Magnetically-driven 2D cells organization on superparamagnetic micromagnets fabricated by laser direct writing.

We demonstrate a proof of concept for magnetically-driven 2D cells organization on superparamagnetic micromagnets fabricated by laser direct writing via two photon polymerization (LDW via TPP) of a photopolymerizable superparamagnetic composite. The composite consisted of a commercially available, biocompatible photopolymer (Ormocore) mixed with 4 mg/mL superparamagnetic nanoparticles (MNPs). The micromagnets were designed in the shape of squares with 70 µm lateral dimension. To minimize the role of topographical cues on the cellular attachment, we fabricated 2D microarrays similar with a chessboard: the superparamagnetic micromagnets alternated with non-magnetic areas of identical shape and lateral size as the micromagnets, made from Ormocore by LDW via TPP. The height difference between the superparamagnetic and non-magnetic areas was of ~ 6 µm. In the absence of a static magnetic field, MNPs-free fibroblasts attached uniformly on the entire 2D microarray, with no preference for the superparamagnetic or non-magnetic areas. Under a static magnetic field of 1.3 T, the fibroblasts attached exclusively on the superparamagnetic micromagnets, resulting a precise 2D cell organization on the chessboard-like microarray. The described method has significant potential for fabricating biocompatible micromagnets with well-defined geometries for building skin grafts adapted for optimum tissue integration, starting from single cell manipulation up to the engineering of whole tissues.

Impact of Hysterectomy on Urethral Suspension with Transobturator Sling for Treatment of Stress Urinary Incontinence in Women.

Urinary stress incontinence is a condition that increases with age, influenced by many factors, mainly anatomical pathological ones, determined by changes of muscular pelvic floor, but also by hormonal and local neurological modifications. The aim of the study was an overview of the efficacy of stress urinary incontinencetherapy with transobturatorurethralsling suspension in patients with or without prior hysterectomy. Additionally, we evaluated the effectiveness and the rate of side effects of this method. The study group consisted of 100 patients and, within it, 32 patients (group A)underwent a hysterectomy.We noticed an improvement of symptoms, a reduced discomfort and an increase of quality of life, more prominent in group B (patients without previous hysterectomy) (p=0,0012, p=0,17 respectively, p=0,03). We consider that transobturator approach in treating stress urinary incontinence is a simple, rapid and effective procedure and, not lastly, modern technique that assures a significant improvement of symptomsand quality of life for the patients; positioning the sling along to the insufficient pubourethral ligaments reinforces the structures supporting the urethra; this approach is minimally-invasive with rapid recovery, reduced hospitalization, is cost effective and involves minimal risks of infection, bleeding, etc. moreover, the transobturator tension free sling has some advantages (no pain or tension). The rate of success of the treatment is significantly lower in patients with prior hysterectomy.

Water quality of Danube Delta systems: ecological status and prediction using machine-learning algorithms.

Environmental issues have a worldwide impact on water bodies, including the Danube Delta, the largest European wetland. The Water Framework Directive (2000/60/EC) implementation operates toward solving environmental issues from European and national level. As a consequence, the water quality and the biocenosis structure was altered, especially the composition of the macro invertebrate community which is closely related to habitat and substrate heterogeneity. This study aims to assess the ecological status of Southern Branch of the Danube Delta, Saint Gheorghe, using benthic fauna and a computational method as an alternative for monitoring the water quality in real time. The analysis of spatial and temporal variability of unicriterial and multicriterial indices were used to assess the current status of aquatic systems. In addition, chemical status was characterized. Coliform bacteria and several chemical parameters were used to feed machine-learning (ML) algorithms to simulate a real-time classification method. Overall, the assessment of the water bodies indicated a moderate ecological status based on the biological quality elements or a good ecological status based on chemical and ML algorithms criteria.

Histopathological Study on Conservatively Operated Breast Carcinomas.

In this histopathological study we looked at 303 cases of breast carcinomas, managed though conservative breast surgery and later analysed with the help of a classical histopathological technique, paraffin embedding. The carcinomas were assessed in terms of tumor size, lymph node status, histological type, correlation between invasive tumors and an situ carcinoma component, resection margins, grading and patients age. Following assessment, we looked at associations between above morphological and clinical parameters and ipsilateral local recurrences. We concluded that more than half of our cases were carcinomas, measuring between 2 cm and 5 cm, with no associated lymph node involvement, in keeping with pTNM criteria for stage II. By far, in our study, the most frequent histopathological type was type NOS (63.37%) followed by invasive lobular carcinoma (10.56%) and mixed ducto-lobular invasive carcinoma (6.27%). Other types of invasive carcinoma were rarer, each representing less than 4% of cases. In regards to in situ carcinomas we noted the most common histological types to be both cribriform intraductal carcinoma and comedocarcinoma, each identified in 1.65% of cases. Amongst invasive breast carcinomas, infiltrating ductal carcinoma not otherwise specified (NOS) was found to be most commonly associated with in situ ductal carcinoma lesions. This was seen in 34.9% of cases, and was the only type associated with an extensive in situ component. Analysing the grading of mammary carcinomas in our study showed that the vast majority of cases (63.04%) were grade 3 tumors. In regards to surgical resection margins, ¾ of cases were noted to have negative margins. Tumor recurrences were noted in 12 cases. These cases were most commonly noted to reoccur following initial poorly differentiated, infiltrating ductal carcinomas, not otherwise specified (NOS), with positive resection margins, measuring less than 2 cm. Patiens tended to be under the age of 40 and had positive lymph nodes. The emergence of local recurrences after conservative surgery for early breast cancer is singnificantly linked to poorly differentiated primary tumors (p <0.05) but not correlated with histological type, presence of extensive intraductal carcinoma component, size of primary breast tumor or lymph node status ( p> 0.05). In terms of increasing the risk of ipsilateral recurrence the most important aspect highlighted in our sudy was the status of the resection margins. Patients with positive resection margins had a significantly high risk to develop recurrences after the conservative surgery, compared to those with negative margins (p <0.001).

Accurate Diagnosis of Sigmoid Colon Endometriosis by Immunohistochemistry and Transmission Electron Microscopy - A Case Report.

Endometriosis is described as the presence of functioning endometrial tissue at sites outside the uterus. Up to 15% ofwomen in their reproductive period are affected by this condition. Endometriosis is mostly foundon the uterosacral ligaments, inside the rectovaginalseptum or vagina, in the rectosigmoid area, ovarianfossa, pelvic peritoneum, ureters, and bladder, causinga distortion of the pelvic anatomy. Colonic involvement is rare but is usually found at the level of the rectum or the sigmoid colon. Acute presentation with intestinal obstruction or perforation is rare. While malignant transformation of endometrial lesions is rare, findings of dysplasia on pathology sections can give rise to questions of management. Immunohistochemistry and electron microscopy can help decision making. We present the case of a 38 year old woman with intestinal obstruction caused by sigmoid colon endometriosis with moderate dysplasia in which transmission electron microscopy was used for postoperative diagnosis. Detailed analysis of these cases, while logistically difficult, can prove useful in understanding the etiology and pathophysiology of the disease.

Investigation of osteoblast cells behavior in polymeric 3D micropatterned scaffolds using digital holographic microscopy.

The effect of micropatterned polymeric scaffolds on the features of the cultured cells at different time intervals after seeding was investigated by digital holographic microscopy. Both parallel and perpendicular walls, with different heights, were fabricated using two-photon lithography on photopolymers. The walls were subsequently coated with polypyrrole-based thin films using the matrix assisted pulsed laser evaporation technique. Osteoblast-like cells, MG-63 line, were cultured on these polymeric 3D micropatterned scaffolds. To analyze these scaffolds with/without cultured cells, an inverted digital holographic microscope, which provides 3D images, was used. Information about the samples' refractive indices and heights was obtained from the phase shift introduced in the optical path. Characteristics of cell adhesion, alignment, orientation, and morphology as a function of the wall heights and time from seeding were highlighted.

Novel method for proliferation of oral keratinocyte stem cells.

BACKGROUND AND OBJECTIVE: Stem cell-based tissue engineering offers clear advantages over conventional normal cell approaches. Owing to their specific characteristics, oral keratinocyte stem cells represent an attractive solution for therapeutic applications. However, when cultured in vitro, these cells lose their unique properties, acquiring a limited capacity for self-renewal, and differentiate rapidly into normal functional keratinocytes. The main aim of the present study was to develop an in-vitro method for the expansion of oral keratinocyte stem cells using a biomaterial approach. MATERIAL AND METHODS: Oral keratinocyte stem cells were isolated based on the identification of two surface markers - integrin α6ß4 and CD71 - using a magnetic method. The cells were cultured on specific substrates formed from blends of polymers: poly(lactide-co-glycolide) (PLGA); poly(lactide-co-glycolide) + polyurethane (PLGA + PU); and poly(lactide-co-glycolide) + extracellular matrix (PLGA + ECM). The polymers were deposited using a laser-based technique - matrix-assisted pulsed laser evaporation. The cells were analyzed for cell size, cell proliferation, colony-forming efficiency, cell adhesion markers (such as E-cadherin and beta 1 integrin), keratinocyte stem cells and differentiation markers. The methods included ELISAs, immunofluorescence and atomic force microscopy imaging. RESULTS: After 14 d in culture, cells seeded on PLGA + PU stained positive for p63, cd44H, cytokeratin 19 and integrin α6ß4 and negative for involucrin, cytokeratin 14 and cytokeratin 10. The levels of adhesion molecules were significantly increased in cells grown on PLGA + PU: at 14 d the E-cadherin levels were 5.4 ± 0.2 ng/mL (for cells grown on PLGA + PU) vs. 4.1 ± 0.4 ng/mL (for cells grown on control medium) (n = 5, p < 0.05 Bonferroni). Oral keratinocyte stem cells grown on PLGA + PU had the highest colony-forming efficiency and proliferation rate, together with the smallest cell size, compared with cells grown on control medium or other polymeric substrates. CONCLUSION: The present study demonstrates that by culturing oral keratinocyte stem cells on PLGA blended with PU it is possible to preserve their phenotype in vitro and to guide their short-term expansion and proliferation. Certain stem-cell characteristics are preserved and their short-term expansion may be enhanced.

Breast-conserving surgery in breast cancer.

The breast cancer treatment is based nowadays on new surgical options: breast-conserving surgery, which applies at least for the first and second stage cancer, with radical intention. We have been practicing breast-conserving surgery for the last 16 years and we have performed 303 breast conserving operations from a total of 673. We recorded 12 local recurrences (3,96%) and 2 deaths due to cancer progression. Our protocol includes removal of the primary tumor with enough surrounding tissue to ensure negative margins of the resectable specimen, associated with axillary lymph-node dissection and postoperative breast irradiation. Our oncologist indicated chemotherapy on different postoperative conditions: tumor size, axillary lymph node involvement, patient's age, etc. The purpose of this paper is to emphasize our modest experience, nevertheless to draw the attention on important results, obtained by long-term monitoring of the patients who underwent breast-conserving surgery, in a two prospective protocols, and demonstrate the importance and applicability of breast conserving therapy. The conclusion of this study is that breast-conserving surgery followed by breast irradiation is reliable, as the results are similar with radical mastectomies; the main objective is to obtain a good cosmetic result, which depends on tumor size / breast size.

[Diseases mimicking advanced-stage epithelial ovarian cancer]. / Afectiuni mimând cancerul ovarian epitelial în stadiu avansat.

This paper draws attention towards 3 cases with different pathologies all of which suggesting however both clinically and by imaging means as the most likely diagnosis advanced-stage epithelial ovarian cancer since all these three postmenopausal women had been admitted to the hospital with ascites, pelvic masses and deterioration of the physical wellbeing (fatigue, decreased appetite, weight loss, pallor). Findings during exploratory laparotomy on all these three pacients included ascites (hemorragic in one case) diffuse tumorous implants throughout the abdominal and pelvic peritoneal surfaces (in two cases) and the ovarian tumour. Postoperatively, the final histopathologic diagnoses consisted of primary peritoneal carcinoma (one pacient), peritoneal tuberculosis (TB, one pacient) and hepatic cirrosis with an incidental benign adnexial mass (one pacient). Moreover, nonmalignant ovarian tumours were certified in all three cases under current presentation. The differential diagnosis of the ovarian cancer and a tailored approach to treatment for each of these three pathologic entities will also be described in detail.

Cell imaging by coherent backscattering microscopy using frequency-shifted optical feedback in a microchip laser.

In this work, we present a new development of the laser optical feedback imaging technique for imaging biological structures with a high resolution. The first results obtained on human red blood cells and mice cerebral and muscular tissues slices are shown. The performances of the system and its future developments are also discussed.

[Stress urinary incontinence. Diagnostic and therapeutic aspects]. / Incontinenta urinara de efort. Aspecte diagnostice si terapeutice.

OBJECTIVE: The paper analyses the incidence, diagnosis and treatment options available for stress urinary incontinence (SUI) in women with pelvic floor dysfunction admitted to Craiova's Surgery Clinic IV. METHODS: This is a retrospective 10-year study comprising a surgical cohort of 420 patients with significant enough to alter quality of life SUI associated to ureterocele and cystocele and in 353 cases with rectocele too. The highest incidence of SUI was encountered between 50 and 59 years of age (range 39 - 81 years). In 21 of this case series the diagnosis of SUI was established soon after the surgical repair of the urethro-cystocele. The diagnosis of SUI was based on careful history and physical examination with emphasis on the gynecologic survey of the abdomen and pelvis but in the absence (for objective reasons) of urodynamic testing which is especially useful for SUI pathophysiological evaluation and thus surgery success rate prediction. All our 420 severe SUI associated with vaginal wall hernias underwent surgical treatment by either open Burch retropubic urethropexy or anterior colporraphy. RESULTS: Among anterior colporraphy treated patients SUI persisted in 19.3% of the cases (33 patients). Complications of Burch urethropexy procedure (despite its high ability for cure) in our case series include: urinary retention, hemorrhage into the space of Retzius, intraoperative injury to the bladder and long-term postoperative incisional hernia. Moreover, 5 patients (2%) of the group who underwent Burch operation were readmitted with recurrent urinary incontinence between 2 and 6 months after the aforementioned surgical intervention despite its good anatomical results in all of these cases. CONCLUSIONS: SUI is a prevalent disorder of women that can be diagnosed easily with history and physical exam. If symptoms persist and severely affect quality of life, despite modern noninvasive treatments, several surgical procedures are now available.

[The mirage of the first lesion (gallstones) and laparoscopic cholecystectomy are able to defer the diagnosis of colon cancer]. / Mirajul primei leziuni (litiaza veziculara) si colecistectomia laparoscopica pot întârzia diagnosticul de cancer colic.

The study's aim was to analyze a series of colon cancer cases in which the mirage of the first (clinically most obvious) lesion (gallstones) along with its minimally invasive approach - that explored only the biliary disease - had contributed to the delay of large bowel malignancy' diagnosis and treatment. 1327 patients aged between 17 and 83 years and diagnosed with cholecystolithiasis were operated upon laparoscopically in the Department of General Surgery of Craiova CFR University Hospital from 2000 through 2004. Four out of these 1327 patients (0,3%) were readmitted with the diagnosis of colon carcinoma between 1 and 16 months after the laparoscopic cholecystectomy. Our retrospective study gives a full report on these 4 cases insisting upon the links between their clinical - laboratory evaluations and final diagnosis. Despite the low laparoscopic cholecystectomy overlooked colon cancer' incidence it seems reasonable to both improve the technique of peritoneal cavity exploration during this type of surgery and extend the preoperative evaluation whenever the slightest suspicion of associated pathology is raised especially in patients over 50 years of age.

[The experience at the 4th Surgery Department of the University Hospital C.F. Craiova in the surgical treatment of inguinal hernia]. / Experienta Clinicii Chirurgicale IV a Spitalului Universitar C. F. Craiova în folosirea plaselor sintetice în tratamentul chirurgical al herniilor inghinale.

This paper aim is to present the experience of Surgery Department IV of University Hospital C. R. Craiova in groin hernias treatment using prosthetic meshes, also describing an original technical procedure of mesh-plasty that we have been practicing successfully in our clinic. The study is based on a number of 1757 groin hernias operated in Surgery Department IV of University Hospital C. F. Craiova during a period of 11 years (1993-2003). There have been used prosthetic meshes in a number of 230 hernias operated for the most part in the last years since mesh repair has become habitually. We have been using so far only prosthetic mesh made in Romania (polyester mesh). We had a single recurrent hernia and the immediate complications were minimal. The study refers us to practice prosthetic mesh repair in an extensive way and offers a technical alternative in using of prosthetic meshes by a simple and efficient procedure with good postoperative results.

[Ovarian cancer--diagnostic dilemmas and unusual therapeutic response]. / Cancerul de ovar între dileme de diagnostic si "capriciile" raspunsului terapeutic.

The aims of this paper are both to highlight some dilemmas concerning the diagnosis of primary epithelial ovarian carcinoma mainly in its early stages and to underline the capricious responses of this type of malignancy to an otherwise well coded modern management. This study is based on the analysis of the records of 78 patients with ovarian carcinoma admitted for diagnosis and surgical treatment to the Department of General Surgery of Craiova C.F.R. Clinic from 1993 through 2003. The results of this analysis are difficult to interpret due to loss to follow up (in terms of response rates) of some of our 78 operated on ovarian carcinoma patients who went on with their platinum-based chemotherapy (following surgical cytoreduction) under the supervision of different Oncology Departments nationwide. Nevertheless, it is worth mentioning that most of this study patients (71.9%) presented with advanced-stage (III and IV) ovarian carcinoma which sometimes seemed quite confusing by its clinical polymorphism but its prognosis was very much related to both the degree of surgical cytoreduction accomplished and tumor sensitivity to chemotherapy. Finally, although this study does not allow us to draw firm conclusions it is an attempt to share out our current perception on the primary epithelial ovarian cancer management.

[Locoregionally advanced rectosigmoid cancer: diagnosis, surgical approach, late results]. / Neoplasmul rectosigmoidian avansat locoregional. Probleme de diagnostic, rezultate la distanta, atitudine terapeutica chirurgicala.

The colorectal cancer continues to be diagnosed in advanced stages in our country, mainly due to unapplying of a programmer of active diagnosis through screening on the population with risk for colorectal cancer, and inefficiency of primary care system. In the department of General Surgery CFR Craiova Hospital between 1991-2001 were operated a number of 231 patients with colon cancer and 104 patients with rectal cancer. The results, showing an increased number of recurrences in cases of resection performed for advanced loco-regional tumors of rectosigmoid, made us to reconsider the attitude of avoiding the abdominoperineal resection even when the distance between the inferior limits of the tumors and the anal edge exceeded the distance considered being standard for a low anastomosis performing. The follow-up of the patients with paraclinic technique that didn't prove efficient led in the most cases to a delaying in diagnosis of local recurrences until the moment of resectability was exceeded. The applying of efficient methods in early diagnosis of colorectal cancer and follow-up could provide in the future better results for anterior resections with low anastomosis.

[Clinical course of an ileal carcinoid with mesenteric metastasis]. / Evolutie particulara a unui carcinoid ileal cu metastaza ganglionara în baza mezenterului.

This paper aim is to present the case of a male patient, age 63 admitted in hospital for non-specific gastroenterologic symptoms easily attributed to colecyst calculosis; the anamnesis and careful clinical examination have avoided a useless cholecystectomy and permitted to establish indication for laparotomy; during that was found an ileal tumor with massive metastasis in the base of mesentery. The surgical attitude was dictated by histological diagnostic difficulties (prostate adenocarcinoma metastasis), so it has given up the idea of tumor excision (unjustified intraoperative risk for a metastasis), performing a bypass of the ileal tumor by an enteroenterostomy, then the patient was sent to the Oncological Department. Postoperative evaluation of the patient finally permitted to establish the neuroendocrine origin of the tumor; after oncological treatment (chemotherapy), about 10 months after operation the patient didn't present any sign of tumor (tumor markers and CT scan within normal). The paper emphasizes the problem of practicing an aggressive surgical treatment for carcinoid intestinal tumors with mesenteric metastasis but also brings attention to nonsymptomatic forms of cholecyst calculosis that may hide associated malignant tumors undetected by laparoscopic cholecystectomy.

[Breast conserving surgery --7 years of experience]. / Chirurgia conservatoare a sânului--7 ani de experienta.

The breast cancer treatment is based nowadays on a new surgical option: breast-conserving surgery, which is reliable at least for the first and second stage of cancer, with radical intention, obviously. We have started to practice the breast-conserving surgery in our surgical clinic (at CFR Hospital, from Craiova) for 7 years; until now we have performed 159 breast-conserving operation and, as results, we have recorded 3 local recurrences (2.12%) and 1 death due to cancer evolution. Our protocol includes removal of the primary tumor with enough surrounding tissue to ensure negative margins of resectable specimen, associated with total axillary lymph-node dissection and postoperative breast irradiation. Our oncologist on different postoperative conditions indicated the chemotherapy: tumor size, axillary lymph node involvement, patient age, etc. The purpose of this paper is to emphasize our unassuming experience but especially to draw attention on important results, obtained by long-term monitoring the patient who underwent breast-conserving surgery, in a two prospective protocols, which demonstrate the importance and applicability of breast-conserving therapy. The conclusion of this study is that breast-conserving surgery followed by breast irradiation is reliable, as the results are similar with the radical mastectomies; the main objective is to obtain a good cosmetic result, which depends on tumor size/size of the breast ratio.

[Tumour-host interaction and prognosis in patients with gastric cancer]. / Aspecte ale relatiei tumora-gazda cu implicatii în aprecierea prognosticului la pacientii cu cancer gastric.

The study analyzed the prognostic significance of a group of three histologic markers belonging to intra- and peritumorous stroma as derived from a series of 66 patients with gastric adenocarcinoma operated upon in Craiova CFR General Surgery Clinic. To this end we attempted to uncover any possible correlation between the three stromal parameters represented by type of angiogenesis, peritumorous inflammatory infiltrate and desmoplastic reaction and both the depth of malignant invasion through the gastric wall and the histologic type of gastric cancer. The results of our investigation highlighted that type A2 angiogenesis is displayed mostly by gastric carcinomas with well-differentiated tubular and secretory structure (class 2 Goseky) whereas type A3, angiogenesis is mainly a feature of gastric cancer with a poorly-differentiated tubular morphology (classes 3 and 4 Goseky). Moreover there was no statistically significant correlation between the intensity of inflammatory infiltrate and both the Goseky classes and the depth of neoplastic spread. Finally a clearly desmoplastic reaction was encountered in less than 50% of our patient series and represented almost a characteristic of both poorly-differentiated tubular structured malignancies and serosa invading tumors.

[Functional secondary megaduodenum]. / Megaduoden secundar functional.

This paper aim is to present the case of a 72 year old male, diagnosed with functional secondary megaduodenum. We intend to discuss the pathogenic and positive diagnostic difficulties (the presence or absence of a mechanical obstructive factor or the participated of the megaduodenum in an intestinal pseudo-obstruction syndrome). Also we outline the rare frequency of this disease and especially the therapeutic difficulties: inefficiency of conservatory treatment, delicate problems of surgical tactics and technique.

[Microbiology of nosocomial infections in a general surgery department]. / Microbiologia infectiilor de spital în clinica de chirurgie generala.

This paper aim is to outline the importance of nosocomial infections, characterized by great incidence, great mortality rate and specific bacteriology, in a surgery clinic. The study include 566 patients that developed 665 nosocomial infections (10.65% incidence), among the 5950 patients that underwent surgical operations in 5 years (1992-1996); 54 patients developed two or three nosocomial infections, which explain the incidence of nosocomial infection greater than number of patients. From bacteriological point of view predominance of Gram negative bacilli (especially E. coli) and pathogen staphylococcus characterized the nosocomial infections. The infection source was the patient himself, previously colonized with hospital specific microorganisms. Every clinical form of nosocomial infections was characterized by the present of specific pathogen microorganisms; the knowledge of these pathogen agents is very important for the antibiotic treatment applied before the bacteriological exam.