Detection of human papillomavirus (HPV) DNA prevalence and p53 codon 72 (Arg72Pro) polymorphism in prostate cancer in a Greek group of patients.

Prostate cancer is the most common neoplasm found in males and the second most frequent cause of cancer-related mortality in males in Greece. Among other pathogens, the detection frequency of human papillomavirus (HPV) has been found to be significantly increased in tumor tissues among patients with sexually transmitted diseases (STDs), depending on the geographical distribution of each population studied. The present study focused on the detection of HPV and the distribution of Arg72Pro p53 polymorphism in a cohort of healthy individuals, as well as prostate cancer patients. We investigated the presence of HPV in 50 paraffin-embedded prostate cancer tissues, as well as in 30 physiological tissue samples from healthy individuals by real-time PCR. Furthermore, the same group of patients was also screened for the presence of the Arg72Pro polymorphism of the p53 gene, a p53 polymorphism related to HPV. Out of the 30 control samples, only 1 was found positive for HPV (3.33 %). On the contrary, HPV DNA was detected in 8 out of the total 50 samples (16 %) in the prostate cancer samples. The distribution of the three genotypes, Arg/Arg, Arg/Pro, and Pro/Pro, was 69.6, 21.7, and 8.7 % in the cancer patients and 75.0, 17.86, and 7.14 % in healthy controls, respectively. No statistically significant association was observed between the HPV presence and the age, stage, p53 polymorphism status at codon 72, or PSA. The increased prevalence of HPV detected in the prostate cancer tissues is in agreement with that reported in previous studies, further supporting the association of HPV infection and prostate cancer.

Effect of Smad pathway activation on podocyte cell cycle regulation: an immunohistochemical evaluation.

BACKGROUND: Mature podocytes are in cell cycle arrest and their inability to proliferate successfully is a consequence of negative cell-cycle regulators' expression, such as p57. Phosphorylated smad2/smad3 (pSmad2/3) is an intracellular heteromeric mediator of transforming growth factor beta (TGF-ß) signals and, together with co-activators such as P300, regulates gene transcription, including cell cycle regulator proteins. METHODS: In order to investigate Smad pathway activation and podocyte cell cycle regulation in glomerular injury, we studied the glomerular immunohistochemical expression of p57, pSmad2/3 and P300 in samples from 67 patients with various types of glomerulonephritis (GN) and 10 normal kidney tissue specimens. RESULTS: pSmad2/3 and p300 expression were found significantly increased in all glomerular cell types in both proliferative and nonproliferative GN, while a significant reduction in p57-positive podocytes was observed when compared to controls. Staining for p57 was found to inversely correlate to pSmad2/3 suggesting that glomerular Smad pathway activation is related to down-regulation of p57 expression in proliferative glomerulonephritis. To our knowledge, this is the first study that indicates a relation between the TGF-beta/Smad signalling pathway and the cell cycle regulatory protein p57 in human GN. CONCLUSION: The increased pSmad2/3 staining together with the reduced p57 expression found in biopsy specimens with intense interstitial inflammation, indicate a possible relation between interstitial inflammation, glomerular Smad pathway activation and podocyte cell-cycle deregulation.

Prognostic significance of phosphorylated STAT-1 expression in premenopausal and postmenopausal patients with invasive breast cancer.

AIMS: STAT-1 is the first member of the family of signal transducers and activators of transcription (STATs). In breast cancer experimental models, an apoptotic and antiproliferative effect has been demonstrated. Our aim was to study the role of phosphorylated STAT-1 (pSTAT-1) in invasive breast carcinoma and its prognostic significance in premenopausal and postmenopausal patients. METHODS AND RESULTS: Immunohistochemistry was performed in 165 patients in order to detect the expression of pSTAT-1 and its correlation with oestrogen receptor (ER), progesterone receptor (PR), caspase-3, and pAkt. pSTAT-1 was immunodetected in the cytoplasm of the malignant cells (11.6%). In premenopausal patients, cytoplasmic pSTAT-1 was positively correlated with stage (P = 0.014), ER (P = 0.008), caspase-3 (P = 0.029), and pAkt (P = 0.045). Univariate analysis showed that cytoplasmic pSTAT-1 was associated with poor overall survival (P = 0.042) and the phenotype of pSTAT-1/ER or PR coexpression with shorter disease-free survival (P = 0.012). In contrast, in postmenopausal patients, no association with clinicopathological parameters and survival was observed, except for the relationship of pSTAT-1/ER or PR coexpression with longer disease-free survival (P = 0.034). CONCLUSIONS: This is the first study examining the role of pSTAT-1 in premenopausal and postmenopausal women with invasive breast cancer. Our results suggest that pSTAT-1 is related to tumour progression in premenopausal patients through the advanced stage and worse survival.

Primary Large Mucosa-Associated Lymphoid Tissue (MALT) Lymphoma of the Gallbladder.

Primary lymphoma of the gallbladder (GB) is a rare condition, and very few cases have been reported so far. Diagnosis is usually made after surgery of suspicious GB mass, which is often difficult to differentiate from GB carcinoma. The GB wall does not contain lymphoid tissue, and tumors arise at the submucosal layer. Stone disease and chronic inflammation may contribute to its pathogenesis. Treatment consists of surgical resection followed by adjuvant therapy in selected cases. We present a case of an unusual, large-sized mucosa-associated lymphoid tissue (MALT) lymphoma of the GB.

A Case Report of Glomus Tympanicum Complicated With Facial Nerve Palsy.

Introduction: Generally, glomus tumors are considered tumors of the autonomic system arising from chromaffin cells of the parasympathetic paraganglia of the skull base and neck. Glomus tympanicum is the most common primary tumor of the middle ear cavity and it arises from the paraganglia of the middle ear. Case Report: We present a case of glomus tympanicum presented in a 70-year-old woman, complicated with facial nerve palsy which at first sight was misdiagnosed as cholesteatoma. Patient presented in our clinic because of otorrhea, pulsatile tinnitus and hearing loss in the right ear. However, facial nerve function was good in the first examination (40 days before the surgery). Eventually, she treated successfully with a canal wall down mastoidectomy. Technique had been chosen because of the mass size and the involvement of external auditory canal, after a discussion with the patient. Conclusions: Although histologically benign, glomus tympanicum is slow growing and destructs adjacent tissues potentially. The two most common complaints are hearing loss (conductive) and pulsatile tinnitus. These neoplasms are more common in women and they can be diagnosed by CT or MRI scan. It is of high importance physicians suspect a glomus tumor when patient 's clinical findings are hearing loss and pulsatile tinnitus and use an intravascular agent in imaging so that the differential diagnosis will be supported.

Immunohistochemical study of PTEN and phosphorylated mTOR proteins in familial and sporadic invasive breast carcinomas.

AIMS: Loss of phosphatase and tensin homologue (PTEN) leads to activation of several kinases, including mammalian target of rapamycin (mTOR), which promotes cell cycle progression. The aim was to study the expression of PTEN and phosphorylated (p)-mTOR in familial and sporadic invasive breast carcinomas and their relation to clinicopathological features, molecular indices (Wnt1) and patients' survival. METHODS AND RESULTS: PTEN and p-mTOR were detected immunohistochemically in 215 sections of invasive breast carcinomas (112 with a familial history of breast cancer). Image analysis was used and univariate and multivariate analyses employed for statistical evaluation of results. PTEN was detected in the nucleus (73.5%) and p-mTOR in the cytoplasm (44.2%) of cancer cells. Loss of PTEN protein was more frequently detected in women with a familial history of breast cancer (72%) (P < 0.0001), while its expression was negatively correlated with Wnt1, in total (P = 0.049). p-mTOR showed a positive association with lymph node status (P = 0.010) and was found to have a negative impact on patients' overall survival (P = 0.016). CONCLUSIONS: Loss of PTEN protein expression appears to occur more frequently in women with a family history of breast cancer, whereas activation of mTOR protein seems to be related to a more aggressive phenotype.

Transcription factor Sp1 expression is upregulated in human glomerulonephritis: correlation with pSmad2/3 and p300 expression and renal injury.

BACKGROUND: Sp1 is a ubiquitous transcription factor that mediates the fibrogenic factor transforming growth factor beta (TGF-beta) signals through cooperation with Smad proteins. The transcriptional coactivator p300 is also suggested to play a role in Smad signal transduction. METHODS: We investigated the immunohistochemical expression of Sp1 as well as the expression of pSmad2/3 and the coactivator p300 in 157 renal biopsy specimens from patients with various types of glomerulonephritis (GN). Correlations between immunohistochemical, clinical, and histologic parameters were performed. RESULTS: Sp1 exhibited an increased glomerular and proximal tubular expression in all forms of GN compared to controls. The proximal tubular expression of Sp1 was significantly increased in proliferative GNs (p = 0.025), whereas in secondary GNs, there was a significant increase in the molecule's glomerular expression (p = 0.008). Sp1 correlated positively with pSmad2/3 and p300 expression in proximal tubules (r = 0.241, p = 0.018 and r = 0.244, p = 0.014, respectively), while in proliferative GNs, its expression correlated positively with pSmad2/3 expression in glomeruli (r = 0.32, p = 0.028). Sp1 glomerular and proximal tubular immunostaining correlated positively with serum creatinine levels (r = 0.265, p = 0.02 and r = 0.306, p = 0.006, respectively), while its proximal tubular expression showed a similar correlation with interstitial fibrosis (r = 0.213, p = 0.025). Sp1 was constantly detected in hyperplastic lesions and cellular crescents (each 100%), and very often in micro adhesions (94%) and segmentally or globally sclerotic areas (each 83%). CONCLUSIONS: This study documents the upregulation of Sp1 expression in glomeruli and proximal tubules of GN specimens. Our findings suggest a possible cooperation of Sp1 with pSmad2/3 and p300 in mediating renal injury as well as a possible role for this molecule in the pathogenesis and the progression of human GN.

Cell adhesion molecules' altered profile in benign and malignant salivary gland tumors. The paradigm of beta4-integrin, desmoglein-2, ICAM-1 and CD44s.

BACKGROUND: Alterations in intercellular and cell-extracellular matrix connections contribute to tumour development. This study investigates the expression of specific cell adhesion molecules (CAMs) in salivary gland tumors (SGTs). METHODS: Formalin-fixed, paraffin- embedded tissue specimens of different types of 34 benign and 31 malignant SGTs and normal salivary glands were studied using Envision/HRP immunohistochemical technique for Desmoglein-2 (Dsg-2), beta4-integrin, CD44s and ICAM-1. Intensity of staining was evaluated in a semi-quantitative manner. Results were analyzed using Kendall's τ and Spearman's ρ as correlation criteria. RESULTS: Dsg-2 in intercellular space, beta4-integrin in cell-basal membrane, and CD44s in both types of contacts were strongly expressed in normal acinar and ductal cells, whereas ICAM-1 was expressed only at the endothelium and sparse stromal cells and monocytes. Strong correlation was found between Dsg-2 expression in adenomas and controls and between adenocarcinomas and controls. In adenomas, a distinct cytoplasmic presence of Dsg-2 was observed in addition to the usual membranous expression, with decreased expression in comparison with normal tissue. In malignant SGTs, Dsg-2 expression was absent. In most SGTs, beta4-integrin was expressed also with a distinct pattern, involving the cytoplasm and the unpolarised membrane, while CD44 was found only on the membrane. Strong correlation between beta4-integrin expression in adenomas and controls was noted, while CD44 expression was found to be correlated significantly between adenocarcinomas and controls (p < 0.001). Regarding ICAM-1, its expression was found increased in adenomas, with non-specific distribution in malignant SGTs and strong correlation between the histological subtypes and controls (p < 0.001). CONCLUSION: The different expression profile of CAMs in SGTs could possibly suggest a role on their pathogenesis, representing a model of how neoplastic cells can take advantage of normal tissue architecture and cell-extracellular matrix interactions.

Treatment with Molgramostim (Recombinant Human Granulocyte-Macrophage Colony Stimulating Factor, Rhugm-Csf, Mielogen) and Lenograstim (Granulocyte-Colony Stimulating Factor) Improves Experimental Colitis in Rats.

BACKGROUND/AIM: Treatment with growth factors could be beneficial in both inflammatory bowel disease and experimental colitis. The aim of this study was to investigate the effect of Colony Stimulating Factor (CSF), and Recombinant Human (rHu) Granulocyte Stimulating Factor (GSF) in experimental colitis in rats. METHODS: Experimental colitis was induced in 62 male Wistar rats, divided into 9 groups, using 2,4,6-trinitrobenzensulfonic acid (TNBS). Group 1: Ten rats with colitis without treatment (control group). Euthanasia after 15 days. Group 2: Ten animals with colitis without treatment (control group). Euthanasia after 30 days. Group 3: Six animals with colitis. Immediate treatment with CSF. Euthanasia after 19 days. Group 4: Six animals with colitis. Treatment started 7 days after the induction of colitis. Animals were kept for 19 days. Group 5: Six animals with colitis. Treatment started 2 weeks after the induction of colitis. Group 6: Six animals with colitis, the same as in group 3. Treatment with GSF. Group 7: Six animals with colitis, the same as in group 4. Treatment with GSF. GROUP 8: Six animals with colitis, the same as in group 5. Treatment with GSF. Group 9: Six animals with colitis. Immediate treatment with prednisolone. Euthanasia after 15 days. RESULTS: CSF and GSF administration significantly improved the histological score (P < 0.05) and reduced malondialdehyde contents (P < 0.05), compared to control groups in all animals. CSF was superior to GSF and to prednisolone. CONCLUSION: Administration of both CSF and GSF could significantly improve the histological score and oxidative stress in experimental colitis in rats.

Statin treated patients have reduced intraplaque angiogenesis in carotid endarterectomy specimens.

BACKGROUND: Treatment with statins is considered a first line therapy in atherosclerotic disease. Intraplaque angiogenesis is involved in plaque progression and instability. It remains unclear whether the beneficial effect of statin treatment in humans is achieved through reduced intraplaque angiogenesis. The aim of this study was to evaluate the capillaries density in carotid plaques removed from patients treated with statin versus untreated patients. METHODS AND RESULTS: We studied 102 patients who underwent carotid endarterectomy: 98 of them met the inclusion criteria and entered the study; 75 men and 23 women; mean age 66+/-8 years (range 42-83 years). Forty-three patients (44%) were on statin treatment at least 3 months before endarterectomy and 55 (56%) had never received statin treatment. The intensity of intraplaque angiogenesis was evaluated with immunohistochemistry using the antibody CD34. The number of capillaries per mm(2) was measured with a custom designed image tool analysis. With the exception of serum total cholesterol levels and serum low-density cholesterol levels, the two groups of patients did not vary significantly in cardiovascular risk factors and in parameters pertaining to the procedure profile. Patients on statin treatment had less capillaries per mm(2) than patients not receiving this kind of drugs (0.97+/-0.61 per mm(2) versus 1.39+/-0.98 per mm(2), p=0.031). Univariate associations between possible explanatory variables and number of capillaries per mm(2) were tested using Spearman rank R. Variables associated with a p-value <0.20 (age, serum creatinine, serum total cholesterol, serum low-density lipoprotein, serum homocysteine, presence of diabetes mellitus and statin treatment) were entered in a multivariable model. Multivariate analysis showed that statin treatment was the only independent predictor (t=-5.39, p<0.001) of intraplaque angiogenesis. CONCLUSIONS: Statin therapy is associated with reduced intraplaque angiogenesis in the carotid arteries. This could provide an explanation for the beneficial effects of this kind of drug on patients with atherosclerotic disease.

Application of alpha-smooth muscle actin and c-kit in the differential diagnosis of adenoid cystic carcinoma from polymorphous low-grade adenocarcinoma.

The expression of vimentin, alpha-smooth muscle actin (alpha-SMA) and c-kit in adenoid cystic carcinomas (AdCCs) and polymorphous low-grade adenocarcinomas (PLGAs) was investigated immunohistochemically to evaluate the application of these markers to distinguish AdCCs from PLGAs when the histological features are equivocal. Tissue specimens of AdCCs and of PLGAs, formalin-fixed and paraffin-embedded were retrospectively studied using vimentin, alpha-SMA and c-kit. Positive staining for alpha-SMA was identified in all AdCCs and 25% of PLGAs. The immunoreactivity of c-kit in all positive cases of AdCCs (83%) and PLGAs (41%) was more than 50% and less than 50% of tumor cells respectively. The expression pattern for both alpha-SMA and c-kit, in tubular structures of AdCCs was different of that seen in the same structures in PLGAs. The results of this study support the potential application of alpha-SMA and c-kit as an adjunctive aid in the differential diagnosis of AdCCs from PLGAs.

Metastatic renal clear cell carcinoma in the parotid gland: a study of immunohistochemical profile and cell adhesion molecules (CAMs) expression in two cases.

Metastasis of renal cell carcinoma (RCC) may involve any organ, including the parotid salivary gland. While the definition of salivary gland neoplasms with clear cell transformation can be concluded by the synchronous presence of areas showing typical morphology, sometimes the definition of a metastatic RCC in the parotid is difficult and the application of immunohistochemistry may support the clinical and radiographic observations in the final diagnosis. The aim of this paper was to describe the heterogeneous immunohistochemical features and, furthermore, to characterize the pattern of expression of cell adhesion molecules (CAMs) E-cadherin, beta4-integrin, desmoglein-2, ICAM-1 and CD44s (HCAM) in two cases of metastatic parotid RCC.

Primary Myxoid Liposarcoma of the Upper Thoracic Spine in an Elderly Patient.

Liposarcoma, one of the most common soft-tissue sarcomas, originates from primitive mesenchymal cells, and its diagnostic criteria have been well established. Myxoid liposarcoma is the second most common histological subtype, occurring more frequently during the fourth and fifth decades of life. There are only a few sporadic published cases of spinal liposarcomas, whatever primary or metastatic. We describe an unusual case of primary myxoid liposarcoma of the thoracic spine in a 79-year-old male, presented with a 2-month history of intractable dorsal pain and progressive weakness of lower limbs. Surgical treatment was performed with wide tumor resection, spinal cord decompression, and posterior instrumentation complemented by radiotherapy. Only one other case has been reported with this spinal localization in the literature. Despite its rarity, myxoid liposarcoma should be considered in the differential diagnosis of primary tumors of the thoracic spine.

Detection of C-KIT (CD117) molecule in benign and malignant salivary gland tumours.

C-KIT (CD117), a tyrosine kinase receptor, is involved in the growth and development of normal tissues and some types of neoplasms. In the present study we analysed the expression of this molecule in salivary gland tumours. Archival formalin-fixed, paraffin-embedded sections of 40 benign and 57 malignant salivary gland tumours were retrieved and retrospectively studied immunohistochemically using a polyclonal C-KIT antibody in an Envision/HRP technique. In addition five samples of chronic submandibular sialadenitis, five normal minor salivary glands and parotid or submandibular gland tissue adjacent to benign tumour were also studied. C-KIT expression was observed in cases of adenoid cystic, acinic cell polymorphous low grade, epithelial-myoepithelial, carcinosarcoma and basal cell adenocarcinomas, as in luminal cells of pleomorphic adenomas, in serous acinar and only in intercalated and a small number of striated ductal cells of inflammatory salivary gland tissue, whereas normal salivary lobules were generally negative except a weak positivity of intercalated cells. Contrary to other reports, this study suggests that, C-KIT protein does not appear to be an exclusively specific marker for benign or malignant salivary gland neoplasms, but may be useful in differential diagnosis of adenoid cystic carcinoma from polymorphous low grade adenocarcinoma. Furthermore its expression in serous acinar cells in sialadenitis and intercalated ductal cells in normal and inflammatory lesions may indicate a possible participation in pathogenesis of both neoplastic and non-neoplastic salivary gland diseases.

Evaluation of the vascular endothelial growth factor (VEGF)-C role in urothelial carcinomas of the bladder.

BACKGROUND: Vascular endothelial growth factor-C (VEGF-C) has been associated with angiogenesis, lymphangiogenesis and regional lymph node metastasis and was reported to have an anti-apoptotic and proliferative role. MATERIALS AND METHODS: An immunohistochemical study was applied to 123 specimens of bladder urothelial carcinoma (BUC) to detect VEGF-C and investigate its clinicopathological and prognostic value. VEGF-C-immunostained BUC specimens (123) were statistically correlated with histological grade and stage, patient overall survival and immuno-expression of Ki-67 and bax proteins. RESULTS: VEGF-C immunopositivity (27/123 BUCs, 22.0%) was inversely correlated with tumor stage and bax immunoexpression (p = 0.007 and p = 0.032, respectively). VEGF-C-positive BUCs tended to have better prognosis (univariate analysis). CONCLUSION: VEGF-C might be associated with an anti-apoptotic phenotype. Our controversial results regarding patient survival suggest that the role of VEGF-C in BUC progression and prognosis remains to be clarified.

Immunohistochemical detection of the expression of the cell adhesion molecules E-cadherin, desmoglein-2, beta4-integrin, ICAM-1 and HCAM (CD44s) in Warthin's tumour of the parotid gland.

The study of the expression of cell adhesion molecules (CAMs), E-cadherin, desmoglein-2, beta4-integrin, HCAM (CD44s) and ICAM-1 in Warthin's tumours. Twenty formalin--fixed, paraffin--embedded parotid Warthin's tumours were studied using an Envision/HRP immunohistochemical technique. Beta4-integrin was strongly expressed in all cell-basement membrane and intercellular contacts of the epithelium, E-cadherin and desmoglein-2 in cell-cell contacts, but not in basal cell-basement membrane connections and on columnar cells' luminal surfaces, HCAM (CD44s) in intercellular contacts of both luminal (mainly), basal cells and also in the periphery of monocytic-lymphocytic stroma, and ICAM-1 was weak to moderate expressed in both luminal and basal epithelial cells and strongly in the germinal lymphocytic centres. CAM expression suggests a bilayered excretory ductal structure of the neoplastic epithelium in Warthin's tumour, as a result of hyperplastic process of the glandular epithelium that interacts with the excessive lymphoid tissue of the stroma.

Effect of BRCA1 immunohistochemical localizations on prognosis of patients with sporadic breast carcinomas.

Our purpose was to investigate the expression pattern of BRCA1 protein in sporadic breast carcinomas, as well as the clinicopathological and prognostic value of its subcellular localizations. Immunohistochemistry was performed on paraffin embedded tissue specimens from 111 sporadic, invasive breast carcinomas to detect the expression of the proteins BRCA1, ER, PR, erbB2, p53 and Ki67. BRCA1 protein was detected in the nuclei and the cytoplasm of the tumor cells. Nuclear BRCA1 immunoreactivity showed no relation with the classic clinicopathological markers and the expression of cerbB2, p53 and Ki67. Reduced expression of nuclear BRCA1 protein was found to exert an independent favorable impact on both the overall and relapse-free (RF) survival of the patients (p=0.019 and p=0.043, respectively). Cytoplasmic BRCA1 was associated with none of the classic histomorphological indices, except from the lymph node metastasis, with which its relation was found to be inverse (p=0.05), prolonging the RF survival of the patients (p=0.05). Our findings suggest that BRCA1 protein depicts different prognostic significance, according to its subcellular distribution. Nuclear detection of the protein was associated with a worse prognosis, while the cytoplasmic one was related to fewer recurrences as a result of fewer lymph node metastases.

Differential effect of the expression of TGF-ß pathway inhibitors, Smad-7 and Ski, on invasive breast carcinomas: relation to biologic behavior.

The aim of our study was to investigate the expression of Smad-7 and Ski proteins in invasive breast carcinomas, to determine their clinicopathological value and their influence on carcinomas biologic behavior. Immunohistochemistry was applied on 150 invasive breast carcinomas to detect the expression of Smad-7 and Ski. Their correlation to clinicopathologic parameters and markers of metastasis was statistically processed using chi-squared test. Overall and disease-free survival was assessed using Kaplan-Meier test and log-rank statistics. Smad-7 was immunodetected in the cytoplasm of cancer cells in 60%, whereas Ski was immunodetected in the cytoplasm and nuclei in 44.5% and 17.6% of the cases, respectively. Smad-7 expression was positively correlated with tumor size, stage, matrix metalloproteinase (MMP)-9, and MMP-14. Cytoplasmic Ski expression was negatively associated with tumor size, stage, and lymph node status, and its nuclear expression was negatively related to histologic grade. Cytoplasmic Ski expression was associated with longer overall and disease-free survival. It appears that two negative regulators of the transforming growth factor-ß pathway, Smad-7 and Ski, behave differentially in invasive breast carcinomas. Smad-7 appears to be related with an aggressive phenotype, whereas Ski expression is related to a less aggressive behavior and positively influences patients' survival.

The effect of negative pressure therapy on the femoral vein blood flow and wall structure.

Negative pressure therapy has been recently used for managing lymphatic or infective groin complications. The aim of this study was to investigate any possible association between application of negative pressure therapy in the groin area and deep-vein thrombosis. Acute surgical wounds were created at the inguinal areas in 7 pigs. Different negative pressures ranging from -50 to -200 mmHg were applied directly over the femoral vessels, and blood flow alterations were studied using a Doppler ultrasound. Femoral vein specimens were also removed for histological examination after 12 hours of therapy. It has been demonstrated that negative pressure therapy does not significantly alter the baseline lower limb venous return. Histology demonstrated several changes, which are associated with vein thrombogenesis. The hemodynamic and pathological findings still leave a potential for thrombogenic effects of negative pressure therapy and warrant care to protect the femoral veins, with the use of thrombosis prophylaxis measures.