Invasive monitoring of the clinical effects of high intra-abdominal pressure for insertion of the first trocar.

Background: To analyze the effects of transitory, high intra-abdominal pressure on clinical, hemodynamic, blood gas and metabolic parameters.

Methods: Sixty-seven laparoscopic patients were divided into groups P12 (n = 30, maximum intra-abdominal pressure of 12 mmHg) and P20 (n = 37, maximum intra-abdominal pressure of 20 mmHg). Through radial artery cannulation, mean arterial pressure (MAP) was assessed and blood gas analysis – pH, arterial oxygen tension (PaO2), arterial carbon dioxide tension (PaCO2), bicarbonate (HCO3) and base excess (BE) – was performed. These parameters were evaluated in both groups at time point zero, before CO2 insufflation; at time point one (TP1), when intra-abdominal pressure of 12 mmHg was reached in both groups; at time point two (TP2), 5 minutes after reaching intra-abdominal pressure of 12 mmHg in group P12 and of 20 mmHg in group P20; and at time point three (TP3), 10 minutes after reaching intra-abdominal pressure of 12 mmHg in group P12 and 10 minutes after TP1 in group P20, when intra-abdominal pressure decreased from 20 mmHg to 12 mmHg. Values out of the normal range or the occurrence of atypical phenomena suggestive of organic disease indicated clinical changes.

Results: Significant variations in MAP, pH, HCO3 and BE were observed in group P20; these changes, however, were within normal limits. Clinical changes were also within normal limits, and no pathological phenomena were observed.

Conclusions: Brief, intra-abdominal hypertension for the insertion first trocar insertion causes variations in MAP, pH, HCO3 and BE without adverse effects, and it may protect from iatrogenic injury

Relationship of arterial and exhaled CO2 during elevated artificial pneumoperitoneum pressure for introduction of the first trocar.

The present study evaluated the correlation between arterial CO2 and exhaled CO2 during brief high-pressure pneumoperitoneum. Patients were randomly distributed into two groups: P12 group (n=30) received a maximum intraperitoneal pressure of 12mmHg, and P20 group (n=37) received a maximum intraperitoneal pressure of 20mmHg. Arterial CO2 was evaluated by radial arterial catheter and exhaled CO2 was measured by capnometry at the following time points: before insufflation, once intraperitoneal pressure reached 12mmHg , 5 minutes after intraperitoneal pressure reached 12mmHg for the P12 group or 20mmHg for the P20 group, and 10 minutes after intraperitoneal pressure reached 12mmHg for the P12 group or when intraperitoneal pressure had decreased from 20mmHg to 12mmHg, for the P20 group. During brief durations of very high intraperitoneal pressure (20mmHg), there was a strong correlation between arterial CO2 and exhaled CO2. Capnometry can be effectively used to monitor patients during transient increases in artificial pneumoperitoneum pressure

Structural changes in intestinal enteroendocrine cells after ileal interposition in normal rats

INTRODUCTION: No therapeutic approach has significantly impacted the progression of diabetes. As early improvement of glicaemic control is observed after bariatric surgeries, there is currently a search for surgical procedures that can promote euglycemia also in non-obese patients. Glicaemic control can be achieved by increasing the blood concentration of GLP-1, a hormone produced by L cells that are more densely concentrated in the terminal ileum. The interposition of ileal segment to a more anterior region (proximal jejunum) can promote a greater stimulation of the L cells by poorly digested food, increasing the production of GLP-1 and reflecting on glicaemic control.
AIMS: To investigate long-term histological modifications of intestinal mucosa of rats submitted to interposition of ileum segment to a proximal region (jejunum).
METHODS: Forty 8-week old male Wistar-EPM1 rats (Rattus norvegicus albinus) were randomly distributed into 3 groups: the Interposition Group (IG) was subjected to ileal interposition, the Sham Group (SG) was subjected to sham operations, and the Control Group (CG) was not subjected to surgery. All animals were followed until the 60th postoperative day (8 postoperative week) when they were euthanized. Segments of jejunum and ileum from all groups were collected and analyzed by optical microscopy and immunohistochemistry.
RESULTS: No structural nor histological changes in intestinal L cells in the interposed intestinal segment and other intestinal segments were noted after ileal interposition surgery. 
CONCLUSION: As L cells endocrine characteristics were likely maintained, the use of metabolic surgical techniques for the treatment of metabolic diseases, especially diabetes, seems to be justified

Organic consequences of ileal transposition in rats with diet-induced obesity

INTRODUCTION: The clinical management of metabolic syndrome - especially diabetes mellitus type 2 - is notoriously complex due to the progressive nature of this disease. At present, there is a need for a surgical procedure that is effective for the treatment of diabetes mellitus type 2, even in non-obese individuals. The isolated ileal transposition theory could lead to an effective alternative therapy. This intervention has not yet been performed in humans, and there are no reports of its use in an experimental model of diet-induced metabolic syndrome.
 OBJECTIVES: The objective of this study is to evaluate the physiological effects of ileal transposition in rats with diet-induced metabolic syndrome. The effects of this procedure on glucose and lipid metabolism will be assessed. 
METHODS: Forty 12-week-old male rats (albino Rattus norvegicus, Wistar, 2BAW, heterogeneous) will be divided into four groups of 10 animals each: the ileal transposition group (TG) comprising animals on a hypercaloric-hyperlipidic diet; the sham group (SG) containing animals that receive the same diet and undergo the sham surgery; control group 1 (CG1), which will receive a hypercaloric-hyperlipidic diet and will not undergo surgery; and control group 2 (CG2), which will consume standard feed and will not undergo surgery. The surgeries will be performed in 20-week-old animals. Blood samples for laboratory testing will be collected from 12-week-old animals on the day of surgery and after eight postoperative weeks, following determination of the weights of the animals and the administration of anesthesia. The levels of serum glucose, insulin, triglycerides, total cholesterol and fractions, glucagon-like peptide-1, C-peptide and glycated hemoglobin will be assessed in all of the animals. The insulin tolerance test will be performed using PRISMA software, and insulin resistance will be calculated by the HOMA-IR indirect test. On specific days, two 20-week-old rats will be separated and randomly distributed in TG and SG. These animals will be followed until the eighth postoperative week. Subsequently, they will be euthanized, and the retroperitoneal and periepididymal fat deposits will be collected and weighed using a precision scale. In addition, the pancreas, liver and intestinal segments will be sent for pathological and immunohistochemical studies.
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Isolated ileal interposition in enteroendocrine L cells differentiation

INTRODUCTION: 
Due to the progressive nature of type 2 diabetes, its complexity and drug treatment perpetuity, there is currently a search for surgical procedures that can promote euglycemia also in non-obese patients. Diabetic patients glycemic control can be achieved by increasing the blood concentration of GLP-1, a hormone produced by L cells that are more densely concentrated in the terminal ileum. Early and extended improvement of diabetes in patients submitted to bariatric surgeries awakened the necessity of investigating the isolated ileal interposition as a surgical alternative for the treatment of diabetes. The interposition of this ileal segment to a more anterior region (proximal jejunum) can promote a greater stimulation of the L cells by poorly digested food, increasing the production of GLP-1 and reflecting on glycemic control. However, in order to consolidate the ileal interposition as a surgical treatment of diabetes it is necessary that the interposed ileum keep the same differentiation rate into L cells for a long period to justify the intervention.

AIMS: 
To investigate the isolated ileal interposition influence on the differentiation of intestinal precursor cells into enteroendocrine L cells over time.

METHODS: 
Twelve 12-week-old male Wistar rats (Rattus norvegicus albinus) of the WAB strain (heterogeneous) will be used. All animals will receive a high-calorie, high-fat diet for 16 weeks or more until they develop glucose dysmetabolism confirmed by glycemic test. They will be divided into two groups of 10 animals each: the isolated ileal interposition group (GI) and the control group (GC), comprising animals that will not be submitted to any surgical intervention. Blood samples will be collected under anesthesia at the weeks 12, 26, 36 and 44 for the determination of serum levels of glucose, insulin, GLP-1, glucagon, C-peptide and glycosilated hemoglobin. The insulin tolerance test will be performed and insulin resistance will be calculated. For the comparative analysis of the ileal precursor cells differentiation into enteroendocrine cells among the two groups, the following intestinal fragments will be collected after euthanasia: interposed ileum and remaining ileum from GI, jejunum and ileum from GC. These fragments will be analyzed by imunofluorescence and also by Real Time PCR using PCR Arrays for target genes including the main ones related to stem cell, stem cell signaling, diabetes, Wnt and Notch signaling pathways and other genes and pathways involved in the differentiation of intestinal precursor cells into enteroendocrine cells, especially GLP-1-producing L cells that play important role in euglycemia

Characterization of HPV16 virus-like particles produced in HEK293T cells

O HPV (Papilomavírus Humano) causa verrugas anogenitais e alguns tipos de câncer, como o câncer de colo do útero. Seu capsídeo é composto pelas proteínas L1 e L2. A L1 se autoestrutura em VLPs, utilizadas nas atuais vacinas comerciais. Para a geração de vacinas com maior espectro de proteção, a L2 é promissora, pois gera proteção cruzada. Para caracterizar VLPs de L1L2 do HPV16, células HEK293T cotransfectadas tiveram a expressão proteica analisada por citometria de fluxo, Western blotting, microscopia confocal a laser e eletrônica de transmissão. As proteínas L1 (60kDa) e L2 (100kDa) estavam presentes no núcleo e no citoplasma celular, formando VLPs de L1L2 de conformação heterogênea, cuja máxima expressão ocorre 12h após a transfecção. As VLPs extraídas estavam em diferentes estágios de estruturação. Foi possível estabelecer um sistema eficaz de produção heteróloga das proteínas de VLPs de L1L2, que poderão ser utilizadas em testes pré-clínicos, na pesquisa básica e contribuir no desenvolvimento de uma vacina profilática de amplo espectro de ação contra o HPV.HPV (Human Papillomavirus) is the causative agent of anogenital warts and several types of cancer, as cervical cancer. The HPV capsid is composed of L1 and L2 proteins. L1 can self-assemble into VLPs (virus-like particles), the basis for HPV commercially available vaccines. To generate broad-spectrum vaccines L2 shows the greatest promise, due to cross-protection. To characterize VLPs composed of HPV16 L1 and L2 proteins, cotransfected HEK293T cells were analyzed by flow cytometry, confocal laser scanning microscopy, transmission electron microscopy and Western blotting. Proteins L1 (60kDa) and L2 (100kDa) were present in cell nucleus and cytoplasm, forming heterologous L1L2 VLPs with highest expression at 12h post-transfection. Extracted VLPs were at different maturation stages. It was possible to establish an efficient system of heterologous L1, L2 and L1L2 VLPs production. After some adjustments in protocols, these particles could be used in preclinical tests, HPV basic research and also in the development of an HPV broad-spectrum vaccine