Intact Glands Research Articles

Composition and transstadial retention of the salivary glands in Ornithodoros hermsi (Acari: Argasidae).

Following several days of blood feeding by larval and nymphal ixodid (hard) ticks, the salivary glands degenerate and are completely replaced in the next life stage. Yet, what happens during the molt of immature argasid (soft) ticks after their rapid and small bloodmeal has remained a mystery. Multiple studies of nymphal Ornithodoros hermsi Wheeler (Acari: Argasidae) ticks infected with the relapsing fever spirochete Borrelia hermsii suggested the salivary glands in these ticks may not disintegrate after feeding. Therefore, cohorts of second-stage O. hermsi nymphs were fed and examined daily after the bloodmeal by fresh dissections and weekly by histological cross-sections of the entire tick. The composition of the salivary glands was typical for argasid ticks in having agranular (Type I) and granular (Type II) acini, the latter being surrounded by a myo-epithelial sheath. In all 197 ticks examined from 1 to 63 days after feeding, morphologically intact salivary glands were present. During apolysis, 5 ticks had extralimital clusters of granular acini adhering to otherwise intact glands. Our observations demonstrate that the salivary glands of nymphal O. hermsi do not disintegrate after feeding and new acini are produced during the molt for incorporation into the existing glands. Cumulatively, these findings suggest a fundamental difference in the transstadial development of argasid and ixodid ticks.

Slow hydrogel matrix degradation enhances salivary gland mimetic phenotype.

We recently developed a salivary gland tissue mimetic (SGm), comprised of salivary gland cells encapsulated in matrix metalloproteinase (MMP)-degradable poly(ethylene glycol) hydrogels within arrays of ∼320 µm diameter spherical cavities molded in PDMS. The SGm provides a functional and physiologically relevant platform well-suited to high-throughput drug screening for radioprotective compounds. However, the utility of the SGm would benefit from improved retention of acinar cell phenotype and function. We hypothesized that tuning biochemical cues presented within the PEG hydrogel matrix would improve maintenance of acinar cell phenotype and function by mimicking the natural extracellular matrix microenvironment of the intact gland. Hydrogels formed using slower-degrading MMP-sensitive peptide crosslinkers showed >2-fold increase in sphere number formed at 48 h, increased expression of acinar cell markers, and more robust response to calcium stimulation by the secretory agonist, carbachol, with reduced SGm tissue cluster disruption and outgrowth during prolonged culture. The incorporation of adhesive peptides containing RGD or IKVAV improved calcium flux response to secretory agonists at 14 days of culture. Tuning the hydrogel matrix improved cell aggregation, and promoted acinar cell phenotype, and stability of the SGm over 14 days of culture. Furthermore, combining this matrix with optimized media conditions synergistically prolonged the retention of the acinar cell phenotype in SGm. STATEMENT OF SIGNIFICANCE: Salivary gland (SG) dysfunction occurs due to off-target radiation due to head and neck cancer treatments. Progress in understanding gland dysfunction and developing therapeutic strategies for the SG are hampered by the lack of in vitro models, as salivary gland cells rapidly lose critical secretory function within 24 hours in vitro. Herein, we identify properties of poly(ethylene glycol) hydrogel matrices that enhance the secretory phenotype of SG tissue mimetics within the previously-described SG-microbubble tissue chip environment. Combining slow-degrading hydrogels with media conditions optimized for secretory marker expression further enhanced functional secretory response and secretory marker expression.

Assessment of bladder filling during prostate cancer radiation therapy with ultrasound and cone-beam CT.

Prostate cancer patients undergoing external beam radiation therapy (EBRT) benefit from a full bladder to decrease bowel and bladder toxicity. Ultrasound may offer a proxy metric for evaluation, sparing CBCT dosing. Patients were prospectively enrolled pre-simulation from January 2017 to February 2018. Bladder volume was evaluated prior to RT using US daily and CBCT for three daily treatments and then weekly unless otherwise indicated. 29 patients completed median 40 days of RT, resulting in 478 CBCT and 1,099 US bladder volumes. 21 patients were treated to intact glands and 8 to the post-prostatectomy bed. Median patient age was 70 years. Bladder volume on CBCT and US positively correlated (r = 0.85), with average bladder volume for all patients of 162 mL versus 149 mL, respectively. Bladder volume during treatment was consistently lower than the volume at CT simulation (153 mL vs 194 mL, p<0.01) and progressively declined during treatment. Patients older than 70 years presented with lower average bladder volumes than those < 70 years (122 mL vs 208 mL, respectively, p<0.01). Patients with the highest agreement between CBCT and US (<10% variability) had higher average bladder volumes (192 mL vs 120 mL, p=0.01). US was found to be an accurate measure of bladder volume and may be used to monitor daily bladder volumes in patients being treated with radiation for prostate cancer.

The effects of Saline Infusion Sonography on the histological quality of endometrial sampling in women with postmenopausal bleeding

BackgroundThe aim of this study is to analyze the histopathological features of endometrial samples obtained by aspiration when performed before or after the saline contrast sonohysterography in women with postmenopausal bleeding and a thickened endometrium. Hypothetically, the saline infusion could disrupt the tissue and therefore affect the quality of the sample. Furthermore, we want to determine which histological features have impact on the quality of the endometrial sample.MethodsWe performed a randomized controlled trial (ESPRESSO trial) in which we analyzed the aspiration samples in two groups. Women were allocated either to saline contrast sonohysterography and subsequent endometrial sampling (SCSH-Sampling group) or to the opposite order (Sampling-SCSH group). Dedicated gyneco-pathologists retrospectively assessed the specimens and recorded the type (blood, mucus, epithelium, intact glands, stroma and tissue context) and quantity (on a scale of 0–3) of material that was found in the specimens.ResultsThis analysis consisted of 197 samples, with 101 women in the SCSH-Sampling group and 96 women in the Sampling-SCSH group. No significant differences were found in the histological features between the two groups. All significant histological features differed significantly in the sufficient samples compared to the insufficient samples: higher amounts of blood, more endometrial epithelium, presence of intact endometrial glands, better stroma and tissue context. Oppositely, a significantly higher amount of mucus was found in the insufficient samples.ConclusionThis study shows that the histological features of the endometrial sample were not affected by the saline contrast sonohysterography, when performed prior to the tissue sampling.Trial registration ESPRESSO TRIAL, NTR5690, registered 16 February 2016, https://trialsearch.who.int/Trial2.aspx?TrialID=NTR5690.

Encapsulation of Primary Salivary Gland Acinar Cell Clusters and Intercalated Ducts (AIDUCs) within Matrix Metalloproteinase (MMP)-Degradable Hydrogels to Maintain Tissue Structure and Function.

Progress in the development of salivary gland regenerative strategies is limited by poor maintenance of the secretory function of salivary gland cells (SGCs) in vitro. To reduce the precipitous loss of secretory function, a modified approach to isolate intact acinar cell clusters and intercalated ducts (AIDUCs), rather than commonly used single cell suspension, is investigated. This isolation approach yields AIDUCs that maintain many of the cell-cell and cell-matrix interactions of intact glands. Encapsulation of AIDUCs in matrix metalloproteinase (MMP)-degradable PEG hydrogels promotes self-assembly into salivary gland mimetics (SGm) with acinar-like structure. Expression of Mist1, a transcription factor associated with secretory function, is detectable throughout the in vitro culture period up to 14 days. Immunohistochemistry also confirms expression of acinar cell markers (NKCC1, PIP and AQP5), duct cell markers (K7 and K5), and myoepithelial cell markers (SMA). Robust carbachol and ATP-stimulated calcium flux is observed within the SGm for up to 14 days after encapsulation, indicating that secretory function is maintained. Though some acinar-to-ductal metaplasia is observed within SGm, it is reduced compared to previous reports. In conclusion, cell-cell interactions maintained within AIDUCs together with the hydrogel microenvironment may be a promising platform for salivary gland regenerative strategies.

Sampling technique biases in the analysis of fruit fly volatiles: a case study of Queensland fruit fly

Diverse methods have been used to sample insect semiochemicals. Sampling methods can differ in efficiency and affinity and this can introduce significant biases when interpreting biological patterns. We compare common methods used to sample tephritid fruit fly rectal gland volatiles (‘pheromones’), focusing on Queensland fruit fly, Bactrocera tryoni. Solvents of different polarity, n-hexane, dichloromethane and ethanol, were compared using intact and crushed glands. Polydimethylsiloxane, polydimethylsiloxane/divinylbenzene and polyacrylate were compared as adsorbents for solid phase microextraction. Tenax-GR and Porapak Q were compared as adsorbents for dynamic headspace sampling. Along with compounds previously reported for B. tryoni, we detected five previously unreported compounds in males, and three in females. Dichloromethane extracted more amides while there was no significant difference between the three solvents in extraction of spiroacetals except for (E,E)-2,8-dimethyl-1,7-dioxaspiro[5.5]undecane for which n-hexane extracted higher amount than both dichloromethane and ethanol. Ethanol failed to contain many of the more volatile compounds. Crushed rectal gland samples provided higher concentrations of extracted compounds than intact rectal gland samples, but no compounds were missed in intact samples. Of solid phase microextraction fibers, polyacrylate had low affinity for spiroacetals, ethyl isobutyrate and ethyl-2-methylbutanoate. Polydimethylsiloxane was more efficient for spiroacetals while type of fiber did not affect the amounts of amides and esters. In dynamic headspace sampling, Porapak was more efficient for ethyl isobutyrate and spiroacetals, while Tenax was more efficient for other esters and amides, and sampling time was a critical factor. Biases that can be introduced by sampling methods are important considerations when collecting and interpreting insect semiochemical profiles.

H2AX Promoter Demethylation at Specific Sites Plays a Role in STAT5-Induced Tumorigenesis.

Deregulated STAT5 activity in the mammary gland of transgenic mice results in parity-dependent latent tumorigenesis. The trigger for cell transformation was previously associated with hyperactivation of the H2AX proximal promoter in a small basal cell population during pregnancy. The current study focuses on the latent activation of tumor development. H2AX was highly expressed in carcinoma and adenocarcinoma as compared to the multiparous mammary gland, whereas pSTAT5 expression decreased in a tumor type-dependent manner. In contrast to the pregnant gland, no positive correlation between H2AX and pSTAT5 expression could be defined in carcinoma and adenocarcinoma. Using targeted methylation analysis, the methylation profile of the H2AX promoter was characterized in the intact gland and tumors. Average H2AX promoter methylation in the tumors was relatively high (~90%), but did not exceed that of the multiparous gland; 5mC methylation was higher in the differentiated tumors and negatively correlated with its oxidative product 5hmC and H2AX expression. Individual analysis of 25 H2AX promoter-methylation sites revealed two consecutive CpGs at positions -77 and - 54 that were actively demethylated in the multiparous gland, but not in their age-matched virgin counterpart. The different methylation profiles at these sites distinguished tumor types and may assume a prognostic role. In-silico and ChIP analyses revealed overlapping methylation-independent SP1-binding and methylation-dependent p53-binding to these sites. We propose that interference with SP1-assisted p53-binding to these sites abrogates H2AX's ability to arrest the cell cycle upon DNA damage, and contributes to triggering latent development of STAT5-induced tumors in estrapausal multiparous mice.

USE OF 29A MICRO-RNA FOR DIAGNOSIS OF PLEOMORPHIC ADENOMAS OF SALIVARY GLANDS

Introduction. Pleomorphic adenoma is known as the most common tumour in salivary glands that makes up 60-90% of all benign tumours of the salivary glands. The modern genetic tendency towards the diagnosis of salivary gland tumours is the study of the role of microRNA molecules, and miRNA-29a in the focus of the great researchers’ interest. It is expressed in 84 % of the pleomorphic adenomas of the salivary glands. Objectives. Determination of expression of miRNA-29a in tissues of pleomorphic adenomas of the large salivary glands that adjacent to the tumour of the tissue of the salivary gland, intact tissue of the salivary gland, was out of touch with the tumour and venous blood. Materials and methods. The study included 20 patients with benign tumours of the large salivary glands (pleomorphic adenomas). The expression of miRNA-29a was evaluated by using reverse transcription and quantitative polymerase chain reaction (qPCR) in real time.&#x0D; Results. The analysis of the level of miRNA-29a expression revealed that among 4 groups of indicators (tumour tissue, tissues adjacent to the tumour salivary gland tissue, intact salivary gland, which was out of touch with the tumour and venous blood) in patients with pleomorphic adenoma of the large salivary gland, the highest expression was noted in the group, where salivary gland tissue was adjacent to the salivary gland tumour (111, 93±56, 97 versus 8,12±4,4). Correlation analysis of patients with pleomorphic adenoma of the large salivary glands with different fragments of tissues samples demonstrated that the expression level of miRNA-29a differed significantly between the groups (adjacent gland - intact salivary tissue).&#x0D; Conclusions. A sufficiently high level of miRNA-29a expression in the tissues of pleomorphic adenoma in the large salivary glands compared with the normal (intact salivary gland tissue), 10 times as much can be used as a genetic marker for verification (identification) of this type of tumours. Studies of biopsy material from patients with pleomorphic adenoma in the large salivary glands at the genetic level (by expression of miRNAs-29a) confirm the need not only in enucleating of the tumour (partial parotidectomy), but also in performing of subtotal resection with removal of salivary gland adjacent to the tumour.

Application of microRNA-34a for diagnosis of pleomorphic adenomas of salivary glands

Objective. Determination of expression of microRNA-34a in tissues of pleomorphic adenomas of large salivary glands, adjacent to tumor salivary gland tissue, intact tissue of a salivary gland, which was not connected with the tumor, and in a venous blood as well.&#x0D; Materials and methods. The investigation was conducted in 20 patients, suffering benign tumors of large salivary glands (pleomorphic adenomas). Expression was estimated, using adverse transcription and quantitative polymerase chain reaction in regime of a real time.&#x0D; Results. Analysis of the expression level for microRNA-34a was conducted in the tumoral tissue, adjacent to the tumor salivary gland tissue, and in intact tissue of salivary gland, which lacked a link with the tumor, the venous blood in patients, suffering pleomorphic adenomas of large salivary glands, and there was revealed, that it have appeared the highest in the salivary gland adjacent to the tumor - (1052.02 ± 367.20, comparing with the same index in the intact gland - 47.72 ± 28.93).&#x0D; Conclusion. Level of expression of microRNA-34a in the tissues of pleomorphic adenomas of salivary glands, which is in 11 times higher, than in norm (in intact tissue of salivary gland), may be applied as a genetic marker for verification of these tumors.

Epithelium segmentation using deep learning in H&E-stained prostate specimens with immunohistochemistry as reference standard

Given the importance of gland morphology in grading prostate cancer (PCa), automatically differentiating between epithelium and other tissues is an important prerequisite for the development of automated methods for detecting PCa. We propose a new deep learning method to segment epithelial tissue in digitised hematoxylin and eosin (H&E) stained prostatectomy slides using immunohistochemistry (IHC) as reference standard. We used IHC to create a precise and objective ground truth compared to manual outlining on H&E slides, especially in areas with high-grade PCa. 102 tissue sections were stained with H&E and subsequently restained with P63 and CK8/18 IHC markers to highlight epithelial structures. Afterwards each pair was co-registered. First, we trained a U-Net to segment epithelial structures in IHC using a subset of the IHC slides that were preprocessed with color deconvolution. Second, this network was applied to the remaining slides to create the reference standard used to train a second U-Net on H&E. Our system accurately segmented both intact glands and individual tumour epithelial cells. The generalisation capacity of our system is shown using an independent external dataset from a different centre. We envision this segmentation as the first part of a fully automated prostate cancer grading pipeline.

DIAGNOSIS OF ENDOCRINE DISEASE: Congenital hypothyroidism: update and perspectives

Congenital hypothyroidism (CH) may be primary, due to a defect affecting the thyroid gland itself, or central, due to impaired thyroid-stimulating hormone (TSH)-mediated stimulation of the thyroid gland as a result of hypothalamic or pituitary pathology. Primary CH is the most common neonatal endocrine disorder, traditionally subdivided into thyroid dysgenesis (TD), referring to a spectrum of thyroid developmental abnormalities, and dyshormonogenesis, where a defective molecular pathway for thyroid hormonogenesis results in failure of hormone production by a structurally intact gland. Delayed treatment of neonatal hypothyroidism may result in profound neurodevelopmental delay; therefore, CH is screened for in developed countries to facilitate prompt diagnosis. Central congenital hypothyroidism (CCH) is a rarer entity which may occur in isolation, or (more frequently) in association with additional pituitary hormone deficits. CCH is most commonly defined biochemically by failure of appropriate TSH elevation despite subnormal thyroid hormone levels and will therefore evade diagnosis in primary, TSH-based CH-screening programmes. This review will discuss recent genetic aetiological advances in CH and summarize epidemiological data and clinical diagnostic challenges, focussing on primary CH and isolated CCH.

Рентгенофлуоресцентный анализ в диагностике рака щитовидной железы

Purpose: To investigate new possibilities of differential diagnosis of benign and malignant thyroid goiter lesions by means of energy dispersive X-ray fluorescence analysis (EDRFA).&#x0D; Material and methods: In the samples of thyroid tissue taken from people with intact thyroid gland (mostly died from trauma, n = 92), as well as in patients with nodular goiter (n = 79) and thyroid cancer (n = 40) bromine (Br), copper (Cu), iron (Fe), iodine (I), rubidium (Rb) and strontium (Sr) were investigated. To determine these elements, the methods of EDRPA have been developed using encapsulated sources with 109Cd and 241Am radionuclides for fluorescence excitation.&#x0D; Results: It is shown that the levels of the content of I, the ratios I / Cu and I / Rb, as well as the products of the ratios I / Cu ∙ (I / Rb) and (I / Br) ∙ (I / Cu) ∙ (I / Rb ) are highly informative markers of thyroid cancer. The accuracy of the developed methods and the reliability of the results obtained were confirmed by measurements of international certified comparison materials.&#x0D; Conclusion: The use of the proposed markers allows in vitro to differentiat thyroid cancer from benign nodes and normal tissue with sensitivity in the range of 86–100 %, specificity of 89–99 %, and accuracy within 90–99 %.

Abstract P276: (Pro)Renin Receptor-dependent Renal Generation of Aldosterone Contributes to ENaC Activation During Angiotensin II-induced Hypertension

(Pro)renin receptor (PRR) has emerged as a novel regulator of ENaC under various physiological and pathophysiological conditions including Ang II-induced hypertension. However, the mechanism for ENaC-activating action of PRR remains incompletely understood. The present study attempted to test renal-derived aldosterone (Aldo) as a potential mediator of PRR signaling in the setting of Ang II treatment. In normal Sprague-Dawley rats with intact adrenal glands, a 7-d minipump infusion of Ang II at 100 ng/kg/min induced a 3.2-fold increase in urinary Aldo excretion and a 1.8-fold increase in plasma Aldo, as assessed by ELISA. Following intramedullary delivery of a PRR decoy peptide PRO20 (120 μg/kg/d) (IM PRO20), Ang II-induced increases in urinary Aldo was reduced by 75% without affecting plasma Aldo, in parallel with attenuated hypertension (radiotelemetry-measured MAP on day 7: 132.6 ± 5.8 mmHg in Ang II+IMPRO20 vs. 157.1 ± 6.2 mmHg in Ang II). Following adrenalectomy (ADX), urinary Aldo was decreased from 9.3 ± 1.2 ng/24h to an unbeatable level under basal condition; it was elevated to 0.96 ± 0.05 ng/24h by Ang II and became undetectable again by IM PRO20, as assessed by mass spectrometry. Similar results were obtained by ELISA. In parallel, urinary Na+/K+ ratio in ADX rats was decreased by Ang II infusion and blunted by IM PRO20. Furthermore, Ang II infusion induced a 1.3-fold increase in urinary Aldo excretion and this increase was reduced by 60% in CD PRR KO mice, correlating to the changes in MAP. In cultured mpkCCD cells exposed to 10 nM prorenin, amiloride-sensitive transepithelial Na+ transport, measured by using epithelial volt-ohmmeter, was increased 2.1-fold at 6 h that sustained at 24 h, which was suppressed by 86% following treatment with the mineralocorticoid receptor antagonist eplerenone. Medium Aldo was increased by 2.6-fold following exposure to 10 nM prorenin for 24 h. Overall, these results suggest that activation of PRR results in local generation of Aldo that likely contributes to increased ENaC activity and thus hypertension during Ang II treatment.

Case report: ACTH-secreting pituitary carcinoma metastatic to the liver in a patient with a history of atypical pituitary adenoma and Cushing\u2019s disease

BackgroundPituitary carcinoma is a rare entity requiring the presence of metastasis to confirm its malignant potential. We report a case of pituitary carcinoma and discuss the diagnosis and management of this lesion in relation to the existing literature.Case presentationThe patient is a 51-year-old woman with Cushing’s disease and intact adrenal glands who was diagnosed with metastatic pituitary carcinoma to the liver, 29 months after initial resection of an ACTH-secreting primary atypical pituitary adenoma (APA). Prior to detection of this metastasis the patient underwent repeat resection and radiotherapy for residual cavernous sinus disease. The metastatic lesion was detected by interval surveillance of serum ACTH and 24-hour urine cortisol, which despite stable pituitary MRI, were significantly elevated. These abnormalities prompted a PET scan that demonstrated hypermetabolic liver parenchyma, which was suspicious for metastasis on abdominal MRI. An ultrasound-guided liver biopsy demonstrated nests of moderately-differentiated cells with intermediate-sized, monotonous nuclei, distinct nucleoli, and abundant basophilic cytoplasm, confirmed by immunohistochemistry to represent metastatic pituitary carcinoma. The liver lesion was subsequently successfully removed by wedge resection. One year later, the patient’s residual cavernous sinus disease grew markedly, and she was placed on dual-agent chemotherapy consisting of oral temozolomide and capecitabine, with stabilization of her intracranial disease to present, although liver metastases recurred.ConclusionsPituitary carcinoma is a rare entity impossible to recognize as a primary tumor because its diagnosis by definition requires the presence of metastasis. Maintaining awareness of the entity and its precursor lesion APA is essential for its accurate pathologic diagnosis and appropriate management.

Uncovering the stem cell hierarchy by genetic lineage tracing in the mammary gland

AbstractThe mammary gland is the distinct feature that gives the name to the class of mammals and distinguishes them from other animals. Functionally, the mammary gland is a secretory organ which main role is to produce milk to nourish the offspring. Organogenesis of the mammary gland starts during embryogenesis but occurs mainly after birth at puberty under the influence of hormonal cues. Throughout the adult life as well as during pregnancy, the mammary gland shows a remarkable regenerative ability, thus constituting an excellent model for studying stem cell biology. Although the mammary gland consists of a relatively simple epithelial structure with a luminal and a basal cell layers, these are indeed composed by distinct subsets of mammary epithelial cells. Flow cytometry and transplantation assay have identified several subpopulations of stem and/or progenitor cells in the mammary gland. Yet, physiological and developmental relevant information can only be obtained when investigating the stem cell hierarchy in the intact mammary gland. Genetic lineage tracing studies have offered unprecedented levels of information regarding the organization of the stem cell compartment and possible role of resident stem and/or progenitor cells at different stages of the mammary gland organogenesis. These studies, although creating a passionate debate, highlight the existence of heterogeneous stem cell compartment, where bipotent as well as unipotent mammary stem cells seems to co-exist. Genetic lineage tracing experiments provide relevant information on stem cells that are key for understanding both normal development as well as associated pathologies in human. It holds the promise of providing new insights into the cell-of-origin and heterogeneity of breast tumorigenesis.

GABAergic signaling in the rat pineal gland.

Pinealocytes secrete melatonin at night in response to norepinephrine released from sympathetic nerve terminals in the pineal gland. The gland also contains many other neurotransmitters whose cellular disposition, activity, and relevance to pineal function are not understood. Here, we clarify sources and demonstrate cellular actions of the neurotransmitter γ-aminobutyric acid (GABA) using Western blotting and immunohistochemistry of the gland and electrical recording from pinealocytes. GABAergic cells and nerve fibers, defined as containing GABA and the synthetic GAD67, were identified. The cells represent a subset of interstitial cells while the nerve fibers were distinct from the sympathetic innervation. The GABAA receptor subunit α1 was visualized in close proximity of both GABAergic and sympathetic nerve fibers as well as fine extensions among pinealocytes and blood vessels. The GABAB 1 receptor subunit was localized in the interstitial compartment but not in pinealocytes. Electrophysiology of isolated pinealocytes revealed that GABA and muscimol elicit strong inward chloride currents sensitive to bicuculline and picrotoxin, clear evidence for functional GABAA receptors on the surface membrane. Applications of elevated potassium solution or the neurotransmitter acetylcholine depolarized the pinealocyte membrane potential enough to open voltage-gated Ca(2+) channels leading to intracellular calcium elevations. GABA repolarized the membrane and shut off such calcium rises. In 48-72-h cultured intact glands, GABA application neither triggered melatonin secretion by itself nor affected norepinephrine-induced secretion. Thus, strong elements of GABA signaling are present in pineal glands that make large electrical responses in pinealocytes, but physiological roles need to be found.

A Renal congenital Anomaly - unilateral extracalyceal Ectopic Right Kidney With Multiple Aberrant Renal Vessels

 Routine dissection of the abdomen of a 63 year old male cadaver in the Dept. of Anatomy Hi- tech medical college Rourkela., revealed the presence of a rudimentary ectopic right kidney fused to the lower pole of the left, with multiple aberrant renal arteries .The ureters were normal, the left kidney (upper) had a normal pelvis while the right (lower) had an extrarenal pelvis (extra calyx). The two ureters entered the bladder normally, Right side showed empty renal fossa with an intact adrenal gland. The possible embryological causes of this anomaly have been discussed here. Even though this type of condition can be presents as a silent renal anomaly (it can be undiagnosed throughout life and revealed through autopsy or dissection) the surgical implications are noteworthy,which too have been highlighted in this report.

Abstract B22: Analysis of Lgr5-positive cells in mouse mammary gland

Abstract Lgr5 has been identified as a stem cell marker in multiple adult tissues, and individual Lgr5+ mammary epithelial cells are capable of regenerating mouse mammary glands in transplantation assays. Here we investigated the proliferation and differentiation capabilities of murine mammary Lgr5+ cells in vitro, and their distribution in mouse mammary glands in vivo. Equal numbers of sorted Lgr5+ and Lgr5- epithelial cells were cultured in Matrigel to evaluate their stem-like properties in vitro. Compared with Lgr5- cells, Lgr5+ cells were more efficient at generating organoids in 3D culture, and the Lgr5+ cell-derived organoids grew to a larger size. Moreover, Lgr5+ cell-derived organoids survived repeated passaging and contained cells of both luminal and basal lineages, as indicated by keratin marker expression. Incidence of Lgr5+ cells in organoids was confirmed by in situ hybridization. These studies indicate that Lgr5+ cells retain the self-renewal and differentiation capabilities of stem cells ex vivo. The in vivo distribution of Lgr5+ cells was investigated using X-Gal-stained mammary glands from Lgr5-lacZ reporter mice ranging from 5 to 32 weeks old. In both adolescent and adult mice, we observed a graded distribution of Lgr5+ cells with greater abundance in the nipple-proximal (ventral) ducts than in the lymph-node proximal central region, and fewest Lgr5+ cells in the distal (dorsal) region of the glands. Lgr5+ cells from all mammary gland regions appeared capable of yielding organoids in culture. These results corroborate in organoid culture the known stem-like properties of Lgr5+ cells defined in intact mouse mammary gland. [Supported by NIH grant CA186981 and the China Scholarship Council] Citation Format: Lixing Zhang, Louise Howe, Richard Kolesnick, Anthony Brown. Analysis of Lgr5-positive cells in mouse mammary gland. [abstract]. In: Proceedings of the AACR Special Conference: Developmental Biology and Cancer; Nov 30-Dec 3, 2015; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Res 2016;14(4_Suppl):Abstract nr B22.

A Case Report of Dramatically Increased Thyroglobulin after Lymph Node Biopsy in Thyroid Carcinoma after Total Thyroidectomy and Radioiodine.

Thyroglobulin (Tg) is an important modality for monitoring patients with thyroid cancers, especially after thyroidectomy followed by radioiodine (RAI). It is also used as a marker for burden of thyroid tissue whether malignant or benign. Although there have been several reports of rising serum Tg transiently after thyroid biopsy in intact glands and following palpation or trauma, there are no reports in the literature of elevation in Tg after biopsy of suspicious lesions in thyroidectomized patients. In this paper we report a fascinating case of a considerable and initially worrying, although ultimately transient, rise in Tg in a patient 2 years after total thyroidectomy and RAI ablation after fine needle aspiration (FNA) of a suspicious thyroid bed nodule that was proven positive.

RARE-15ACTH-SECRETING PITUITARY CARCINOMA METASTATIC TO THE LIVER IN A PATIENT WITH A HISTORY OF ATYPICAL PITUITARY ADENOMA AND CUSHING'S SYNDROME

Pituitary carcinoma is a rare entity requiring the presence of metastasis to confirm its malignant potential. Most examples secrete prolactin or adrenocorticotropic hormone (ACTH). Reported sites of metastasis include to the cerebral cortex, cerebellum, spinal cord, leptomeninges, eyes, heart, lung, lymph nodes, pancreas, liver, kidney, ovary, myometrium and bone. Liver metastasis is most common with prolactin-secreting tumors. In ACTH-secreting tumors it is frequently associated with Nelson's syndrome (rapid enlargement of a pituitary adenoma after bilateral adrenalectomy to palliate Cushing's syndrome). Here we report a case of an ACTH-secreting atypical pituitary adenoma metastasizing to the liver in a 51-year-old woman with Cushing's syndrome and intact adrenal glands 2.5 years after resection of the primary pituitary lesion with subsequent radiotherapy for residual cavernous sinus disease. Surveillance of ACTH and 24 hour urine cortisol levels revealed that they were significantly elevated to 142 pg/mL (normal range 10-60 pg/mL) and 541 µg/day (normal range 3.5-45 µg/day), respectively. These abnormalities prompted a PET scan which demonstrated a hypermetabolic liver lesion with a maximal SUV of 3.3, which was suspicious for metastasis on MRI. An ultrasound-guided liver biopsy demonstrated involvement by a moderately differentiated carcinoma consistent with a primary pituitary lesion. Nests of tumor cells showed a monotonous epithelioid appearance, abundant cytoplasm and small nucleoli. Immunostains for AE1/3, synaptophysin, and ACTH diffusely highlighted the lesional cells. This case thus represents an additional rare example of an ACTH-producing pituitary carcinoma metastatic to the liver in a patient with Cushing's syndrome and intact adrenal glands.