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Biomedical subjects

L Bosco

Publications and source records attributed to L Bosco.

At least 19 recordsLinked to original sources

Expression of substance P, neurokinin 1 receptors (NK1) and neurokinin 3 receptors in the developing mouse retina and in the retina of NK1 knockout mice.

To complete a series of studies on the expression of substance P and neurokinin receptors in mammalian retinas, we investigated the occurrence of these molecules in developing mouse retinas and in retinas of mice with genetic deletion of the neurokinin 1 receptor, the preferred substance P receptor. Using semi-quantitative reverse transcription-polymerase chain reaction, we measured detectable levels of the gamma isoform of preprotachykinin A (a substance P precursor) mRNA at postnatal day 4. Neurokinin 1 receptor and neurokinin 3 receptor mRNAs were also detected at postnatal day 4. While gamma preprotachykinin A and neurokinin 1 receptor mRNA levels significantly increased up to eye opening (postnatal day 11), neurokinin 3 receptor mRNA levels remained constant throughout development. Substance P, neurokinin 1 receptor and neurokinin 3 receptor immunoreactivities were present at postnatal day 5. Substance P was in amacrine cells, neurokinin 1 receptor in developing amacrine and bipolar cells and neurokinin 3 receptor in OFF-type cone bipolar cells. Interestingly, a transient increase in the density of neurokinin 1 receptor immunoreactive processes was observed at eye opening in lamina 3 of the inner plexiform layer, suggesting a role of substance P and neurokinin 1 receptor in this developmental phase. However, in neurokinin 1 receptor knockout retinas, besides a significant increase of the gamma preprotachykinin A mRNA levels, no major changes were detected: neurokinin 3 receptor mRNA levels as well as substance P and neurokinin 3 receptor immunostainings were similar to wild types. Together with previous studies, these observations indicate that there are major differences in neurokinin 1 receptor expression patterns among developing mammalian retinas. The observations in neurokinin 1 receptor knockout mice may not be applicable to rats or rabbits, and substance P and neurokinin 1 receptor may play different developmental roles in different species.

Aging↗

Glutamic acid decarboxylase and GABA immunoreactivities in the cerebellar cortex of adult rat after prenatal exposure to a low concentration of carbon monoxide.

Glutamic acid decarboxylase and GABA immunoreactivities were qualitatively and quantitatively evaluated in the cerebellar cortex of adult rats prenatally exposed to a low concentration of carbon monoxide (75 parts per million). Carbon monoxide-exposed and control rats were perfused with modified Bouin's fluid and their cerebella were embedded in paraffin. Sections from the vermis of each cerebellum were stained with Toluidine Blue or assayed with anti-glutamic acid decarboxylase 65/67 or with anti-GABA antisera. In the Toluidine Blue-stained sections, no differences were observed in the microscopic structure of the cerebellar cortex between carbon monoxide-exposed rats and controls. The distribution patterns of glutamic acid decarboxylase and GABA immunoreactivities in the cerebellar cortex of the treated animals were qualitatively comparable to those of the controls, and in accordance with previous descriptions of glutamic acid decarboxylase and GABA immunoreactivities in the rat cerebellar cortex. However, quantitative analyses demonstrated a significant reduction of immunoreactivities to both substances in the exposed rats in comparison with the controls. The reduction regarded: in the molecular layer, the number of glutamic acid decarboxylase/GABA-immunoreactive neuronal bodies and of axon terminals and the area they covered; in the Purkinje neuron layer, the number and the area covered by glutamic acid decarboxylase/GABA immunoreactive axon terminals. The differences detected in the prenatally exposed adult rats could be due to carbon monoxide-induced impairment of the differentiation of cerebellar GABA synthesizing neurons. A consequently diminished synthesis of GABA might account for some behavioral disorders detected in adult rats submitted to the same experimental procedure.

Animals↗

Neurokinin 1 receptor expression and substance P physiological actions are developmentally regulated in the rabbit retina.

We investigated the expression of the substance P (SP) receptor (the neurokinin 1 receptor, NK1 receptor) and SP functional effects in developing rabbit retinas. NK1 receptors in adult retinas were in a population of cone bipolar cells and in dopaminergic amacrine cells, as previously described. In contrast, at birth and at postnatal day (PND) 6, NK1 receptors were exclusively expressed by cholinergic amacrine and displaced amacrine cells. NK1 receptor expression in cholinergic cells was still observed at PND10 (eye opening), while at PND21 it was confined to cholinergic cells of the inner nuclear layer. Starting at PND10, NK1 receptors were also in bipolar cells and in dopaminergic amacrine cells. A fully mature NK1 receptor expression pattern was observed at PND35. Dopamine release was assessed in isolated retinas in the presence of SP, the NK1 receptor agonist GR73632 or the NK1 receptor antagonist GR82334. At PND35, extracellular dopamine was significantly increased by 10 microM SP or 0.01-100 microM GR73632, and it was decreased by 0.01-10 microM GR82334. No effects were detected in developing retinas up to PND21. Ca2+ imaging experiments were performed in single cholinergic cells identified by their "starburst" morphology in perinatal retinas. Intracellular Ca2+ levels were significantly increased by 1 microM SP or GR73632. This effect was reversibly inhibited by 1 microM GR82334. These data demonstrate that both NK1 receptor expression and SP physiological actions are developmentally regulated in the retina. SP neurotransmission in the immature retina may subserve developmental events, and SP is likely to represent an important developmental factor for the maturation of retinal neurons and circuitries.

Acetylcholine↗

Expression of the neurokinin 1 receptor in the mouse retina.

Previous studies have revealed that the expression pattern of the neurokinin 1 receptor (the preferred receptor for substance P, SP) varies in different mammalian retinas. We investigated NK1 receptor expression in the mouse retina to provide background information for future studies in transgenic mice on SP functional roles in the retina. Mouse retinal sections were treated for single and double-label immunofluorescence. NK1 receptor immunoreactivity was in bipolar cells and in numerous amacrine cells. Double-label studies showed that NK1 receptor-expressing bipolar cells constituted a population of ON-type cone bipolar cells, since they were distinct from rod bipolar cells and contained glycine. They were nonrandomly distributed with highest density in central retina. These cells were similar and may correspond to the population of NK1 receptor-expressing bipolar cells of the rabbit retina. Different subsets of NK1 receptor-expressing amacrine cells were identified on the basis of the expression of selected neurotransmitter substances: i) about 23% of NK1 receptor-expressing amacrine cells also contained glycine; ii) the remaining 77% were likely to be GABAergic, although some inconsistency was observed in the GABA immunostaining obtained with two different GABA antibodies; iii) all dopaminergic amacrine cells also expressed NK1 receptors; iv) about one third of SP-containing amacrine cells also expressed NK1 receptors. These findings confirm and expand previous observations in rat and rabbit retinas. In particular, common to all three species is the expression of NK1 receptors in dopaminergic amacrine cells, indicating that SP neurotransmission may be a universal feature of the circuitry of the dopaminergic amacrine cell. Peculiar to the mouse retina is the presence of putative NK1 autoreceptors expressed by SP-containing amacrine cells.

Amacrine Cells↗

Expression of the neurokinin 1 receptor in the rabbit retina.

Substance P is the preferred ligand for the neurokinin 1 (NK1) receptor. In vertebrate retinas, substance P is expressed by amacrine, interplexiform and ganglion cells. Substance P influences the activity of amacrine and ganglion cells and it is reported to evoke dopamine release. We investigated NK1 receptor expression in the rabbit retina using affinity-purified NK1 receptor antibodies. NK1 receptors were expressed by two distinct populations of retinal neurons. One is a population of ON-type bipolar cells characterized by axonal arborizations that ramified in the inner plexiform layer near the ganglion cell layer. Double-label studies showed that NK1 receptor-expressing bipolar cells were distinct from rod bipolar cells and from other immunocytochemically identified types of cone bipolar cells. Their density was about 2250 cells/mm2 in the visual streak and 1115 cells/mm2 in ventral mid-periphery. They were distributed in a non-random pattern. In the outer plexiform layer, the dendrites of these bipolar cells converged into heavily immunostained clusters having a punctate appearance. The density of these clusters in mid-peripheral ventral regions (about 13000 clusters/mm2) was similar to the reported cone density [Famiglietti and Sharpe (1995) Vis. Neurosci. 12, 1151-1175], suggesting these dendrites contact all cone photoreceptors. The second NK1 receptor expressing cell population corresponds to the tyrosine hydroxylase-containing amacrine cell population. NK1 receptor immunostaining was localized to the cell body and processes, but not to the processes that form the 'rings' that are known to encircle somata of AII amacrine cells. These findings show that NK1 receptor immunoreactivity is localized to a population of ON-type cone bipolar cells and to dopaminergic amacrine cells, suggesting that substance P acting on NK1 receptors influences multiple retinal circuits in the rabbit retina.

Amacrine Cells↗

Developmental expression of neurokinin-1 and neurokinin-3 receptors in the rat retina.

Tachykinin (TK) peptides act on retinal neurons through neurokinin (NK) receptors. We examined the expression of neurokinin-1 (NK1; the substance P receptor), NK3 [the neurokinin B (NKB) receptor], and TK peptides in developing rat retinas. NK1 immunolabeling was found in newborn retinas in rare amacrine cells and in putative ganglion cells. At postnatal day 2 (PND 2), NK1 immunostaining was reduced greatly among ganglion cells, and it appeared in many amacrine cells and in fibers in the inner plexiform layer (IPL), with the highest density in laminae 1, 3, and 5. A similar pattern was found at PND 7. At PND 12, interplexiform NK1-immunoreactive (-IR) cells were detected, and NK1-IR fibers in the IPL were concentrated in lamina 2, similar to what was seen in adults. NK3 was expressed mainly by OFF-cone bipolar cells, and the developmental pattern of NK3 was compared with that of cone bipolar cells that were labeled with antibodies to recoverin. Immature recoverin-IR cone bipolar cells were seen at PND 2. NK3 immunolabeling was detected first in the outer plexiform layer and in sparse bipolar cell somata at PND 10, when recoverin-IR cone bipolar cells are nearly mature. By PND 15, both the NK3 immunostaining pattern and the recoverin immunostaining pattern were similar to the patterns seen in adults. TK immunoreactivity was present at PND 0 in amacrine cells and displaced amacrine cells. By PND 10, the morphologic maturation of TK-IR cells was complete. These findings indicate that, in early postnatal retinas, substance P may act on NK1 receptors, whereas NKB/NK3 interactions are unlikely, suggesting that there are different levels of importance for different TK peptides in the developing retina.

Animals↗

Epiluminescence microscopy as a useful approach in the early diagnosis of cutaneous malignant melanoma.

The aim of this study was to evaluate the role of epiluminescence microscopy (ELM) in the differential diagnosis of cutaneous pigmented lesions in order to improve the detection of cutaneous malignant melanoma (CMM) at earlier stages of the disease. In total, 3865 pigmented lesions from 2121 selected patients were evaluated using ELM with a hand-held video microscope imaging system (MS 500B Micro-Scopeman, Moritex). Comparison with histology was performed on the 476 surgically excised lesions. ELM sensitivity, specificity, positive and negative predictive values as well as agreement for the different risk levels of the lesions were determined. Of the 476 cutaneous pigmented lesions removed and histologically examined, 101 (21.2%) were non-melanocytic lesions and 375 (78.8%) were melanocytic lesions. Overall agreement was 83.4% (93.1% and 80.8% for non-melanocytic and melanocytic lesions, respectively). Sensitivity and specificity of ELM in the analysis of melanocytic lesions with a pigment network were both very high (92.3% and 91.2%, respectively). Sixty new cases of CMM were identified. A high proportion of melanoma at stage AJCC IA (23 out of 32; 71.8%) was diagnosed exclusively by ELM (four of these were in situ CCM lesions). ELM is therefore a powerful tool to discriminate between melanocytic and non-melanocytic lesions in order to avoid inopportune surgical treatments for low risk lesions. Unfortunately, ELM did not show 100% sensitivity in diagnosing CMM and therefore ELM features should be integrated with data from both the history and clinical evaluation. However, ELM is much more accurate than clinical examination in detecting thin CMM.

Adolescent↗

Apoptosis in sea urchin embryos.

It is demonstrated by DNA electrophoresis analysis, morphological observations and TdT in situ reaction, that Paracentrotus embryos if treated with TPA plus heat undergo an apoptotic reaction. Indication is also obtained that non treated embryos undergo spontaneous apoptosis at the early pluteus stage, especially in the districts of arms and intestine. The possible meaning of this latter observation is discussed.

Animals↗

Sea urchin mitochondrial matrix contains a 56-kDa chaperonine-like protein.

Paracentrotus lividus mitochondrial matrix contains a constitutive hsp of 56-KDa which cross reacts with a serum anti-hsp-60 chaperonine from yeast mitochondria. The localization of hsps preexisting or newly synthesized in different subcellular fractions of gastrula embryos is also analyzed by two-dimensional electrophoresis.

Animals↗

Lens fibre transdifferentiation in cultured larval Xenopus laevis outer cornea under the influence of neural retina-conditioned medium.

The outer cornea of larval Xenopus laevis can reprogram cell differentiation when cultured in medium conditioned by X. laevis neural retina (XRCM) or by Rana esculenta neural retina (RRCM). Under these experimental conditions corneal cells showed the same series of cytological changes of fibre cell differentiation observed during ontogenesis and in vivo lens regeneration: enlargement of nuclei and nucleoli, increase of ribosomal population (cytoplasm-basophilia), cell elongation gradual loss of basophilic properties and acquisition of acidophilic properties for crystallin synthesis and accumulation. These events were completely dependent on XRCM or RRCM, suggesting that the neural retina secretes a factor(s) which initiates and sustains lens fibre transdifferentiation of the corneal epithelial cells. This culture system appears to be a suitable one for investigating the control of lens fibre transdifferentiation in vitro.

Animals↗

In vitro lens transdifferentiation of Xenopus laevis outer cornea induced by Fibroblast Growth Factor (FGF).

It has been shown that lens regeneration from outer cornea of larval Xenopus laevis is dependent on neural retina both in vivo and in tissue culture. The isolated outer cornea cultured in the presence of bovine brain-derived acidic Fibroblast Growth Factor (aFGF) is able to reprogram the differentiation into lens fibers, although this transdifferentiative process is not coupled with the formation of a normally organized lens. The capacity of aFGF to promote lens differentiation from cornea is not linked to its mitogenic activity. The cultured corneal cells can transdifferentiate into lens fibers in the presence of aFGF when DNA replication and cell proliferation are prevented by addition of aphidicolin, a specific inhibitor of DNA polymerase in eukaryotes, to the culture medium.

Animals↗

[Laparoscopic approach excision of mesenteric cysts].

The authors reports a case of a young woman suffering from mesenteric cyst. The incidence of these lesions is 1 to 100,000 admissions in adult patients and 1 to 20,000 admissions in pediatric age. These cysts, with retroperitoneal and omental cysts, have a similar etiopathogenesis as lymphatic ectopic tissue. In the majority of cases these cysts are asymptomatic and often the diagnosis is intraoperative. In this case the cyst was removed successfully with laparoscopic approach. This mini-invasive method allows a complete and radical excision of the cyst and reduces postoperative hospital stay and morbidity.

Adult↗

Clustering of adverse drug events: analysis of risk factors for cerebellar toxicity with high-dose cytarabine.

BACKGROUND: Cerebellar toxicity is a severe, therapy-limiting adverse reaction of cytarabine given in high doses. The Food and Drug Administration received a report of an increased frequency of cerebellar toxicity at the University of Wisconsin Hospital and Clinics after a switch from the product (Cytosar-U) manufactured by The Upjohn Co., Kalamazoo, Mich., to the generic form made by Quad Pharmaceuticals, Inc., Indianapolis, Ind. PURPOSE: To compare the incidence of cerebellar toxicity in Quad-treated patients with Upjohn-treated patients, a record-based cohort study was conducted at the University of Wisconsin Hospital and Clinics between January 1986 and August 1989. METHODS: The incidence of cerebellar toxicity was studied in 63 leukemia patients according to the manufacturer of the product received (34 Upjohn only, 25 Quad only, and four both manufacturers). The relative risk of cerebellar toxicity was adjusted for other known risk factors. RESULTS: Patients in the manufacturer-defined treatment groups did not differ significantly with respect to age, sex, type of leukemia, disease stage, calculated creatinine clearance, presence of abnormal liver function tests, or total dose received. The crude relative risk of cerebellar toxicity comparing the Quad product with the Upjohn product was 5.0 (95% confidence interval = 1.8-13.7). Adjustment for potential confounders did not alter the association. Other risk factors for cerebellar toxicity, independent of manufacturer, were age greater than 50 years, type of leukemia, disease stage, total dose greater than or equal to 20 g/m2, abnormal pretreatment liver function, and reduced creatinine clearance. CONCLUSION: This study found a significantly higher incidence of cerebellar toxicity with high-dose cytarabine manufactured by Quad Pharmaceuticals when compared with the incidence of cerebellar toxicity with the Upjohn product. Further research at independent institutions would be necessary to allow generalization of this finding. In addition, our findings suggest that a dose reduction in high-dose cytarabine therapy may be indicated for patients with reduced glomerular filtration rates.

Adolescent↗

[Use of mechanical staplers in Billroth II gastrectomy].

Some considerations on the use of staplers, use currently classified as "elective" and "of convenience", are shown. The Author wishes a change in this classification especially for what elective subtotal gastrectomy is concerned. In fact, the use of staplers in subtotal gastrectomy for tumors of the pyloric antrum and for complicated ulcer assures considerable advantages in terms of quickness, safety and sterility. Finally, the Author describes in detail a Billroth II subtotal gastrectomy performed with staplers.

Gastrectomy↗

[Histochemical study of rabbit duodenal mucosa carried out using lectins].

The Authors report histochemical findings about rabbit's duodenal mucosa. The present study has been carried out using five different lectins (Peanut Agglutinin (PNA), Dolichos Biflorus Agglutinin (DBA), Wheat Germ Agglutinin (WGA), Soybean Agglutinin (SBA), Ulex Europaeus Agglutinin I (UEA-I). These lectins have been labelled with Horseradish Peroxidase and binding sites have been stained with 3-3' Diaminobenzidine, according to Farragiana et al. The PNA reacted with the glandular cells, while the reaction was negative in the superficial cells. The DBA reacted exclusively with the glandular cells. The superficial and the glandular cells showed strong positive binding sites to the WGA and slight positive binding sites to the SBA. The UEA-I did not react with the epithelial cells. The presence of binding sites for the lectins we have used in the present study, shows a different glycoprotein composition of the cellular secretion, in comparison with the other animals we have already studied. In addition, these lectins can not be used as cellular differentiation markers in the epithelial cells of the rabbit's duodenal mucosa.

Animals↗