RNA-binding protein TIAR is essential for primordial germ cell development.

Primordial germ cells (PGCs) give rise to both eggs and sperm via complex maturational processes that require both cell migration and proliferation. However, little is known about the genes controlling gamete formation during the early stages of PGC development. Although several mutations are known to severely reduce the number of PGCs reaching and populating the genital ridges, the molecular identity of only two of these genes is known: the c-kit receptor protein tyrosine kinase and the c-kit ligand (the steel factor). Herein, we report that mutant mice lacking TIAR, an RNA recognition motif/ribonucleoprotein-type RNA-binding protein highly expressed in PGCs, fail to develop spermatogonia or oogonia. This developmental defect is a consequence of reduced survival of PGCs that migrate to the genital ridge around embryonic day 11.5 (E11.5). The numbers of PGCs populating the genital ridge in TIAR-deficient embryos are severely reduced compared to wild-type embryos by E11.5 and in the mutants PGCs are completely absent at E13.5. Furthermore, TIAR-deficient embryonic stem cells do not proliferate in the absence of exogenous leukemia inhibitory factor in an in vitro methylcellulose culture assay, supporting a role for TIAR in regulating cell proliferation. Because the development of PGCs relies on the action of several growth factors, these results are consistent with a role for TIAR in the expression of a survival factor or survival factor receptor that is essential for PGC development. TIAR-deficient mice thus provide a model system to study molecular mechanisms of PGC development and possibly the basis for some forms of idiopathic infertility.

Acid-induced laryngospasm in a canine model.

A canine model was used to investigate the efferent laryngeal responses to stimulation by topically applied acid and pepsin. Five adult mongrel dogs were studied. Electromyographic recordings from the thyroarytenoid muscle were measured with hooked-wire electrodes as an acid solution (normal saline/hydrochloric acid at pH 6.0, 5.0, 4.0, 3.0, 2.5, 2.0, 1.5, and 1.0) was sequentially instilled into the larynx. Laryngospasm (tonic, sustained contraction of the thyroarytenoid muscle) occurred in all animals at pH 2.5 to 2.0 or less. Control substances such as neutral pH isotonic saline, hypotonic saline, hypertonic saline, water, and pepsin alone failed to produce laryngospasm. Next, solutions containing both acid (in the same pH range) and pepsin were tested. The laryngeal responses were similar to those of acid alone. The superior laryngeal nerves were sectioned bilaterally and the above experiments repeated. None of the test solutions produced laryngospasm; however, when capsaicin (1%) was instilled into the subglottis, laryngospasm occurred. Thus, chemoreceptors in the subglottis (supplied by the recurrent laryngeal nerves) appear to be responsive to capsaicin stimulation but not to acid stimulation. The data suggest that pH-sensitive chemoreceptors in the canine larynx cause laryngospasm (when the pH of the test solution is 2.5 or less) and that these acid receptors are supplied by the superior laryngeal nerves.

Home hemodialysis: patient outcomes during a 24-year period of time from 1970 through 1993.

In the United States, from 1983 to 1993, home hemodialysis use has decreased from 6% to 1.3% of the dialysis population, whereas continuous ambulatory peritoneal dialysis (CAPD) has increased to 20%. Most home hemodialysis programs have withered away because of current patient mix, increase in CAPD, proliferation of outpatient centers, disinterest in nephrologists, and fear of self-cannulation by patients. From 1970 through 1993, 896 patients began dialysis at North Shore and were followed up through 1994. During this period, 687 patients were on in-center hemodialysis, 95 on CAPD, 74 on home hemodialysis, and 40 on in-center peritoneal dialysis. The home hemodialysis patients were younger, with a median age of 44 versus 59 years for in-center hemodialysis patients, and had less comorbidity. The home hemodialysis group had fewer diabetic patients and no renal vascular patients. The 5-year and median survival estimates were significantly better for the home hemodialysis patients versus other dialysis modalities. More home hemodialysis patients received transplants. Compared with the other dialysis modalities, home hemodialysis patients showed significantly improved survival rates. When matched by age, sex, and end-stage renal disease (ESRD) diagnosis to corresponding in-center hemodialysis, the home hemodialysis patients still had significantly better survival rates, but the home hemodialysis patients had less comorbidity. In conclusion, home hemodialysis patients survive longer and have better rehabilitation than other dialysis patients. Reasons for better survival in addition to a younger age and more favorable ESRD diagnosis may include less comorbidity, more patient involvement, and longer dialysis time. Because of these better outcomes, home hemodialysis should be offered to more ESRD patients.

Structural analysis of the polysaccharide from Pachymenia lusoria (Cryptonemiaceae, Rhodophyta).

The highly complex polysaccharide extracted from the New Zealand red alga Pachymenia lusoria (Grev.) J. Ag. has been characterised and certain structural features defined. A reductive hydrolysis procedure was used for constituent sugar and linkage analyses, with trideuteriomethylation being employed to enable the location of natural methyl ether groups to be determined. A reductive partial-hydrolysis procedure allowed agarobiosyl constituent residues to be identified. The analytical results are consistent with the polymer having a linear backbone of 3-linked D-galactopyranosyl alternating with 4-linked D- or L-galactopyranosyl residues. The 3-linked residues are nearly all 2-sulfated, with 1 in 3 also being 6-O-methylated and 1 in 5 also bearing a 4,6-pyruvic acetal residue. About one-third of the polymer is comprised of blocks of agarobiosyl repeat units that are 2-sulfated on the beta-D-galactopyranosyl and one-third 2-O-methylated on the 3,6-anhydro-L-galactosyl constituents. Of the remaining 4-linked residues, half are 2-O-methyl-D-galactopyranosyl residues and half are galactopyranosyl residues, of which approximately half are in the L configuration.

Effects of chorda-lingual nerve injury and repair on human taste.

Citric acid detection threshold and magnitude response were measured on the anterior tongue in 10 patients with unilateral chorda-lingual nerve transections before and after repair. Fungiform taste buds were analysed by videomicroscopy. Preliminary data suggests that humans can regenerate fungiform taste buds and recover some taste sensitivity after repair.

PTC/PROP tasting: anatomy, psychophysics, and sex effects.

Taste worlds of humans vary because of taste blindness to phenylthiocarbamide (PTC) and its chemical relative, 6-n-propylthiouracil (PROP). We review early PTC studies and apply modern statistical analyses to show that a higher frequency of women tasted PTC crystals, and were tasters (threshold classification). In our laboratory, scaling of PROP bitterness led to the identification of a subset of tasters (supertasters) who rate PROP as intensely bitter. Supertasters also perceive stronger tastes from a variety of bitter and sweet substances, and perceive more burn from oral irritants (alcohol and capsaicin). The density of taste receptors on the anterior tongue (fungiform papillae, taste buds) correlate significantly with perceived bitterness of PROP and support the supertaster concept. Psychophysical data from studies in our laboratory also show a sex effect; women are supertasters more frequently. The anatomical data also support the sex difference; women have more fungiform papillae and more taste buds. Future investigations of PTC/PROP tasting and food behaviors should include scaling to identify supertasters and separate sex effects.

A novel, erythroid cell-specific murine transcription factor that binds to the CACCC element and is related to the Krüppel family of nuclear proteins.

We describe a novel erythroid cell-specific cDNA (EKLF [erythroid Krüppel-like factor]) isolated by enriching for genes expressed in a mouse erythroleukemia cell line but not expressed in a mouse monocyte-macrophage cell line. The complete cDNA sequence is predicted to encode a protein of approximately 38,000 Da that contains a proline-rich amino domain and three TFIIIA-like zinc fingers within the carboxy domain. Additional sequence analyses reveal that the EKLF zinc fingers are most homologous to the Krüppel family of transcription factors and also allow us to predict potential DNA-binding target sites for the EKLF protein. On the basis of this prediction, we show that EKLF is able to bind the sequence CCA CAC CCT, an essential element of the beta-globin promoter. Its tissue distribution establishes that the EKLF transcript is expressed only in bone marrow and spleen, the two hematopoietic organs of the mouse, and analysis of murine cell lines indicates that EKLF expression is limited to erythroid and mast cell lines. Cotransfection assays establish that EKLF transcriptionally activates a target promoter that contains its DNA-binding site. The tissue expression pattern of EKLF, in conjunction with its function as a transcriptional activator, strongly suggests that the EKLF protein may be intimately involved in establishment and/or maintenance of the erythroid cell phenotype.

Comparative lectin histochemistry on taste buds in foliate, circumvallate and fungiform papillae of the rabbit tongue.

Taste buds (TB) in the foliate, circumvallate and fungiform papillae of the rabbit tongue were examined with lectin histochemistry by means of light (LM) and electron (EM) microscopy. Biotin- and gold-labeled lectins were used for the detection of carbohydrate residues in TB cells and subcutaneous salivary glands. At the LM level, the lectins of soybean (SBA) and peanut (PNA) react with material of the foliate and circumvallate taste pores only after pretreatment of the section with neuraminidase. This indicates that the terminal trisaccharide sequences are as follows: Sialic acid-Gal-GalNAc in O-glycosylated glycoproteins or Sialic acid-Gal-GlcNAc in N-glycosylated glycoproteins. In fungi-form taste buds the lectins of Dolichos biflorus (DBA) and Helix pomatia (HPA), also specific to GalNAc residues, are reactive without preincubation with neuraminidase. Wheat germ agglutinin (WGA), specific to GlcNAc, reacts with TBs of all papillae; and the lectin from Ulex europaeus (UEA I), specific to fucose, binds to individual TB cells. The presence of sialic acid may protect mucus or other glycoproteins in TB cells and inside the taste pore from premature enzymatic degradation. In a post-embedding EM procedure on LR-White-embedded tissue sections, only gold-labeled HPA was found to bind especially on membrane surfaces of the microvilli which protrude into the taste pore; however HPA did not bind to the electron-dense mucus inside the taste pore. The mucus situated in the trough and at the top of the adjacent epithelial cells also is strongly HPA-positive, but is of different origin and composition than that found in the taste pore. These results demonstrate distinct carbohydrate histochemical differences between fungiform and circumvallate/foliate taste buds. The different configuration of galactosyl residues and the occurrence of mannose in circumvallate and foliate TBs leads to the suggestion that the lectin reactivities of TBs are not only due to the presence of mucins, but also to N-linked glycoproteins, possibly with a hormone-like paraneuronal function. A possible relationship to v. Ebner glands in these papillae is discussed.

Variations in human taste bud density and taste intensity perception.

Some variations in human taste sensitivity may be due to different numbers of taste buds among subjects. Taste pores were counted on the tongue tips of 16 people with videomicroscopy, and the subjects were divided into two groups (N = 8) by the rank order of their taste bud densities. The "higher" density group averaged 374 +/- 134 taste pores/cm2, while the "lower" density group averaged 135 +/- 43 tp/cm2. The higher density group had an average fungiform papilla density which was 1.8 times greater than the lower density group and an average of 1.5 times more taste pores/papilla. The subjects also rated the intensity for 4 suprathreshold concentrations of 5 taste stimuli placed on the same region of the tongue where taste pores were counted. The group with higher taste bud densities gave significantly higher average intensity ratings for sucrose (196%), NaCl (135%) and PROP (142%), but not for citric acid (118%) and quinine HCl (110%) than the lower density group. Thus, the subjects with higher fungiform taste bud densities also reported some tastes as more intense than subjects with fewer fungiform taste buds.

The therapeutic empathic communication (TEC) process.

While the communication of empathy is thought to be a curative factor in psychotherapy, current conceptualizations usually focus on empathy as only an attitude or state of mind of the therapist. A five-stage model of therapeutic empathic communication is presented. The model is applied to analyzing the therapeutic impact of empathic communication in dynamic psychotherapy. Its usefulness is demonstrated in a case example.

Sucrose octaacetate-taster mice have more vallate taste buds than non-tasters.

Taste buds were counted in two strains of mice which have been characterized in terms of their taste avoidance of the bitter-tasting substance, sucrose octaacetate (SOA). One strain (SWR/J) avoids SOA and is referred to as "taster' while the other strain (C57BL/6J) does not avoid SOA at the same concentration and is termed "non-taster'. The taster-strain contains a significantly greater number of taste buds in its vallate papillae than the non-tasters do. The relative number of taste buds which individual mice and humans possess probably contributes to the relative differences in their sensitivity and preference behaviors.

Gustatory responsiveness of fibers in the hamster glossopharyngeal nerve.

1. Mammalian taste receptors are distributed within separate subpopulations, innervated by branches of cranial nerves VII, IX, and X. Most gustatory electrophysiology has focused on input from the fungiform papillae on the anterior portion of the tongue, carried by the chorda tympani branch of the VIIth nerve. However, only a small percentage of the taste buds are located in the fungiform papillae (approximately 18% in the hamster). There have been no studies on the hamster's IXth nerve, which innervates greater than 50% of its taste buds, and most other studies of IXth nerve function have employed only whole-nerve recording. 2. Action potentials were recorded from 83 individual fibers in the IXth nerve of the hamster. Stimuli were five concentrations each of sucrose, NaCl, HCl, and quinine hydrochloride (QHCl), all presented to every fiber at 37 degrees C. Responses were quantified as the number of impulses in 10 s minus the preceding 10 s of spontaneous activity. 3. Across these concentration series, HCl and QHCl were by far the most excitatory stimuli, with mean responses across all cells three to four times greater than those evoked by sucrose or NaCl. The order of effectiveness of the stimuli was H greater than Q much greater than N greater than S. 4. Of the 83 fibers, 56 were stimulated via the foliate papillae and 27 via the single vallate papilla. No fibers responded to both of these fields. There were generally no differences in the sensitivity of these two subpopulations of taste buds, except that QHCl was more effective when applied to the foliates. 5. A "total" response measure was derived by summing the excitatory responses to each stimulus across the entire concentration series. The fibers were then classified according to the best total response, resulting in 52 HCl-, 19 QHCl-, 8 sucrose- and 4 NaCl-best cells. Considering the slope of the concentration-response functions as a criterion for classification produced very similar results. The fiber classification varied somewhat with concentration, with more fibers categorized as HCl- and QHCl-best at the higher concentration levels. 6. Breadth of responsiveness was measured using the equation developed by Smith and Travers. At the concentrations used to examine hamster chorda tympani fibers, IXth nerve fibers were not very responsive and were quite narrowly tuned to the four taste qualities. At higher concentrations the fibers became more broadly responsive across the four stimuli.(ABSTRACT TRUNCATED AT 400 WORDS)

A comparison of the effects of bilateral sections of the chorda tympani nerve and extirpation of the submaxillary and sublingual salivary glands on the eating and drinking patterns of the rat.

The chorda tympani nerve (CT) innervates the fungiform papillae on the tip of the tongue and has been considered an important nerve for the sense of taste. The CT also contains the parasympathetic supply to the submaxillary and sublingual salivary glands. Therefore, changes in taste or feeding behavior following bilateral sections of CT are caused by both degeneration of fungiform papillae and the inevitable partial desalivation of the rat. In the present experiments we compared the effects of bilateral chorda tympani nerve sections with extirpation of submaxillary and sublingual glands on daily home cage eating and drinking patterns in the rat. Before and after surgery we analyzed the daily eating and drinking patterns, including such measures as intake, bout number, bout length, interbout interval and rate of consumption during bouts. The results of desalivation and bilateral CT sections were indistinguishable. The most profound change was that eating bout duration was increased following surgery. Since food intake did not increase, the results indicate a marked loss in eating efficiency over the daily ingestion periods. Although the eating patterns of desalivated and chorda tympani sectioned rats are quite similar, the evidence is not compelling that they have the same physiological basis. A second experiment was designed to test the hypothesis that the atypical eating patterns observed following bilateral sectioning of CT were the direct result of partial desalivation resulting from the denervation of the salivary glands. In this experiment a unilateral section was made of one CT and it was shown that the eating behavior was not affected. Then the contralateral submaxillary and sublingual salivary glands were removed. This resulted in a six-fold increase in feeding bout length. In all cases a unilateral CT section combined with extirpation of the contralateral salivary glands resulted in rats whose eating behavior was indistinguishable from the earlier data following either the bilateral CT sections or bilateral desalivations. The conclusion is drawn that the eating irregularities noted following bilateral CT sections result from this partial desalivation. CT sections were verified by taste bud counts in the fungiform papillae and histological examinations were made of salivary glands in rats receiving CT sections.

Proliferation of taste buds in the foliate and vallate papillae of postnatal hamsters.

The growth of the taste system in the hamster is considered in comparison to the postnatal development of other organ systems and the entire animal. No taste buds are present in vallate or foliate papillae of the hamster at birth, but they attain both the appearance and numbers of adult taste buds within 5 weeks of age. The most rapid increase in the number of taste buds occurs within the first 10 days of life, and this proliferation anticipates the weaning of hamsters which occurs by about three weeks of age. Foliate taste buds reach a maximal number within two months, but vallate taste buds continue to increase in number through 4 months of age. Taste bud proliferation and development occur earlier and more rapidly than in other organ systems. This early development of taste buds may protect the weanling hamster against accidental poisoning by noxious plants, and it may also reinforce the food intake which is required for normal growth.

Human taste bud density across adult age groups.

Some of the subjective variability attributed to taste experience could be related to wide variations of taste bud density. Studies of taste perception show a direct relationship between sensation and the number of receptors. Taste bud densities are quantified in this study using light microscopy to reconstruct two regions of 18 human cadaver tongues. Specimens came from male and female cadavers representing three age groups: young adults, middle-aged adults, and older adults. The results show a range of more than 100-fold in taste bud density that is evenly distributed among age groups and sexes. The disparity is not attributable to the state of health of the adults prior to death, and it is corroborated in the literature. Differences in taste bud density that extend across age groups probably confound some inferences about the effects of aging on taste sensitivity that are derived from cross-sectional studies of human populations. It is not clear from the data whether or not human taste bud density in individuals and in populations is stable or changing with time.

The effect of bilateral sectioning of the chorda tympani and the greater superficial petrosal nerves on the sweet taste in the rat.

The effects of bilateral deafferentation of the greater superficial petrosal (GSP) and the chorda tympani (CT) nerves on the ingestion of sucrose solutions were studied in rats. The rats received five daily sequential 30 second exposures for each sucrose concentration, and the average number of licks per exposure was calculated. Sucrose concentrations of 0.01, 0.03, 0.10, 0.32, and 1.00 M were presented in ascending order across days, both before and after bilateral sectioning of both the CT and the GSP nerves, the CT alone, the GSP alone, or a sham surgery. Prior to surgery, mean lick rate increased with increasing concentrations of sucrose. Following surgery, the rats with combined GSP and CT nerve sections showed a significant decrease in mean rate of licking to the sucrose solutions. The rats with GSP sections showed a similar decrease in mean lick rate to the sucrose solutions. Animals with sections of the CT nerve and sham-operated animals showed no significant reduction in mean lick rate to the sucrose solutions. These results indicate that the GSP nerve is important to the rat in reinforcing high lick rates to sucrose.

Variation in human fungiform taste bud densities among regions and subjects.

Taste sensitivity is known to vary among regions of the tongue and between subjects. The distribution of taste buds on the human tongue is examined in this report to determine if interregional and intersubject variation of taste bud density might account for some of the variation in human taste sensitivity. The subjects were ten males, aged 22-80 years, who died from acute trauma or an acute cardiovascular episode. Specimens were obtained as anatomical gifts or from autopsy. A sample of tissue about 1 cm2 was taken from the tongue tip and midlateral region; frozen sections were prepared for light microscopy; and serial sections were examined by light microscopy to count the taste buds. The average taste bud (tb) density on the tongue tip was 116 tb/cm2 with a range from 3.6 to 514 among subjects. The number of gustatory papillae on the tip averaged 24.5 papillae/cm2 with a range from 2.4 to 80. Taste bud density in the midregion averaged 25.2 tb/cm2 (range: 0-85.9), and the mean number of gustatory papillae was 8.25/cm2 (range: 0-28). The mean number of taste buds per papilla was 3.8 +/- 2.2 (s.d.) on the tip and 2.6 +/- 1.5 (s.d.) on the midregion. Subjects with the highest taste bud densities on the tip also had the highest densities in the midregion and the highest number of taste buds per papilla. Taste bud density was 4.6 times higher on the tip than the midregion, which probably accounts for some of the regional difference in taste sensitivity.(ABSTRACT TRUNCATED AT 250 WORDS)