution/dispersion causes a decrease in the solvent–gelator affinities, such that at the gelation temperature, the surfactant molecules self-assemble into toroidal inverse vesicles. Further cooling results in the conversion of the toroids into rod-shaped tubules. Once formed, the tubules associate with others, and a three-dimensional network is formed which immobilizes the solvent. An organogel is thus formed. Sorbitan monostearate gels are opaque, thermoreversible semisolids, and they are stable at room temperature for weeks. The gels are affected by the presence of additives such as the hydrophilic surfactant, polysorbate 20, which improves gel stability and alters the gel microstructure from a network of individual tubules to star-shaped “clusters” of tubules in the liquid continuous phase. Another solid monoester in the sorbitan ester family, sorbitan monopalmitate, also gels organic solvents to give opaque, thermoreversible semisolids. Like sorbitan monostearate gels, the microstructure of the palmitate gels comprise an interconnected network of rodlike tubules. Unlike the stearate gels, however, the addition of small amounts of a polysorbate monoester causes a large increase in tubular length instead of the “clustering effect” seen in stearate gels. The sorbitan stearate and palmitate organogels may have potential applications as delivery vehicles for drugs and antigens.

Flow injection potentiometric determination of saccharin in dietary products using a tubular ion-selective electrode

The construction and analytical evaluation of a tubular ion-selective electrode coated with an ion pair formed between saccharinate anion and toluidine blue O cation incorporated on a poly(vinylchloride) (PVC) matrix was constructed and adapted in a flow-injection (FI) system. The optimum experimental conditions found were an analytical path of 120 cm, an injection sample volume of 500 μL, a pH of 2.5, a flow rate of 2.3 mL min−1, and a tubular electrode length of 2.5 cm. The linear response to saccharin concentration ranged from 1.0 × 10−4 to 2.0 × 10−2 mol L−1, with a slope of −53.1 ± 0.4 mV/decade and a detection limit of 8.0 × 10−5 mol L−1. The analytical frequency was 40 determinations h−1, and this system was used with success for saccharin determination in several dietary products. © 1999 John Wiley & Sons, Inc. Lab Robotics and Automation 11: 234–239, 1999

Testing the potential of sodium fluoride to affect spermatogenesis: a morphometric study

This study provides quantitative information on the effect of sodium fluoride (NaF) on the testes of F 1 generation male rats exposed in utero and during lactation to NaF at one of four concentrations (25, 100, 175, 250 ppm). At weaning, the F 1 generation males were exposed to NaF in their drinking water for 14 weeks, after which time testicular tissues were perfusion-fixed with glutaraldehyde and observed after being embedded in plastic. The seminiferous tubules comprised 89%, 87%, 88%, 88% and 88% of the total testis volume while the interstitial space occupied 9.3%, 11.2%, 10.2%, 9.8% and 9.9% of the total testis volume for the 0, 25, 100,175 and 250 ppm NaF treatment groups, respectively. Statistically significant differences between control and NaF-treated rats were not observed with respect to absolute volume of the seminiferous tubules, interstitial space, Leydig cells, blood vessels boundary layer, lymphatic space, macrophages, tubular lumen or absolute tubular length and absolute tubular surface area, mean Sertoli cell nucleoli number per tubular cross-section, mean seminiferous tubule diameter and the mean height of the seminiferous epithelium. A statistically significant decrease in the absolute volume and volume percent of the lymphatic endothelium was observed in the 175 and 250 ppm NaF-treated groups and in the testicular capsule in the 100 ppm NaF-treated groups. The significance of this finding is unknown at the present time. Overall, the quantitative information obtained suggests that exposure to NaF at the doses used in the present study does not adversely affect testis structure or spermatogenesis in the rat.

Quantitative Morphology of Renal Cortical Structures during Compensatory Hypertrophy

The compensatory hypertrophy in different renal cortical structures was studied in rats 10 and 21 days after unilateral nephrectomy (UNX). Quantitative morphological/stereological analysis revealed significant increases in total renal cortical volume – 33% on day 10 and 48% on day 21 – after UNX. These changes were paralleled by significant increments in the volumes of proximal convoluted tubule (PCT, 55%), distal convoluted tubule (DCT, 114%), and cortical collecting duct (CCD, 106%) segments on day 10. The corresponding changes on day 21 were 76, 122, and 212%, respectively. These alterations were accompanied by increases in segment length; 3% PCT, 23% DCT, and 50% CCD on day 10 and 9% PCT, 30% DCT, and 142% CCD on day 21 after UNX. The total luminal and basolateral cell membrane surface areas also exhibited a time-dependent increase after UNX. The increments in both luminal and basolateral membrane domains in PCT and DCT after 10 days were not significant, but reached significance after 21 days (PCT: luminal membrane 21%, basolateral membrane 63%; DCT: luminal membrane 98%, basolateral membrane 63%). In contrast, CCD membrane areas had increased substantially already 10 days after UNX (luminal membrane 92%, basolateral membrane 71%). It declined subsequently by day 21 (luminal membrane 57%, basolateral membrane 32%). The cell rubidium concentration after a 30-second rubidium infusion, an index of Na-K-ATPase activity, as well as sodium concentrations were unaltered in cells of all nephron segments investigated. Altogether the stereological analysis shows that the compensatory increase in organ volume can be attributed primarily to an increase in nephron epithelial volume. The PCT responds with ‘radial’ hypertrophy (thickening of the tubular epithelial wall), while the DCT undergoes ‘length’ hypertrophy (increase of tubular length without thickening of the tubular wall and without an increase in number of cells). This type of hypertrophy is especially prominent on day 21 after UNX for the CCD which doubles in length. Only on day 10 does the CCD seem to respond with hyperplasia. Adaptive changes in response to UNX develop gradually. Only a few of the morphological parameters studied had completed their change by 10 days, the majority required longer.

In situ characterization of oxalate transport across the basolateral membrane of the proximal tubule.

Oxalate transport across the contraluminal membrane of the proximal tubule was studied in vivo using the "capillary stopped flow microperfusion method" (Pflügers Arch 400:250-256, 1984). Cellular uptake of oxalate was characteristic of a carrier-mediated transport process (Jmax = 1.6 +/- 0.6 pmol/s per cm proximal tubular length, Km = 2.03 +/- 0.77 mmol/l). Sulphate inhibited oxalate transport in a dose-dependent manner (Ki-value = 1.53 +/- 0.38 mmol/l). Sulphate transport across the basolateral membrane was also characteristic of a carrier- mediated transport process (Jmax = 1.83 +/- 0.56 pmol/s per cm proximal tubular length, Km = 1.37 +/- 0.57 mmol/l). Oxalate inhibited the sulphate transport in a dose-dependent manner (Ki = 2. 06 +/- 0.82 mmol/l). No significant differences were found between the Ki values and the Km values of the two substances, indicating that oxalate and sulphate are transported by the same carrier across the basolateral membrane of the proximal tubule. Oxalate transport was not dependent on the extracellular sodium or potassium concentration. Bicarbonate competitively inhibited the oxalate transport. Chloride significantly inhibited the oxalate transport, but not dose dependently. It is, therefore, suggested that oxalate is transported into the cell of the proximal tubule in exchange for sulphate or bicarbonate. The dose-independent inhibition by chloride is suggested to be mediated by the coupling of the sulphate (bicarbonate)/oxalate exchanger with the chloride/bicarbonate exchanger at the basolateral membrane of the proximal tubule. This, furthermore, suggests that the transport of oxalate or sulphate across the basolateral membrane might be indirectly coupled with the reabsorption of chloride at this membrane side.

BMP-2 and OP-1 exert direct and opposite effects on renal branching morphogenesis.

The bone morphogenetic proteins, BMP-2 and OP-1, are candidates for growth factors that control renal branching morphogenesis. We examined their effects in embryonic kidney explants and in the mIMCD-3 cell model of collecting duct morphogenesis (mIMCD-3 cells are derived from the terminal inner medullary collecting duct of the SV40 mouse). Osteogenic protein-1 (OP-1), at a dose of 0.25 nM, increased explant growth by 30% (P = 0.001). In contrast, 100-fold greater concentrations of OP-1 (28 nM) decreased explant growth by 10% (P < 0.001). BMP-2 was entirely inhibitory (maximum inhibition of 7% at 5 nM, P < 0.0004). In an in vitro model for branching morphogenesis utilizing the kidney epithelial cell line, mIMCD-3, low doses of OP-1 (< 0.5 nM) increased the number of tubular structures formed by 28 +/- 5% (P = 0.01), whereas concentrations > 0.5 nM decreased that number by 22 +/- 8% (P = 0.02). All concentrations of BMP-2 (0.05-10 nM) were inhibitory (maximum inhibition at 10 nM of 88 +/- 3%, P < 0.0001). Stimulatory doses of OP-1 increased tubular length (P = 0.003) and the number of branch points/structure (3.2-fold increase, P = 0.0005) compared with BMP-2. To determine the molecular basis for these effects, we demonstrated that BMP-2 is bound to mIMCD-3 cells by the type I serine/threonine kinase receptor, ALK-3, and that OP-1 bound to an approximately 80-kDa protein using ligand-receptor affinity assays. To demonstrate that OP-1 can exert both stimulatory and inhibitory effects within a developing kidney, embryonic explants were treated with agarose beads saturated with 2 microM OP-1. OP-1 decreased the number of ureteric bud/collecting duct branches adjacent to the beads by 58 +/- 1% (P < 0.0001). In contrast, the number of branches in tissue distal to the OP-1 beads was enhanced, suggesting a stimulatory effect at lower doses of OP-1. We conclude that OP-1 and BMP-2 directly control branching morphogenesis and that the effects of OP-1 are dependent on its local concentration within developing kidney tissue.

Localization of mRNAs Encoding Ca2+-inhibitable Adenylyl Cyclases along the Renal Tubule: FUNCTIONAL CONSEQUENCES FOR REGULATION OF THE cAMP CONTENT

Expression of Ca2+-inhibitable types V and VI adenylyl cyclases was studied by reverse transcription-polymerase chain reaction in rat renal glomeruli and nephron segments isolated by microdissection. Quantitation of each mRNA was achieved using a mutant cRNA which differed from the wild type by substituting two bases to create a new restriction site in the corresponding cDNA. Type VI mRNA was present all along the nephron but was more abundant in distal than in proximal segments. The expression of type V mRNA was restricted to the glomerulus and to the initial portions of the collecting duct. Expression of the Ca2+-insensitive type IV mRNA studied on the same samples was evidenced only in the glomerulus. The functional relevance of the expression of Ca2+-inhibitable isoforms was studied by measuring cAMP content in the microdissected outer medullary collecting duct which expressed both type V mRNA (2367 +/- 178 molecules/mm tubular length; n = 8) and type VI mRNA (5658 +/- 543 molecules/mm, n = 8). Agents known to increase intracellular Ca2+ in this segment induced a Ca2+-dependent inhibition on either arginine vasopressin- or glucagon-stimulated cAMP level. The characteristics of these inhibitions suggest a functional and differential expression of types V and VI adenylyl cyclases in two different cell types of the rat outer medullary collecting duct.

Efficiency of spermatogenesis.

Spermatogenesis is a process of division and differentiation by which spermatozoa are produced in seminiferous tubules. A measure of efficiency of spermatogenesis is the estimated number of spermatozoa produced per day per gram of testicular parenchyma. This measure is not influenced by species differences in testicular size; however, it is influenced by species differences in the numerical density of germ cells and in the life spans of these cells. Seminiferous tubules are composed of somatic cells (myoid cells and Sertoli cells), and germ cells (spermatogonia, spermatocytes, and spermatids). Activity of these three germ cells divide spermatogenesis into spermatocytogenesis, meiosis, and spermiogenesis, respectively. Spermatocytegenesis involves mitotic cell division to increase the yield of spermatogenesis and to produce stem cells and primary spermatocytes. Meiosis involves duplication and exchange of genetic material and two cell divisions that reduce the chromosome number and yield four spermatids. Spermiogenesis is the differentiation of spherical spermatids into mature spermatids which are released at the luminal free surface as spermatozoa. The spermatogenic cycle is superimposed on the three major divisions of spermatogenesis. Spermatogenesis and germ cell degeneration can be quantified from numbers of germ cells in various steps of development throughout spermatogenesis, and quantitative measures are related to number of spermatozoa in the ejaculate. Germ cell degeneration occurs throughout spermatogenesis; however, the greatest impact occurs during spermatocytogenesis and meiosis. Efficiency of spermatogenesis is related to the amount of germ cell degeneration, pubertal development, season of the year, and aging of humans and animals. Number of Sertoli cells and amount of smooth endoplasmic reticulum of Leydig cells (but not Leydig cell number) are related to efficiency of spermatogenesis. In humans, efficiency of spermatogenesis is reflected in number of spermatogenic stages per cross-section and number of missing generations within each stage; however, the arrangement of stages along the tubular length does not reflect differences in the efficiency of spermatogenesis. In short, spermatogenesis involves both mitotic and meiotic cell divisions and an unsurpassed example of cell differentiation in the production of the spermatozoon, and daily sperm production per g parenchyma is a measure of its efficiency.

Quantitative RT-PCR analysis of calcitonin receptor mRNAs in the rat nephron.

A quantitative assay based on the method of reverse transcription and polymerase chain reaction (RT-PCR) was developed to study the expression of calcitonin (CT) receptors in microdissected rat nephron segments. Steady-state mRNA levels of two CT-receptor spliced variants (CT1a and CT1b) were measured using a mutant cRNA as internal standard. CT1a, but not the CT1b isoform, was detected in the kidney cortex, outer medulla, and papilla. Among the tested segments, predominant expression of CT1a mRNA was found in the cortical thick ascending limb of Henle's loop (754 +/- 87 mRNA molecules/mm tubular length; n = 8). Lower expression levels were measured in the medullary thick ascending limb (460 +/- 62 molecules/mm tubule length; n = 7) and in the cortical collecting duct (327 +/- 61 molecules/mm tubule length; n = 6). A weak expression was also detected in the outer medullary collecting duct and the glomerulus. No expression was found in the proximal convoluted tubule, pars recta, and thin descending and thin ascending limb of Henle's loop. We conclude that only the CT1a-receptor mRNA is present in the rat kidney, with a significant level of expression in the cortical and medullary thick ascending limb and in the cortical collecting duct.

Molecular analysis of beta-adrenergic receptor subtypes in rat collecting duct: effects on cell cAMP and Ca2+ levels.

Expression and functional properties of beta-adrenergic receptors (beta-ARs) were studied in rat collecting tubules isolated by microdissection. Reverse transcription-polymerase chain reaction experiments demonstrated that the beta 1- and beta 2-AR mRNAs, but not the beta 3-subtype, are expressed in the cortical collecting duct (CCD). Quantitation of mRNAs, carried out using mutant RNAs as internal standards, further showed that beta 1- and beta 2-ARs transcripts are present at comparable amounts in CCD (3,000-4,000 copies/mm of tubular length), but reach 6-8 times lower levels in the outer medullary collecting duct (OMCD: beta 1, 480 +/- 180; beta 2, 590 +/- 110 copies/mm of tubular length). Functional studies, carried out in CCD, corroborated the expression of these two receptor subtypes. The rank order of potency of beta-agonists for stimulating adenosine 3',5'-cyclic monophosphate (cAMP) accumulation was isoproterenol > norepinephrine = epinephrine, and similar efficiencies were found for a beta 1- and a beta 2-antagonist to inhibit isoproterenol-dependent cAMP formation. Fura 2 fluorescence measurements revealed that isoproterenol (10 microM) induces a biphasic rise of intracellular free Ca2+ concentration ([Ca2+]i), consisting of an initial fast increase (delta [Ca2+]i = 122 nM) followed by a plateau phase (delta [Ca2+]i = 58 nM). In the absence of basolateral Ca2+, the initial peak was still observed, suggesting intracellular Ca2+ release. Norepinephrine and epinephrine, as well as selective beta 1- and beta 2-agonists, also increased [Ca2+]i in CCD. Only slight [Ca2+]i variations were produced by isoproterenol in the OMCD (delta [Ca2+]i = 21 nM) and the cortical thick ascending limb (delta [Ca2+]i = 25 nM). These results show that both beta 1- and beta 2-ARs are expressed in the collecting tubule and that they predominate in the CCD. The two receptor subtypes contribute to cAMP accumulation induced by beta-agonists. They also trigger [Ca2+]i variations, indicating their possible coupling to several transduction pathways in the rat CCD.

Molecular analysis of vasopressin receptors in the rat nephron. Evidence for alternative splicing of the V2 receptor.

Expression and regulation of vasopressin V2 and V1a receptors were studied at the mRNA level in the rat kidney. Two V2 mRNA variants were identified and shown to arise from a single gene by alternative splicing using one donor and two different acceptor sites. The long (V2L) form encodes the adenylyl cyclase-coupled receptor. The short (V2S) form lacks the nucleotide sequence encoding the putative seventh transmembrane domain and undergoes a frame shift in its 3'end coding region; it is inactive on the cyclase pathway in transfected cells. Measurement of mRNAs, carried out by quantitative reverse transcription-polymerase chain reaction (RT-PCR) on microdissected nephrons, demonstrated that neither V2L, V2S nor V1A mRNAs are expressed in glomeruli and proximal tubules (< 100 mRNA copies/glomerulus or mm of tubular length), whereas they are present in the ascending limb of Henle's loop and in the collecting tubule. The V2L mRNA, which is always predominant in these structures, is expressed throughout the collecting tubule at 10 times higher levels (30,000 copies/mm) than in the thin and thick ascending limbs. The ratio of the V2S over V2L mRNA is constant (15%) in all nephron segments; hence high V2S levels are only observed in the collecting tubule. The V1A mRNA is slightly expressed in the thin ascending limb, absent in the thick ascending limb and reaches its maximum in the cortical collecting duct (4,000 copies/mm), before gradually decreasing to undetectable levels in the terminal collecting duct. Finally, in vivo administration of a vasopressin V2 agonist decreased by 50% V2L and V2S mRNAs, but did not alter the V1A mRNA level. We conclude that this study provides the quantitation, on a molar basis, of vasopressin receptor mRNAs in kidney tubules and demonstrates the occurrence of two V2 mRNA spliced variants which are similarly down-regulated.

Dolichoectatic aneurysm of common carotid artery: an animal model with histological correlation.

There is a limited understanding of the pathogenesis of dolichoectatic (dolicho = long; ectatic = dilated) aneurysms of elastic arteries. Therefore, we developed a model of dolichoectatic changes in elastic arteries by injecting porcine elastase into the media of the carotid artery of male New Zealand white rabbits. After 3 months, gross examination of the carotid arteries in vivo revealed dilated and tortuous vessels. The carotid arteries were then harvested, and cross-sections of the vessels were stained by the Verhoff-van Giesson stain. Histologically, the internal elastic lamina was dissolved in the most dilated areas. The elastic lamellae of the media were also digested and there was reorientation of the innermost medial smooth muscle layer. These gross and histologic changes were present in 80% of the treated carotid arteries and in none of the contralateral control vessels. Our study suggests the importance of the elastic lamellae for the maintenance of tubular shape and length of the carotid artery and describes a new chronic animal model of dolichoectatic aneurysm of the common carotid artery.

Architectural arrangement of stages of the spermatogenic cycle within human seminiferous tubules is related to efficiency of spermatogenesis

Stages of the spermatogenic cycle in human seminiferous tubules were evaluated in men with varied efficiencies of spermatogenesis to determine if the architectural arrangement of stages or the atypical cell types contributed to variation in sperm production rates. Testes were selected from men with low, intermediate, and high daily sperm production per g parenchyma (DSP/g). Round tubular cross sections were photographed by bright-field microscopy. Stages were identified for each cross section by two observers and the number of stages represented in each cross section was recorded. Number of stages per cross section in men with low efficiency of spermatogenesis were significantly (P < 0.05) fewer than men with intermediate and high efficiency of spermatogenesis. Further, the percentage of stages with atypical cell types in men with high DSP/g was significantly (P < 0.05) higher than men with low DSP/g. There was a significant relationship (P < 0.01) between the percentages of stages with atypical cell types per stage and number of stages per cross section. The atypical cell types appear to result from high density of stages per cross section in men with high DSP/g. There was no significant difference observed between groups for tubular volume, diameter, length, volume density, and volume density of seminiferous epithelium. However, a significant (P < 0.05) positive correlation between percent seminiferous epithelium per testis with DSP/g or with the number of stages per cross section was found.(ABSTRACT TRUNCATED AT 250 WORDS)

Predominant expression of beta 1-adrenergic receptor in the thick ascending limb of rat kidney. Absolute mRNA quantitation by reverse transcription and polymerase chain reaction.

Beta 1- and beta 2-adrenergic receptor (beta-ARs) expression in the thick ascending limb of rat kidney was studied at the level of mRNA and receptor coupling to adenylyl cyclase. Absolute quantitation of beta 1- and beta 2-AR mRNAs in microdissected nephron segments was performed with an assay based on reverse transcription and polymerase chain reaction, using in vitro transcribed mutant RNAs as internal standards. In the cortical thick ascending limb (CTAL), the number of mRNA molecules/mm of tubular length was 2,806 +/- 328 (n = 12) for beta 1-AR and 159 +/- 26 for beta 2-AR (P < 0.01). Lower levels were obtained in the medullary thick ascending, beta 1-AR mRNA still being predominant. The pharmacological properties of beta-ARS was also studied in the CTAL. Cyclic AMP accumulation was stimulated by beta-agonist with a rank order of potency of isoproterenol > norepinephrine > epinephrine. This observation, and the higher efficiency of a beta 1 than of a beta 2 antagonist to inhibit isoproterenol-induced cAMP accumulation, establish the typical beta 1-AR sensitivity of the CTAL. No detectable contribution of atypical or beta 3-ARs to adenylyl cyclase stimulation could be found. In conclusion, this study, which shows markedly different levels of beta 1- and beta 2-AR mRNAS in the CTAL, provides a molecular basis for the predominant expression of the beta 1 receptor subtype in this nephron segment.

Assessment of testicular cytology by fine needle aspiration as a diagnostic parameter in the evaluation of the oligospermic subject

To investigate whether testicular cytology by fine needle aspiration may be considered a diagnostic parameter in the evaluation of the oligospermic subject. Cytologic smears were obtained using a 23-gauge needle, stained with May-Grünwald-Giemsa stain and examined under a light Orthoplan microscope (Wild Leitz, Wetzlar, Germany) for qualitative and quantitative analysis. One hundred sixty-six oligospermic patients were analyzed, and the findings were compared with those obtained from 40 normozoospermic infertile subjects used as controls. At least 200 spermatogenic cells were counted per slide and classified at the various steps of spermatogenesis. Because the number of Sertoli cells may be considered as a constant per unit of tubular length, Sertoli cells:spermatogenic cells ratio (termed Sertoli index) provided further elucidation and more comprehensible results. The procedure provided no sign of traumatization. The cytologic analysis permitted identification of different classes of oligospermic subjects, characterized by the following specific cytologic pictures: [1] bilateral or [2] unilateral germ depopulation (hypospermatogenesis), associated with maturation abnormalities of the first steps of spermatogenesis on both testes; [3] difficult maturation of the immature germ cells (spermatogonial or spermatocytic arrest); [4] ineffective spermiogenesis (spermatidic arrest); [5] normal maturation of the germ line in the presence of oligospermia, resulting from a transient acute damage on the spermatogenic line. Plasma levels of FSH and the testicular volumes agree with the cytologic picture of each group of patients. The results support use of fine needle aspiration of the testis as a minimally invasive diagnostic parameter for the assessment of oligospermic subjects.

Macro-orchidism: Light and electron microscopic study of four cases

A hormonal and quantitative light microscopy study of one man with macro-orchidism associated with mental retardation and fragile X chromosome (case no. 1) and three men with idiopathic macro-orchidism (cases no. 2 to 4) is reported. Hormonal study revealed slightly increased follicle-stimulating hormone serum levels in cases no. 1 to 3. The testes from cases no. 1 (orchidoepididymoectomy specimen) and 2 (testicular biopsy) presented interstitial edema and three different tubular patterns that were arranged in a mosaic-like manner. Type I tubules had an increased diameter (<220 μm), dilated lumen, and thin seminiferous epithelium usually consisting of Sertoli cells, spermatogonia, primary spermatocytes, and sometimes a few spermatids. Type II tubules had a normal diameter (180 to 220 μm) and germ cell development varied between complete spermatogenesis and Sertoli-cell-only tubules. Type III tubules had decreased diameter (<180 μm), atrophic seminiferous epithelium, and thickened tunica propria. The appearance of the nuclei of the Sertoli cells in the three types of tubules could be either mature or immature. Some of the mature Sertoli cells presented a granular cytoplasm. A few of these granular cells grouped together, forming nests that protruded into the tubular lumen. The testicular biopsies from cases no. 3 and 4 only presented type II tubules that contained both mature and immature Sertoli cells. Quantitative study revealed that the large testicular size was principally due to an increased tubular length in all four cases. Although the seminiferous tubule lesions and interstitial edema suggest an obstructive process, the testicular excretory ducts (studied in case no. 1) appeared normal or only slightly dilated. It is possible that the seminiferous tubule lesions (dilated lumen and germ cell depletion) might be secondary to the Sertoli cell lesions (granular cytoplasm and nuclear immature-like pattern).

Assessment of testicular cytology by fine needle aspiration as a diagnostic parameter in the evaluation of the azoospermic subject

To investigate whether testicular cytology by fine needle aspiration (FNA) may be considered a diagnostic parameter in the evaluation of the azoospermic subject. Cytologic smears were obtained using a 23-G needle, stained with May-Grünwald-Giemsa stain and examined under a light Orthoplan microscope (Wild, Leitz, Germany) for qualitative and quantitative analysis. Fifty-four azoospermic patients were analyzed, and the findings were compared with those obtained from 40 normozoospermic infertile subjects used as controls. Two hundred spermatogenic cells were counted and classified at the various steps of spermatogenesis. Spermatic index and Sertoli index provided further elucidations and more comprehensible results. No sign of traumatization was observed. Cytologic analysis was proved to have high statistical reproducibility (P less than 0.01 for spermatogonia and secondary spermatocytes and P less than 0.001 for the other cell types, when compared between differential counts) and permitted identification of different situations associated with azoospermia: Sertoli cell-only syndrome, germ depopulation (hypospermatogenesis), spermatogonial arrest, spermatidic arrest, and obstructive azoospermia. These findings agreed with clinical and hormonal parameters and with the results of bilateral surgical biopsies, when performed. The results support use of FNA of the testis as a noninvasive diagnostic parameter for the assessment of azoospermic subjects.

Tubular sodium handling and tubuloglomerular feedback in compensatory renal hypertrophy.

Tubular sodium handling and tubuloglomerular feedback (TGF) activity were assessed in established compensatory renal hypertrophy in Sprague Dawley rats. Hyperfiltration at the level of the single nephron was confirmed 4-6 weeks following a reduction in renal mass. TGF activity, determined as the difference between late proximal and early distal measurements of single-nephron glomerular filtration rate (SNGFR), was significantly increased in compensatory renal hypertrophy, being 7.8 +/- 1.0 vs 23.3 +/- 1.9 vs 25.5 +/- 2.6 nl/min (P for analysis of variance less than 0.05) following sham operation, unilateral nephrectomy, and 1 1/3 nephrectomy, respectively. Enhanced net tubular Na transport was also observed, with total Na reabsorption up to the late proximal site being 1.8 +/- 0.2 vs 2.7 +/- 0.1 vs 3.1 +/- 0.3 nmol/min (P less than 0.05), and to the early distal site being 3.4 +/- 0.5 vs 5.8 +/- 0.6 vs 7.9 +/- 0.8 nmol/min (P less than 0.05) in the three animal groups respectively. Comparison of proximal tubular length demonstrated a 71.9 +/- 8.1% increase in uninephrectomised vs sham-operated animals. This increase was proportionately greater than the increase in proximal Na reabsorption (50.0 +/- 4.0%) observed in the corresponding animal groups. Concurrent electron microprobe experiments in uninephrectomised and sham-operated animals demonstrated that the proximal tubular intracellular Na concentration was significantly lower following uninephrectomy (16.8 +/- 0.6 vs 18.9 +/- 0.5 mmol/kg wet weight, P less than 0.01), in association with evidence of reduced basolateral Na/K-ATPase activity.(ABSTRACT TRUNCATED AT 250 WORDS)

Normal values for metacarpal and phalangeal lengths in Nigerian children.

Metacarpal and phalangeal lengths were measured on 1290 hand radiographs of Nigerian children, aged 3-16 years. The radiographs were obtained during a combined cross-sectional and longitudinal study of growth and development. There is a linear increase in tubular bone length with age in both sexes. The girls have higher values for all the bones up to the age of 13 years when the boys overtake them. Comparison of our data with those from North American children shows that the values amongst Nigerian children are higher than White, Black American and Mexican American children. Of particular note is the difference between Black American and Nigerian figures. It is postulated that the decreased metacarpophalangeal lengths in Black Americans compared with Nigerians may be due to 'gene dilution'.

Morphometric study of the prepubertal rabbit testis: Germ cell numbers and seminiferous tubule dimensions

Testes from rabbits aged 1-9 weeks were examined by light microscopy. Changes in seminiferous tubule dimensions, testicular volume, and volume fraction of tubules were assessed. Germ cells and Sertoli cells were counted in round tubular cross sections and total germ cell number in each testis was estimated. Mitotic, meiotic, and degenerative activities of germ cells as well as their basal or central positions within tubules were quantified. A marked, steady increase in testis volume and in tubular length and volume occurred over the prepubertal period; but diameter underwent no significant increase and in fact decreased until week 4. Overall, tubules lengthened 40-fold and testis volume increased 25-fold; the percentage volume of the testis occupied by tubules rose from one-third neonatally to three-fifths at the onset of spermatogenesis. The ratio of germ cells to total tubular (germ and Sertoli) cells was lowest at 3 weeks. However, the total number of germ cells increased little until 3 weeks, after which it rose at a sharp rate commensurate with testis volume. Percentage of germ cells in mitosis peaked sharply at 3 weeks, dropped in subsequent weeks, and then rose at 7 weeks at the initiation of spermatogenesis. Importantly, the surge in mitosis at 3 weeks was followed by a redistribution of germ cells to a predominantly basal location from 3 to 7 weeks. Meiotic activity was sparse at 7 weeks and became abundant by 9 weeks. Germ cell degeneration remained relatively constant during weeks 1 through 6, with an increase at 7 weeks.