Differential cytokine and chemokine expression during rejection and infection following intestinal transplantation.

BACKGROUND/OBJECTIVES: Rejection and infectious enteritis in intestinal transplant (ITx) patients present with virtually identical symptoms. Currently, the gold standard for differentiating between these two conditions is endoscopy, which is invasive and costly. Our primary aim was to identify differences in peripheral blood cytokines during episodes of acute cellular rejection (ACR) and infectious enteritis in patients with intestinal transplants. METHODS: This was a prospective, cross-sectional study involving ITx patients transplanted between 2000 and 2016. We studied 63 blood samples collected from 29 ITx patients during periods of normal (n = 24) and abnormal (n = 17) allograft function. PBMCs from whole blood samples were cultured under unstimulated or stimulated conditions with phytohemagglutinin (PHA). The supernatant from these cultures were collected to measure cytokine and chemokine levels using a 38-plex luminex panel. RESULTS: Our study found that cytokines and chemokines are differentially expressed in normal, ACR, and infectious enteritis samples under unstimulated conditions based on heatmap analysis. Although each cohort displayed distinctive signatures, only MDC (p = 0.037) was found to be significantly different between ACR and infectious enteritis. Upon stimulation of PBMCs, patients with ACR demonstrated increased immune reactivity compared to infectious enteritis; though this did not reach statistical significance. CONCLUSIONS: To our knowledge, this is the first comprehensive study comparing cytokine expression during acute rejection and infectious enteritis in intestinal transplant recipients. Our results suggest that cytokines have the potential to be used as clinical markers for risk stratification and/or diagnosis of ACR and infectious enteritis.

Understanding fragility fracture patients' decision-making process regarding bisphosphonate treatment.

We aimed to understand how patients 50 years and older decided to persist with or stop osteoporosis (OP) treatment. Processes related to persisting with or stopping OP treatments are complex and dynamic. The severity and risks and harms related to untreated clinical OP and the favorable benefit-to-risk profile for OP treatments should be reinforced. INTRODUCTION: Older adults with fragility fracture and clinical OP are at high risk of recurrent fracture, and treatment reduces this risk by 50 %. However, only 20 % of fracture patients are treated for OP and half stop treatment within 1 year. We aimed to understand how older patients with new fractures decided to persist with or stop OP treatment over 1 year. METHODS: We conducted a grounded theory study of patients 50 years and older with upper extremity fracture who started bisphosphonates and then reported persisting with or stopping treatment at 1 year. We used theoretical sampling to identify patients who could inform emerging concepts until data saturation was achieved and analyzed these data using constant comparison. RESULTS: We conducted 21 interviews with 12 patients. Three major themes emerged. First, patients perceived OP was not a serious health condition and considered its impact negligible. Second, persisters and stoppers differed in weighting the risks vs benefits of treatments, where persisters perceived less risk and more benefit. Persisters considered treatment "required" while stoppers often deemed treatment "optional." Third, patients could change treatment status even 1-year post-fracture because they re-evaluated severity and impact of OP vs risks and benefits of treatments over time. CONCLUSIONS: The processes and reasoning related to persisting with or stopping OP treatments post-fracture are complex and dynamic. Our findings suggest two areas of leverage for healthcare providers to reinforce to improve persistence: (1) the severity and risks and harms related to untreated clinical OP and (2) the favorable benefit-to-risk profile for OP treatments.

Successful term pregnancy in an intestine-pancreas transplant recipient with chronic graft dysfunction and parenteral nutrition dependence: a case report.

Pregnancy after solid organ transplantation is becoming more common, with the largest recorded numbers in renal and liver transplant recipients. Intestinal transplantation is relatively new compared to other solid organs, and reports of successful pregnancy are far less frequent. All pregnancies reported to date in intestinal transplant recipients have been in women with stable graft function. The case reported here involves the first reported successful term pregnancy in an intestine-pancreas transplant recipient with chronic graft dysfunction and dependence on both transplant immunosuppression and parenteral nutrition (PN) at the time of conception. Pregnancy was unplanned and unexpected in the setting of chronic illness and menstrual irregularities, discovered incidentally on abdominal ultrasound at approximately 18 weeks' gestation. Rapamune was held, tacrolimus continued, and PN adjusted to maintain consistent weight gain. A healthy female infant was delivered vaginally at term. Medical complications during pregnancy included anemia and need for tunneled catheter replacements. Ascites and edema were improved from baseline, with recurrence of large volume ascites shortly after delivery. Successful pregnancy is possible in the setting of transplant immunosuppression, chronic intestinal graft dysfunction, and long-term PN requirement, but close monitoring is required to ensure the health of mother and child.

Diastereoselective synthesis of propargylic N-hydroxylamines via NHC-copper(I) halide-catalyzed reaction of terminal alkynes with chiral nitrones on water.

The first NHC-copper(I)-halide catalyzed addition of terminal alkynes to enantiomerically pure nitrones on water is described. This reaction provides a straightforward access to propargylic N-hydroxylamines in excellent yield and with excellent stereoselectivity (up to 97%). The presented methodology was applied as a key step in the formal synthesis of (-)-lentiginosine.

A common peripheral blood gene set for diagnosis of operational tolerance in pediatric and adult liver transplantation.

To identify biomarkers of operational tolerance in pediatric and adult liver transplant recipients, transcriptional profiles were examined from 300 samples by microarrays and Q-PCR measurements of blood specimens from pediatric and adult liver transplant recipients and normal tissues. Tolerance-specific genes were validated in independent samples across two different transplant programs and validated by Q-PCR. A minimal set of 13 unique genes, highly expressed in natural killer cells (p = 0.03), were significantly expressed in both pediatric and adult liver tolerance, irrespective of different clinical and demographic confounders. The performance of this gene set by microarray in independent samples was 100% sensitivity and 83% specificity and the AUC was 0.988 for only three genes by Q-PCR. 26% of adults and 64% of children with excellent liver allograft function, on minimal or dual immunosuppression, showed high prediction scores for tolerance. Novel peripheral transcriptional profiles can be identified in operational tolerance in pediatric and adult recipients of liver allografts, suggesting a high incidence of a pro-tolerogenic phenotype in stable patients on chronic immunosuppression. Given the high incidence of viral infections and malignancies in liver transplant recipients, this gene set provides an important monitoring tool that can move the field toward personalized and predictive medicine in organ transplantation.

A review of natural and synthetic antioxidants important for health and longevity.

Reactive oxygen species (ROS) generated in the presence of O(2) by mitochondria, phagocytic cells, peroxisomes, and cytochrome P450 enzymes under physiological conditions, may play a dual function in the human organism. On the one hand, they participate in cell signal transduction cascades, leading to the activation of some transcription factors responsible for regulating of the expression of genes relevant for cell growth and differentiation. On the other hand, they cause oxidative damage of cellular DNA, protein and lipids, resulting in the initiation or development of numerous diseases such as cancer, cardiovascular diseases, type 2 diabetes mellitus, cataract, rheumatoid arthritis, or different neurodegenerative diseases. Both endogenous compounds (glutathione, ubiquinol, urate, bilirubin) and enzymes (superoxide dismutase, catalase, glutathione peroxidase) are engaged in the detoxification of ROS. In addition, numerous dietary components such as vitamin C, vitamin E, carotenoids, and polyphenols are thought to be involved in the antioxidant defense system. The present review article is focused on the summary and the assessment of research on the impact of dietary antioxidants in the prevention of chronic diseases, particularly cancer and cardiovascular diseases.

Physiological and pathophysiological functions of SIRT1.

The human SIRT1 is a nuclear enzyme from the class III histone deacetylases (HDACs) which is widely distributed in mammalian tissues. A variety of SIRT1 substrates hints that this protein is involved in the regulation of diverse biological processes, including cell survival, apoptosis, gluconeogenesis, adipogenesis, lipolysis, stress resistance, muscle differentiation, and insulin secretion. This review emphasizes catalytic properties of SIRT1 and its role in apoptosis, insulin pathway, and neuron survival.

The cardiothoracic ratio in AGA and SGA very low birth weight newborn infants.

BACKGROUND: Cardiothoracic (CT) ratio is a common measurement used to assess heart size in chest radiographs of pediatric patients, but no recent studies have analyzed the standards for CT ratios in very low birth weight (VLBW) infants. OBJECTIVE: The aim of this study was to provide improved standards for CT ratios measured from chest radiographs of VLBW (<1500 g) infants, and to compare CT ratios between small for gestational age (SGA) and appropriate for gestational age (AGA) infants in this population. DESIGN/METHODS: Among VLBW infants admitted to the Jacobi Medical Center NICU from 2002 to 2004, CT ratios were calculated from anteroposterior supine chest radiographs taken of 54 VLBW infants (18 SGA and 36 AGA group-matched on the basis of birthweight and sex) during the first 24 h of life. RESULTS: There were no significant differences between the two groups with respect to birthweight, sex, 1-min Apgar score, 5-min Apgar score, intubation status and degree of inspiration. Median GA of the SGA infants was significantly greater than the AGA infants (30 and 27 weeks, respectively; P<0.001). CT ratios among SGA infants were significantly larger than those among AGAs. Using the widest internal width of the bony thorax, the mean CT ratio among SGA and AGA infants was 0.523 and 0.479, respectively (P=0.00102). CONCLUSIONS: VLBW SGA infants have larger CT ratios than VLBW AGA infants, suggesting that existing standards for normal CT ratios may be inappropriate for use among SGA infants.

Oligomeric building block approach to the synthesis of diastereomerically pure pentathymidine 3',5'-methanephosphonates.

[formula: see text] A method for a large-scale synthesis of stereodefined oligo(nucleoside 3',5'-methanephosphonates) has been developed, based on transient 3'-O protection, which allows for the conversion of the protecting chirally defined methanephosphonanilidate group, located at the 3' end of a stereoregular oligomer, into diastereomerically pure "oligomeric building blocks" for stereospecific coupling with the 5'-OH group of another oligonucleotide.

p53 mutations in sweat gland carcinomas.

Sweat gland carcinomas are rare skin tumours and little is known about their etiology and molecular basis. In this study, we analyzed p53 mutations in 16 sweat gland carcinomas with different histologic types, including 2 spiradenocarcinomas, 1 composed adnexal carcinoma, 5 porocarcinomas, 2 eccrine hidradenocarcinomas, 2 syringocystadenocarcinomas, 1 sclerosing sweat gland carcinoma, 1 adenoid cystic carcinoma, 1 cylindrocarcinoma and 1 apocrine adenocarcinoma. Single-stranded conformation polymorphism (SSCP) analyses followed by direct DNA sequencing revealed that 5 carcinomas (31%) contained a p53 mutation, 4 of which were G:C-->A:T transition mutations and 1 of which was a deletion. Three G:C-->A:T mutations were located at dipyrimidine sequences on the antisense strand (2 spiradenocarcinomas, 1 eccrine hidradenocarcinoma), suggesting that UV light may play a role in the development of sweat gland carcinomas. In 2 spiradenocarcinomas, p53 mutations were present in the carcinoma but not in the adenoma portions, suggesting that p53 mutations may be associated with malignant progression in these rare adnexal tumours.

Potassium peroxymonosulfate (oxone)--an efficient oxidizing agent for phosphothio compounds.

Potassium peroxymonosulfate (oxone) is demonstrated as a versatile chemoselective and stereospecific oxidizing agent for phosphothio compounds. Its application in nucleotide chemistry is presented.

Novel cost-effective methanephosphonoanilidothioate approach to the stereoselective synthesis of dinucleoside (3',5')-methanephosphonates.

A new method of stereoselective preparation of di(2'-deoxy or 2'-OMe)ribonucleoside (3',5')-methanephosphonate 5 is presented. The DBU/LiCl-assisted reaction of 5'-O-DMT-(2'-deoxy or 2'-OMe)ribonucleoside 3'-O-(S-alkyl methanephosphonothioate) 9 with 5'-OH nucleosides proceeds with full stereospecificity, giving 5 in moderate to good yield. The conversion of 5'-O-DMT-(2'-deoxy or 2'-OMe) ribonucleoside 3'-methanephosphonoanilidothioates 8 and 3'-O-methanephosphonoanilidates 10 by means of NaH/CX2 (X = O,S) followed by S-alkylation leads to monomers 9, with the possibility of use of both separated diastereomers of 8 for the preparation of one selected diastereomer of 5. The relative configuration at the P atom in 2'-OMe and deoxynucleoside derivatives of compounds 9 was established by means of stereoselective degradation of nucleoside 3'-O-methanephosphonothioates 11 (precursors of 9) with nuclease P1.

Phenotypic heterogeneity of nodular hidradenoma. Immunohistochemical analysis with emphasis on cytokeratin expression.

We studied the immunohistochemical phenotype in 13 cases of the nodular hidradenoma (NH), with special emphasis on the expression of different types of keratins (cytokeratins, 7, 10, 6/18, 8/18, and 10/17/18 and their distribution in normal sweat glands. Variable reactions with keratins, alpha-smooth muscle actin, and epithelial membrane antigen (EMA) were found, as these markers were present in different cellular components of the tumors. The most constant finding was almost complete absence of cytokeratins (all but keratin 10/17/18, which was positive in two of 13 cases) in clear cells, which yet were positive for EMA. The tumors expressed mostly cytokeratin 6/18, 7, 8/18, and 10/17/18, which were found in 11, 13, 11, and 12 cases, respectively. The cellular distribution and quantity of stained cells differed, as keratins 6/18, 8/18, and 7 produced the most abundant staining and were predominantly localized in small squamoid cells and the cells lining the tubular and cystic spaces. Cytokeratin 10/17/18 was expressed in smaller or larger clusters of squamoid cells and rarely in clear cells. Cytokeratins 10, 19, and 20 were found sporadically in single cells or small cellular clusters. alpha-Smooth muscle actin was expressed in four cases, whereas we did not find reactivity of S-100 protein. Comparing these results with the pattern of keratin distribution and antigenic reactivity in eccrine sweat glands, we conclude that NH presents cellular heterogeneity of its elements and differentiation toward different parts of the sweat gland.

Carcinosarcoma of the skin. Case report and literature review.

A case of carcinosarcoma arising in the skin of the left arm of a 69-year-old woman is reported with a review of the literature. The tumor was composed of low-differentiated squamous cell carcinoma, which was intermingled with a pleomorphic sarcoma. The carcinomatous component had keratin and lacked vimentin, whereas the phenotype of the sarcomatous portion was the reverse. The former presented additionally focal expression of S-100 protein, which was lacking in other portions of the carcinoma. The phenotypic data, supplemented by p53 immunostaining, which was present in both components, suggest their common origin in this tumor.

An unusual case of giant dermatofibroma in a patient with diabetes mellitus and necrobiosis lipoidica.

A case of nine dermatofibromata, including a giant one, associated with necrobiosis lipoidica and diabetes mellitus type II is reported. This case is unusual because of the number and size of the tumors and their association with the above-mentioned pathologic conditions.

Expression of the latent membrane protein of Epstein-Barr virus in Hodgkin's disease. Age and subtype distribution in Polish patients.

We examined immunohistochemically 135 cases of HD (4, lymphocytic predominance (LP); 34, mixed cellularity (MC); 90, nodular sclerosis (NS) and 7, lymphocytic depletion (LD) for the presence of EBV latent membrane protein (LMP). Ten patients were younger than 14 years, 55 were young adults (15-34 years), 45 were 35-49-years old and 26 were older than 50 years. The average LMP-immunopositivity was 33% of all HD cases. The highest proportion of LMP-immunopositivity was found in the MC subtype (61.8%), followed by the NS (NS 1-25%, NS 2-26%) and the LD subtype (14%), but none in the LP subtype of HD. The differences between LMP-immunopositivity in the MC and NS subtypes were statistically significant in children and young adults (p < 0,01). The LMP-positivity was mainly associated with the MC subtype which occurred predominantly in children, while the lowest proportion of LMP-positive cases was found in the young adult group. This report supports a notion that there are regularities in age and subtype distribution of EBV in HD.

p53 expression in sweat gland tumors.

We analyzed the expression of p53 in 74 cutaneous adnexal tumors, with enhancement of the detection by incubation of the slides in the microwave. The immunostaining in benign tumors was almost uniformly negative as we found p53-positivity only in one poroma, one nodular hidradenoma, and one case of syringocystadenoma papilliferum (amongst 13 spiradenomas, 9 cylindromas, 12 nodular hidradenomas, 7 poromas, 6 syringomas, 7 syringocystadenomas papilliferum, 2 papillary tubular adenomas and 4 chondroid syringomas). These results contrasted with the widespread p53 overexpression, which was revealed in the sweat gland carcinomas. All spiradenocarcinomas (3), malignant nodular hidradenoma (1), apocrine hidradenocarcinoma (1), and malignant syringoadenoma (1) showed a strong reaction to anti-p53 antibody. Two of three eccrine hidradenocarcinomas, and two of three porocarcinomas presented p53 overexpression, whereas in one case of malignant cylindroma and adenoid cystic carcinoma we did not find p53-positivity. The results of the study indicate an important role, that p53 protein plays in the malignant sweat gland tumors in comparison to their benign counterparts, but reveal that its overexpression may also occur in the reactive and benign neoplastic processes.

Diagnostic significance of the myoepithelial cells in the benign and malignant sweat gland tumors.

We studied the presence of myoepithelial cells (MCs) in the benign and malignant sweat gland adenomas by immunostaining with antibody to alpha-smooth muscle actin. We found peripheral arrangement of MCs in the neoplastic nests in cutaneous cylindromas (9), spiradenomas (13), syringoadenomas (7), and one case of adenoid cystic carcinoma. Dispersed MCs were identified in some cases of nodular hidradenoma (7/13) and malignant portion of spiradenocarcinoma (2/3). The remnants of the peripheral arrangement of MCs were retained in the benign part in cylindrocarcinoma and every case of spiradenocarcinoma, likewise in composed malignant adnexoma (malignisation within cylindroma/spiradenoma). We could not find MCs in papillary eccrine adenoma (2), poromas (7) and porocarcinomas (3), syringomas (6), chondroid syringomas (3), malignant nodular hidradenoma (1) and malignant portion of syringoadenoma (1). In one of three cases of eccrine hidradenocarcinoma focal myoepithelial differentiation could be identified at the periphery of the epithelial nests. These results confirm heterogeneity of the differentiation in sweat gland tumors, emphasizing the validity of division of those tumors into those with the differentiation towards secretory and ductal portion of the gland. The former group demonstrates the variability of arrangement of myoepithelial cells depending on the degree of dedifferentiation of the developing carcinoma. These cells may be replaced by outgrowing tumor, but may also accompany the neoplastic growth. Less significant is the presence of the MCs in the adenomas with ductal differentiation, which either in benign and malignant tumors may be lacking. The variability of MCs occurrence in cutaneous adenomas and carcinomas precludes its significance as a solitary factor in the differentiation between benign and malignant proliferations.

Over-expression of p53 protein as an indicator of the malignant transformation in spiradenoma.

Malignant spiradenomas (spiradenocarcinomas) are exceedingly rare tumours of cutaneous adnexal origin, consisting of two components: benign--the pre-existent adenoma, and malignant--developing from the former part. We studied p53 protein expression in both compartments of three cases of malignant spiradenoma and compared these results with results obtained with eight cases of spiradenoma. Nuclear staining was consistently negative in all benign tumours, whilst in the cases of malignant transformation within spiradenoma p53 protein was present in the carcinomatous component, but the immunostaining remained negative in the benign counterpart of the tumour. In the zone of transition between both components of the spiradenocarcinomas p53 expression was positive in the cells with morphological atypia, providing clear discrimination. Thus, we conclude that the accumulation of p53 protein, which results from alterations in its turnover, accompanies the process of malignant transformation within long-standing spiradenomas.