Expression of Pancreatic Endocrine Markers by Mesenchymal Stem Cells From Human Umbilical Cord Vein

Background

Mesenchymal stem cells (MSCs) from human umbilical cord vein have great potential for use in cell therapy because of their ease of isolation, expansion, and differentiation, in addition to their relative acceptance from the ethical point of view. Obtaining the umbilical cord at birth does not present any risk to either mother or child.

Objective

To isolate and promote in vitro expansion and differentiation of MSCs from human umbilical cord vein into cells with a pancreatic endocrine phenotype.

Methods

Mesenchymal stem cells obtained from human umbilical cord vein via collagenase digestion were characterized at cytochemistry and fluorescent-activated cell sorting, and expanded in vitro. Differentiation of MSCs into an endocrine phenotype was induced using high-glucose (23 mmol/L) medium containing nicotinamide, exendin-4, and 2-mercaptoethanol. Expression of insulin, somatostatin, glucagon, and pancreatic and duodenal homeobox 1 was analyzed using immunofluorescence.

Results

Cells isolated from the umbilical cord vein were MSCs as confirmed at cytochemistry and fluorescent-activated cell sorting. Expression of somatostatin, glucagon, and pancreatic and duodenal homeobox 1 by differentiated cells was demonstrated using immunofluorescence. Insulin was not expressed.

Conclusions

The MSC differentiation protocol used in the present study induced expression of some endocrine markers. Insulin was not produced by these cells, probably because of incomplete induction of differentiation.     

Clinical characteristics and outcomes of renal transplantation in elderly recipients

Background

Elderly renal transplant candidates constitute one the fastest-growing populations among end-stage renal disease patients. Since the impacts of advanced recipient age have not yet been fully defined, we evaluated the clinical characteristics and outcomes of elderly renal transplant recipients.

Methods

Among 564 adult renal transplant recipients, at our center between 2000 and 2009, 64 were at least 60 years of age (Elderly group), and 500 were younger than 60 years (Young group) at the time of the procedure. We compared their clinical features and surgical management.

Results

There were significant differences in mean donor age (55.6 years vs. 53.2 years, P = .030) and gender mismatch (77.0% vs. 63.4%, P = .035). However, there were no significant differences between the two groups in patient and graft survivals (P = .177 and P = .365, respectively). Malignancy after transplantation was a significant risk factor upon univariate evaluation but only ABO incompatibility upon multivariate analysis of patient and graft survival. The main cause of graft loss among the Elderly group was death with a functioning graft due to heart failure.

Conclusions

Renal transplantation is a feasible, safe option for the elderly and should be actively implemented. However, screening for cancer and heart disease should be mandatory to improve outcomes.     

A novel Czech kindred with familial medullary thyroid carcinoma and Hirschsprung's disease

Purpose

The RET proto-oncogene is involved in neural crest disorders. Activating germline mutations in the RET proto-oncogene cause the development of familial medullary thyroid carcinoma (FMTC) or medullary thyroid carcinoma (MTC) as a part of multiple endocrine neoplasia type 2 (MEN2) syndrome. Inactivating germline mutations in the RET proto-oncogene are detected in Hirschsprung's disease (HSCR). Only in a very small number of families are these 2 diseases expressed together.

Methods

This study presents a novel Czech kindred with FMTC-HSCR phenotype. Two family members (mother and daughter) were tested for RET germline mutations in exons 10, 11, 13, 14, 15, and 16.

Results

Direct fluorescent sequencing of genomic DNA revealed a heterozygous mutation in the RET proto-oncogene in exon 10 at codon C609Y in both persons tested. This family was reclassified, thanks to genetic screening from the apparently sporadic MTC-HSCR to FMTC-HSCR.

Conclusion

The germline mutation was detected because of the systematic genetic screening of the RET proto-oncogene, which is useful for genetic counseling of potential risk of HSCR and MTC in other family members. This family could be added to the small worldwide cohort of families with MEN2A/FMTC-HSCR.     

Effect of systemic corticosteroids on elective left-sided colorectal resection with colorectal anastomosis

Background

The impact of systemic steroid therapy on surgical outcome after elective left-sided colorectal resection with rectal anastomosis is not well known.

Methods

We compared 606 consecutive patients including 53 patients who were on steroids and undergoing surgery between 1995 and 2005.

Results

Postoperative mortality and anastomotic leakage rates were equivalent. The postoperative complications rate, especially infections, was higher in steroid-treated patients than in non-steroid-treated patients: 38% (20 of 53 patients) versus 25% (139 of 553 patients), respectively (P = .046). In the steroid group, univariate analysis revealed 3 significant risk factors for postoperative complications: blood transfusion, preoperative anticoagulation, and chronic respiratory failure. In a multivariate analysis, blood transfusion and chronic respiratory failure remained independent factors for postoperative complications.

Conclusion

Patients on steroids have a higher incidence of postoperative complications after elective left-sided colorectal resection with rectal anastomosis.     

Hepato-pancreato-biliary training in general surgery residency: is it enough for the real world?

Background

Advanced training in hepato-pancreato-biliary (HPB) surgery is available at select centers. No approved fellowships have yet been established.

Objective

To determine the level of training in HPB surgery during general surgery residency and to assess the need for additional training.

Method

All general surgical residency programs in the United States were surveyed. Resident Review Committee (RRC) and International Hepato-Pancreato-Biliary Association (IHPBA) requirements were compared to Accreditation Council of Graduate Medical Education (ACGME) data.

Results

Eighty of 250 general surgical residency programs (32%) responded to the survey. Eighty percent felt their graduating residents had sufficient HPB training. The average number of pancreatic cases per graduating resident was 10.2 ± 7.3. The average number of hepatic resections was 8.6 ± 5.1, and for complex biliary cases, 5.3 ± 1.3.

Conclusions

A significant portion of HPB surgery is performed at transplant centers or by HPB surgeons. Guidelines must be established to assure adequate training. When HPB surgery is the main focus of the future practice, residents should seek additional training.     

Bipolar radiofrequency tonsillotomy compared with traditional cold dissection tonsillectomy in adults with recurrent tonsillitis

Objective

To compare short- and long-term results of radiofrequency tonsillotomy and traditional cold dissection tonsillectomy in adult patients with recurrent tonsillitis.

Study Design

A randomized clinical trial.

Setting

A tertiary referral university hospital.

Subjects and Methods

Of 62 adults with recurrent tonsillitis, 24 patients were treated with cold dissection tonsillectomy, while radiofrequency tonsillotomy (RF) by use of radiofrequency induced thermotherapy probes was performed in the remaining 38 patients. Duration of surgery, amount of intraoperative bleeding, recovery time, postsurgical pain (measured by use of visual analogue scale on days 1, 3, 5, and 10 after surgery), dysphagia, weight loss, and time of return to normal diet and activity were measured. All the patients were followed for 12 to 24 months for recurrence of tonsillitis episodes.

Results

Comparing the radiofrequency tonsillotomy group to the cold dissection tonsillectomy group, mean duration of surgery was 16.89 versus 45.04 minutes, recovery time was 14.32 minutes versus 17.08 minutes, and amount of intraoperative bleeding was 15 to 20 cc versus 250 to 300 cc, respectively (P < 0.005). There was no difference between the two groups in the recurrence of tonsillitis episodes after 24 months.

Conclusion

Tonsillotomy with radiofrequency is a simple, rapid, and effective method in adult patients with recurrent tonsillitis.     

The effect of extent of local resection on patterns of disease progression in malignant pleural mesothelioma

Background

We sought to determine whether or not there are differences in disease progression after radical or nonradical (debulking) surgical procedures for malignant pleural mesothelioma.

Methods

Over a 49-month period, 132 patients with malignant pleural mesothelioma underwent surgery. Fifty-three underwent extrapleural pneumonectomy and 79 underwent nonradical procedures. Time to evidence of clinical disease progression was recorded, as was the site(s) of that disease.

Results

One-hundred nineteen patients were evaluable, of which 59% (22 radical; 48 nonradical) had disease progression. Overall 30-day mortality was 8.5% (7.5% radical; 9% nonradical). The median time to overall disease progression was considerably longer after extrapleural pneumonectomy than debulking surgery (319 days vs 197 days, p = 0.019), as was the time to local disease progression (631 days vs 218 days, p = 0.0018). There was no preponderance of earlier stage disease in the radical surgery group. There was a trend toward prolonged survival in those undergoing radical surgery, but no significant difference between the groups (497 days vs 324 days, p = 0.079). In those who had extrapleural pneumonectomy, time-to-disease progression significantly decreased with N2 disease compared with N0/1 involvement (197 days vs 358 days, p = 0.02).

Conclusions

Extrapleural pneumonectomy may be preferable to debulking surgery in malignant pleural mesothelioma to delay disease progression and give greater control of local disease. Involvement of N2 nodes is associated with accelerated disease progression and is therefore a contraindication to extrapleural pneumonectomy.     

Preincisional local infiltration of tramadol at the trocar site versus intravenous tramadol for pain control after laparoscopic cholecystectomy

Study Objective

To compare the effects of preoperative intravenous (IV) tramadol and preoperative tramadol infiltration of trocar sites on postoperative pain and postoperative nausea and vomiting (PONV) after laparoscopic cholecystectomy.

Design

Prospective, randomized study.

Setting

Operating room, recovery room, and surgical ward.

Patients

70 ASA physical status 1 and 2 patients, aged 20-70 years, scheduled for elective laparoscopic cholecystectomy.

Interventions

In Group I, patients received IV 2.0 mg/kg of tramadol; in Group II, trocar insertion points were infiltrated with 2.0 mg/kg of tramadol in 20 mL of 0.9% NaCl.

Measurements

Pain scores, sedation scores, postoperative analgesic requirement, and PONV were recorded at 0 and 30 minutes and one, three, 6, 12, and 24 hours. At 30 minutes and one hour, pain localization (incisional or diffuse abdominal) was also recorded.

Main Results

Visual analog scale scores at 30 minutes were significantly lower in Group II [3 (0-7)] than Group I [6 (3-8)] (P < 0.001). In Group I, 91.4% of patients received sodium diclofenac, while 68.6% of Group II patients received sodium diclofenac (P = 0.002). The time to first analgesic requirement was significantly lower in Group II (P = 0.004). At the 30-minute measurement time, a significant difference was recorded between the groups in incisional pain (P < 0.001). There was also a significant difference between groups in the frequency of PONV.

Conclusions

Trocar site infiltration of tramadol improves early postoperative pain and decreases PONV.     

Detection of Potential Organ Donors: 2-Year Analysis of Deaths at a German University Hospital

Objective

In 2006, inhouse coordinators were introduced in all hospitals with intensive care units in Baden-Württemberg to improve organ donation. At our university hospital with a neurosurgery and a transplantation unit, we analyzed whether brain death certification and donation requests were always initiated (if possible).

Materials and Methods

We retrospectively reviewed all 1312 hospital deaths from 2006-2007 by studying medical records and consulting with physicians. The possibility of organ donation was questioned.

Results

Donation was requested among 68/702 deaths due to cerebral complications. A request was impossible in 8 cases. Consent for donation was obtained in 29 cases, and it was realized in 24 cases. In 14 cases of resuscitation from shock or cardiac failure, therapy was not continued because of questionable hemodynamic stability. In 17 cases admitted to peripheral wards and 19 to intensive care units death due to cerebral complications occurred within 48 hours, but medical records were not relevant for exclusion criteria for organ donation.

Conclusions

The detection of donors was not acceptable. In patients without a prognosis after resuscitation, further hemodynamic stabilization was frequently omitted, because organ donation had not been considered. In cases of donation requests the refusal rate was high. We initiated specific training.     

Comparison of bacteria isolated from urinary tract infections in patients on transplant versus urologic wards

Objective

Urinary tract infections (UTI) are the most common hospital infections. Complications include sepsis and shock. Immunosuppressed transplant surgery patients may experience loss of the graft due to UTI. The purpose of this study was to determine the main microorganisms responsible for UTIs among patients of transplant wards compared to urologic wards. Additionally, drug susceptibility profiles of the most frequent microorganisms were analyzed.

Materials and Methods

We analyzed the results of positive urine cultures from patients on 2 transplant versus 1 urologic ward in 2010.

Results

The most common pathogen in urine samples from all 3 wards was Escherichia coli. Often, other Gram-negative bacilli of the genus Klebsiella spp, were cultured as well as Gram-positive cocci (Enterococcus spp). Yeasts of the genus Candida were only found in urine of patients of transplant wards. The percentage of resistant bacteria was much higher among bacteria from transplant patients.

Conclusions

The high level of antimicrobial resistance of microorganisms isolated from the urine of transplant patients and the relatively high incidence of fungal infections, demand an especially quick, accurate microbiological diagnosis for this group of patients.     

Mitochondria-initiated apoptosis triggered by oxidative injury play a role in total parenteral nutrition-associated liver dysfunction in infant rabbit model

Purpose

The aim of the study was to investigate oxidative injury and apoptosis as the mechanisms underlying total parenteral nutrition (TPN)-associated liver dysfunction.

Methods

Twenty New Zealand rabbits (2 weeks old) were divided into 2 groups as follows: 10 in the control group (maternal feed) and 10 in the TPN group. The rabbits in the TPN group received continuous PN infusion through a silastic catheter inserted in the right jugular vein.

Results

After 10 days of treatment, the serum levels of total bilirubin and bile acid were significantly higher in the TPN group than in the control group (P < .01, respectively). The light microscopic findings in the TPN rabbits included inflammatory cell infiltration and hepatic steatosis. Electron microscopy showed change in the cytosolic vacuoles and rare microvilli in the microbile duct. Moreover, 10 days of treatment resulted in an inhibition of the superoxide dismutase (SOD) activity in hepatocytes, an increase of the malondialdehyde level, a significant increase in cytochrome c release from the mitochondria, a significant increase in caspase 3 activity, and increased apoptosis (P < .01, individually).

Conclusions

Oxidative damage may be one of the essential mechanisms of TPN-associated liver dysfunction. Moreover, mitochondria-initiated apoptosis triggered by oxidative damage may play an important role in this process.     

Adverse events after radiofrequency ablation of unresectable liver tumors: a single-center experience

Background

Radiofrequency ablation (RFA) is a relatively new modality to treat liver tumors that is being incorporated into practice despite the fact that its risk profile has not been well described.

Methods

A retrospective cohort study, using structured chart review, on patients with liver tumor(s) was conducted from August 1998 to November 2006. Univariate and multivariate exploratory analyses were used to evaluate factors associated with adverse events.

Results

RFA procedures were performed on 196 patients (58% primary tumors, 24% colorectal metastases, and 18% other metastases). Twenty-three patients (12%) experienced serious adverse events. Multivariate analysis showed advanced age (≥55 y), underlying liver disease, large tumor size (>4 cm), and concomitant procedure were associated with an increased risk of adverse events (P = .01, P < .01, P = .01, and P = .01, respectively). There were no in-hospital deaths.

Conclusions

RFA was associated with acceptable morbidity and mortality. Factors associated with adverse events should be considered when counseling patients regarding RFA procedures.     

Human epidermal growth factor receptor signaling contributes to tumor growth via angiogenesis in her2/neu-expressing experimental Wilms' tumor

Background:

The human epidermal growth factor family (HER) members play a significant role in the mesenchymal-to-epithelial transition during renal tubulogenesis. HER misexpression has been linked also to loss of growth control, invasiveness, and promotion of angiogenesis in breast cancers and other human malignant tumors

Methods:

The authors screened Wilms’ tumor samples and derived cell lines for expression of her2/neu, which was detected in both unfavorable and favorable histology tissues. Xenografts were implanted in mice using her2/neu(+) and her2/neu(−) cell lines and the effect of specific blockade tested using monoclonal anti-her2/neu antibody.

Results:

Blocking antibody suppressed tumor growth in her2/neu(+) but not her2/neu(−) experimental Wilms’ tumor. In addition, antibody exposure resulted in suppression of tumor angiogenesis but no decrease in tumor cell proliferation in her2/neu(+) xenografts.

Conclusions:

Her2/neu contributes to the growth of some Wilms’ tumors, and an important mechanism of its action is promotion of angiogenesis.     

Effects of bone marrow stromal cells and umbilical cord blood-derived stromal cells on daunorubicin-resistant residual Jurkat cells

Objective

To observe the effects of the hematopoietic inductive microenvironment (HIM) simulated by stromal cells of different origins on daunorubicin-resistant residual Jurkat cells (Jurkat/DNR cells).

Methods

Jurkat/DNR cells were cultured and identified. Human umbilical cord blood-derived stromal cells (UCBDSCs) and normal human bone marrow stromal cells (BMSCs) were isolated and cocultured with Jurkat/DNR cells. Jurkat/DNR cells were collected after 14 days of coculture and analyzed with regard to cell proliferation and differentiation abilities, apoptosis, drug sensitivity, and MRD1 multidrug resistance gene mRNA expression.

Results

UCBDSC-simulated HIM suppressed proliferation and promoted apoptosis, differentiation, and drug sensitivity of Jurkat/DNR cells more significantly than BMSC-simulated HIM.

Conclusions

Both BMSCs and UCBDSCs reconstruct the leukemic HIM and reverse drug resistance in Jurkat/DNR cells. UCBDSCs reconstruct the leukemic HIM and reverse drug resistance more significantly than BMSCs.     

Hepatocellular Carcinoma and Liver Transplantation: A 12-Year Experience

Background

Orthotopic liver transplantation (OLT) for patients with cirrhosis and concomitant hepatocellular carcinoma (HCC) in early stages is the treatment of choice, with an acceptable recurrence rate and excellent survival.

Aim

We sought to evaluate (1) the accuracy of preoperative imaging; (2) the impact of pre-OLT treatments on survival and recurrence; and (3) the influence of beyond Milan criteria selection on global outcomes.

Methods

We studied a cohort of 65 patients with HCC among 300 consecutive OLTs over a single 12-year experience. We analyzed the overall outcomes of survival and recurrence, the accuracy of preoperative diagnosis and staging the influence of neoadjuvant treatment prior to OLT, and the effect on overall outcomes beyond the Milan criteria in our series.

Results

The 65 transplants were performed for HCC, mostly in association with hepatitis C virus and alcoholic cirrhosis with HTP. At a mean follow-up of 40.32 months, the recurrence rate was 5.7% among the 61 HCC confirmed by histopathology. The overall survival was 30.07. Actuarial survivals at 1, 5, and 10 years were 82%, 77%, and 62%, respectively. Six retransplants occurred among the seven graft losses albeit with poor survival after the second graft. Most explants showed low pTNM stages with favorable microscopic features. Preoperative imaging tests failed to achieve an accurate diagnosis in 15.38% of the series. The role of alpha-fetoprotein (AFP) and hepatic biopsy was irrelevant. Unfavorable histopathologic factors predicted a greater recurrence rate, but had no influence on survival. Neither recurrence nor survival were modified by pre-OLT therapy.

Conclusions

In our series, AFP, hepatic biopsy, and pre-OLT treatment had limited roles. Radiological imaging techniques underestimated HCC staging and lead to a misdiagnosis to an expected degree. Despite these findings, this single institution experience with OLT for HCC showed excellent survivals with a low recurrence rate including cases of patients beyond the Milan criteria.     

Late hormonal function after testicular torsion

Introduction

Testicular torsion may be an important cause of male infertility. We aimed to investigate the late hormonal function in patients with testicular ischemia/reperfusion injury of the testis after orchidectomy or detorsion.

Methods

Twenty patients (mean age, 13.6 years) were prospectively evaluated at a mean of 5 years after testicular torsion. The serum follicle-stimulating hormone, luteinizing hormone (before and after gonadotropin-releasing hormone stimulation), testosterone, and inhibin B were measured. Fifteen age-matched adolescents without evidence of endocrine disease were used as controls for inhibin B values. Data are quoted as mean ± SEM.

Results

Twelve patients were treated with detorsion and orchidopexy, and 8 underwent orchidectomy. Serum follicle-stimulating hormone, luteinizing hormone, and testosterone were all within the reference range. Inhibin B levels were significantly reduced in the 2 groups compared with the controls (34.5 ± 5.2 vs 63.9 ± 12.8 pg/mL, P = .02), but were not significantly different between the orchidectomy group and the group that underwent detorsion (41.3 ± 9.7 vs 30.4 ± 5.9 pg/mL, P = .41).

Conclusion

Hormonal testicular function can be compromised after testicular torsion, although the type of surgery (orchidectomy or orchidopexy) does not seem to change the effect of this ischemia/reperfusion injury.     

Different pattern of matrix metalloproteinases expression in alveolar versus embryonal rhabdomyosarcoma

Background/purpose

The matrix metalloproteinases (MMPs), a family of enzymes that degrade the extracellular matrix (ECM), are important in neoplastic cell invasion and metastasis. Data for rhabdomyosarcoma (RMS), the most frequent soft tissue sarcoma of childhood, are lacking. The aim of this study was to assess their expression in this tumor and to evaluate the correlation with clinicopathologic parameters.

Methods

Immunohistochemical expression of MMP-1, MMP-2, MMP-3, MMP-7, MMP-9, TIMP-1, and TIMP-2 was investigated in 33 human RMSs, 12 alveolar, and 21 embryonal histologic subtypes (12 high risk and 9 low/standard risk). Evaluation of the results was based on the percent of positive neoplastic cells and on staining intensity (negative, moderate, and strong). In situ zymography was carried out on 4 frozen RMS samples (2 alveolar and 2 high-risk embryonal).

Results

Alveolar type showed a stronger MMP-1, -2 and -9 expression in comparison with embryonal (P = .006, P <.001, and P <.001, respectively). Intratumoral vessels and perivascular ECM were positive for MMP-9 in the majority of RMSs. Both TIMPs had negative results.

Conclusions

Gelatinases MMP-2 and MMP-9 and collagenase MMP-1 overexpression seem to contribute to the more aggressive phenotype of alveolar rhabdomyoblastic cells. Further characterization of the expression of MMPs and consequent utilization of their inhibitors in aggressive alveolar RMSs might lead to the development of novel anticancer therapies.     

Antenatal diagnosis of bowel dilatation in gastroschisis is predictive of poor postnatal outcome

Purpose

Although gastroschisis infants usually have a good outcome, there remains a cohort of babies who fare poorly. We inquired whether the presence of bowel dilatation in utero is predictive of postnatal course in infants with gastroschisis.

Methods

We compared the clinical course of infants who had bowel dilatation with those who did not. Bowel dilatation was defined as more than 20 mm in cross-sectional diameter on ultrasound at any gestational age. Outcome measures used were length of time of parenteral nutrition, death, and surgery for intestinal failure.

Results

A review of 170 infants with gastroschisis identified 74 who had dilatation of more than 20 mm (43.5%). There was no significant difference in the incidence of intestinal atresia in those with bowel dilatation and those without (P = .07). Those with bowel dilatation spent a longer period on parenteral nutrition. There were significantly more deaths in the group with bowel dilatation (P = .01). There was no significant difference in the number of infants requiring surgery for intestinal failure between the 2 groups (P = .47).

Conclusions

We found that sonographically detected bowel dilatation more than 20 mm in utero in fetuses with gastroschisis may have value in predicting clinically significant adverse postnatal outcomes.