Cytoreductive surgery and hyperthermic intraperitoneal chemotherapy combined with liver resection for concurrent peritoneal and hepatic metastases of gastrointestinal and gynecological primary tumors.

AIM OF THE STUDY: Cytoreductive surgery including liver resection and hyperthermic intraperitoneal chemotherapy provide survival benefit to selected patients but is associated with relevant morbidity and mortality rates. We aimed to report morbidity and mortality rates and parameters linked to increased morbidity. PATIENTS AND METHODS: Retrospective analysis of 37 patients who underwent liver resection and cytoreductive surgery plus hyperthermic intraperitoneal chemotherapy between 2006 and 2016. From a prospectively collected database the morbidity and mortality rates and survival data were analyzed. RESULTS: The mortality rate was 0% and grade III-IV morbidity was 42%. Re-operation rate was 27%. Patients with complications tended to have a higher peritoneal cancer index (16 vs. 13; P=0.23). The performance of rectal resections was statistically significantly associated with morbidity (P=0.02). Neither performance of other type of resections nor the hyperthermic intraperitoneal chemotherapy compound nor the completeness of cytoreduction score was associated with elevated morbidity. No complications related to liver resections were observed. Furthermore, origin of peritoneal metastases did not impact on occurrence of complications. Median overall survival for colorectal primaries was 22 months (range, 9-60 months) and 30 months (range, 12-58 months) for ovarian cancer. CONCLUSION: Simultaneous resection of hepatic and peritoneal metastases seems to provide a survival benefit for selected patients and is associated with acceptable morbidity and mortality rates. Knowledge of patients and operative factors linked to morbidity will help to provide a strict selection process and a safer surgical procedure.

[Oligometastases in gastric and esophageal cancer : Current clinical trials and surgical concepts]. / Oligometastasierung beim Magen- und Ösophaguskarzinom : Aktuelle Studienlage und chirurgische Konzepte.

There is an increasing role of surgery even in metastatic tumor stages. In colorectal cancer patients with limited metastases, complete cytoreduction is able to offer long-term survival in synchronic as well as metachronic situations. In tumors of the upper gastrointestinal tract (UGIT), however, the impact of surgery in metastatic disease remains to be clarified. Even though several retrospective studies suggest prolonged survival, surgery is not recommended by the current German S3 guidelines for tumors of the UGIT. Appropriate selection of these particular patients that show less aggressive tumor biology will be the primary goal of preoperative evaluation. The results of the prospective randomized RENAISSANCE/FLOT-5 and GASTRIPEC studies are urgently awaited in order to gain more evidence on the impact of surgery in the management of oligometastases of the UGIT and especially on survival.

Abdominal Wall Morbidity Following Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy.

BACKGROUND: Incisional hernia formation has been reported as high as 20% within 1 year following midline laparotomy. Since hyperthermic intraperitoneal chemotherapy is likely to impair wound healing, we sought to investigate the incidence of incisional hernia formation and abdominal wall rupture following cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. METHODS: Consecutive patients with radiographic evidence of peritoneal metastases were scheduled for cytoreductive surgery and hyperthermic intraperitoneal chemotherapy at the Comprehensive Cancer Center, University Hospital Tuebingen, Germany. Clinical data were retrospectively analyzed. RESULTS: Between May 2005 and May 2014, 271 patients underwent cytoreductive surgery and hyperthermic intraperitoneal chemotherapy. Within follow-up, 19 (7%) incisional hernias and 11 (4%) abdominal wall ruptures were observed. Age ⩾70 years, cardio-pulmonary comorbidity, the presence of pseudomyxoma peritonei or mesothelioma, and postoperative abdominal wall rupture were detected as risk factors for hernia formation. However, Cox multivariate analysis only confirmed the presence of pseudomyxoma peritonei or mesothelioma and postoperative abdominal wall rupture as independent risk factors. CONCLUSION: Our data do not suggest that cytoreductive surgery and hyperthermic intraperitoneal chemotherapy is necessarily associated with a higher incidence of incisional hernia formation. However, patients suffering from pseudomyxoma peritonei or mesothelioma and patients with postoperative abdominal wall rupture seem to be at risk for developing incisional herniation.

Pancreas-preserving surgical management of grade-C pancreatic fistulas after pancreatic head resection by external wirsungostomy.

PURPOSE: Completion pancreatectomy for grade-C pancreatic fistula is associated with unacceptably high mortality and therefore this strategy should be reassessed. This study presents an update of our experience with a pancreas-preserving technique in the course of salvage re-laparotomy in terms of closure of the open jejunum via segmental resection and external drainage of the pancreas. METHODS: Between April 2004 and January 2015, 292 pancreaticoduodenectomies (PD) with pancreaticojejunostomy (PJ) were performed. Thirteen patients (5 %) underwent salvage re-laparotomy for symptomatic grade-C fistulas, and clinical data were retrospectively analyzed. RESULTS: In all patients, the preservation of the pancreas remnant and external drainage of the pancreatic juice was feasible. Median hospital stay was 58 days (range, 21-142 days). In 4/13 patients (31 %), further reoperations were necessary. In-hospital mortality was 15 % (2/13). 3/13 patients (23 %) were readmitted and two received inpatient non-surgical treatment. To date re-pancreaticojejunostomy was performed in seven of the remaining 11 patients (63 %) after 168 days in median. In 1/7 patients (14 %), a re-operation after re-PJ was necessary. In one patient, externalization of the pancreas juice was chosen as a definite option. In another patient, secretion ceased spontaneously without stasis and normal endocrine function. Neither before nor after re-anastomosis impairment of endocrine function was observed. CONCLUSIONS: Closure of the intestinum and preservation of the pancreas remnant in grade-C pancreatic fistula is easy to perform and can be categorized as a life-saving procedure. Prevention of total pancreatectomy associated with high morbidity and mortality was achieved in all cases.

[Surgical Management of Peritoneal Surface Malignancy with Respect to Tumour Type, Tumour Stage and Individual Tumour Biology]. / Chirurgische Therapie der peritonealen Metastasierung in Abhängigkeit von Tumorentität, -stadium und -charakteristik(a).

Peritoneal tumour dissemination is still considered as a terminal disease. For the last two decades, cytoreductive surgery (CRS) combined with intraoperative hyperthermic chemotherapy (HIPEC) has been popularised by Paul Sugarbaker almost doubling survival in selected patients compared with systemic chemotherapy alone. Nowadays, this particular treatment protocol is available in comprehensive cancer centres with reasonable mortality and morbidity. However, patient selection is still challenging. In general, CRS and HIPEC is indicated in primary peritoneal tumours such as mesothelioma and pseudomyxoma peritonei as well as in peritoneal metastases derived from gastrointestinal malignancies and ovarian cancers. Since systemic tumour spread is uncommon in patients with peritoneal metastases, peritoneal tumour dissemination was defined as localised disease within the "compartment abdomen". However, CRS and HIPEC are only beneficial as long as complete cytoreduction is achieved (CC-0 or CC-1). Histopathological parameters, the Sugarbaker peritoneal carcinomatosis index (PCI) and general condition of the patient have been established as patient selection criteria. In primary peritoneal cancers, individual tumour biology is the predominant criterium for patient selection as opposed to intraabdominal tumour load in peritoneal metastases derived from gastrointestinal cancers. In gastric cancer, CRS and HIPEC should be restricted to synchronous limited disease because of its biological aggressiveness. In patients with free floating cancer cells without macroscopic signs of peritoneal spread, however, CRS and HIPEC following preoperative "neoadjuvant" chemotherapy preserves chances for cure. So far, there is no general recommendation for CRS and HIPEC by clinical practice guidelines. In the recent S3 guideline for treatment of colorectal cancer, however, CRS and HIPEC have been included as possible treatment options.

[Recurrent Peritoneal Metastases after Cytoreductive Surgery and Hyperthermic Intraperitoneal Chemotherapy (HIPEC) in Colorectal Cancer]. / Rezidive nach zytoreduktiver Chirurgie und hyperthermer intraperitonealer Chemotherapie (HIPEC) beim kolorektalen Karzinom.

BACKGROUND: Peritoneal recurrences after cytoreductive surgery and hyperthermic intraperitoneal chemotherapy (HIPEC) in colorectal cancer are challenges in conservative and surgical oncology. METHODS: Surgical concepts are discussed in this review on the basis of the current literature. RESULTS: The majority of patients suffer from recurrent disease after cytoreductive surgery and hyperthermic intraperitoneal chemotherapy in colorectal cancer. In most cases, a palliative chemotherapy is performed. Repeated surgery with the aim of complete cytoreduction may be done in selected cases. CONCLUSIONS: New concepts are necessary to treat patients with recurrent disease as early as possible.

[Cytoreductive surgery and HIPEC for peritoneal malignancies in children]. / Zytoreduktive Chirurgie und HIPEC bei peritoneal-metastasierten Tumoren im Kindesalter.

Peritoneal tumour dissemination represents an advanced tumour stage and survival rates are usually low. In the past, cytoreductive surgery (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) has been established in adults leading to increased survival rates in comparison to chemotherapy alone. CRS and HIPEC are indicated in primary peritoneal tumours such as mesothelioma and pseudomyxoma peritonei, and in peritoneal metastases from gastrointestinal and ovarian cancers in adults. The incidence of peritoneal surface malignancies in children seems to be lower than in adults, but the incidence is unknown. Nevertheless, peritoneal carcinomatosis/sarcomatosis may occur in patients suffering from desmoplastic small round cell tumour (DSRCT), soft tissue sarcoma (rhabdomyosarcoma, leiomyosarcoma, GIST or liposarcoma), as well as in patients with gastrointestinal cancers. CRS and HIPEC have been established as a novel treatment option in children suffering from peritoneal carcinomatosis/sarcomatosis in very few centres worldwide. This paper reviews the indications, treatment regimens, and pitfalls of this approach in children.

Assessment of relapse in patients with peritoneal carcinomatosis after cytoreductive surgery and hyperthermic intraperitoneal chemotherapy using F-18-FDG-PET/CT.

PURPOSE: In patients with peritoneal carcinomatosis (PC), cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy (HIPEC) is an evolving therapeutic approach with curative intention. The differentiation between posttherapeutic findings after HIPEC and relapse of PC is challenging. We evaluated the diagnostic value of F-18-FDG-PET/CT in patients with relapse of PC after HIPEC. MATERIALS AND METHODS: 36 patients with recurring PC after HIPEC were examined on a whole-body PET/CT system (44 examinations). The examination included 3 D F-18-FDG-PET and contrast-enhanced CT. Images were assessed by two experienced readers regarding the presence and the extent of PC using the peritoneal carcinomatosis index (PCI). Imaging results were correlated with surgical findings or follow-up. RESULTS: Relapse was suspected in 40 of 44 examinations. Relapse was missed by F-18-FDG PET/CT in 4 patients and significantly underestimated in 8 patients. The diagnostic accuracy for the detection of PC on a patient basis was 91 %, the sensitivity was 91 % and the positive predictive value was 100 %. The mean PCI was 11.4 ±â€Š11.9 for PET/CT, 8.4 ±â€Š10.3 for CT and 16.6 ±â€Š15.0 in the case of surgical exploration. The extent of PC was underestimated by PET/CT and even more by CT alone (p < 0.05). CONCLUSION: The diagnostic value of F-18-FDG PET/CT after cytoreductive surgery and HIPEC in the detection of recurring PC is superior to contrast-enhanced CT. However, the quantification of the extent of PC is limited due to post-therapeutic tissue alterations. KEY POINTS: • Imaging of recurrent PC after HIPEC is challenging due to posttherapeutic tissue alterations.• The extent of recurrent PC after HIPEC is systematically underestimated by F-18-FDG PET/CT.• F-18-FDG PET/CT provides improved sensitivity for recurrent PC compared to contrast-enhanced CT.• The correlation of the extent of recurrent PC depicted by F-18-FDG PET/CT and surgical exploration is better than that of contrast-enhanced CT and surgical exploration.

Correlation of preoperative magnetic resonance imaging of peritoneal carcinomatosis and clinical outcome after peritonectomy and HIPEC after 3 years of follow-up: preliminary results.

PURPOSE: In patients with peritoneal carcinomatosis, cytoreductive surgery (CRS) and hyperthermic intraperitoneal chemotherapy (HIPEC) is an evolving approach with curative intention. Previous studies indicate a correlation between preoperative magnetic resonance imaging (MRI) and surgical findings regarding the extent of peritoneal carcinomatosis. The aim of this study was to assess retrospectively whether preoperative MRI can predict the outcome and is therefore a suitable tool for patient selection. MATERIALS AND METHODS: Fifteen patients with laparoscopically proven peritoneal carcinomatosis were preoperatively examined using a 1.5-T whole-body MRI system. Results were correlated with surgical exploration. Follow-up was done by contrast-enhanced abdominal computed tomography and, if suspicious for recurring disease, laparoscopy or laparotomy. Survival time and interval to recurring disease were correlated with the preoperative peritoneal carcinomatosis index (PCI) on MRI (Spearman's rank correlation). RESULTS: In five patients radical resection could not be achieved (PCI 34 ± 6.9); survival time was 78.2 ± 54.1 days. In seven patients recurring disease was found 430 ± 261.2 days after initial complete cytoreduction (PCI 11.6 ± 6.9); survival time was 765.9 ± 355 days. Two patients are still alive after 3 years. Two patients with initially complete cytoreduction are without recurring disease after 3 years (PCI 5 and 12). One patient was lost for follow-up. CONCLUSIONS: Results of the preoperative MRI correlate well with the surgical PCI, postoperative resection status, and survival time. MRI might be a suitable technique for patient selection when considering peritonectomy and HIPEC. In our patients the outcome seems to correlate well with the extent of peritoneal carcinomatosis found by the preoperative MRI.

Factor XIII Val34Leu polymorphism is associated with increased factor XIII activation and decreased transcutaneous oxygen readings in patients with diabetic foot ulcers.

AIM: To investigate the relationship between the factor XIII Val34Leu polymorphism, plasma factor XIII activation and transcutaneous oxygen readings in patients with diabetic foot ulcers. Methods Ninety-two consecutive patients with Type diabetes and active foot ulcers were investigated. Plasma factor XIII activation was assessed by ELISA. Genetic polymorphism was detected using the ABI PRISM® SNaPshotTM Multiplex Kit. Results are expressed as median (minimum-maximum). Differences between groups were calculated by Mann-Whitney U-test or χ(2) -test where appropriate. A P-value < .05 was considered significant. RESULTS: e Val34Leu polymorphism was found in 50 subjects (54%) with 42 being heterozygous (Val/Leu) and eight being homozygous (Leu/Leu). The rate of plasma factor XIII activation was increased in patients with Val/Leu or Leu/Leu [Val/Leu or Leu/Leu 152 (55-283) % vs. Val/Val 103 (33-282) %; P < 0.0001]. Transcutaneous oxygen readings were lower in the Val/Leu or Leu/Leu group [Val/Leu or Leu/Leu 16 (0-58) mmHg vs. Val/Val 35 (1-65) mmHg; P = 0.008]. Similarly, plasma factor XIII activation negatively correlated with transcutaneous oxygen readings (r(2) = -0.314; P = 0.014). CONCLUSIONS: Val34Leu polymorphism is associated with increased plasma factor XIII activation and seems to be linked with impaired cutaneous microcirculation in patients with diabetic foot ulcers.

[Peritoneal carcinomatosis of colorectal cancer: cytoreductive surgery and hyperthermic intraperitoneal chemotherapy]. / Peritoneal Metastasiertes Kolorektales Karzinom: Zytoreduktive Chirurgie und Hypertherme Intraperitoneale Chemotherapie.

Peritoneal carcinomatosis caused by colorectal carcinoma is still considered as the end-stage of disease. A multi-modal therapeutic concept including maximal cytoreduction followed by intraperitoneal hyperthermic chemotherapy (HIPEC) has the potential to cure selected patients. In case of peritoneal carcinomatosis palliative systemic treatment is no longer the state of the art. This article addresses aspects of the disease, the rationale behind peritonectomy with HIPEC, and the surgical management of peritoneal carcinomatosis.

Rapamycin-induced impaired wound healing is associated with compromised tissue lactate accumulation and extracellular matrix remodeling.

INTRODUCTION: Extracellular matrix (ECM) remodeling involving matrix metalloproteinases (MMPs) and wound lactate accumulation are essential elements of tissue repair. The aim of this study was to investigate whether rapamycin-induced impaired healing is associated with compromised wound fluid lactate accumulation and altered ECM remodeling. METHODS: Polyvinyl alcohol sponges were subcutaneously implanted in male C57/BL6 mice. Animals were randomized to daily intraperitoneal treatment with either vehicle or 1.5 mg/kg rapamycin. After 7 or 14 days, sponges were harvested to collect wound fluid for subsequent analyses. Wounds were excised for assessment of tensile strength. RESULTS: After 7 days, wound hydroxyproline content was significantly decreased due to rapamycin therapy, whereas the observed difference in tensile strength marginally failed to show statistical significance. In addition, rapamycin reduced wound lactate accumulation and enhanced MMP-2 protein expression, and both MMP-2 and MMP-9 activity. At day 14, wound tensile strength and hydroxyproline content were significantly lower along with an increase in MMP-2 and MMP-9 activity in rapamycin-treated mice. Similarly, wound fluid lactate concentration and MMP-2 protein expression were found to be persistently decreased and increased, respectively. CONCLUSIONS: Rapamycin affects tissue repair by interfering with fundamental mechanisms involved in healing, namely lactate accumulation and ECM remodeling.

[Peritoneal carcinomatosis]. / Peritonealkarzinose.

Cytoreductive surgery followed by hyperthermic intraperitoneal chemotherapy (HIPEC) is now an established therapy with a curative option for patients with gastrointestinal and gynecological peritoneal carcinomatosis as well as for primary peritoneal carcinomatous tumors. Decisive for the prognosis is a complete cytoreduction, which in most cases necessitates multi-organ resection in addition to a partial or subtotal parietal peritonectomy (PE). The highest priority is given to maintain an adequate quality of life for the patient while performing maximum tumor resection. The morbidity following PE and HIPEC in experienced centers lies between 25% and 35% with a mortality risk of <5%. Consideration must be given not only to the technical surgical aspects and the intraoperative decision-making but also to the intraoperative management, intensive care therapy, pain therapy, management of complications, physiotherapy and many more. The greatest challenge in the management of peritoneal carcinomatosis is still patient selection. Computed tomography imaging together with (18)fluorodeoxyglucose positron emission tomography (FDG-PET) plays an important role in the assessment of operability.

Wound fluid lactate concentration: a helpful marker for diagnosing soft-tissue infection in diabetic foot ulcers? Preliminary findings.

AIMS: To investigate the impact of wound fluid lactate concentration on diagnosing soft-tissue infection in diabetic foot ulcers. METHODS: Lactate concentration in wound fluid obtained from diabetic foot ulcers was determined using a lactate analyser and compared with clinical examination findings. RESULTS: Overall median wound fluid lactate concentration was 21.03 mm (5.58-80.40 mm). Wound lactate levels were significantly higher in infected compared with non-infected diabetic foot ulcers (P=0.001). Non-infected diabetic foot ulcers that healed within 6 months of treatment showed a significantly lower wound fluid lactate concentration at baseline as opposed to those that did not heal (P=0.007). CONCLUSIONS: Non-healing diabetic foot ulcers are characterized by high wound fluid lactate levels. Assessment of wound fluid lactate concentration might be helpful for confirming the suspicion of soft tissue infection, particularly when clinical signs are atypical.

Haemodialysis is associated with changes in cutaneous microcirculation in diabetes mellitus.

AIMS: To examine the cutaneous microcirculation on the dorsum of the foot before, during and after haemodialysis in diabetic and non-diabetic patients. METHODS: Fourteen age-matched patients (seven diabetic, seven non-diabetic) without active foot ulceration were studied. Cutaneous microcirculation was assessed using a micro-lightguide spectrophotometer to measure venous oxygen saturation and relative blood flow determined at two tissue depths: 2 and 6 mm. Cumulative relative changes of each parameter during haemodialysis were calculated as area under the curve. Differences between and within the groups were calculated by Mann-Whitney U-test and anova following post hoc testing, respectively. RESULTS: At baseline, relative blood flow at 6 mm tissue depth was significantly greater in diabetic patients (P = 0.048). Thirty minutes after the end of dialysis, relative blood flow at 2 and 6 mm tissue depth was significantly higher in non-diabetic patients (P = 0.048 and P = 0.001). Mean cumulative relative changes in venous oxygen saturation and relative blood flow at 2 mm as well as 6 mm tissue depth were positive for non-diabetic subjects and negative for diabetic patients. CONCLUSIONS: Haemodialysis is associated with changes in cutaneous microcirculation, which differ between people with and without diabetes. In those without diabetes, we found an increase in blood flow during haemodialysis, whereas blood flow was reduced in diabetic patients. This may be the result of abnormal vasomotor regulation due to distal neuropathy.

[Selection criteria for peritonectomy with hyperthermic intraoperative chemotherapy (HIPEC) in peritoneal carcinomatosis]. / Selektion zur Peritonektomie mit hyperthermer intraoperativer Chemotherapie (HIPEC) bei Peritonealkarzinose.

BACKGROUND: Cytoreductive peritonectomy with hyperthermic intraoperative chemotherapy (HIPEC) is an established therapy for patients with gastrointestinal, gynaecological metastasised peritoneal carcinomatosis as well as primary peritoneal carcinomatous tumours. METHODS: On the basis of a literature review and our personal experience, selection criteria for peritonectomy are discussed. RESULTS: Computed tomography (CT) scans and diagnostic laparoscopy are not sufficient for the diagnosis of peritoneal carcinomatosis. The combination of fluorodeoxyglucose positron emission tomography (FDG-PET) and CT seems to be the most reliable diagnostic imaging method. In our institution, all patients undergo PET / CT prior to peritonectomy. CONCLUSION: The PET / CT scan may play an important role in forecasting the operability of patients with peritoneal carcinomatosis.

[The physiology of wound healing]. / Die Physiologie der Wundheilung.

Wound healing is a complex biological phenomenon. A variety of cellular and biochemical events take place designed to achieve tissue integrity following injury. Even though hypoxia caused by the damaged microvasculature is an important initial stimulus of the healing cascade, adequate tissue perfusion and oxygenation is an absolute pre-requisite for a successful repair since essential wound healing mechanisms such as collagen deposition and bactericidal defence are oxygen dependent reactions. Based upon these findings, there are several ways to overcome the obstacle of tissue hypoxia in clinical practice. Supplemental oxygen is capable of increasing tissue oxygen tension. Following surgery, pain, cold and to little fluids are the main issues of vasoconstriction, impaired tissue perfusion and oxygenation as well. However, all these parameters must be corrected at the same time because any one is sufficient to cause maximal vasoconstriction. A well hydrated, pain free and warm patient should be the main goal of peri- and postoperative surgical care.

Increased translocator protein (TSPO) mRNA levels in colon but not in rectum carcinoma.

BACKGROUND: The peripheral-type benzodiazepine receptor or translocator protein (TSPO) is an 18-kDa protein involved in cell proliferation and apoptosis. TSPO was shown to be overexpressed in malignant tumors and cancer cell lines, correlating with enhanced malignant behavior. The present study analyzed the role of TSPO in patients with colorectal carcinomas. METHODS: Tumor tissues and corresponding normal mucosa from 55 patients who underwent resection for colorectal carcinomas were analyzed for TSPO expression in correlation to GAPDH expression(glyceraldehyde-3-phosphate dehydrogenase) using a multiplex RT-PCR assay. RESULTS: TSPO was overexpressed in 67% of the tumors in comparison to corresponding normal mucosa, and positivity as well as expression levels in colon carcinomas were significantly higher than in the rectum carcinomas. In contrast, TSPO expression was not different in intermediate versus high-grade tumors or in lymph node-positive versus -negative patients. CONCLUSION: The differences in TSPO expression between colon and rectum carcinoma may imply that these tumors are of different biological behavior.

Lactate down-regulates cellular poly(ADP-ribose) formation in cultured human skin fibroblasts.

BACKGROUND: Polyadenosine diphosphate-ribose (poly(ADP-ribose)) is a nuclear polymer which is derived from nicotinamide adenine dinucleotide (NAD(+)) catalysed by poly(ADP-ribose) polymerase 1 (PARP-1). Aside from the well known role of poly(ADP-ribosyl)ation (pADPR) in DNA repair, pADPR is also involved in other cellular processes such as apoptosis and gene expression. However, the factors that regulate the level of pADPR are not fully elucidated. In view of the fact that healing wounds contain high concentrations of lactate (10-15 mM) and exogenous lactate reduce the NAD(+) pool in cultured fibroblasts, we propose that high lactate lowers the level of nuclear pADPR. MATERIALS AND METHODS: Neonatal human dermal fibroblasts (NHDF) were plated to subconfluence and allowed to adhere. Cells were treated with 15 mM l-lactate and pADPR production was assessed by immunofluorescence analysis using 10H antibody. Difference in pADPR production was determined by calculation of positively stained cells compared to total cell numbers. Inhibition of PARP activity was tested by treatment with 100 microM 3-aminobenzamide (3-AB). Specificity of the lactate effect on pADPR synthesis was verified by using the analogue d-lactate. The contents of nicotinamide adenine dinucleotide (NAD(+)) and its reduced form (NADH) in lactated and non-lactated cell cultures were quantified by the enzymatic cyclic assay. RESULTS: We found that exogenous l-lactate (15 mM) can significantly depress pADPR content in cultured fibroblasts. PARP-1 activity was inhibited by 3-AB and analogue d-lactate showed no effect on pADPR synthesis. NAD(+)/NADH ratio was significantly lowered in lactated compared to non-lactated cell culture. CONCLUSIONS: Exogenous l-lactate (15 mM) can depress pADPR content in cultured fibroblasts. In view of the fact that healing wounds contain such high concentrations of lactate, we propose that down regulation of pADPR is associated with elevated tissue repair via pADPR dependent gene expression. This observation is important in understanding the stimulation of lactate-mediated protein expression during wound healing.

Strong genetic evidence for association of TOR1A/TOR1B with idiopathic dystonia.

Recently, association of a TOR1A(DYT1)/TOR1B risk haplotype with common forms of idiopathic dystonia has been reported in the Icelandic population. Here we report a strong association of two single nucleotide polymorphisms within or in close proximity to the TOR1A 3'UTR, with the lowest p value being 0.000008, in a larger cohort of German and Austrian patients with predominantly focal sporadic dystonia.