Ghrelin suppresses insulin secretion in human islets and type 2 diabetes patients have diminished islet ghrelin cell number and lower plasma ghrelin levels.

It is not known how ghrelin affects insulin secretion in human islets from patients with type 2 diabetes (T2D) or whether islet ghrelin expression or circulating ghrelin levels are altered in T2D. Here we sought out to identify the effect of ghrelin on insulin secretion in human islets and the impact of T2D on circulating ghrelin levels and on islet ghrelin cells. The effect of ghrelin on insulin secretion was assessed in human T2D and non-T2D islets. Ghrelin expression was assessed with RNA-sequencing (n = 191) and immunohistochemistry (n = 21). Plasma ghrelin was measured with ELISA in 40 T2D and 40 non-T2D subjects. Ghrelin exerted a glucose-dependent insulin-suppressing effect in islets from both T2D and non-T2D donors. Compared with non-T2D donors, T2D donors had reduced ghrelin mRNA expression and 75% less islet ghrelin cells, and ghrelin mRNA expression correlated negatively with HbA1c. T2D subjects had 25% lower fasting plasma ghrelin levels than matched controls. Thus, ghrelin has direct insulin-suppressing effects in human islets and T2D patients have lower fasting ghrelin levels, likely as a result of reduced number of islet ghrelin cells. These findings support inhibition of ghrelin signaling as a potential therapeutic avenue for stimulation of insulin secretion in T2D patients.

Maternal thyroid hormone is required for parvalbumin neurone development in the anterior hypothalamic area.

Thyroid hormone (TH) is crucial for brain development and function. This becomes most evident in untreated congenital hypothyroidism, leading to irreversible mental retardation. Likewise, maternal hypothyroxinaemia, a lack of TH during pregnancy, is associated with neurological dysfunction in the offspring, such as autism and reduced intellectual capacity. In the brain, TH acts mainly through TH receptor α1 (TRα1). Consequently, mice heterozygous for a dominant-negative mutation in TRα1 display profound neuroanatomical abnormalities including deranged development of parvalbumin neurones. However, the exact timing and orchestration of TH signalling during parvalbumin neurone development remains elusive. In the present study, we dissect the development of parvalbumin neurones in the anterior hypothalamic area (AHA) in male mice using different mouse models with impaired pre- and postnatal TH signalling in combination with bromodeoxyuridine birth dating and immunohistochemistry. Our data reveal that hypothalamic parvalbumin neurones are born at embryonic day 12 and are first detected in the AHA at postnatal day 8, reaching their full population number at P13. Interestingly, they do not require TH postnatally because their development is not impaired in mice with impaired TH signalling after birth. By contrast, however, these neurones crucially depend on TH through TRα1 signalling in the second half of pregnancy, when the hormone is almost exclusively provided by the mother. For the first time, our findings directly link a maternal hormone to a neuroanatomical substrate in the foetal brain, and underline the importance of proper TH signalling during pregnancy for offspring mental health. Given the role of hypothalamic parvalbumin neurones in the central control of blood pressure, the present study advocates the inclusion of cardiovascular parameters in the current discussion on possible TH substitution in maternal hypothyroxinaemia.

Protection against maternal infection-associated fetal growth restriction: proof-of-concept with a microbial-derived immunomodulator.

Infection-associated inflammatory stress during pregnancy is the most common cause of fetal growth restriction and/or miscarriage. Treatment strategies for protection of at-risk mothers are limited to a narrow range of vaccines, which do not cover the bulk of the common pathogens most frequently encountered. Using mouse models, we demonstrate that oral treatment during pregnancy with a microbial-derived immunomodulator (OM85), currently used clinically for attenuation of infection-associated airway inflammatory symptoms in infants-adults, markedly reduces risk for fetal loss/growth restriction resulting from maternal challenge with bacterial lipopolysaccharide or influenza. Focusing on LPS exposure, we demonstrate that the key molecular indices of maternal inflammatory stress, notably high levels of RANTES, MIP-1α, CCL2, KC, and G-CSF (granulocyte colony-stimulating factor) in gestational tissues/serum, are abrogated by OM85 pretreatment. Systems-level analyses conducted in parallel using RNASeq revealed that OM85 pretreatment selectively tunes LPS-induced activation in maternal gestational tissues for attenuated expression of TNF, IL1, and IFNG-driven proinflammatory networks, without constraining Type1-IFN-associated networks central to first-line antimicrobial defense. This study suggests that broad-spectrum protection-of-pregnancy against infection-associated inflammatory stress, without compromising capacity for efficient pathogen eradication, represents an achievable therapeutic goal.

Factor H uptake regulates intracellular C3 activation during apoptosis and decreases the inflammatory potential of nucleosomes.

Factor H (FH) binds apoptotic cells to limit the inflammatory potential of complement. Here we report that FH is actively internalized by apoptotic cells to enhance cathepsin L-mediated cleavage of endogenously expressed C3, which results in increased surface opsonization with iC3b. In addition, internalized FH forms complexes with nucleosomes, facilitates their phagocytosis by monocytes and induces an anti-inflammatory biased cytokine profile. A similar cytokine response was noted for apoptotic cells coated with FH, confirming that FH diminishes the immunogenic and inflammatory potential of autoantigens. These findings were supported by in vivo observations from CFH(-/-) MRL-lpr mice, which exhibited higher levels of circulating nucleosomes and necrotic cells than their CFH(+/+) littermates. This unconventional function of FH broadens the established view of apoptotic cell clearance and appears particularly important considering the strong associations with genetic FH alterations and diseases such as systemic lupus erythematosus and age-related macular degeneration.

Novel Striatal GABAergic Interneuron Populations Labeled in the 5HT3a(EGFP) Mouse.

Histological and morphological studies indicate that approximately 5% of striatal neurons are cholinergic or γ-aminobutyric acidergic (GABAergic) interneurons (gINs). However, the number of striatal neurons expressing known interneuron markers is too small to account for the entire interneuron population. We therefore studied the serotonin (5HT) receptor 3a-enhanced green fluorescent protein (5HT3a(EGFP)) mouse, in which we found that a large number of striatal gINs are labeled. Roughly 20% of 5HT3a(EGFP)-positive cells co-express parvalbumin and exhibit fast-spiking (FS) electrophysiological properties. However, the majority of labeled neurons do not overlap with known molecular interneuron markers. Intrinsic electrical properties reveal at least 2 distinct novel subtypes: a late-spiking (LS) neuropeptide-Y (NPY)-negative neurogliaform (NGF) interneuron, and a large heterogeneous population with several features resembling low-threshold-spiking (LTS) interneurons that do not express somatostatin, NPY, or neuronal nitric oxide synthase. Although the 5HT3a(EGFP) NGF and LTS-like interneurons have electrophysiological properties similar to previously described populations, they are pharmacologically distinct. In direct contrast to previously described NPY(+) LTS and NGF cells, LTS-like 5HT3a(EGFP) cells show robust responses to nicotine administration, while the 5HT3a(EGFP) NGF cell type shows little or no response. By constructing a molecular map of the overlap between these novel populations and existing interneuron populations, we are able to reconcile the morphological and molecular estimates of striatal interneuron numbers.

[Delayed gastric emptying after partial duodenopancreatectomy]. / Porucha evakuace zaludku po parciální duodenopankreatektomii.

INTRODUCTION: Delayed gastric emptying (DGE) is a relatively common complication in patients after partial duodenopancreatectomy (PDPE) and significantly contributes to their postoperative morbidity. There has only been a small amount of interest attributed to DGE in Czech literature. The aim of this article is to present and analyze our own experience with the occurrence of DGE after PDPE. MATERIALS AND METHODS: Retrospective analysis of prospectively collected data from 106 patients who underwent a PDPE at the Department of Surgery, University Hospital Motol and 2nd Medical Faculty, between 2001 and 2011. The data were statistically analyzed using the Chi-square test with statistical significance set at 5% of probability value (p < 0.05). RESULTS: During the reporting period, PDPE was performed in 106 patients with a pathological process in the pancreatic head. 4 patients died in the postoperative period (30-day mortality 3.8%). Major postoperative complications occurred in 31 patients (morbidity 29%), with clinically significant DGE (grade B and C) in almost half of them (16 patients). Along with the DGE, 4 patients suffered from other major complications (2 pancreatic anastomotic insufficiency, 1 bile fistula and 1 external pancreatic fistula). We did not prove a statistically significant difference in the incidence of DGE depending either on the type of resection or pancreatic anastomosis type. CONCLUSION: DGE contributes significantly to postoperative morbidity in patients after PDPE and is associated with other serious postoperative complications in a considerable number of cases.

Joint kinematics following bi-compartmental knee replacement during daily life motor tasks.

In many cases knee osteoarthritis leads to total knee replacement surgery (TKR) even if the lateral compartment is not involved. More recently, a bicompartmental knee replacement system (BKR) (Journey Deuce, Smith & Nephew Inc., Memphis, TN, USA) has been developed that only replaces the medial tibiofemoral and the patellofemoral compartments, thus preserving both cruciate ligaments with its associated benefits. However information on the effect of BKR on in vivo knee joint kinematics is not widely available in the literature. Therefore, this study analyzed full three-dimensional knee joint kinematics in 10 postoperative BKR-subjects for a broad spectrum of relevant daily life activities: walking, walking followed by a cross-over or sidestep turn, step ascent and descent, mild squatting and chair rise. We analyzed to what extent normal knee motion is regained through comparison with their non-involved limb as well as a group of matched controls. Furthermore, coefficients of multiple correlation were calculated to assess the consistency of knee joint kinematics both within and between subject groups. This analysis demonstrated that, despite the presence of differences indicative for retention of pre-operative motion patterns and/or remaining compensations, knee joint kinematics in BKR limbs replicate, for a large range of daily-life motor tasks, the kinematics of the contra-lateral non-affected limbs and healthy controls to a similar extent as they are replicated within both these control groups.

[Minimal node affection in gastric carcinoma--pilote multicentric study results]. / Minimální uzlinové postizení u karcinomu zaludku--pilotní výsledky multicentrické studie.

INTRODUCTION: Lymphadenectomy is an essential part of surgical treatment of gastric cancer. In the part of hematoxylin-eosin negative nodes, small foci of tumour cells (micrometastastasis and isolated tumour cells) can be found using immunostaing or RT-PCR. The aim of this study is to asses clinical and prognostic relevance of these findings. MATERIAL AND METHODS: Multicenter, prospective, non-randomised clinical trial running in four Czech centres. All lymphatic nodes from patients after radical resection are stained using standard hematoxylin-eosin technique, all negative nodes are further processed with immunostaining employing cytokeratin antibody. RESULTS: In the period of two years (1st January 2009 - 31st December 2010), 73 patients (100%) were included into the study from four Czech centers. All patients underwent radical resection for gastric cancer. Subtotal resection was performed in 33 patients (45%), total gastrectomy in the remaining 40 patients (55%). Total number of acquired lymphatic nodes (LN) reached 1245, average number of nodes per one patient was 17.3. H-E metastasis were disclosed in 364 LN (29%). All H-E negative nodes were further processed using immunohistochemical staining. Lymph node micrometastasis (MM) were discovered in 35 LN (3%), isolated tumour cells (ITC) in another 72 LN (6%). Clinical and prognostic relevance of lymph node MM and ITC was assessed based on the patients' survival data. CONCLUSION: Preliminary results of this study indicate that presence of the lymph node MM and ITC in gastric cancer patients is not linked to worse oncological outcome. Based on our results we can conclude, that expensive, time consuming and technically demanding immunostaining technique could not yet be recommended as a routine part of histological investigation of lymphatic nodes.

[Cystic neoplasms of the pancreas]. / Cystické neoplazie slinivky brisní.

INTRODUCTION: Neoplastic changes represent an important part of cystic deposits in pancreas. It is morphologically non-homogenous group of neoplasms with different occurrence depending on sex and age, different localization and different biologic properties. The aim of the paper is to remind current knowledge of this topic and confront it with our own experience. MATERIAL AND METHODS: Together 13 patients with histologically proved cystic neoplasm of pancreas underwent surgery during the period of ten years from 1997 to 2007. They represent 6% of all patients operated for pancreatic tumor (213 patients). Women (9 patients) represented more than two thirds of all operated patients and deposits were more often localized in the head of pancreas (8). RESULTS: The most frequent operation was partial duodenopancreatectomy (7) and most frequently cystadenocarcinoma was identified histologically (5 times). Median survival of these patients is 54 months. Left sided resection, done in 5 cases, identified benign tumor in all patients; no recurrence was found in 2 years follow-up. CONCLUSION: Cystic neoplasms localized in the pancreatic head are more frequent in men than in women and predominantly malignant, on the contrary localization in the tail of pancreas is particularly in younger women linked with benign tumor. Prognosis after resection is markedly better in malignant processes even in comparison with ductal carcinoma.

[Surgical treatment of bleeding gastroduodenal ulcer]. / Chirurgické lécení krvácení z gastroduodenálního vredu.

THE AIM OF THE STUDY: Based on the evaluation of results we prove authorization of our elected approach to the treatment of bleeding gastroduodenal ulcer with the restrained and careful choice of patients for surgeries. MATERIAL AND METHODS: The authors analyse in a retrospective study a group of 45 patients operated on for bleeding gastroduodenal ulcer in the period from 2001 to 2005 year. Only patients with actual conservatively or endoscopically ensolvable bleeding were consistenly indicated for surgery. We prefered the local surgical hemostasis procedure. RESULTS: In the period from 2001 to 2005 year 45 patients were operated on. Altogether 9 patients died (20.0%). It was needed reoperated 8 patients (17.8%) for bleeding recurrence. In most cases (39 i.g. 86.7%) local surgical procedures were done. CONCLUSION: Our reached results give clear warrant to the conservative surgical treatment, when only patients with actual endoscopically ensolvable bleeding from gastroduodenal ulcer are indicated for surgery and the aim of local surgical procedure is safe hemostasis. Our results are comparable with data in literature.

[Long-term results after radical resections for pancreatic ductal adenocarcinoma--10 years experience]. / Dlouhodobé výsledky po radikálních resekcích pro duktální adenokarcinom pankreatu--desetileté zkusenosti.

INTRODUCTION: Pancreatic ductal adenocarcinoma is the most often and the most malignant type of pancreatic tumor. Effective systemic anticancer treatment is still missing and only radical resection can potentially lead to the life prolongation. TARGET: Long-term therapeutic outcomes evaluation in patients after radical resections due to the pancreatic ductal adenocarcinoma during the 10 years period. MATERIAL AND METHODS: Population included 42 patients after resection of pancreas due to ductal adenocarcinoma realized during the period from 1995 to 2005. Therapeutic outcomes including long-term survival in different stages of the disease were compared with data collected from patients with another histological type of periampullar tumor by statistical analysis. RESULTS: 48 radical resections of pancreas due to ductal adenocarcinoma were realized during the 10 years period. Six patients were excluded from the follow up. Median of survival with the minimum 6 months of follow up has reached 14 months and the maximal survival time was 35 months. None of the patients has survived 5 years. Five patients were alive after the end of follow up period. There were no statistical difference in survival when particular disease stages were compared (p = 0.3226). Survival of female patients in this population was statistically lower in comparison to male patients (p = 0.0222). Significantly lower survival of patients with ductal adenocarcinoma in comparison to the patients with other types of carcinoma in periampullar localization was demonstrated (p = 0.0234). CONCLUSION: Achieved results proved that pancreatic carcinoma is solid tumor with the worst long-term prognosis. Long-term survival in this population did not exceed 35 months and was independent on per-operative staging. Long-term prognosis of ductale adenocarcinoma is significantly worse in comparison to other types of carcinoma in periampullar localization.

Strictures of the terminal bile duct--who profits from the surgery?

BACKGROUND: Elucidation of the etiology of terminal bile duct strictures is oftentimes challenging. In choosing the optimal treatment method, a multidisciplinary approach is necessary. AIMS AND METHODS: To demonstrate the advantages and indications of surgical management, 153 patients with terminal bile duct strictures were retrospectively analysed. All patients had been treated during a period of eight years (1st January 1995 - 31st December 2002) in same department. The principal datas studied were surgical morbidity, thirty-days' mortality rates and recurrence of jaundice. RESULTS: During a period of eight years, 153 patients (100%) underwent laparotomy. Radical surgery (duodenopancreatectomy) was performed in 79 patients (51.6%), while 74 others (48.4%) benefited from bypass procedures. Postoperative morbidity reached 13.1%; nine patients (5.9%) died within the thirty days of surgery. Recurrence of jaundice was observed in three patients (1.9%). CONCLUSION: Surgical management of terminal bile duct strictures is indicated in most patients with benign strictures, strictures of unknown origin and with malignant strictures, where the patients are in a reasonably satisfactory clinical condition. The surgical approach provides the potential advantage to obtain a definite histological diagnosiscare and to remove the tumour in many patients. Patients with non resectable tumours could benefit from bypass procedures ensuring long-term and effective drainage of the bile.

[Carcinoma of the pancreatic head and periampular carcinomas. 10-year results of their resection management]. / Karcinom hlavy pankreatu a periampulární karcinomy. Výsledky resekcní lécby za 10 let.

INTRODUCTION: Parcial duodenopancreatectomy (DPE) remains the principal treatment method of carcinomas located in the region of the pancreatic head. Although several surgical clinics use this method, assessment of, mainly the long-term, treatment results is virtually absent in our literature. AIM: Presentation of early and late results of the procedure collected in a clinic, managing the procedures with "medium frequency". The study is aimed at patients with histologically confirmed carcinomas of the pancreatic head. MATERIAL AND METHODS: Prospective data from a group of patients who had undergone the DPE during the 10-year study in the Surgical Clinic of the IInd Medical Faculty of the Charles University in Prague-Motol, have been collected. RESULTS: From January 1995 to the end of 2004, 121 DPEs were completed, the 30-day mortality rate was 4.9% and the morbidity rate reached 17.5%. The long-term results were assessed in 71 cancer patients - the median survival rate was 18 months following the procedure (1-121 months), depending on the tumor type. CONCLUSIONS: The collected results are similar to those presented by foreign clinics (but conducted with lower frequencies here) and fall within a European better average.

[Spontaneous rupture of a splenic pseudocyst]. / Spontánní ruptura pseudocysty sleziny.

In the course of chronic pancreatitis diverse complications may arise. Intraparenchymal splenic complications are very rare. In the following case the authors describe rupture of splenic pseudocyst with acute intraabdominal bleeding as a complication of chronic pancreatitis.

[Gastric carcinoma and D2 lymphadenectomy--technique and possible complications]. / Karcinom zaludku a D2 lymfadenektomie--technika, mozné komplikace.

Lymphadenectomy in gastric cancer became vital and obligatory part of staging procedure. Therapeutic effect of lymphadenectomy with patient's prognosis improvement still remains uncertain. In subgroups, extensive lymphadenectomy enhance radicality of the procedure, enables to reach R0 resection in the area of tumour bed and hopefully improves survival. D2 lymphadenectomy is now becoming as a standard also in our country. Technique and possible complications of D2 lymphadenectomy are the matter of this article.

Gastric cancer in young patients--is there any hope for them?

Gastric cancer usually affects people older than sixty years. This type of cancer is very rare in adults under thirty years of age. In addition, the prognosis in this part of population is grave due to the high incidence of undifferentiated tumours and advanced stage at time of diagnosis. Radical surgery affords the only chance for long term survival, but even this option is often limited upon finding locally advanced disease or peritoneal seeding. The following are case studies of three young adults from a group of 45 patients, who were treated between January 1st, 2000 and December 31st, 2003.

[Massive hemorrhage from a pancreatic pseudocyst into the duodenum]. / Masivní krvácení z pseudocysty pankreatu do duodena.

Pancreatic pseudocyst or pseudoaneurysm bleeding related to chronic pancreatitis is associated with high mortality. Diagnostics includes angiography and contrast CT; therapy is based mainly on percutaneous transvascular embolisation (PTE) and surgery. Mortality burden related to conservative approach is up to 90%. Case report describes massive pancreas pseudocyst bleeding from erosion-affected arteria gastroduodenalis in 44-years-old patient with chronic pancreatitis. Spontaneous pseudocyst perforation into the duodenum caused hemorrhagic shock with necessity of urgent surgery. Identification of bleeding vessel was enabled thanks to longitudinal gastroduodenotomy and pseudocyst dissection through duodenum wall. Article reminds some diagnostics and therapeutics issues related to chronic pancreatitis complications with bleeding as well as information published in recent literature.

Sentinel node biopsy in gastric cancer: preliminary results.

BACKGROUND: Extent of lymphadenectomy in gastric cancer is one of the still unresolved and ongoing questions. Whether extensive procedures lead to better survival remains controversial. The detection of sentinel node could become a helpful method to define the desirable extent of lymph node dissection for each particular patient. MATERIAL AND METHODS: Prospective, single-centre, clinical trial. Sentinel node is identified using vital blue dye. After performance of D2 lymphadenectomy, the pathologist minutely investigates all lymphatic nodes by histological and immunohistochemistry techniques. RESULTS: During a period of 36 months, an attempt to localise the sentinel node for biopsy was made in 22 patients. The successful rate of sentinel node detection with a good relation between metastatic involvement of the sentinel node and other nodes was 56% (13 out of 22 patients). All these patients suffered from small tumours and early stage of the disease. In large tumours and advanced stages, blue dye stained the tissue around the tumour diffusely rendering the identification of true sentinel node impossible. Fresh frozen section of the sentinel node resulted in two patients in a false negative outcome because of micrometastatic involvement. CONCLUSIONS: Sentinel node biopsy in gastric cancer using vital staining is a feasible method. Reliable results were seen in early stage of the disease. Fresh frozen section of sentinel node has probably a low sensitivity for detection of micrometastases.

[Lymphadenectomy in gastric carcinoma--present status, new trends and personal experience]. / Lymfadenektomie u karcinomu zaludku--soucasný stav, nové trendy, vlastní zkusenosti.

The only hope for long time survival in gastric cancer is radical surgery with complete tumour removal. Lymphadenectomy as a part of surgical procedure helps to remove tumour completely. The value of lymphadenectomy for staging is clear and surgeons have no doubt about it. Therapeutic effect of lymphadenectomy with improvement of the prognosis remains uncertain. In this work we would like to discuss current trends in lymphadenectomy in gastric cancer, especially evaluating the value and extent of the procedure.

[Stenoses of the terminal choledochus--surgical treatment]. / Stenózy terminálního choledochu--chirurgické lécení.

INTRODUCTION: Obstructive jaundice caused by stenosis of the terminal choledochus remains even nowadays a serious differential diagnostic and therapeutic problem. OBJECTIVE: The objective is to draw attention to possible contemporary treatment and its indications. MATERIAL AND METHODS: The authors evaluated retrospectively the results achieved by surgical treatment in 145 patients operated during the last 8 years on account of benign or malignant obstruction of the terminal choledochus. RESULTS: Within 30 days after surgery 9 patients (6%) died. A relapse of the obstructive jaundice occurred in three instances (2%). The morbidity amounted to a total of 14% and is comparable with data in the contemporary literature. CONCLUSIONS: The presented results support the inclusion of surgical treatment as the method of first choice in patients with stenosis of the terminal choledochus of uncertain biological nature. For surgery also patients are indicated with obstruction of malignant origin who cannot be treated radically (M1), provided they are in a satisfactory general condition (ASA I-III). Surgical therapy is suitable also for treatment of the majority of patients with a benign stenosis or stricture of the terminal choledochus.