Alterations in regulatory T cells and immune checkpoint molecules in pancreatic cancer patients receiving FOLFIRINOX or gemcitabine plus nab-paclitaxel.

PURPOSE: This pilot study aimed on generating insight on alterations in circulating immune cells during the use of FOLFIRINOX and gemcitabine/nab-paclitaxel in pancreatic ductal adenocarcinoma (PDAC). PATIENTS AND METHODS: Peripheral blood mononuclear cells were isolated before and 30 days after initiation of chemotherapy from 20 patients with advanced PDAC. Regulatory T cells (FoxP3+) and immune checkpoints (PD-1 and TIM-3) were analyzed by flow cytometry and immunological changes were correlated with clinical outcome. RESULTS: Heterogeneous changes during chemotherapy were observed in circulating T-cell subpopulations with a pronounced effect on PD-1+ CD4+/CD8+ T cells. An increase in FoxP3+ or PD-1+ T cells had no significant effect on survival. An increase in TIM3+/CD8+ (but not TIM3+/CD4+) T cells was associated with a significant inferior outcome: median progression-free survival in the subgroup with an increase of TIM-3+/CD8+ T cells was 6.0 compared to 14.0 months in patients with a decrease/no change (p = 0.026); corresponding median overall survival was 13.0 and 20.0 months (p = 0.011), respectively. CONCLUSIONS: Chemotherapy with FOLFIRNOX or gemcitabine/nab-paclitaxel induces variable changes in circulating T-cell populations that may provide prognostic information in PDAC.

Development and validation of a novel SPR-based assay principle for bispecific molecules.

Surface plasmon resonance (SPR) is increasingly applied in drug discovery, early development and production. However, there are remarkably few reports describing the application of SPR in a regulated environment. Here, we describe a novel SPR-based assay, which enables us to assess the binding activity of a bivalent-bispecific anti-Ang-2/anti-VEGF antibody to both targets in a single setup. Validation of the assay revealed a high level of precision, accuracy, linearity and specificity. Upon analysis of temperature stressed samples it could be shown that firstly, the assay is able to indicate function-loss and secondly, it allows the parallel analysis of an additional interaction. Therefore, the described assay is highly suitable for quality assessment of the Ang-2/VEGF CrossMab. Additionally, the use of SPR in the context of assay development and routine use in a GMP environment is discussed.

Malaria parasite liver stages render host hepatocytes susceptible to mitochondria-initiated apoptosis.

Intracellular eukaryotic parasites and their host cells constitute complex, coevolved cellular interaction systems that frequently cause disease. Among them, Plasmodium parasites cause a significant health burden in humans, killing up to one million people annually. To succeed in the mammalian host after transmission by mosquitoes, Plasmodium parasites must complete intracellular replication within hepatocytes and then release new infectious forms into the blood. Using Plasmodium yoelii rodent malaria parasites, we show that some liver stage (LS)-infected hepatocytes undergo apoptosis without external triggers, but the majority of infected cells do not, and can also resist Fas-mediated apoptosis. In contrast, apoptosis is dramatically increased in hepatocytes infected with attenuated parasites. Furthermore, we find that blocking total or mitochondria-initiated host cell apoptosis increases LS parasite burden in mice, suggesting that an anti-apoptotic host environment fosters parasite survival. Strikingly, although LS infection confers strong resistance to extrinsic host hepatocyte apoptosis, infected hepatocytes lose their ability to resist apoptosis when anti-apoptotic mitochondrial proteins are inhibited. This is demonstrated by our finding that B-cell lymphoma 2 family inhibitors preferentially induce apoptosis in LS-infected hepatocytes and significantly reduce LS parasite burden in mice. Thus, targeting critical points of susceptibility in the LS-infected host cell might provide new avenues for malaria prophylaxis.

Malignant risk and surgical outcomes of presacral tailgut cysts.

BACKGROUND: Presacral tailgut cysts are uncommon and few data exist on the outcomes following surgery. METHODS: Patients undergoing tailgut cyst resection at the Mayo Clinic between 1985 and 2008 were analysed retrospectively. Demographic data, clinicopathological features, operative details, postoperative complications and recurrence were reviewed. RESULTS: Thirty-one patients were identified (28 women), with a median age of 52 years. Seventeen patients were symptomatic and 28 had a palpable mass on digital rectal examination. Median cyst diameter was 4.4 cm. Four patients had a fistula to the rectum. Complete cyst excision was achieved in all patients; eight underwent distal sacral resection or coccygectomy. Postoperative complications occurred in eight patients but without 30-day mortality. Malignant transformation was present in four patients: adenocarcinoma in three and carcinoid in one. The cyst recurred in one patient after surgery for a benign lesion. CONCLUSION: Presacral tailgut cysts should be removed due to the risk of malignant transformation.

Tuberculosis control in Pakistan: reviewing a decade of success and challenges.

Pakistan has shown drastic expansion of tuberculosis (TB) care during the past 10 years, increasing case notifications from 11 050 in 2000 to 248 115 in 2008. Over 1 million TB patients have been cared for since 2000, with a treatment success rate of 91% in 2007. This paper examines the strategic decisions and infrastructure improvements underlying this achievement, such as the implementation of universal DOTS coverage, expansion of the laboratory network, effective drug management systems, improved communication strategies, and inclusion of private practitioners, laboratories and hospitals in the TB control programme through the public-private mix strategy. The paper also outlines challenges faced in further expanding TB control within the private sector and parastatal health care institutions; strengthening the laboratory network for diagnosis of drug-resistant TB; and ensuring uninterrupted supply of quality anti-TB drugs, all requiring continued and coordinated technical and donor support.

Tuberculosis control in Pakistan: reviewing a decade of success and challenges

Pakistan has shown drastic expansion oftuberculosis [TBTcareduring the past 10 years, increasing case notifications from 11 050 in 2000 to 248115 in 2008. Over 1 million TB patients have been cared for since 2000, with a treatment success rate of 91% in 2007. This paper examines the strategic decisions and infrastructure improvements underlying this achievement, such as the implementation of universal DOTS coverage, expansion of the laboratory network, effective drug management systems, improved communication strategies, and inclusion of private practitioners, laboratories and hospitals in the TB control programme through the public-private mix strategy. The paper also outlines challenges faced in further expanding TB control within the private sector and parastatal health care institutions; strengthening the laboratory network for diagnosis of drug-resistant TB; and ensuring uninterrupted supply of quality anti-TB drugs, ail requiring continued and coordinated technical and donor support

Variants in Apaf-1 segregating with major depression promote apoptosome function.

APAF1, encoding the protein apoptosis protease activating factor 1 (Apaf-1), has recently been established as a chromosome 12 gene conferring predisposition to major depression in humans. The molecular phenotypes of Apaf-1 variants were determined by in vitro reconstruction of the apoptosome complex in which Apaf-1 activates caspase 9 and thus initiates a cascade of proteolytic events leading to apoptotic destruction of the cell. Cellular phenotypes were measured using a yeast heterologous expression assay in which human Apaf-1 and other proteins necessary to constitute a functional apoptotic pathway were overexpressed. Apaf-1 variants encoded by APAF1 alleles that segregate with major depression in families linked to chromosome 12 shared a common gain-of-function phenotype in both assay systems. In contrast, other Apaf-1 variants showed neutral or loss-of-function phenotypes. The depression-associated alleles thus have a common phenotype that is distinct from that of non-associated variants. This result suggests an etiologic role for enhanced apoptosis in major depression.

Botryococcus braunii: a rich source for hydrocarbons and related ether lipids.

This paper presents a review on Botryococcus braunii, a cosmopolitan green colonial microalga characterised by a considerable production of lipids, notably hydrocarbons. Strains like wild populations of this alga differ in the type of hydrocarbons they synthesise and accumulate: (1) n-alkadienes and trienes, (2) triterpenoid botryococcenes and methylated squalenes, or (3) a tetraterpenoid, lycopadiene. In addition to hydrocarbons and some classic lipids, these algae produce numerous series of characteristic ether lipids closely related to hydrocarbons. This review covers the algal biodiversity, the chemical structures and biosynthesis of hydrocarbons and ether lipids and the biotechnological studies related to hydrocarbon production.

Anatomic variations of the frontal sinus.

INTRODUCTION: Minimally invasive approaches are increasing in popularity. We have undertaken an anatomic and radiological study of the frontal sinus that is frequently involved in the supercilliary minicraniotomy used to access the parasellar region. OBJECTIVE: We wanted to evaluate the prevalence and morphological characteristics of the frontal sinus in a certain population sample. METHODS: 98 randomly assigned individuals were subjected to CT examinations. The antero-posterior diameter, sagittal diameter, transverse diameter and total volume of the frontal sinus were calculated. RESULTS: The frontal sinus tended to be larger in males than in females with the exception of the transverse diameter (p < 0.10). CONCLUSION: Knowledge of the anatomic variations of the frontal sinus is important in surgical approaches through the superciliary arc in order to avoid complications such as infections and CSF fistula.

The influence of intraoperative complications on adhesion formation during laparoscopic and conventional cholecystectomy in an animal model.

BACKGROUND: The aim of this study was to evaluate the extent of postoperative adhesion formation after laparoscopic and open cholecystectomy. MATERIALS AND METHODS: Qualified surgeons performed 60 experimental laparoscopic cholecystectomies (LC) in dogs with the aim to acquire the laparoscopic technique. To assess the relation between the complications during the operation (bleeding, laceration of the liver bed, or gallbladder perforation) and the formation of adhesions, surviving animals were divided into four groups according to the type of complication occurred. Assessment of the results was made by second-look laparoscopy 4 weeks after LC using the adhesion index (AI; score range, 0-4). The animals then were killed so the extent of adhesion formation could be measured. As a control, open cholecystectomy was performed in 15 dogs without intraoperative complications. The Mann-Whitney rank-sum test and Dunn's method were used for statistical analysis. RESULTS: No adhesion formation or intraoperative complications were registered in the laparoscopic group I. In all the cases wherein bleeding or laceration of the liver bed occurred and was managed with electrocoagulation, adhesions formed. Adhesion formation in these groups was significantly higher than in "ideal LC" or cases of gallbladder perforation alone (p < 0.01). All the animals in the control group developed significantly more adhesions than those in the experimental group (p < 0.05). CONCLUSIONS: It seems that LC has a lower rate of adhesion formation than the conventional open technique. Complications such as bleeding or laceration of the liver bed during LC can enhance adhesion formation. No adhesion formation can be mentioned in relation to gallbladder perforation during LC.

[Laparoscopic colon surgery]. / Laparoszkópos vastagbélsebészet.

We review our experience in laparoscopic colorectal surgery, with indications, technical aspects and results. Between 1992 and 31/12/2000, we performed 113 laparoscopic or laparoscopically assisted colorectal operations. Of 79 malignant cases, 37 operations were oncologically radical and therapeutic, 42 were palliative. During the immediate postoperative period two deaths occurred (2.8%), the causes of death were not related to surgery. Port site metastasis developed in one patient (1.4%). Postoperative complications developed in 18 patients (14.5%). Only one patient required conversion to laparotomy. We emphasize the importance of hand assisted laparoscopic surgery (HALS) in laparoscopic colorectal surgery, because it can increase the number of laparoscopic colorectal operations. Based on our results and experience, we recommend the routine use of laparoscopic technique in colorectal surgery.

[How do intraoperative complications effect adhesion formation after laparoscopic cholecystectomy: a comparative animal study]. / Az intraoperatív szövódmények hatása a posztoperatív hasúri adhesiók kialakulására laparoszkópos cholecystectomiát követóen: összehasonlító állatkísérlet.

BACKGROUND: The aim of this study was to evaluate the extent of postoperative formation of adhesions following laparoscopic and open cholecystectomy. MATERIAL AND METHODS: 60 experimental laparoscopic cholecystectomies (LC) were performed in dogs by qualified surgeons to learn laparoscopic technique. To assess the relationship between complications occurred during the operation (bleeding, laceration of the liver bed or gallbladder perforation) and the formation of adhesions surviving animals were divided into 4 groups according to the type of complication. We assessed the results during second-look laparoscopy 4 weeks following LC using the adhesion index (AI: 0-4 score). Animals were then sacrificed to measure the extent of adhesions. As a control group open cholecystectomy was performed in 15 dogs without intraoperative complications. Mann-Whitney Rank Sum test and Dunn's Method were used for statistical analysis. RESULTS: No adhesions were observed in the laparoscopic group without intraoperative complications. In all dogs with bleeding or laceration of the liver bed maintained by electrocoagulation, adhesions developed. Formation of adhesion in these groups was significantly higher than in "ideal LC" or in case of gallbladder perforation (P < 0.01). All animals in the control group developed significantly more adhesions compared to the experimental group (p < 0.05). CONCLUSION: LC produces less adhesion compared to conventional open technique. Complications such as bleeding or laceration of the liver bed during LC can increase the formation of adhesions. No formation of adhesions can be related to gallbladder perforation during LC.

Percutaneous video choledochoscopic treatment of retained biliary stones via dilated T-tube tract.

BACKGROUND: Retained biliary stones is a common clinical problem in patients after surgery for complicated gallstone disease. When postoperative endoscopic retrograde cholangiopancreatography (ERCP) and endoscopic sphincterotomy are unsuccessful, several percutaneous procedures for stone removal can be applied as alternatives to relaparotomy. These procedures are performed either under fluoroscopic control or with the use of choledochoscopy, but it is also possible to combine these methods. METHODS: Since 1994, we have used the percutaneous video choledochoscopic technique for the removal of difficult retained biliary stones via dilated T-tube tract in 17 patients, applying the technique of percutaneous stone extraction used in urology. While waiting for the T-tube tract to mature and after the removal of the T-tube, the dilatation of its tract was 26-30 Fr. Stone removal was carried out using a flexible video choledochoscope and a rigid renoscope under fluoroscopic control, with the aid of Dormia baskets, rigid forceps, and high-pressure irrigation. RESULTS: We performed 23 operative procedures, and the clearance of the biliary ducts was successful in all cases. There were no major complications or deaths. CONCLUSION: Percutaneous video choledochoscopic-assisted removal of large retained biliary stones via the T-tube tract is a highly effective and safe procedure. Its advantages over other procedures include the ability to visualize the stones and noncalculous filling defects; it also guarantees that the stones can be removed under visual video endoscopic control. It has no problems related to tract or stone size.

Six novel tetraterpenoid ethers, lycopanerols B-G, and some other constituents from the green microalga Botryococcus braunii.

Six novel tetraterpenoid ethers, lycopanerols B-G, were isolated from lipidic extracts of the green microalga Botryococcus braunii (L race), along with a series of phytyl esters and alpha- and beta-tocopherols. The structures of the compounds were determined by means of spectral analyses including 2D NMR techniques. A biogenetic relationship is proposed between lycopanerols and lycopadiene, the acyclic diunsaturated tetraterpenoid hydrocarbon synthesized by the alga.

The use of a thrombus-specific ultrasound contrast agent to detect thrombus in arteriovenous fistulae.

RATIONALE AND OBJECTIVES: To evaluate the use of a new thrombus-specific ultrasound contrast agent, MRX-408, in the ultrasonic detection of thrombus in arteriovenous (AV) fistulae. METHODS: Six purpose-bred mongrels with two AV fistulae each were imaged with gray-scale ultrasound 7 weeks after graft implantation before and after the intravenous bolus injection of MRX-408 (a GPIIb receptor-targeted ultrasound contrast agent). Pre- and postcontrast videotaped segments were randomized and reviewed by four radiologists blinded to the presence of thrombus in the grafts. RESULTS: After the use of MRX-408, there was improved visualization of thrombus within the grafts (P < 0.0001). This was due to the enhancement of the thrombus (P < 0.0001). The improved visualization and contrast enhancement were more marked in the grafts that contained thrombus nonhyperechoic to surrounding soft tissues. CONCLUSIONS: MRX-408 demonstrated better visualization of thrombus within AV fistulae. This was shown in both patent and occluded grafts. These results are encouraging and suggest that this contrast agent merits further development.

Clostridium difficile infection: risk factors, medical and surgical management.

BACKGROUND: Clostridium difficile has become recognized as a cause of nosocomial infection which may progress to a fulminant disease. METHODS: Literature review using electronic literature research back to 1966 utilizing Medline and Current Contents. All publications on antibiotic-associated diarrhea, antibiotic-associated colitis, and pseudomembranous colitis as well as C. difficile infection were included. We addressed established and potential risk factors for C. difficile disease such as an impaired immune system and cost benefits of different diagnostic tests. An algorithm is outlined for diagnosis and both medical and surgical management of mild, moderate and severe C. difficile disease. RESULTS: Diagnosis of C. difficile infection should be suspected in patients with diarrhea, who have received antibiotics within 2 months or whose symptoms started after hospitalization. A stool specimen should be tested for the presence of leukocytes and C. difficile toxins. If this is negative and symptoms persist, stool should be tested with 'rapid' enzyme immunoabsorbent and stool cytotoxin assays, which are the most cost-effective tests. Endoscopy and other imaging studies are reserved for severe and rapidly progressive courses. Oral metronidazole or vancomycin are the antibiotics of choice. Surgery is rarely required for selected patients refractory to medical treatment. The threshold for surgery in severe cases with risk factors including an impaired immune system should be low. CONCLUSION: C. difficile infection has been recognized with increased frequency as a nosocomial infection. Early diagnosis with immunoassays of the stool and prompt medical therapy have a high cure rate. Metronidazole has supplanted oral vancomycin as the drug of first choice for treating C. difficile infections.

The judgement of adhesion formation following laparoscopic and conventional cholecystectomy in an animal model.

INTRODUCTION: The development of postoperative adhesions remains an almost inevitable consequence of visceral and gynaecologic surgery, appearing in 50-95% of all patients. Although decreased adhesion formation is one of the accepted advantages of laparoscopic surgery, only a small number of prospective studies have been done to support this claim. AIMS OF THE STUDY: To evaluate the extent of postoperative adhesion formation after laparoscopic and open cholecystectomy. MATERIAL AND METHOD: 60 experimental laparoscopic cholecystectomies (LC) were performed by qualified surgeons in dogs with the aim to acquire the laparoscopic technique. To assess the relation between the complications during the operation (bleeding, injury to the liver substance or gallbladder perforation) and the formation of adhesions, the surviving animals were divided into 4 groups according to the complications occurred. The assessment of the results was made by second--look laparoscopy 4 weeks following LC using the adhesion index. As a control group open cholecystectomy was then performed in 5 dogs without intraoperative complications. RESULTS: No adhesion formation was observed in the groups where no intraoperative complications occurred. In all the cases where bleeding or injury to the liver bed occurred adhesion formation occurred. No adhesion formation was observed in case of gallbladder perforation. In all the animals of the control group adhesion formation was observed. CONCLUSION: It seems that LC has a reduced rate of adhesion formation when compared with the open technique. Complications such as bleeding or injury to the liver substance during LC can enhance adhesion formation. No adhesion formation can be mentioned in relation with gallbladder perforation when the laparoscopic technique is applied.

The history and the future of teaching and training of laparoscopic surgery in Hungary.

INTRODUCTION: Laparoscopic biliary surgery was introduced in Hungary at the end of the 1990. A variety of experimental training and teaching courses had been performed in basic techniques and the human field, which was followed by laparoscopic biliary surgery and various advanced fields. AIMS OF THE STUDY: To review the history of teaching and training of laparoscopic surgery in Hungary in both the experimental and the human field, and to draw the consequences of this experience. MATERIAL AND METHODS: In a period of 6 years 704 qualified surgeons received a full hands--on hands experimental training in laparoscopic biliary surgery, laparoscopic advanced surgery, laparoscopic gynaecologic and laparoscopic urology surgery. DISCUSSION: The courses performed in the first and the second phase were of theoretical and practical components. The theoretical knowledge was based with emphasise to the new instruments and equipment, the indications, the new surgical technique. The practical knowledge gave every participant the full time to acquire this new type of surgery. At the end of the courses the successful participants received certificates to shift for training to the human field. Each institution needs to wrestle with issues concerning credentialling in advanced laparoscopic surgery. Like many technical skills, proficiency is maintained through repetition. A philosophy must be developed to determine whether each surgeon will perform this highly specialised type of surgery or whether it will be considered a general skill.