Alterations of aorta intima and media transcriptome in swine fed high-fat diet over 1-year follow-up period and of the switch to normal diet.

BACKGROUND AND AIM: We previously showed that 12-month high-fat diet (HFD) in pigs led to fattening and increased artery intima-media-thickness, which were partly reversed after 3-month return to control diet (CD). The aim of this study was to decipher underlying mechanism of action by using transcriptomic analyses of intima and media of aorta. METHODS AND RESULTS: Thirty-two pigs were divided into three groups: CD for 12 months; HFD for 12 months; switch diet group (regression diet; RD): HFD for 9 months followed by CD for 3 months. After 12 months, RNA was isolated from aorta intima and media for nutrigenomic analyses. HFD significantly affected gene expression in intima, while RD gene expression profile was distinct from the CD group. This suggests that switch to CD is not sufficient to correct gene expression alterations induced by HFD but counteracted expression of a group of genes. HFD also affected gene expression in media and as for intima, the expression profile of media of pigs on RD differed from that of these on CD. CONCLUSIONS: This study revealed nutrigenomic modifications induced by long-term HFD consumption on arterial intima and media. The return to CD was not sufficient to counteract the genomic effect of HFD.

Prevalence of Sleep Apnea in Patients with Carotid Artery Stenosis.

Obstructive sleep apnea (OSA) is a common disease affecting about 13% of men and 6% of women, usually having severe cardiovascular sequalae. OSA is responsible for the systemic inflammatory response and oxidative stress and results in endothelial injury being a risk factor for atherosclerosis. The aim of this study was to estimate the prevalence of OSA among patients with severe carotid artery stenosis. Fifty-five patients (F/M-24/31, mean age 70 ± 7 years, body mass index 28.3 ± 6.3 kg/m2) were enrolled into the study. The patients were qualified for elective surgical treatment of carotid artery stenosis. Polysomnography was performed in all patients the night before surgery. Thirty-six patients underwent surgical endarterectomy and nineteen patients underwent carotid artery stenting. Sleep apnea was diagnosed in 44 (80%) of all patients. The mean apnea-hypopnea index (AHI) was 14.5 ± 12.9. The OSA severity distribution was as follows: 22 mild, 16 moderate, and 6 severe cases. We found that the percentage of carotid endarterectomies was the greatest in mild and moderate OSA. It was approximately twofold greater than that in non-OSA patients. Carotid artery stenting was performed in nearly half of the patients in each of these groups. We conclude that OSA is highly prevalent in patients with carotid artery stenosis scheduled for carotid surgery. Nonetheless, the exact pathogenetic mechanisms underlying mutual interaction between OSA and vascular wall damage remain elusive. OSA is not routinely diagnosed among patients with advanced atherosclerosis. The study results might be an argument for performing polysomnography in patients with carotid artery stenosis.

Upper Extremity Lymphedema After Axillary Lymph Node Dissection: Prospective Lymphoscintigraphic Evaluation.

This prospective study was designed to evaluate changes in upper extremity lymphatic drainage after ALND in comparison to the preoperative status using lymphoscintigraphy. The study enrolled 44 women (mean age: 57.95; range: 35-80) with a new diagnosis of unilateral invasive breast carcinoma who had been scheduled to undergo ALND. This was a substudy of the physiotherapeutic project, in which subjects after ALND were randomized into 4 groups treated with: 1) rehabilitation exercises; 2) manual lymphatic drainage; 3) pneumatic compression pump; and 4) education only. Clinical evaluation which included arm measurements and lymphoscintigraphy was performed in every subject before surgery and 3 times after surgery (1-6 weeks, 1 and 2 years after ALND). Follow-up was completed in 44 subjects at 1 year and in 32 subjects at 2 years. Lymphedema diagnosis was made in 4 subjects 1 year after ALND (9%) and in 8 subjects 2 years after ALND (25%). Among them, respectively, only 50% and 62% noticed and reported lymphedema. Quantitative analysis of lymphoscintigrams and photoplethysmography results did not reveal upper extremities lymphatic transport and/or venous function impairment after the ALND procedure. Qualitative analysis of lymphoscintigrams revealed most commonly disappearance of previously functional lymph nodes and appearance of dermal backflow in subjects who developed lymphedema. Conversely, appearance of functional lymph nodes in different locations after ALND may indicate protection from development of upper extremity lymphedema.

Low-dose computer tomography as a screening tool for lung cancer in a high risk population.

The aim of the study was to evaluate the usefulness of low-dose computer tomography as a screening tool for early stage lung cancer. The study was performed in 332 individuals aged 55-70 who were asymptomatic, who had not previously suffered from cancer, and who smoked at least ten packs of cigarettes a year. Baseline and repeated LD-CT scans were performed. Pulmonary nodules were classified according to the size and morphology, and the results were categorized as negative (no nodules observed), semi-positive (nodules of 4 mm or smaller in diameter) and positive (nodules 5 mm or larger). Based on the category of the patient, either a repeat low-dose CT, a bronchoscopy with or without a biopsy, or a PET-CT was performed. The baseline screening showed 59 positive results. Eighteen patients were hospitalized and underwent bronchoscopy and biopsy. One of these patients had Stage I non small cell lung carcinoma (NSCLC) and a lobectomy was performed. Three patients had Stage IV NSCLC and were referred for chemotherapy. We identified 103 semi-positive results. Only 25 of those patients had a repeat scan because of noncompliance. We observed no significant growth of diagnosed nodules in a semi-positive group. Low-dose CT can be used as a screening tool for early stage lung cancer. A high percentage of false-positive results are observed. There are difficulties in diagnosing nodules in patients with post-tuberculosis changes. A high rate of noncompliance was noticed.

Development, reproducibility and validity of the food frequency questionnaire in the Poland arm of the Prospective Urban and Rural Epidemiological (PURE) study.

BACKGROUND: A food frequency questionnaire (FFQ) is the most commonly used method in large epidemiological studies. The validation of an FFQ is essential for specific populations because foods are culture-dependent. The present study aimed to develop an FFQ and evaluate its validity and reproducibility in estimating the intake of nutrients in urban and rural areas of Poland. METHODS: Adult participants (n = 146) in the Polish arm of the ongoing Prospective Urban and Rural Epidemiological (PURE) study completed FFQs on two occasions, as well as four 24-h dietary recalls (DRs) during a 12-month period. Correlation coefficients (r) and de-attenuated correlation coefficients between dietary recalls and both FFQs were calculated for selected macro- and micronutrients. Agreement between the two methods was evaluated by classification into quartiles and the Bland-Altman method. Reproducibility was assessed by the intra-class correlation coefficient (ICC). RESULTS: The final food list contained 134 food items. For urban participants, FFQ2 generally underestimated energy, protein and fat compared to the FFQ1 and mean of DRs. In rural areas, compared to DRs, both FFQs overestimated energy and macronutrients. For both urban and rural settings, de-attenuated correlation exceeded 0.4 for almost all nutrients and the exact agreement in quartile categorisation was >66%. When assessing repeatability, ICC varied from 0.39-0.63 in an urban setting and 0.19-0.45 in a rural setting. CONCLUSIONS: This 134-item FFQ has good validity and reproducibility in relation to the reference method and can be used to rank individuals based on their macro- and micronutrient intake.

Efficacy of manual lymphatic drainage in preventing secondary lymphedema after breast cancer surgery.

This study evaluated the effectiveness of manual lymphatic drainage (MLD) in the prevention of secondary lymphedema after treatment of breast cancer. The study consisted of 67 women, who underwent breast surgery for primary breast cancer. From the second day of surgery, 33 randomly chosen women were given MLD. The control group consisted of 34 women who did not receive MLD. Measurements of the volumes of both the arms were taken before surgery and on days 2, 7, 14, and at 3 and 6 months after surgery. At 6 months after breast cancer surgery, among the women who did not undergo MLD, a significant increase in the arm volume on the operated side was observed (p=0.0033) when compared with the arm volume before surgery. At this time, there was no statistically significant increase in the volume of the upper limb on the operated side in women who underwent MLD. This study demonstrates that regardless of the surgery type and the number of the lymph nodes removed, MLD effectively prevented lymphedema of the arm on the operated side. Even in high risk breast cancer treatments (operation plus irradiation), MLD was demonstrated to be effective against arm volume increase. Even though confirmatory studies are needed, this study demonstrates that MLD administered early after operation for breast cancer should be considered for the prevention of lymphedema.

Chyloperitoneum, chylothorax and lower extremity lymphedema in woman with sporadic lymphangioleiomyomatosis successfully treated with sirolimus: a case report.

Lymphangioleiomyomatosis (LAM) is a rare disease characterized by diffuse thin-walled cysts throughout the lungs on computed tomography and diffuse proliferation of abnormal smooth muscle-like cells (LAM cells) on lung biopsy. LAM affects women almost exclusively, predominantly in their reproductive age. The most typical presenting symptoms include dyspnea, spontaneous pneumothorax, cough and chylothorax. Abdominal findings represent less common initial manifestations of the disease and may pose diagnostic difficulties. The treatment of LAM has not been fully established. Recent studies report effectiveness of sirolimus in LAM patients. We report the case of a 45-year-old woman with sporadic LAM, successfully treated with sirolimus, in whom the first manifestation of the disease was chyloperitoneum and after three and nine years, respectively, lymphedema of the left lower extremity and right sided chylothorax occurred.

Adventitial lymphatics and atherosclerosis.

Lymphatic vessels are important in reverse cholesterol transport and play a crucial role in regression of atherosclerotic plaque in experimental animal models. Therefore, we attempted to analyze adventitial microcirculation including lymphatic vessels and adventitial macrophages in large human arteries in various stages of atherosclerosis. Eighty-one arterial segments of large arteries (iliac arteries and abdominal aortas) were obtained from deceased organ donors. Lymphatic vessels were identified using anti-LYVE-1 and anti-D2-40/podoplanin immunohistochemical staining. Adventitial blood vessels and macrophages were visualized using anti-CD-31 and anti-CD-68. Intimal thickness was measured under 100x magnification with an Olympus BX 41 light microscope using the visual mode analySIS 3.2 software. Lymphatic vessels were counted in each cross section of the examined arteries, and adventitial blood vessels (CD31+) were counted using the "hot spot" method. Statistical analysis was performed with Statistica 9.1 PL software (StatSoft, Cracow, Poland). Mann-Whitney, F-Cox, Chi-square, and Spearman's correlation tests were performed and the differences were considered significant at p < 0.05. Lymphatic and blood vessels in the adventitia of examined arteries were identified and quantified. Significant positive correlations were found between the number of adventitial lymphatics (LYVE-L +) and intimal thickness (r = 0.37; p < 0.05) as well as with age of the subjects (r = 0.3; p < 0.05). Thus, lymphatic vessels are present in the adventitia of large arteries in humans and the number of adventitial lymphatic vessels increases with progression of atherosclerosis as assessed by intimal thickness.

Axillary lymph nodes and arm lymphatic drainage pathways are spared during routine complete axillary clearance in majority of women undergoing breast cancer surgery.

Alterations in axillary lymph nodes (ALNs) after complete axillary lymph node dissection (ALND) in comparison to the preoperative status were evaluated using lymphoscintigraphy performed preoperatively and 1-6 weeks after surgery in 30 women with a new diagnosis of unilateral, invasive breast carcinoma. Analysis of lymphoscintigrams revealed that ALNs after surgery were present in 26 of 30 examined women. In comparison to preoperative status, they were visualized in the same location (12 women), in the same and additionally in different locations (9 women), or only in different locations (4 women). No lymph nodes were visualized in one woman and lymphocoele were in 4 women. Thus, after ALND, a variable number of axillary lymph nodes remain and were visualized on lymphoscintigraphy in the majority of women. The classical ALND, therefore, does not allow complete dissection and removal of axillary nodes with total disruption of axillary lymphatic pathways, accounting in part for the variable incidence and severity of lymphedema after the procedure.

Popliteal artery thrombosis secondary to a tibial osteochondroma.

Popliteal artery thrombosis may present as a complication of an osteochondroma located in the vicinity of the knee joint. This is a case report of a 26-year-old man with symptoms of the right lower extremity ischaemia without a previous history of vascular disease or trauma. Plain radiography, magnetic resonance angiography and Doppler ultrasonography documented the presence of an osteochondrous structure of the proximal tibial metaphysis, which displaced and compressed the popliteal artery, causing its occlusion due to intraluminal thrombosis..The patient was operated and histopathological examination confirmed the diagnosis of osteochondroma.

Identification of linear epitopes on the flagellar proteins of Clostridioides difficile.

Clostridioides difficile (C. difficile) is an opportunistic anaerobic bacterium that causes severe diseases of the digestive tract of humans and animals. One of the possible methods of preventing C. difficile infection is to develop a vaccine. The most promising candidates for vaccine antigens are the proteins involved in the adhesion phenomena. Among them, the FliC and FliD are considered to be suitable candidates. In this paper, the FliC and FliD protein polypeptide epitopes were mapped in silico and by using PEPSCAN procedure. We identified four promising epitopes: 117QRMRTLS123, 205MSKAG209 of FliC and 226NKVAS230, 306TTKKPKD312 of FliD protein. We showed that 117QRMRTLS123 sequence is not only located in TLR5-binding and activating region, as previously shown, but forms an epitope recognized by C. difficile-infected patients' antibodies. 205MSKAG209 is a C. difficile-unique, immunogenic sequence that forms an exposed epitope on the polymerized flagella structure which makes it a suitable vaccine antigen. 226NKVAS230 and 306TTKKPKD312 are well exposed and possess potential protective properties according to VaxiJen analysis. Our results open the possibility to use these epitopes as suitable anti-C. difficile vaccine antigens.

Different lymphscintigraphic patterns in patients with lymphedema distichiasis.

Mutation of the transcription factor FOXC2 gene has been identified as the cause of lymphedema-distichiasis syndrome (LD). Subjects with LD usually present with lower extremity lymphedema and distichiasis--an additional row of eyelashes. Typically, lymphscintigrams of patients with LD show good transport of the radiotracer from the feet to the inguinal lymph nodes accompanied by reflux of tracer to the skin of the lower extremities ("dermal backflow"). We have examined two patients with LD syndrome and were able to demonstrate two different distinct lymphscintigraphic patterns: lymphatic hyperplasia with reflux and obstructive.

Melkersson-Rosenthal syndrome: lymphoscintigraphy-documented impairment and restoration of facial lymphatic drainage in the course of disease.

Melkersson-Rosenthal syndrome (MRS) is an idiopathic, rare disorder manifested by facial swelling, congenital plicated tongue and recurrent peripheral facial nerve palsy. Labial involvement alone is referred to as cheilitis granulomatosa. Differential diagnosis of MRS includes allergic angioedema, bacterial, viral or filarial infections as well as autoimmunological inflammation in the course of systemic lupus erythematosus, dermatomyositis, and others. We present 4 patients who experienced periodically painless edema of the face and/or lips. Lesions were diagnosed as recurrent Quincke's edema and were treated with antihistamine agents and glucocorticoids without improvement. In all four cases of MRS, we were able to document impaired lymphatic drainage from the swollen area using lymphoscintigraphy. We also documented in follow-up lymphoscintigraphy a restoration of lymphatic flow in three of the four patients with MRS and these results corresponded with clinical improvement. We have demonstrated that lymphatic pathology plays an important role in pathophysiology of chronic facial swelling in patients with Melkersson-Rosenthal syndrome.

Effect of selected drugs on plasma asymmetric dimethylarginine (ADMA) levels.

Asymmetric dimethylarginine (ADMA) is an endogenous methylated amino acid derived from arginine which can inhibit the activity of nitric oxide synthases. In various pathological states such as hypercholesterolemia, hyperglycemia, hyperhomocysteinemia, hypertension, coronary artery disease, heart failure, and stroke, plasma levels of ADMA may be increased and lead to inhibition of NO synthesis and endothelial dysfunction. Inhibition of ADMA synthesis or intensification of metabolism of this compound might indirectly lower ADMA. Antioxidants, estrogen, vitamin A, angiotensin converting enzyme inhibitors, angiotensin AT1 receptor antagonists, and also some hypolipemic, hypoglycemic and beta-adrenoreceptor blocking drugs decrease ADMA levels. In some situations like neurological disorders, decreased plasma levels of ADMA are noticed and drugs increasing the concentration of this compound could exert protective effects. It is reasonable to explore which drugs can increase or decrease ADMA levels and what their mechanism of that action is.

Skin lymphatic system in the pathogenesis of arterial hypertension - review and critique.

Although numerous studies have confirmed the relationship between high salt intake and elevated blood pressure, the exact molecular mechanisms of this relationship are still unclear. There is growing evidence that skin interstitium, as well as the skin lymphatic system, are important regulators of both sodium (Na+) balance and blood pressure. Skin is in itself a large reservoir of Na+ ions which are stored in an osmotically inactive form on glycosaminoglycans (GAGs). Local hypertonicity due to extensive accumulation of Na+ within the skin as a result of a high-salt diet was demonstrated to induce macrophages to express a transcription factor termed tonicityresponsive enhancer binding protein (TonEBP) and subsequently to secrete vascular endothelial growth factor-C (VEGF-C), activating lymphangiogenesis within the skin. This regulatory axis seems to be adaptive in maintaining blood pressure in high salt-load states. Recent studies have added new insights into the functioning of lymphatic vessels and the pathogenesis of salt-sensitive hypertension as well as questioned the classic view of Na+ homeostasis. This review aims to summarize recent findings pertaining to the involvement of the skin lymphatic system in Na+ and blood pressure regulation.

Influence of compression cycle time and number of sleeve chambers on upper extremity lymphedema volume reduction during intermittent pneumatic compression.

The cycle time and number of chambers in the pneumatic sleeve may influence the outcome of lymphedema therapy with intermittent compression devices. The aim of our study was to assess efficacy of several commonly used different IPC protocols on edema volume reduction in women with postmastectomy lymphedema. Fixty-seven (57) women with secondary arm lymphedema (age 39-80) were selected to the study. Women were randomly assigned to two study groups with different IPC cycle times: I--90:90s and II--45:15s. Both groups were then randomly divided into two subgroups with different sleeves: A--1 chamber sleeve (28 women) and B--3 chamber sleeve (29 women). All women underwent IPC treatment for 5 weeks, 5 times a week for 1 hour (25 sessions). Arm volume measurements were performed before and after each IPC session. Significant reduction of edema volume was observed in all therapeutic subgroups, regardless of cycle times and number of chambers. In the group with short IPC cycle, better efficacy was noticed with 3-chamber sleeve. IPC is an effective method of volume reduction in women with postmastectomy arm lymphedema regardless of cycle times and number of sleeve chambers.

The effect of different compression pressure in therapy of secondary upper extremity lymphedema in women after breast cancer surgery.

This study evaluated how different levels of pressure under inelastic multi-layer bandages affect the reduction of secondary arm lymphedema and patient comfort throughout therapy. Ninety-six women with lymphedema after breast cancer treatment were randomized into three groups of 32 patients depending on level of applied pressure in compression therapy: group I (control) at 21-30 mmHg; study groups II A at 31-40 mmHg and group II B at 41-60 mmHg. All patients were treated with complex decongestive therapy (CDT) and intermittent pneumatic compression (IPC) before compression. Fixed points of bilateral arms were measured at the start (first visit), after 24 h, 7 days, and 14 days of therapy. Edema severity was measured by summary calculation. Results were presented as a relative metric coefficient of arm lymphedema (RMCAL) which is the difference between sums of circumferences of the edematous arm and the contralateral side expressed in percents. In order to evaluate the patient comfort after finishing a two-week therapy all patients assessed the level of accompanying pain using the numeric pain rating scale (NRS). At the start of therapy median arm circumference difference (RMCAL) was 18.60%, 18.51%, and 19.05% in groups I-II B, respectively. After 24 h the median RMCAL was reduced to 14.49%, 12.13%, and 12.64%. This was further reduced to 10.77%, 6.98%, and 8.48% at one week and 10.28%, 5.75%, and 7.20% in each group, respectively. There was no statistically significant difference between RMCAL values in group II A and II B throughout the therapy. In group II A (NRS = 2), applied bandages were better tolerated than in both II B (NRS = 5) and control groups (NRS = 8). These results demonstrate that inelastic multi-layer bandages applied in groups II A and II B (41-60 mmHg) led to the same reduction of swollen arm circumference with group II being better tolerated. The lowest compression (control at 21-30 mmHg) produced the smallest reduction. In addition, since the greatest reduction was seen in the first week of therapy while the second week served to maintain the reduction, compression garments may be able to be ordered after one week of therapy for more efficient patient care.

Presence of functional axillary lymph nodes and lymph drainage within arms in women with and without breast cancer-related lymphedema.

This pilot study was designed to investigate lymphatic transport in the arms of women after breast cancer treatment without lymphedema and with mild and severe arm lymphedema. Nineteen breast cancer survivors [6 without (Group 1), 6 with mild (Group 2) and 7 with severe (Group 3) lymphedema] were examined. Lymphatic transport in the arm and to axillary lymph nodes were evaluated using quantitative radionuclide lymphscintigraphy. The radioactivity ratio between the operated and unoperated axillae (axillary ratio), and both lymphatic transport and tracer disappearance rates (TDR) from the injection sites were calculated. We found that axillary ratio and lymphatic transport were significantly higher in Groups 1 and 2 compared to 3 and that TDR was not affected in any Group. Additionally, lymphscintigraphy revealed presence of functional axillary lymph nodes within the operated axilla in women without or with mild lymphedema, while in women with severe lymphedema, no axillary lymph nodes were visualized. We conclude from our subjects that breast cancer survivors who did not develop or had mild arm lymphedema have functional lymphatic transport and lymph nodes in the axillary area on the operated side. This suggests that efficient collateral circulation and/or peripheral lymphovenous communications has developed in these subjects which may be preventative for the development of severe lymphedema.

Can obstructive intralymphatic granulomas be the cause of cheilitis granulomatosa?

Cheilitis granulomatosa (ChG), also known as Miescher's cheilitis, is an uncommon, immunologically mediated nonnecrotizing granulomatous inflammatory disease characterized by recurrent, painless swelling of the lips. The aim of the study was a pathomorphological and immunohistochemical assessment of cases clinically classified as ChG to investigate potential pathological mechanisms of ChG symptoms and to verify the hypothesis of intravascular granulomas as a cause of lymphatic vessel obstruction and localized edema in ChG. We report 6 patients with ChG who clinically presented localized edema of the lips. Lip biopsy with pathomorphological and immunohistochemical examination was performed in all cases. We found discrete, non-necrotizing granulomas which were adjacent to numerous blood and lymphatic vessels. The lumen of lymphatic channels was dilated and was either empty or contained lymph and few macrophages or was completely occluded by nearby granulomas. All patients demonstrated a characteristic pattern of lymphangiectasia and perivascular lymphatic aggregates with evidence of non-necrotizing granulomas. None manifested intralymphatic granulomas. These results do not support the view that lymphatic vessel obstruction is caused by intravascular histiocytic granulomas described as the main part in the etiology of lymphatic edema in ChG. However, perivascular granulomas and dilation of lymphatic vessels confirm presence of inflammatory lymphostasis in all studied cases of ChG.

Disability, psychological distress and quality of life in breast cancer survivors with arm lymphedema.

The aim of this study was to assess disability, psychological distress and quality of life in Polish breast cancer survivors with arm lymphedema. One thousand sets of questionnaires consisting of WHO-DAS II, GHQ-30, EORTC QLQ-C30 and QLQ-BR23 were sent to members of the Polish Federation of Breast Cancer Survivors Clubs "Amazonki." The response rate was 28.3% of whom 31.70% reported arm lymphedema. The WHO-DAS II survey showed that patients with arm lymphedema had a higher overall disability score (45.04 versus 38.80 in group without arm lymphedema; p=0.01) and higher mean values in the scales of understanding and communicating, getting around, life activities at home, getting along with people, participating in society. The EORTC QLQ-C30 survey showed that patients with lymphedema had lower mean values in physical (0.55 versus 0.65; p=0.001), emotional (0.47 versus 0.57; p=0.01), social (0.59 versus 0. 73; p=0.002), cognitive and role functioning, increased fatigue, pain, insomnia, dyspnea, nausealvomiting and financial problems. The EORTC QLQ-BR23 data demonstrated worse future perspectives and an increase in breast and arm symptoms, and the GHQ-30 survey produced higher psychological distress (scores 15.18 versus 11.24; p=0.004). In conclusion, breast cancer survivors with arm lymphedema were more disabled, experienced a poorer quality of life and had increased psychological distress in comparison to survivors without this condition.