A small scale oxygenator for cardiopulmonary bypass in rats.

Cardiopulmonary bypass (CPB) devices replace transiently the function of both heart and lungs, allowing the investigator to work safely on a stopped heart. Although this technology has greatly improved since its first applications in the early 1950's there is still no definitive rat CPB model, various experiments reporting drawbacks like pulmonary edema, large priming volumes,etc. We present a new oxygenator that can be used in cardiopulmonary bypass experiments in rats, simple in design and efficient in function,in which the process of blood oxygenation takes place in a vertical cylinder filled up with air or oxygen, with blood being spread onto the wall and then trickling down in a thin layer that facilitates the oxygen transfer. The oxygenation is efficient, the pO2 reaches levels of almost 150 mmHg (physiological level is around 100 mmHg) in conditions of oxygen saturation of over 99% (normal levels 95%-98%).

Identification of a novel subset of T lymphocytes in patients with Balkanic nephropathy.

Balkanic Nephropathy (BN) is characterized by: an incidence limited to a geographic area: a familial character and a slow progressive evoluation towards chronic renal failure associated with the symmetrical reduction of the kidney size. The etiology of BN is unknown. The aim of our study was to find out the immune alterations in BN pathology. In the BN patients we identified a novel subset of the CD3+ CD16+ and CD56+ T cells expressing the phenotypic characteristics of both T lymphocytes and NK cells. The analysis of various subpopulations of lymphocytes, however, showed no quantitative differences in comparison with healthy subjects and healthy subjects from the endemic area.

P-glycoprotein mediated multidrug resistance assessment by flow-cytometry in malignant hemopathies.

A relatively common and frequent form of multidrug resistance(MDR) in cancer cells is due to membrane overexpression of P-glycoprotein. Mdr phenotype was investigated by flow-cytometry in several types of malignant hemopathies -chronic lymphocytic leukemia, non-Hodgkin lymphomas, acute lymphoblastic and myeloblastic leukemias. We used daunomycin and fluo-3 as fluorochromes, and verapamil as reversor agent. The method is lacking unitary clinical parametrization and in order to improve it, we tried to establish an optimal concentration of verapamil, which was shown to be 14.92 micrograms/ml. The reliability of results obtained with fluo-3 in culture media containing Ca2+ is questionable, as low variations in the intracellular level of this ion dramatically influences light emission by the fluorochrome and possibly the function of P-gp. To avoid such fluorescence intensity variations, Ca(2+)-free cell culture medium for fluo-3-based flow-cytometric assay is suggested to be used.

Balkan endemic nephropathy, the haematopoietic system and the environmental connection.

We previously reported the detection of an increased subpopulation of cytotoxic T lymphocytes in patients with Balkan (endemic) nephropathy (BEN) and in area controls (individuals free of clinical syndrome but born in a BEN endemic area and having a family history of BEN). Extending the flow-cytometric analyses to other populations of peripheral blood leucocytes, we found a decrease in the proportion of B lymphocyte subset and an increased proportion of eosinophils in BEN patients and in area controls. Although these numerical alterations cannot be categorically linked to the aetiopathogeny of the disease, it is presumed that they can be induced by the same factor(s) causing the kidney damage, through a direct haemato- and lymphotoxic effect.

Surgical treatment of pericyst in liver hydatid cysts.

The results of the surgical treatment of the pericyst in 94 patients with liver hydatid cyst are analyzed. Hepatic resections were performed in 2 patients, subtotal pericystectomy in 2 patients, partial pericystectomy in 65 patients, resection of the cyst dome in 9 patients and pericystodigestivostomy in 16 patients. The postoperative complications were statistically significant in complicated cysts (p < 0.001), in those with rigid walls (p < 0.001) and when lacking the adequate drainage (p < 0.01). The mean period of hospitalization was of 14 days in patients with supple pericyst and 40.1 days in those with rigid pericyst. Only one patient died of pulmonary embolism. In the absence of modern means of investigations and technical equipment in the treatment of the liver hydatid cyst, which is a benign disease, we have applied much simpler methods, but without vital risks.

DNA topology and V(D)J recombination: a tentative connection.

Gene recombination is the fundamental basis of immunoglobulin (Ig) and T cell receptor (TCR) diversity. Although several specific and nonspecific enzymatic equipments were revealed to be necessary for Ig and TcR gene assembly, almost nothing is known about the developmental and tissue specific control of recombination and the individual functions of the heptameric and nonameric signals and 12/23 spacers in this process. According to certain conformational and functional respects, we consider the nonamer a DNA insertion site to the nuclear scaffold, in relation with its structural homology to the satellite (5'-ACAAACC-3') and microsatellite repetitions, involved in DNA-nucleoskeleton impact. A topological control for V(D)J recombination is proposed, through different accessibilities of the substrates in the catalytic site, defined by a specific nonamer-mediated insertion to the nuclear scaffold. Recognition of heptamer and nonamer sequences by RAG proteins is followed by the assembly of an asymmetric recombinant complex. Even more important in this assembly may be the role of nonamer which, through DNA flexibilization and bending, could participate at the formation of the enzyme core. This core with the attached DNA could have a nucleosome-like geometry, a motif present in certain DNA processing enzymatic systems. Such an assumption emerges from the close homology of the nonamers with the DNA mobilization intergenic sequences (CA5-6T), found in many eukaryotic organisms.

[Predictive value of diagnostic tests in polytrauma patients with abdominal and craniocerebral injuries]. / Valoarea predictiva a testelor de diagnostic la politraumatizatii cu leziuni abdominale si cranio-cerebrale.

In a study on 1024 polytrauma patients with craniocerebral injury, abdominal injury was present in 206 (20.11%) of them. In order to identify the intraabdominal organs injury, physical examination and specific diagnostic tests will be used. Their value was calculated by means of informational indices. The main indices, sensitivity, positive predictive value and accuracy were: for physical examination = 81.06%, 90.3%, 94.72%; for peritoneal lavage = 94.17%, 95.12%, 97.94%; for abdominal computed tomography = 98.44%, 97.65% and 99.25%. As one might expect the abdominal computed tomography is the most accurate method for the diagnosis of intraabdominal organs injuries.

[Risk factors for the immediate outcome of gastrointestinal bleeding in patients with cirrhosis]. / Factori de risc in evolutia imediata a hemoragiei digestive la cirotici.

The risk factors in the immediate outcome of gastrointestinal bleeding were examined in a prospective study of 134 patients with liver cirrhosis. The hemorrhagic episode has a negative prognosis in the immediate outcome (p < 0.01), recording 48 (35.82%) deaths. The deaths occurred in the acute phase of bleeding (n = 18; 13.40%), after recurrent hemorrhage (n = 24; 17.23%) and in 6 other cases (4.47%) after sclerotherapy or surgical treatment of varicosities. After varicose veins sclerotherapy (n = 8), 2 deaths were recorded and 4 other after surgery (n = 22). The advanced stage of cirrhosis, Child C stage, (n = 62) was accompanied by most of deaths (n = 38; 61.29%) (p < 0.01). Among the risk factors responsible for death, there were recorded: encephalopathy (p < 0.05), jaundice (p < 0.01), altered general status ((p < 0.01) and increased seric level of bilirubin over 3 mg% (p < 0.001). Jaundice is among the risk factors with the highest sensitivity (83.33%) and seric bilirubin over 3 mg% has the highest positive predictive value (64.28%).

Molecular mechanisms in the pathogenesis of sepsis.

Innate immune system is a universal form of host defense against infections. The recognition of the innate immunity is based on a limited number of encoded receptors that have evolved to recognize microbial metabolism products. The recognition of these molecular structures allows the immune system to distinguish its own infectious components from non-communicable structures. The immune suppression is a hallmark of sepsis. The complement system is activated in the early stages of sepsis, generating large amounts of anaphylatoxin C5a. Complement and TLRs (toll-like receptors) family are two major upstream sensors and effectors systems of innate immunity. It was found that TLR4 and complement system are involved in the initiation of the inflammatory response in sepsis. Clinical studies in which TLR4 was blocked have not shown beneficial effects. TLRs, that are a subfamily of PRRs (pattern recognition receptors), have emerged as the crucial receptors for the recognition of DAMPs (Damage-associated molecular pattern molecules). Recently, a special form of non-coding genetic material called microRNA has been highlighted in the complex cascade of sepsis. The individual role of every microRNA and the exact role of microRNA network are under investigation. Currently, studies are performed in order to find micro RNA to be used as biomarkers of sepsis. Researches are performed to determine microRNA, small fragments of non-coding RNA, in order to distinguish between patients with sepsis and healthy patients, and if the plasma levels of microRNA correlate with the severity of the disease. Recent researches report that the regulation of gene expression through microRNA plays a very important role in the following cellular processes, for example: apoptosis, the differentiation process, and the cell cycle.

Proinflammatory cytokines in peritonitis.

RATIONALE: We performed this study with the purpose of revealing different aspects of the systemic inflammatory response syndrome in peritonitis. OBJECTIVES: The aim of the presentation was to make a research on some of the immune response mediators in secondary peritonitis and to observe their capacity to anticipate the evolution towards septic complications. METHODS AND RESULTS: We have undertaken a study on a group of 100 patients with acute diffuse peritonitis, between 2009 and 2011, in which we have accomplished the dosage of IL-1 beta, IL-6 and TNF alpha cytokines in the serum of patients, in dynamics, for 7 days by using the Elisa method. Subsequently, we have compared the results to the ones of a control group. The data obtained indicated high levels of proinflammatory cytokines in the patients who subsequently suffered an unfavorable evolution towards septic complications. DISCUSSION: The study of IL-1 beta, IL-6 and TNF alpha blood dynamics, offers valuable information about the severity of a systemic inflammatory response syndrome in peritonitis. They can be valuable biomarkers in establishing the unfavorable evolution of patients, helping the physician to establish a sustained and specific treatment, even from the early phases of the illness.

Endogenous phospholipid metabolite containing topical product inhibits ultraviolet light-induced inflammation and DNA damage in human skin.

BACKGROUND: N-palmitoylethanolamine (PEA) and organic osmolytes are endogenous components of the human epidermis and are generated from phospholipids in the stratum granulosum. PEA has been shown to exert potent antioxidant and anti-inflammatory activities. The endogenous organic osmolytes such as betaine and sarcosine control skin humidity, but have also been shown to inhibit ultraviolet (UV) light-induced oxidative stress in keratinocytes. OBJECTIVES: To investigate the effect of a PEA- and organic osmolyte-containing topical product (Physiogel AI) on the development of UV light-induced erythema, thymine dimer formation and p53 tumor suppressor gene activation, as well as intercellular adhesion molecule 1 (ICAM-1) and Ki67 expression in normal human skin. METHODS: The UV-induced erythema was measured by a spectrofluorometric method. Thymine dimers, p53, ICAM-1 and Ki67 were detected in skin biopsies using immunohistochemistry. RESULTS: Physiogel AI cream significantly inhibited the development of UV light-induced erythema and thymine dimer formation in normal human skin, but did not alter the number of Ki67+ proliferating keratinocytes and the expression of p53 and ICAM-1. CONCLUSIONS: Our results suggest that PEA and organic osmolytes might represent a new generation of compounds which suppress UV-induced photodamage.

A pressure-controlled rat ventilator with electronically preset respirations.

Major experimental surgery on laboratory animals requires adequate anesthesia and ventilation to keep the animal alive throughout the procedure. A ventilator is a machine that helps the anesthesized animal breathe through an endotracheal tube by pumping a volume of gas (oxygen, air, or other gaseous mixtures), comparable with the normal tidal volume, into the animal's lungs. There are two main categories of ventilators for small laboratory rodents: volume-controlled and pressure-controlled ones. The volume-controlled ventilator injects a preset volume into the animal's lungs, no matter the airways' resistance (with the peak inspiratory pressure allowed to vary), while the pressure ventilator controls the inspiratory pressure and allows the inspiratory volume to vary. Here we show a rat pressure ventilator with a simple expiratory valve that allows gas delivery through electronic expiration control and offers easy pressure monitoring and frequency change during ventilation.

Biliary involvement in the evolution of gastric and duodenal ulcer.

The complex clinical, biological and radiologic study of 1000 patients with gastric ulcer (GU) and duodenal ulcer (DU) showed biliary involvement (BI) in 141 (14.10%) cases, more frequently in women (57.44%) in the 30 to 39 years age group (35.76%). With reference to the onset or the clinical form, the frequency of BI increases with the duration of the ulcer. After the development of biliary distress, the clinical picture changes rendering the diagnosis more difficult. Ulcer being the cause of BI by functional, inflammatory and adhesion factors, a pathogenetic treatment is obligate.

Factors of prediction of immediate postoperative evolution in cancer of the colon.

A study carried out in 152 patients operated on for cancer of the colon showed that the early postoperative evolution of disease depends on the therapeutic method chosen according to the clinical stage of the tumor. The 31 deaths recorded in the first 30 postoperative days occurred mostly in the first week (45.1%) and were due either to hydro-electrolytic disturbances (69%) or to intraperitoneal septic complications (65%) with 0.66 and respectively 0.60 positive predictive value. Advanced forms of disease, emergency surgeries and presence of associated diseases are statistically significant risk factors.

The prevention of postoperative thromboembolic complications with small heparin doses.

For the prevention of postoperative thromboembolic complications (TEC) in patients with thromboembolic risk, heparin was administered subcutaneously in small doses, adapted to each patient. The efficiency of each method was confirmed clinically by the absence of TEC, verified by the administration of labelled fibrinogen and by the anatomopathologic examination. In contrast, in the control group there were 3.93% cases of TEC with 0.97% fatal pulmonary embolism. The local complications of heparin treatment depend upon the type of product administered. The method has advantages but also certain drawbacks that require the taking of all pre-, intra- and postoperative measures for prevention of TEC.

Immediate and late results in the surgical treatment of gastric cancer.

The study of 248 patients mostly males, aged over 50 years, operated on for gastric cancer (GC) has shown that the majority were in stages III and IV of disease. The antral localization of the tumor was the most frequent. In 6.45% of the cases acute complications occurred during the evolution of gastric cancer. Association with other diseases was found in proportion of 5.64%. The surgical treatment was differentiated according to the site and evolutive stage of the tumor but the most frequent was radical subtotal gastrectomy. The postoperative complications were either general or local. Postoperative mortality was higher in cancers situated in the upper third of the stomach, after total gastrectomy, and in stage IV of disease. The 5-year survival was 66.66% for stage II and 10.18% for stage III and the 10-year survival decreased to 40 and respectively 1.85%.

Factors predicting postoperative morbidity in biliary calculi.

Statistical analysis of a group of 1,114 patients operated for biliary lithiasis shows that the gravity of the disease, the nature of the complications that accompany it, the need to approach the common bile ducts (CBD) and age, stand at the origin of postoperative complications. To reduce postoperative morbidity elective surgery should be performed in cases of biliary lithiasis with paucity of symptoms during periods free of an attack and before the patient ages.

Immediate and late results in the surgical treatment of gastric ulcer.

The study of 205 patients showed that complicated gastric ulcer (GU) (55.57%) is followed by a high postoperative morbidity (8.78%) and mortality (1.95%). The good and very good results obtained by gastric resection (89.84%) confirm the value of this treatment in GU. The satisfactory (8.40%) and unsatisfactory (1.69%) results arise from errors of indications or deviation from the operative approach and technique. To obtain the best results the surgical act must be performed irreproachably and the patient must comply with protection prescriptions.

Influence of age on the surgical treatment of cancer of the colon.

The study of 152 patients with cancer of the colon (CC) showed that age represents a factor of risk in the evolution of disease having a decisive influence on the immediate and delayed results of the surgical treatment. Over two thirds of the cases with advanced clinical stages were detected in subjects over the age of 50. In young agers the evolution of CC was severe and metastases appeared early. In old agers the incidence of disease increased and postoperative mortality was maximum. The mean postoperative survival time in the extreme age groups (young and old agers) was short in all the clinical stages.

[Possibilities of early detection of postoperative venous thromboses]. / Posibilitati de depistare precoce a trombozelor venoase postoperatorii.

The use of labelled fibrinogen in 106 patients with thromboembolic risk led to the detection of thrombosis of deep veins of lower limbs in 25 patients (23,58%) as compared with a single patient (0,94%) who showed all the clinical signs. The thromboses were identified more frequently (in 60% of the cases) in patients whose age was above 60 years. In 80% of the cases the thromboses were detected in the first 24 h after surgery. Most frequently involved were the veins of the leg (64%), and especially in the IV-th area, corresponding to the upper third of the leg (23,80%). Surgery performed in the pelvic area gave a high percentage of thromboses. Advanced age, the existence of varicose veins, the presence of diabetes, of obesity, as well as previous surgical interventions, increase the risk of thrombosis and of embolies. Due to existing possibilities for an early diagnosis of thromboses in deep veins following surgery, for detecting latent clinical thrombosis, as well as for assessing the evolution of an already formed thrombus, it appears that the test with labelled fibrinogen is a highly useful clinical investigation.