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Purpose Laboratory, imaging, and pathological features of Graves’ disease (GD), although well characterized, have been barely correlated each other. Aim of the study was to link laboratory and ultrasound characteristics of GD with its pathological features. Methods We correlated laboratory and ultrasound data at the time of diagnosis in 28 consecutive GD patients who underwent thyroidectomy with their pathological features, i.e., lymphocytic infiltration and follicular hyperplasia (both classified as mild or severe). Results Thyroid volume correlated positively with the levels of FT4 (P = 0.002, r2 = 0.42), FT3 (P = 0.011, r2 = 0.22), autoantibodies to thyroglobulin (TgAbs) (P = 0.016, r2 = 0.32), autoantibodies to thyroid peroxidase (TPOAbs) (P = 0.011, r2 = 0.34) and the extent of lymphocytic infiltration (P = 0.006 comparing mild to severe lymphocytic infiltration) but not with the levels of autoantibodies to the thyrotropin receptor (TRAbs) and to follicular hyperplasia. Compared to subjects with mild lymphocytic infiltration, those with severe lymphocytic infiltration showed higher levels of TgAbs (316 vs 0.0 IU/mL, P P P = 0.68). Compared to patients with mild, those with severe follicular hyperplasia had similar levels of TgAbs (76 vs 30 IU/mL, P = 0.31) and TPOAbs (251 IU/mL vs 45 IU/mL, P = 0.26) but higher levels of TRAbs (39 vs 7.2 IU/mL, P Conclusion In GD, TgAbs and TPOAbs levels correlate with the extent of lymphocytic infiltration, TRAbs levels with the degree of follicular hyperplasia. Thyroid volume, the main factor influencing the severity of hyperthyroidism, is related to lymphocytic infiltration and not to follicular hyperplasia.

Riassunto La chirurgia della tiroide si e evoluta dalla sua descrizione iniziale di Kocher alla fine del XIX secolo. Alla fine del XX secolo, e apparsa la tiroidectomia endoscopica, chiamata anche videoassistita. All’inizio del XXI secolo, gli sviluppi tecnologici hanno permesso la realizzazione della tiroidectomia robotica. Se la tiroidectomia convenzionale tramite cervicotomia rimane la tecnica piu ampiamente utilizzata in tutto il mondo, essa ha visto la sua pratica implementata dal monitoraggio dei nervi ricorrenti e vago. La tiroidectomia ha le sue indicazioni per il cancro della tiroide e per le patologie benigne della tiroide. In caso di cancro, comprende un trattamento del tumore tiroideo e delle aree linfonodali. Le raccomandazioni internazionali hanno tentato di standardizzare le indicazioni chirurgiche per le lesioni maligne. Questo articolo descrive la tecnica della tiroidectomia convenzionale tramite cervicotomia.

Resumen La cirugia tiroidea ha evolucionado desde su descripcion inicial por Kocher a finales del siglo XIX. A finales del siglo XX, surgio la tiroidectomia endoscopica, tambien denominada videoasistida. A comienzos del siglo XXI, los avances tecnologicos han permitido la realizacion de la tiroidectomia robotica. Aunque la tiroidectomia convencional por cervicotomia sigue siendo la tecnica mas utilizada en todo el mundo, su tecnica se ha complementado con la monitorizacion de los nervios recurrentes y vago. La tiroidectomia tiene sus indicaciones en caso de cancer de glandula tiroidea y para patologias tiroideas benignas. En caso de cancer, consta del tratamiento del tumor tiroideo y de las areas ganglionares. Varias recomendaciones internacionales han intentado estandarizar las indicaciones quirurgicas para las lesiones malignas. En este articulo se describe la tecnica de la tiroidectomia abierta convencional por cervicotomia.

Riassunto La chirurgia tiroidea si e evoluta dalla sua descrizione iniziale da parte di Kocher alla fine del XIX secolo. Alla fine del XX secolo, e apparsa la tiroidectomia endoscopica, definita anche videoassistita. All’inizio del XXI secolo, gli sviluppi tecnologici hanno permesso la realizzazione della tiroidectomia robotica. Se la tiroidectomia convenzionale mediante cervicotomia rimane la tecnica piu utilizzata al mondo, questa ha visto la sua pratica implementata dal monitoraggio dei nervi ricorrenti e vaghi. La tiroidectomia ha le sue indicazioni in caso di cancro alla tiroide e di alcune patologie tiroidee benigne. In caso di cancro, comprende il trattamento del tumore tiroideo e delle aree linfonodali. Alcune raccomandazioni internazionali hanno tentato di standardizzare le indicazioni chirurgiche per le lesioni maligne. Questo articolo descrive la tiroidectomia videoassistita nota anche come MIVAT (mini-invasive video-assisted thyroidectomy) e la tiroidectomia robotica.

Aims Leucocyte esterase (LE) has been shown to be an accurate marker of prosthetic joint infection (PJI), and has been proposed as an alternative to frozen section (FS) histology for intraoperative diagnosis. In this study, the intraoperative assessment of LE was compared with FS histology for the diagnosis of prosthetic hip infection. Patients and Methods A total of 119 patients undergoing revision total hip arthroplasty (THA) between June 2015 and December 2017 were included in the study. There were 56 men and 63 women with a mean age of 66.2 years (27 to 88). Synovial fluid was collected before arthrotomy for the assessment of LE using enzymatic colourimetric strips. Between five and six samples were stained with haematoxylin and eosin for FS histology, and considered suggestive of infection when at least five polymorphonuclear leucocytes were found in five high-power fields. Results The sensitivity and specificity of the LE assay were 100% and 93.8%, respectively; the positive (PPV) and the negative (NPV) predictive values were 79.3% and 100%, respectively. The mean time between the collection of the sample and the result being known was 20.1 minutes (sd 4.4). The sensitivity and specificity of FS histology were 78.3% and 96.9%, respectively; the PPV and the NPV were 85.7% and 94.9%, respectively. The mean time between the collection of the sample and the result being known was 27.2 minutes (sd 6.9). Conclusion The sensitivity of LE assay was higher, with similar specificity and diagnostic accuracy, compared with FS histology. The faster turnaround time, its ease of use, and low costs make LE assay a valuable alternative to FS histology. We now use it routinely for the intraoperative diagnosis of PJI. Cite this article: Bone Joint J 2019;101-B:372–377.

Minimally invasive video-assisted thyroidectomy (MIVAT) and endoscopic thyroidectomy through areola (ETA) access are among the endoscopic approaches for thyroidectomy that have been perfected, but reports comparing the two are lacking. This study evaluated the safety, postoperative recovery, and patient satisfaction of MIVAT compared with ETA.This study included 119 patients undergoing MIVAT and 42 patients undergoing ETA from January 2006 to October 2009. Operative time, complications, postoperative recovery, cosmetic result, and patient satisfaction were analyzed.The ETA group had a higher percentage of women (100% vs. 78.2%, P0.05) and was younger (33.5 vs. 39 years, P0.05). Thyroid volume (24.5 vs. 14.5 mL, P0.001) and nodular diameter (26 vs. 22 mm, P0.05) were larger in ETA group. The MIVAT group had a shorter operative time (28.2 vs. 112.8 minutes) and a lower rate of intraoperative (7.2 vs. 21.2 mL) and postoperative (0 vs. 80 mL, P0.0001) blood loss. Rates of conversion and complications were similar. Postoperative pain at 12 hours was 1.9 in MIVAT vs. 3.1 in ETA (P0.0001). Hospitalization was 1 day in the MIVAT group vs. 3.5 days in ETA (P0.0001). Patient satisfaction was similar.MIVAT and ETA showed similar results for safety, although ETA might be considered more invasive than MIVAT. Patients of both groups were equally satisfied with the cosmetic result. Differently from ETA, MIVAT allows total thyroidectomy to be performed also for malignant diseases. Although different, the two approaches were safe and reliable and both are valid options. Choice might depend on the surgeon's preference, thyroid size, type of disease, and the patient's expectation about cosmesis.

Data on the cardiovascular middle-term follow-up of patients with primary aldosteronism (PA) are scanty.To detect the cardiovascular effects of surgery in patients with aldosterone (ALD)-producing adenoma (APA) and of pharmacotherapy in those with bilateral adrenal hyperplasia (BAH), a prospective study involving 60 consecutive patients with PA was performed. MATERIAL/ METHODS: Clinical, biochemical, and cardiovascular assessment was obtained before and after (31.5±4.4 months) surgery or proper medical treatment (32.1±5.0 months) in 19 and 41 patients, respectively.As expected, plasma ALD normalized in all operated patients, while in the other group it did not change. Systolic and diastolic blood pressure decreased (p0.001) after both treatments. However, absolute and percentage reduction was significantly more pronounced (p0.01) in operated than in non-operated patients. Left ventricular (LV) mass showed significant reduction after surgery (LV mass g/m(2), p0.0007; LV mass g/m(2.7), p0.01), but no change after medical treatment, so that the differences between absolute and percentage values at follow- up were statistically significant (p0.01) between groups. Basal LV mass/m(2.7) was positively associated with age (p0.009), body mass index (p0.0008), drug number (p0.03), and ALD/plasma renin activity ratio (p0.01). Allocating the patients according to plasma ALD and cardiac parameters, patients who presented ALD reduction during the study also had a decrement in cardiac mass (p0.04).Our data indicate that in patients with PA the removal of ALD excess by surgery in APA is effective in reducing blood pressure and in improving cardiac parameters, while anti-hypertensive therapy in BAH shows less positive impact on cardiovascular system.

This study was aimed to demonstrate the clinical benefits of rearranged during transfection (RET) genetic screening in patients with apparently sporadic medullary thyroid cancer (MTC) not only to identify the hereditary nature of the disease in the index case but also to discover family members harbouring the same germline mutations (i.e. gene carriers) who are unaware of their condition.RET genetic screening allowed the identification of germline RET mutations in apparently sporadic MTC resulting in their re-classification as hereditary forms.RET genetic screening was performed in 729 apparently sporadic MTC patients by direct sequencing RET exons 5, 8, 10, 11 and 13-16. Clinical and biochemical evaluation of gene carriers was also performed.We discovered an unsuspected germline RET mutation in 47 of 729 (6·5%) apparently sporadic MTC who were re-classified as hereditary. We found 60 of 146 (41·1%) gene carriers, 35 of whom had biochemical or clinical evidence of MTC. Thirty gene carriers underwent total thyroidectomy and 27 of 30 (90%) were persistently cured after a mean follow-up of 6·0 years. As a further result of RET genetic screening, we observed a significantly higher prevalence of familial medullary thyroid cancer (FMTC) in our series with respect to the largest series of the International RET Consortium (P = 0·0002).RET genetic screening of patients with apparently sporadic MTC represents a major tool for the preclinical diagnosis and early treatment of unsuspected affected family members and allows the identification of a relevant percentage of hidden FMTC.

The onset of cervicoscopy dates back to the first endoscopic parathyroidectomy in 1996. This operation, along with its several variants, has become a valid option widespread in many important centres. Later on, endoscopic or video assisted thyroidectomy was introduced in spite of the limits imposed by the mass of the gland to remove. It is indicated for a minority of patients for this reason but both parathyroidectomy and thyroidectomy showed some important advantages with respect to conventional surgery, advantages demonstrated also in prospective studies. They are mainly represented by a better cosmetic outcome and a less distressful postoperative course. These approaches proved to be safe and feasible in any surgical background: their complication rate is the same as traditional open surgery in the neck. Very promising seems to be the videoscopic access to neck lymph nodes (central and lateral compartments) whereas other fields of application such as carotid artery surgery and spine surgery still remain object of experimental studies. As far as the lateral neck dissection is concerned the technique is going to be standardized in our centre as a variant of the well known video assisted approach adding a 5 mm trocar placed in the supraclavicular space. By consequence, cervicoscopy has to be considered an important surgical tool which can be further improved but which also has an excellent potentiality.

Sialoderivatives expressed in the rabbit oviduct under hormonal treatment have been investigated in situ by lectin histochemistry with a view to specifying further regional and temporal specializations which enable ampulla and isthmus to play distinct roles in the reproductive events. Application of MAL II and SNA lectins allowed sialoglycoconjugates containing Sia(alpha2,3)Gal and Sia(alpha2,6)Gal groups to be discriminated. Sialic acid residues linked to Gal(beta1,3)-D-GalNAc sequences were identified using PNA combined with sialidase digestion. Information on structural features of sialic acids were acquired by deacetylation and differential oxidation pretreatments. In both oviductal portions, Sia(alpha2,6) groups were restricted to the luminal surface of the lining epithelium while Sia(alpha2,3) groups were specifically located in the secretory, non-ciliated cells. In the ampullary epithelium, non-acetylated sialic acids alpha2,3-linked to Gal(beta1,3)-D-GalNAc sequences were largely present. Only at ovulation time were sialic acid residues containing acetyl substituents on C4 also found. A great variety of sialic acids were found in the isthmic epithelium which showed the highest expression of acetylated forms at the first hours after the hormonal treatment. The heterogeneity of sialoderivatives differently expressed in the ampulla and isthmus as well as their distinct cycle-dependent modulation suggest that sialylated components may contribute to the molecular and functional specificities within the oviductal epithelium.

Lectin cytochemistry was performed in vitro on primary cultures from the rat submandibular gland. For this purpose, prepubertal rats (17, 27, 33 days old) of both sexes were used. Several types of medium supplements were tested and it was found that cells survived until 15 days in presence of all medium supplements and extracellular matrix gel. The binding patterns of all FITC/TRITC-labeled lectins, with and without prior sialidase digestion and deacetylation, were analyzed in a confocal laser scanning microscope. In particular, the occurrence of C4 acetylated sialic acid linked to beta-galactose at day 27 and the presence of fucose residues at day 33 indicated that lectin probes applied to cultured cells give results similar to those obtained in intact tissues and can be used as markers of growth and differentiation.

Direct and indirect staining procedures were developed to characterize sialoglycoconjugates in developing rat submandibular gland. Lectin histochemistry, with and without prior sialidase digestion, combined with differential oxidation and deacetylation procedures was performed in situ. This allowed the expression of sialic acids to be followed during acinar cell development. It was found that terminal periodate-labile sialic acids linked to beta-galactose occurred early. In contrast, the terminal disaccharide sialic acid-N-acetylgalactosamine was only detectable at the adult stage and so was considered to be a good marker of the full maturity of this gland. The developing acinar cells were mainly characterized by C4-acetylated sialic acids belonging to short side-chains. Dimorphic expression of sialoglycoconjugate components was evident by postnatal day 44.

Sialoglycoconjugates in rat sublingual gland acinar cells, at different stages of pre- and post-natal development, were investigated in situ with specific lectins and by the selective removal of terminal sialic acids. Cleavage of acetyl substituents sited in the pyranose ring and/or polyhydroxyl side chain was used as an additional means of characterising the glycoconjugates. The first expression of terminal sialic acid linked to beta-galactose was found at gestational day 17 and progressive different derivatives were observed. The terminal disaccharide sialic acid-N-acetylgalactosamine was constantly visualized in the sublingual gland from gestational day 18. In both terminal disaccharides, sialic acids were characterized by variable degrees of acetylation and were found to be highly packaged and responsible for the hydration coat. The complex data obtained indicated that the sublingual gland is characterized by a marked fluctuation of complex sialoglycoconjugates that differ from those in the submandibular gland of the same species.

The research was planned to study the subcellular distribution of enzymatic secretory products within the secretory structures of the mouse major salivary glands at light and electron microscopy level by immunogold silver stain (IGSS) technique and double-sided post-embedding immunogold binding and silver amplification in order to speculate about their compartmentation. In particular, we experimented the above immunogold labeling approaches to localize the lysozyme and to verify its distribution patterns in relation to another secretion enzyme, alpha-amylase. Co-presence of lysozyme and alpha-amylase was observed in the convoluted granular tubule cells of the submandibular gland and in the demilunar cells of the sublingual gland as well as in the electron-dense regions of the mottled secretory granules in the parotid gland. Exclusive binding patterns of lysozyme were observed in the acinar cells of the submandibular and sublingual glands where alpha-amylase did not occur.

Zwischen 02/97 und 04/00 wurde bei 140 Pat. mit Nebenschilddrusenadenom eine minimal invasive Parathyreoidektomie (MIVAP) und bei 64 Pat. mit kleinen Knotenstruma eine minimal invasive Thyreoidektomie (MIVAT) vorgenommen. Bei der MIVAP betrug die mittlere OP-Dauer 54 (15–180)Min; 12 Falle (8.5%) mussten konvertiert werden; eine Recurrensparese trat 2x (1.4%) auf, eine Hypokalzamie 6x (4.3%); 115 Pat. (82%) wurden am 1. postop. Tag entlassen. Bei der MIVAT betrug die mittlere OP-Dauer 80 (35–130)Min; 3 Falle (4.7%) mussten konvertiert werden; eine Recurrensparese trat 1x (1.6%) auf; 56 Pat. (88%) wurden am 2. postop. Tag entlassen. Die MIVAP und die MIVAT erfreuen sich fur spezielle Indikationen sehr guter funktioneller und kosmetischer Ergebnisse.

The novel combination of sialidase digestion with simultaneous PNA and DBA binding yielded marked differences on sialoglycoconjugate occurrence and distribution in the mouse submandibular gland acinar cells of the two sexes. Striking differences in the structure of terminal disaccharides within stored secretory sialoglycoconjugates were also found. High content of sialic acid, characterized by the terminal sequence sialic acid-alpha-N-acetylgalactosamine, was established to only occur in the male acini where secretory cells appeared to be differently stained; indeed, some cells exhibited codistribution of sialic acid-alpha-N-acetylgalactosamine and sialic acid-beta-galactose terminal disaccharides, whereas other ones exclusively contained one of the two kinds of terminal sequences. In the female acinar cells, the secretory products were found to be almost exclusively composed by glycoconjugates having sialic acid subtended to beta-galactose without appreciable differences between acinar cells. Our finding of such extensive differences in the acinar cells of male and female mice adds new insights into the submandibular gland sexual dimorphism, commonly attributed to the androgen responsiveness of the granular convoluted tubule portion of the gland.

In the present study we report the development of an ultrastructural electron microscopic double-sided staining technique that, using gold probes of 10 nm and enhancement of the gold signal by silver amplification, allows the demonstration of two antigenic sites on the same section. The labeling was carried out in the following manner: one face of uncoated floating grids was incubated with an antibody directed to alpha-amylase, followed by a secondary gold-labeled antibody, amplification of gold particles, drying and carbon coating; subsequently, the reverse face of the same grid, was processed for lectin cytochemistry, with and without sialidase digestion, and it was incubated with HRP-conjugated lectins, anti-HRP antibody and protein-A gold. Also the reverse sequence of steps and amplification of gold signal after the first or second labeling were experimented. The resultant small and large particles revealed different distributional patterns of antigenic sites on the opposite faces of the same tissue section. The transparency of the resin-embedded ultrathin sections in the electron beam allowed the simultaneous visualization of the gold probes of different sizes present on the two faces. The analysis of immunolabeling revealed that the alpha-amylase is chiefly secreted by the parotid and submandibular glands. The application of this double-sided staining technique also indicated that, when present in glycosylated form, the alpha-amylase enzyme does not contain sialic acid in the submandibular and sublingual glands; conversely, its location on the electron-dense areas of target granules in the parotid acinar cells seems to suggest that a sialylated isoenzymatic form can occur within these granule regions where sialic, acid linked to beta-galactose, was found to be located.

After 3 years from the introduction of laparoscopic adrenalectomy in an endocrine surgery unit the results are retrospectively compared with those achieved by traditional techniques with the aim of comparing the respective advantages.During this period 68 laparoscopic adrenalectomies have been performed. The main pre-, intra- e postoperative parameters of the adrenalectomies for benign neoplasm have been examined. Mean follow-up was 51 months (65.3 for open adrenalectomy and 18.8 for laparoscopic).Statistical studies were homogeneous between the two groups. The laparoscopic adrenalectomy--with the same effectiveness--thanks to less peritoneum and parietal stress, is followed by fewer postoperative complications, faster resumption of biological functions, earlier return to work and better cosmetic results.On the basis of our personal experience laparoscopic adrenalectomy is to be considered the treatment of choice in the majority of adrenal benign neoplasms.

The effect of repeated stress (1 h of daily immobilization for seven consecutive days) on the adrenal cortex of young adult male albino rats was evaluated by morphohistochemical methods and plasma assays; at the same time, testes and major salivary glands, as steroid-producing and -depending organs, respectively, were examined. Morphological and histochemical changes were found in the adrenal cortex, testis and submaxillary gland, though varying in degree and extent depending on the gland examined. Corticosterone and progesterone plasma levels increased, in agreement with the lipid depletion observed in the zona fasciculata, while testosterone and androstenedione decreased, as confirmed by the less marked enzymatic activity in the Leydig cells. The study thus proves that repeated stress, even of temporary duration, is able to influence directly or indirectly the morphofunctional state of the three examined glands, suggesting a functional linkage.

Double lectin labeling for confocal microscopy and lectin-protein A-gold binding for electron microscopy were applied to the mouse sublingual gland in order to study surface and cytoplasmic sialoglycoconjugates. For this purpose, serially cut sections were submitted to sialidase followed by incubation with lectins recognizing usually acceptor sugars for terminal sialic acids. At the electron microscope level, the residues subtended to sialic acid were individually identified on adjacent sections by an indirect technique of labeling, whereas with confocal microscopy the above sugars were simultaneously visualized on the same section by a double staining method using fluorescein isothiocyanate (FITC)- and tetramethylrhodamine isothiocyanate (TRITC)-conjugated lectins. Acinar cells were found to contain the terminal sequence sialic acid-beta-galactose in abundance while the sequence sialic acid-alpha-N-acetylgalactosamine appeared to be present in modest amounts. Both sialoglycoconjugates were homogeneously codistributed inside acinar cells. The combination with a saponification method also allowed the occurrence of C4 acetylated sialic acids linked to beta-galactose to be discovered, at the electron microscope level, on acinar cell secretory products.

An ultrastructural analysis of lectin receptors on the submandibular glands from mice of both sexes was performed utilizing horseradish peroxidase-labelled lectins in conjunction with antiperoxidase antibody and protein A-gold. Both qualitative and quantitative sex-related differences in terminal sugar expression within secretory granules were detected. Following sialidase digestion, also subterminal acceptor sugars for terminal sialic acids, proved to be differentially expressed in the submandibular glands of males and females. Heterogeneous distribution of sialoglycoconjugates characterized by the terminal disaccharide sialic acid-beta-galactose was found to occur in female acinar cells. Also DBA reactive sites indicating the presence of terminal alpha-N-acetylgalactosamine discriminated between male and female acinar secretory glycoconjugates. This difference was emphasized by sialidase pretreatment that evidenced a marked occurrence of sialic acid subtended to alpha-N-acetylgalactosamine in males in contrast to a modest presence in females. The different sialylation patterns of acinar cell secretory products, probably related to a different expression of O- and N-linked sialoglycoconjugates, give insight into the sexual dimorphism of the mouse submandibular gland known until recently for the convoluted granular tubules.

An electron microscopic investigation was performed to examine the ultrastructural changes occurring in the rabbit oviductal epithelium after human chorionic gonadotropin (HCG) administration. Mainly, the non-ciliated secretory cells proved to be affected by the hormonal treatment which resulted in qualitative and quantitative modifications of the secretory patterns differently expressed in the ampulla and isthmus. Thus, morphological evidence of intense secretion was observed in both the oviduct regions at preovulatory stages. Following ovulation, timing of expression of active secretory patterns in the ampulla and isthmus correlated well with the rate of gamete transport and relative functional roles of the oviductal regions in the reproductive process. At present, HCG-induced changes concerning the ciliated cells seem to consist of the occurrence of secretory granules responsible for the appearance of "mixed cells".

The localization of individual glycosidic residues in the mouse and rat submandibular glands was examined using confocal laser scanning microscopy (CLSM). For these organs we tested some procedures of fixation and embedding to better understand the distribution of some lectin-probes inside well preserved secretory cells and observed that fixation and inclusion steps did not influence appreciably the location and intensity of the reactive sites. The fixation mixture of 4% paraformaldehyde, 1% glutaraldehyde and 0.2% picric acid produced the most satisfactory results. In specimens labeled with PNA-, Con A-, LTA-FITC and WGA-, DBA-TRITC lectins, the convoluted granular tubules (CGT) proved to be composed of secretory granules with high-density lectin labeling. The complex organization of secretory glycocomponents within the granule matrix was further resolved by double labeling and dual scanning experiments. Some lectins exhibited colocalization while others displayed differential localization providing information about the occurrence of O- and N-linked glycoconjugates. The CLSM technique applied to fluorochrome-conjugated lectins also revealed a more marked dimorphism in the rat rather than in the mouse submandibular gland. In particular, the male rat submandibular gland was found to consist of CGT heterogeneous cell populations, while the mouse submandibular gland did not show glycochemical differences between cells. Female rats exhibited a lectin profile very different from that of female mouse.

Sections from the major salivary glands of rats and mice were used to locate, characterize and compare sialoglycoconjugates by means of lectin histochemistry, sialidase digestion, periodate oxidation and potassium hydroxide deacetylation. The gland sialylated macromolecules contained the terminal dimers sialic acid-beta-galactose and sialic acid-alpha-N-acetyl-galactosamine but differed in the varieties of sialic acids and the linkages of sialic acids to penultimate sugars. Indeed, the submandibular and parotid glands exhibited a notable occurrence of periodate labile sialic acids with C7 and/or C8 and/or C9 acetyl groups in their polyhydroxyl chains. In particular, C9 acetylated sialic acids were mostly linked alpha 2-6 to beta-galactose. The sublingual glands, instead, were strongly characterized by a presence of C9 acetylated sialic acids bound alpha 2-3 to beta-galactose. Also, sialic acids with O-acetyl substituents at C4 were evident in the mouse parotid gland and in the rat submandibular and sublingual glands. The great variety of sialoderivatives expressed by the rodent salivary glands was correlated with the differential involvement of these compounds in lubricating and defensive processes. Sex-related differences regarding the sialic acid location, acetylation degree and linkage were shown in the submandibular glands of both species.

In this study colloidal gold methodologies that allow the lectin affinity patterns in the mouse parotid gland to be studied by postembedding techniques were addressed. DBA, Con A, PNA, WGA and LTA lectins conjugated with gold particles or horseradish peroxidase for a direct or indirect technique of binding were used. Thin sections were obtained from tissue embedded in the acrylic hydrophilic resins Bioacryl and Lowicryl K4M. The most satisfactory results originated from sections infiltrated in Bioacryl and submitted to the indirect technique of binding. The mouse parotid gland appeared to be composed of polymorphous secretory granules differentially stained by lectins. The glycosylated components of acinar cells reacted intensely, and gold particles specified the ultrastructural distribution of lectin affinity sites reflecting the location of beta-galactose, alpha-N-acetylgalactosamine, beta-N-acetylglucosamine, alpha-D-mannose, and alpha-L-fucose in the electron-dense or electron-lucent areas of bizonal, mottled, and target granules.

This review was focused on the salivary gland oligosaccharide chains studied by lectin histochemistry combined with exoglycosidase digestion. Glycoconjugates play an important role in many biofunctions and, generally, salivary mucins, which consist of numerous oligosaccharide chains attached at closely spaced intervals to a peptide backbone, serve as lubricants and protective agents, but in many instances we are ignorant about the role of biochemically identified oligosaccharides. Lectin histochemistry represents the greatest analytic tool to study carbohydrates in situ; in addition, there is availability of selective enzymes, so glycosidase degradation is useful to both investigate the structure of a given oligosaccharide and verify the influence of neighbouring sugars on the affinity towards the respective specific lectins. Using stepwise digestion of samples, followed by lectin labelling, the structure of terminal short oligosaccharides with blood-group activity was also elucidated. Additional histochemical methodologies were developed to establish the presence of acetylated groups in sialic acid residues, and the position of the linkage to the underlying monosaccharide. Sequencing approaches by exoglycosidases and lectins were also seen to be particularly useful when substantial differences did not emerge in lectin affinity, glycoconjugate composition and complex carbohydrate cytochemistry.

An investigation by electron microscope was performed on the submandibular gland of unweaned cats for studying its morphology and carbohydrate histochemistry. The cytoarchitecture was comparable with that of adult animals with the exception of the degree of membrane merging in both demilunar and acinar cell secretory granules. Conversely, the submandibular gland of unweaned cats showed a peculiar HID-TCH-SP reactivity at acinar cell level.

Background: The advantage of minimally invasive parathyroidectomy and the cost saving associated with outpatient surgery has led to an increased interest in local or local/regional anesthesia.

In this study we investigated immunohistochemically the distribution of VIP in ampullary and isthmic segments of the rabbit oviduct at different hormonal stages in order to correlate the presence of VIP with the morphological and functional differences between the two oviduct segments. Noticeable changes in the VIP-positive sites emerged between corresponding oviduct segments during the stages examined. A reduction in VIP presence was noted in oestrus and the lowest VIP values occurred one and three days after mating in the ampulla and the isthmus respectively. In the subsequent stages an increase of VIP was noted in both tracts which showed features comparable to that described in anoestrus. The levels and distribution of VIP is correlated with the functions of the oviduct segments concerned with egg transport and formation of a mucopolysaccharide envelope around blastocysts.

A histochemical study has been carried out by light and electron microscopy on the localization of the activity of arylsulphatases A and B in the salivary glands of rat. Results obtained indicate that in the submandibular gland a greater activity is present in the cells of the convoluted granular tubules than in the acina cells, whereas in the sublingual gland the cells of the preterminal tracts appear to be more reactive than those of the terminal tracts. The probable role played by these sulphatases is discussed.

SUMMARY The placenta and the foetal membranes of rats (pregnant 5–21 days) have been assayed by histochemical methods for 3β-hydroxysteroid dehydrogenase (3β-HSDH), NAD-dependent, and 17α- and 17β-hydroxysteroid dehydrogenases (17α- and 17β-HSDH), both NAD and NADP-dependent. 3β-HSDH was limited to the trophoblastic giant cells of the first generation, whereas 17α- and 17β-HSDH were present in both the first- and second-generation giant cells and also in the endodermal cells of the inverted yolk sac placenta.

The effect of cyanoketone on some steps of steroidogenesis has been studied on testicular and adrenal tissue of male Wistar rats. Pregnenolone, dehydroepiandrosterone and progesterone (4-C14) have been used as substrates. The metabolites have been separated by thin layer chromatography and identified by crystallization to constant specific activity. The results have shown that cyanoketone inhibits the activity of 3Β-hydroxysteroid dehydrogenase; 17Α-hydroxylase is not inhibited in adrenal tissue, while in testicular tissue the metabolic pathway pregnenolone →17Α-hydroxypregnenolone – > dehydroepiandrosterone is altered and testosterone biosynthesis is almost completely inhibited.

Some important steps of steroidogenesis have been studied during the breeding season in the testicular tissue of Esox lucius using histochemical and biochemical techniques. Incubations of acetone powder from testicular tissue with several substrates yielded the following metabolites: pregnenolone was converted to dehydroepiandrosterone and progesterone, dehydroepiandrosterone was converted to androstenediol and testosterone, androstenediol was converted to testosterone and dehydroepiandrosterone, and androstenedione was only slightly converted to testosterone. These steroids were identified and quantified through gas-liquid chromatography before and after derivative formation. These findings support the view that androstenediol is an important intermediate in the biosynthesis of testosterone in the testis of Esox lucius. Histochemical reactions for 3β- and 17β-hydroxysteroid dehydrogenases with all the substrates used and for cholesterol were negative for both seminiferous lobules and “lobule boundary cells.”

In Phallusia mamillata and Ascidia malaca the cells containing sulfated acid mucopolysaccharides at the sides of the intestine originate from the A 4.1 blastomeres of stage 8, or, more accurately, from the blastomeres A 6.3, A 6.1, A 6.1, A 6.3, of stage 32.

In the cells of the gastric glands of Salmo irideus, the activity of carbonic anhydrase enzyme, localized at the ultrastructural level, appears in the course of development in the microvilli. In later stages, the reaction is extended to the cytoplasmatic evaginations of the intercellular spaces and to the mitochondrial crystae. The contemporary presence, in the same secreting element, of carbonic anhydrase and of secretion granules of a proteic nature points to the conclusion that these cells are of a "oxyntic-peptic" type.

Binding patterns of lectins conjugated with horseradish peroxidase were studied on rabbit oviduct after mating and during pregnancy. The results of this research demonstrated beyond any doubt that the oviduct in the first days after mating and during the gestation period undergoes enormous changes at both surface and cytoplasmic carbohydrates. All the lectins used displayed modifications in the intensity of their reactivity, but the most interesting information was that concerning PNL, SBL and Con A lectins which showed remarkable differences in the behaviour of glycosidic residues in ampulla and isthmus.

The development of the submandibular gland has been examined in rabbit embryos from the 22nd day to the birth. The 2 main types of secretory cells, which synthetize dense serous granules or large pale mucous granules can be identified from the 24th day of development. A third type of cells, characterized by granules having different densities, has been identified in the preterminal tracts. The possibility that these cells are similar to some small cell with an highly dilated endoplasmic reticulum found in adult rabbit submandibular glands is discussed.

Oviducts play a central role in many reproduction functions. We investigated, by means of biochemical analyses, the chemical composition of glycopeptides extracted from ampulla and isthmus of hare and rabbit. Our results showed that both Lagomorphs present a different qualitative and quantitative composition of the 2 oviductal segments. We conclude that the considerably diverse glycoconjugate composition would be related to marked diversities occurring during the fertilization process, the post-ovulatory phases and the pregnancy length.

The PAS and protein reaction positive secretion can be found in the parotid and inframandibular gland of rabbit on the 22nd pregnancy day only in the perinuclear zone. On the contrary during the last developmental phase it is localized in the apical zone of the cells and after the birth in all the cytoplasm with the exception of the terminal tracts of the inframandibular gland, which are always PAS negative but positive to the reaction to MPS sulfurated acids. Assumptions are made about the presence of the secretion in the gland cells during the fetal period.