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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.

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.

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.

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.

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.

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.

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 the salivary glands of young rats in different experimental conditions, we have found glycoconjugates containing hexosamines, hexoses, fucose and sialic acid in lower percentages than those found in adult rats. In the submandibular gland, the presence of sialic acid can be demonstrated only after the second week, whereas in the sublingual gland sialic acid is present in an amount roughly equal to half of what has been found in adult animals. The appearance of sialic acid in the submandibular gland is due to the development of the animals and not, as we had initially surmised to their diet.

The secretory granules of the terminal and pre-terminal tracts of the rabbit submandibular gland display distinct features, while the cell morphology is quite similar. Among these cells, some smaller ones presenting an highly dilated endoplasmic reticulum have been identified in the pre-terminal tracts. Some hypothesis have been formulated concerning the function of these cells.

A histochemical study was effected, by light and electron microscopy, on the activity of arylsulphatases A and B in the submandibular and sublingual glands of male and female rabbits, distinguishing, among the latter, between the physiological estrus and anestrus condition. The results obtained demonstrated that the enzymatic activity is more intense in the sublingual gland and that the estrus condition causes an increase in this activity. No correlation was found between the topochemical distribution of the soluble arylsulphatases and that of sulphates.

Control of postoperative pain is one of the most important concerns for both the patients and the surgical team. In this regard the efficacy of wound infiltration with local analgesia and the most proper drugs to be used are not settled. We conducted our study trying to investigate this point in a prospective randomized double blinded manner.With inclusion criteria of: age (18-65 y), volume90 mL, first time, no lymph node enlargement and exclusion criteria of: duration90 min, incision length10 cm, neck dissection decided during surgery and a postoperative complication within the first 24 hours. Sixty patients planned for total thyroidectomy under general anesthesia were enrolled in the study in three groups with twenty patients assigned randomly to each one: group (A) a control group in which no wound infiltration was done, group (B) in which preoperative wound infiltration with 10 mL bupivacaine 0.5% was done, and group (C) in which preoperative wound infiltration with 10 mL ropivacaine 0.75% was done. Postoperative pain was evaluated by Visual Analogue Score (VAS) with a scale of (0-10) at 1 hour, 4 hours, 8 hours and 16 hours.The postoperative pain experienced by all the patients reached a maximum point at 1 hour postoperatively then started to decrease to be minimal at 8 hours and almost negligible at 16 hours. Ropivacaine group showed a statistically significant decrease in pain perception at 1 hour postoperatively (P=0.028), bupivacaine group showed also a decrease in pain perception at 1 hour but it was not statistically significant. At 4 hours of operation and after; neither ropivacaine nor bupivacaine showed an effect on pain perception.The benefit of local wound infiltration with local analgesia in decreasing postoperative pain is limited to a short period after surgery in which the use of ropivacaine 0.75% is recommended over pubivaccaine 0.5%.