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International Institute of Anticancer Research

Publications in this journal

• Effects of Culture Supernatant from Lactobacillus pentosus Strain S-PT84 on Autonomic Nerve Activity in Rats.

  Authors: Yoshinori Beppu, Takayuki Izumo, Yuko Horii, Jiao Shen, Yoshiyuki Fujisaki, Toshihiro Nakashima, Nobuo Tsuruoka, Katsuya Nagai

  In vivo (Athens, Greece). 26(3):355-9.

  Intestinal administration of various lactobacilli has been reported to affect autonomic neurotransmission, blood pressure, blood glucose, and body weight in rats, however, the mechanisms of action ofIntestinal administration of various lactobacilli has been reported to affect autonomic neurotransmission, blood pressure, blood glucose, and body weight in rats, however, the mechanisms of action of the lactobacilli remain to be clarified. Therefore, the effect of the culture supernatant of Lactobacillus pentosus strain S-PT84 on the autonomic nerve activity in urethane-anesthetized rats was investigated. Intraduodenal injection of the low-molecular-weight (LMW) fraction (molecules less than 10,000 Da) of the S-PT84 culture supernatant elevated the brown adipose tissue sympathetic nerve activity and reduced the gastric vagal nerve activity. Moreover, intraoral administration of this LMW fraction increased the body temperature of rats above the interscapular brown adipose tissue. These results suggest that the LMW fraction of the S-PT84 culture supernatant affects the autonomic nerve activity and thermogenesis, and that the change in thermogenesis may be caused by the change in the sympathetic nerve activity of brown adipose tissue.

• In vitro effects of doxycycline on inflammatory cytokines and gelatinases in chronic rhinosinusitis.

  Authors: J Ulrich Sommer, Johannes D Schultz, Judith Grossbaier, Jens Stern-Straeter, Karl Hörmann, Alexander Sauter

  In vivo (Athens, Greece). 26(3):369-74.

  Background/Aim: The pathophysiology of chronic rhinosinusitis (CRS) is unknown, but the majority of patients suffer from eosinophilic infiltration. We hypothesised that doxycycline might alter theBackground/Aim: The pathophysiology of chronic rhinosinusitis (CRS) is unknown, but the majority of patients suffer from eosinophilic infiltration. We hypothesised that doxycycline might alter the eosinophil-associated expression of interleukin-5 (IL-5) and eotaxin-3 in CRS and also the expression of matrix metalloproteinase 9 (MMP-9), being involved in the tissue-remodelling in CRS. After obtaining samples from 10 CRS patients with and without nasal-polyposis undergoing functional endoscopic sinus surgery and two healthy individuals, the expression of IL-5, eotaxin-3 and MMP-9 were evaluated by an ELISA technique. The tested agent was doxycycline at 0.1 or 1 mg/ml. IL-5 levels remained unchanged, but eotaxin-3 levels actually increased under doxycycline treatment. The only marker showing a slight drop was MMP-9, albeit not significant. As first clinical trials with doxycycline in the treatment of CRS produced reasonable results we could demonstrate that the underlying pathology is more complex, and doxycycline affects only a part of the factors believed to support the chronic infection of the respiratory mucosa.

• The role of nonsteroidal antiinflammatory drugs and cyclooxygenase-2 inhibitors on experimental colitis.

  Authors: Ana Paula Ribeiro Paiotti, Patrícia Marchi, Sender Jankiel Miszputen, Celina Tizuko Fujiyama Oshima, Marcello Franco, Daniel Araki Ribeiro

  In vivo (Athens, Greece). 26(3):381-93.

  The nonsteroidal anti-inflammatory drugs (NSAIDs) have been widely used in the management of pain and inflammation. Unfortunately, they are associated with dose-dependent gastrointestinal (GI)The nonsteroidal anti-inflammatory drugs (NSAIDs) have been widely used in the management of pain and inflammation. Unfortunately, they are associated with dose-dependent gastrointestinal (GI) adverse events ranging from dyspepsia to symptomatic and complicated ulcers. The mechanism of NSAID action is attributed to the cyclooxygenase (COX) inhibition. New anti-inflammatory drugs have been synthesized, such as selective COX-2 inhibitors, however, these drugs may present side effects, such as modification of the epithelial barrier. Inflammatory bowel disease (IBD) is a common chronic gastrointestinal disorder characterized by alternating periods of remission and active intestinal inflammation. A possible association between the use of NSAIDs and the relapse of IBD has been repeatedly suggested. For this reason, many studies are conducted with the use of COX-2 in experimental models. The objective of this review is to describe the role of NSAIDs and COX-2 inhibitors in different experimental models of colitis. We reviewed controlled trials, original articles, case reports and reviews. The role of selective inhibition of COX-2 in the inflammatory process and the course of experimental and human colitis is controversially discussed. In conclusion, the relative role of COX-2 selective inhibitors on human and experimental colitis remains to be explored. Thus, the use of COX-2 inhibitors in IBD should be considered with caution.

• The Cytotoxic Protein Can Induce Autophagocytosis in Addition to Apoptosis in MCF-7 Human Breast Cancer Cells.

  Authors: Giou-Teng Yiang, Yung-Luen Yu, Pei-Lun Chou, Hsiu-Feng Tsai, Li-Ann Chen, Yen Hsieh Chen, Kuo-Jung Su, Jaang-Jiun Wang, DA-Tian Bau, Chyou-Wei Wei

  In vivo (Athens, Greece). 26(3):403-409.

  Phagocytic clearance of dying cells is found in many phagocytes. It has been shown that dying cells can be phagocytosed by other phagocytic cells through autophagic or apoptotic cellular death. ToPhagocytic clearance of dying cells is found in many phagocytes. It has been shown that dying cells can be phagocytosed by other phagocytic cells through autophagic or apoptotic cellular death. To date, whether cancer cells have such phagocytic activity has not been studied. In this study, our data shows that RC-RNase can trigger cell death in human breast cancer MCF-7 cells through the apoptotic pathway. Interestingly, when treated with cytotoxic protein, the remaining MCF-7 cells can phagocytose the dying MCF-7 cells via autophagocytic activity, demonstrated directly by real-time image observation and electron microscopy analysis. To sum up, this study demonstrates for the first time that RC-RNase can trigger apoptosis and autophagocytosis in MCF-7 cancer cells.

• Efficacy of a Proapoptotic Peptide towards Cancer Cells.

  Authors: Clara E Jäkel, Karoline Meschenmoser, Young Kim, Hans Weiher, Ingo G H Schmidt-Wolf

  In vivo (Athens, Greece). 26(3):419-26.

  Conventional cancer therapies are associated with severe side-effects and the development of drug resistance. Therefore, new strategies to specifically target tumor cells leaving healthy tissueConventional cancer therapies are associated with severe side-effects and the development of drug resistance. Therefore, new strategies to specifically target tumor cells leaving healthy tissue unaffected are of great interest. On this respect, we tested the antimicrobial peptide (KLAKLAK)(2). This peptide exhibits cytotoxicity against human breast cancer and other tumor cells, while healthy cells remain unaffected. Moreover, treatment with this cationic amphipathic peptide results in slower tumor growth and longer overall survival in vivo. Our data suggest a potential use of (KLAKLAK)(2) peptide for patients with breast and other types of cancer.

• Evaluation of Genotoxicity and Subclinical Toxicity of Agaricus blazei Murrill in the Ames Test and in Histopathological and Biochemical Analysis.

  Authors: Jin-Biou Chang, Hsu-Feng Lu, Nien-Chieh Liao, Ching-Sung Lee, Ming-Yang Yeh, Chi-Ming Liu, Ming-Teng Chung, Au Man-Kuan, Jen-Jyh Lin, Ming-Fang Wu, Jing-Gung Chung

  In vivo (Athens, Greece). 26(3):437-45.

  This study was conducted in order to assess the safety and tolerability of Agaricus blazei Murrill (ABM) in general toxicological studies by Ames tests in vitro and in 28-day feeding toxicityThis study was conducted in order to assess the safety and tolerability of Agaricus blazei Murrill (ABM) in general toxicological studies by Ames tests in vitro and in 28-day feeding toxicity experiments. There were no dose-dependent increases or decreases in the number of revertant colonies both with and without metabolic activation in Ames tests. Doses of 10, 5 and 0.1 mg/per mouse of ABM daily were administered by oral gavage to mice (n=10) for 28 days. The effects on clinical observations, clinical pathology, and histopathology were evaluated. There were no significant changes in the brain, heart, kidney, liver, spleen, adrenal gland, testes or ovaries visually. With increasing doses, male and female treated mice did not show any gradual elevation of serum concentration in any of the nine items we examined, except for aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in females. The AST levels of the treatment by medium or high dose and the ALT levels of the treatment by high dose in females were abnormal in comparison to those of the baseline control group, with significant differences. On studying the histological changes in mice, tissue sections of negative control and experimental groups exhibited no apparent pathological alterations. In summary, the Ames test, pathology determinations, biochemical analysis and routine blood parameters were all normal, except for AST and ALT in females. Results showed that the statistical differences observed in one sex were not observed in the other and were not dose dependent.

• Nasolabial cyst: case report with respect to immunohistochemical findings.

  Authors: Reinhard E Friedrich, Hanna A Scheuer, Alexandre T Assaf, Jozef Zustin

  In vivo (Athens, Greece). 26(3):453-8.

  The nasolabial cyst is a rare, usually unilateral lesion arising in the soft tissues adjacent to the alveolar process of the anterior maxilla, above the apices of frontal teeth and below the alarThe nasolabial cyst is a rare, usually unilateral lesion arising in the soft tissues adjacent to the alveolar process of the anterior maxilla, above the apices of frontal teeth and below the alar base. The typical clinical features of nasolabial cysts are: swelling between the upper lip and nasal aperture caused by a smooth and fluctuant, well defined space-occupying lesion, elevation of the nasal ala and obliteration of the nasolabial fold. This report describes some clinical, radiological and morphological findings in a nasolabial cyst. The cyst was lined up with bilayered epithelium showing scattered goblet cells. The immunohistochemical analysis revealed that the basaloid epithelial cells exhibited nuclear positive reactions for p63. The proliferative activity of the epithelial cells was low (<5%). Reaction for podoplanin was only discretely positive in basal cells within the non-inflamed portions but was enhanced in areas with inflammatory changes of the cyst wall. Cytokeratin subtyping showed a distinct expression of intermediate filaments in the nasolabial cyst. Nasolabial cysts are developmental cysts that can be cured by adequate surgical techniques. The expression pattern of podoplanin in this entity points to an association of this protein expression with inflammatory reactions to the cyst.

• Local Tolerance of a Sublingual Nicotine Tablet, an Open Single-centre Study.

  Authors: Lars Sand, Ann-Kristin Lekerud, Mats Wallström, Jan M Hirsch

  In vivo (Athens, Greece). 26(3):463-8.

  Nicotine replacement therapy (NRT) is now widely used in various forms of administration to aid cessation of tobacco use. In this smoking cessation programme, a new tablet formulation has beenNicotine replacement therapy (NRT) is now widely used in various forms of administration to aid cessation of tobacco use. In this smoking cessation programme, a new tablet formulation has been compared to and found bioequivalent to the existing one. The present trial was performed in order to investigate the local tolerance of the new sublingual tablet. The study was performed as a prospective follow-up study of 16 weeks' duration on smokers using the new tablet, Nicorette® Microtab, over a period of 12 weeks. Fifty smokers were included in the study and the oral mucosa was inspected and photographed at each visit. At 12 weeks, participants were asked for their consent to take a biopsy from the site of application. Compliance with tablet use was high, with participants using an average of 12 tablets/day throughout the 12 week treatment period. Adverse events related to treatment were mild and tolerable. The changes observed were classified as being induced by frictional stimuli, with no changes to the deeper layers of the epithelium, and no thickening of the basal layers of the epithelium. The new tablet was considered well tolerated during the course of this study, with a benign local effect on the mucous membrane that was consistent with lesions that are reversible in nature.

• Heterogeneity of DNA Ploidy in Endometrial Carcinoma: Comparison of Different Tissue Samples Obtained during Diagnosis and Treatment.

  Authors: Pavla Libalova, Zdenka Vernerova, Lucie Hubickova-Heringova, Daniela Pinterova, Katarina Kolostova, Vladimir Bobek, Karel Tikovsky, Daniel Housa, Martina Kubecova, Ladislav Pecen, Bohuslav Svoboda

  In vivo (Athens, Greece). 26(3):473-80.

  Aim: Comparison of DNA ploidy status of different tumour tissue samples (fresh/frozen vs. paraffin-embedded; curettage vs. hysterectomy samples) obtained during diagnosis and treatment of patientsAim: Comparison of DNA ploidy status of different tumour tissue samples (fresh/frozen vs. paraffin-embedded; curettage vs. hysterectomy samples) obtained during diagnosis and treatment of patients with endometrial carcinoma. DNA ploidy status and conventional prognostic parameters were recorded for 74 patients with endometrial carcinoma prospectively. In 59 (79.7%) patients the DNA status was described as diploid in all analyzed tissue samples. The remaining 15 (20.3%) cases were described as DNA aneuploid in at least one of the corresponding tissue samples. The concordance between DNA ploidy status in fresh vs. paraffin-embedded hysterectomy samples as well as curettage vs. hysterectomy paraffin-embedded samples was high (kappa coefficient κ=0.6348, 95% confidence interval CI=0.3673-0.9023, and p=0.6408, 95% CI=0.3977-0.8838), however, the methods are not interchangeable. The DNA ploidy discordance observed in our study group seems to document intratumoral heterogeneity that should be expected when applying DNA ploidy status in the clinical management of endometrial carcinoma.

• Ionizing Irradiation Protection and Mitigation of Murine Cells by Carbamazepine is p53 and Autophagy Independent.

  Authors: Hyun Kim, Mark E Bernard, Amy Farkas, Julie Goff, Ronny Kalash, Frank Houghton, Donna Shields, Darcy Franicola, Tracy Dixon, Xichen Zhang, Michael Epperly, Hong Wang, Murat Can Cobanoglu, Joel S Greenberger

  In vivo (Athens, Greece). 26(3):341-54.

  Carbamazepine, a sodium channel blocker and pro-autophagy agent used in the treatment of epilepsy and trigeminal neuralgia, is also an ionizing radiation mitigator and protector. We measured theCarbamazepine, a sodium channel blocker and pro-autophagy agent used in the treatment of epilepsy and trigeminal neuralgia, is also an ionizing radiation mitigator and protector. We measured the effect of carbamazepine, compared to other pro-autophagy drugs (i.e. lithium and valproic acid), on irradiation of autophagy incompetent (Atg5(-/-)) and competent (Atg5(+/+)) mouse embryonic fibroblasts, p53(-/-) and p53(+/+) bone marrow stromal cells, and human IB3, KM101, HeLa, and umbilical cord blood cell and in total body-irradiated or orthotopic tumor-bearing mice. Carbamazepine, but not other pro-autophagy drugs, was a radiation protector and mitigator for mouse cell lines, independent of apoptosis, autophagy, p53, antioxidant store depletion, and class I phosphatidylinositol 3-kinase, but was ineffective with human cells. Carbamazepine was effective when delivered 24 hours before or 12 hours after total body irradiation of C57BL/6HNsd mice and did not protect orthotopic Lewis lung tumors. Carbamazepine is a murine radiation protector and mitigator.

• Human Chorionic Gonadotropin Inhibits N-Methyl-N-nitrosourea-induced Mammary Carcinoma Growth in Female Lewis Rats.

  Authors: Takashi Yuri, Yen-Chang Lai, Katsuhiko Yoshizawa, Airo Tsubura

  In vivo (Athens, Greece). 26(3):361-7.

  Human chorionic gonadotropin (hCG), a hormone produced by the placenta, is essential for maintaining pregnancy. The tumor-suppressive property of hCG against palpable N-methyl-N-nitrosoureaHuman chorionic gonadotropin (hCG), a hormone produced by the placenta, is essential for maintaining pregnancy. The tumor-suppressive property of hCG against palpable N-methyl-N-nitrosourea (MNU)-induced rat mammary carcinomas was examined. A single intraperitoneal injection of 60 mg/kg MNU was administered to female Lewis rats. When the MNU-induced mammary tumors reached a palpable size (≥1 cm in diameter), 0, 100, or 300 IU hCG were intraperitoneally injected 5 times per week for 4 weeks (a total of 20 injections). At the end of the treatment period, the 300 IU hCG treatment had significantly suppressed the growth of MNU-induced mammary carcinomas as compared to the sham treatment. The lower dose of hCG (100 IU) did not exert a significant effect. Final tumor volume and tumor wet weight, respectively, were as follows: sham-treated, 8151.3±1367.1 mm(3) and 6011.3±1042.2 mg; 100 IU hCG, 7480.6±2011.2 mm(3) and 5613.5±1142.0 mg; and 300 IU hCG, 3925.0±875.3 mm(3) and 3482.4±817.3 mg. Serum estrogen and progesterone levels were markedly elevated to pregnancy levels in 300 IU hCG-treated animals, while serum hCG levels remained low, resulting in significantly increased ovarian and uterine weights with multiple corpora lutea in the ovary and cystically dilated endometrial glands. Immunohistochemical studies revealed that almost all hCG-treated mammary carcinoma cells expressed estrogen and progesterone receptors, whereas luteinizing hormone/chorionic gonadotropin receptor was barely detectable. In conclusion, 300 IU hCG treatment for a short duration (4 weeks) suppressed the growth of overt and palpable MNU-induced mammary carcinomas. The mechanism of action may be through accelerated ovarian steroid secretion with the elevation of estrogen and progesterone levels to those in pregnancy.

• Canine malignant mammary gland neoplasms with advanced clinical staging treated with Carboplatin and cyclooxygenase inhibitors.

  Authors: Gleidice Eunice Lavalle, Cecília Bonolo DE Campos, Angélica Cavalheiro Bertagnolli, Geovanni Dantas Cassali

  In vivo (Athens, Greece). 26(3):375-9.

  Surgery remains the treatment of choice for female dogs with mammary gland tumors. Chemotherapy is not commonly used as an adjuvant therapy. Cyclooxygenase 2 (COX-2) has been related to angiogenesisSurgery remains the treatment of choice for female dogs with mammary gland tumors. Chemotherapy is not commonly used as an adjuvant therapy. Cyclooxygenase 2 (COX-2) has been related to angiogenesis development in tumors, disease progression and worse prognosis. The aim of this prospective study was to compare overall survival periods of female dogs diagnosed with advanced mammary tumors submitted to different treatment protocols, including surgery, chemotherapy and cyclooxygenase inhibitors. Twenty-nine female dogs were evaluated and treated with four different protocols. The overall survival of patients with low COX-2 scores was longer when compared to patients with high COX-2 scores. Different proposed adjuvant treatments associated with surgery led to a statistically significant longer overall survival when compared to surgical treatment alone. Canine patients presenting malignant mammary gland neoplasms with advanced clinical staging should be submitted to complementary therapeutic medication based on clinical staging and immunophenotypical characteristics of the disease.

• Oxidative Stress-mediated Biomolecular Damage and Inflammation in Tumorigenesis.

  Authors: Fabiola Sesti, Ourania E Tsitsilonis, Athanassios Kotsinas, Ioannis P Trougakos

  In vivo (Athens, Greece). 26(3):395-402.

  At the cellular level, free radicals are tightly controlled by an inducible antioxidant program, since at low non-hazardous amounts they contribute to physiological signalling and homeostasis.At the cellular level, free radicals are tightly controlled by an inducible antioxidant program, since at low non-hazardous amounts they contribute to physiological signalling and homeostasis. However, high levels of oxidative stress promote the accumulation of damaged biomolecules, the impairment of cell signalling pathways and the increase of oncogenic hits. As the intracellular and extracellular levels of oxidative stress increase during ageing or in various diseases, so does the amount of damaged biomolecules, since the repair mechanisms are also targets of oxidative damage and thus become gradually ineffective over time. Depending on the severity of the biomolecular damage, the responses of normal human cells to oxidants may range from transient growth arrest to premature senescence, and even to cell death. Although some responses are clearly tumour suppressing (apoptosis), others may be potentially oncogenic as they combine damage accumulation with a retained ability for proliferation (transient growth arrest) or with inflammation (senescence, necrosis). This array of events significantly increases the likelihood of the appearance of tumour-initiating cells, which may then give rise to pre-neoplastic focal lesions and eventually to neoplasia. In the present manuscript, we will focus on the role of free radical-mediated biomolecular damage and inflammation in tumorigenesis.

• Biological Activity of SE-10, Granulated Powder of Sasa senanensis Rehder Leaf Extract.

  Authors: Hiroshi Sakagami, Tomohiko Matsuta, Kazue Satoh, Sumiko Ohtsuki, Chiyako Shimada, Taisei Kanamoto, Shigemi Terakubo, Hideki Nakashima, Yurika Morita, Atsuko Ohkubo, Tadashi Tsuda, Katsuyoshi Sunaga, Jun Maki, Tomoko Sugiura, Madoka Kitajima, Hiroshi Oizumi, Takaaki Oizumi

  In vivo (Athens, Greece). 26(3):411-8.

  We have previously reported that alkaline extract of Sasa senanensis leaves (SE) showed potent anti-HIV, anti-UV and radical scavenging activity. In the present study, we investigated the biologicalWe have previously reported that alkaline extract of Sasa senanensis leaves (SE) showed potent anti-HIV, anti-UV and radical scavenging activity. In the present study, we investigated the biological activities of SE-10, a granulated powder of SE supplemented with lactose, lactitol, trehalose and tea extract. Cell viability of mock-infected, HIV-infected, and UV-irradiated cells was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. Scavenging activity of superoxide anion and hydroxyl radicals was determined by electron-spin resonance spectroscopy. Cytochrome P-450 (CYP)3A4 activity was measured by β-hydroxylation of testosterone in human recombinant CYP3A4. SE-10 had slightly higher anti-HIV and anti-UV activities, but slightly lower radical-scavenging and CYP3A4-inhibitory activities, as compared with SE. The present study demonstrates that the biological activities of SE were well preserved during the manufacturing process of SE-10.

• Retinal and Renal Vascular Permeability Changes Caused by Stem Cell Stimulation in Alloxan-induced Diabetic Rats, Measured by Extravasation of Fluorescein.

  Authors: Z Doczi-Keresztesi, J Jung, I Kiss, T Mezei, L Szabo, I Ember

  In vivo (Athens, Greece). 26(3):427-35.

  Aim: To determine whether treatment with the stem cell stimulator Olimpiq® Stem×Cell prevents increase of retinal and renal vascular permeability in alloxan-induced diabetic rats. Two groups ofAim: To determine whether treatment with the stem cell stimulator Olimpiq® Stem×Cell prevents increase of retinal and renal vascular permeability in alloxan-induced diabetic rats. Two groups of Wistar rats were made diabetic by single intraperitoneal injection of Alloxan. The third, the control group, received vehicle alone. One diabetic group received Olimpiq® Stem×Cell treatment for 4 weeks. The permeability of the blood-retinal barrier (BRB) and renal vessels were measured by the extravasation of fluorescein-labeled bovine serum albumin. Six weeks subsequently to Alloxan injection, significantly elevated the tissue fluorescence, the renal vascular leakage and BRB breakdown was demonstrated in the diabetic group, compared to the nondiabetic group. Olimpiq® Stem×Cell treatment significantly reduced the BRB breakdown, tissue fluorescence, and vascular leakage. Olimpiq® Stem×Cell would be a useful choice of treatment for complications associated with increased vascular permeability of diabetes, such as retinopathy or nephropathy.

• Venous Graft for Reconstruction of Neoplastic and Post-traumatic Eyelid Defects.

  Authors: G M Tomassini, V Ansidei, A L Ricci, C Cagini, F Cencetti, S Simonetti, P Covarelli

  In vivo (Athens, Greece). 26(3):447-51.

  Aim: The repair of an eyelid-wide full-thickness defect is a challenging procedure, mostly for the tarso-conjunctival layer reconstruction. The Authors illustrate their own experience inAim: The repair of an eyelid-wide full-thickness defect is a challenging procedure, mostly for the tarso-conjunctival layer reconstruction. The Authors illustrate their own experience in reconstructing eyelid-wide defects with a composite venous wall and skin graft to repair both neoplastic and post-traumatic injuries, aiming to reach both functionally and cosmetically satisfactory results. Eight patients were treated with this procedure; six of them were affected by a local invasive tumor, two had a wide defect following a trauma. Most of the patients had good functional and cosmetic results after a median follow-up of 51 months; only one had a minor complication. Eyelid reconstruction with a venous wall and skin graft is a recently introduced technique that represents a reliable alternative to traditional procedures, granting esthetically and functionally good results.

• Angiomyolipomatous hamartoma of the inguinal lymph node - report of two cases and literature review.

  Authors: Tihana Dzombeta, Mirela Francina, Kristijan Matkovic, Ivan Markovic, Zoran Jukic, Cvjetko Lez, Bozo Kruslin

  In vivo (Athens, Greece). 26(3):459-62.

  Angiomyolipomatous hamartoma is a variant of angiomyomatous hamartoma (AMH), a rare nodal smooth muscle proliferation, first identified as a distinct entity by Chan et al. in 1992. To date, severalAngiomyolipomatous hamartoma is a variant of angiomyomatous hamartoma (AMH), a rare nodal smooth muscle proliferation, first identified as a distinct entity by Chan et al. in 1992. To date, several cases have been described, mostly involving inguinal lymph nodes. We present two cases of angiomyolipomatous hamartoma, in a 52-year-old male and 67-year-old female patient. Both patients were surgically treated. Microscopically, in the affected nodes, the parenchyma was mostly replaced with bundles of smooth muscle cells, fibrous tissue and lobules of mature adipocytes. Only a few atrophic lymphatic follicles were maintained in the subcapsular area. The presence of smooth muscle cells and endothelial cells was confirmed immunohistochemically by staining for smooth muscle actin, desmin and CD31. The hilus contained numerous thick-walled vessels extending to the medulla. Pleomorphism, mitoses and necrosis were absent. Considering there are no reported recurrences of AMH, it probably has benign behaviour; thus extensive resection may not be needed. Nevertheless, we believe that recognition of AMH is important in the differential diagnosis of other pathological conditions that may affect lymph nodes.

• An Antibiotic Care Bundle Approach Based on Results of Rapid Molecular Screening for Nasal Carriage of Methicillin-resistant Staphylococcus aureus in the Intensive Care Unit.

  Authors: Paola Stano, Manuela Avolio, Rita DE Rosa, Maria Luisa Modolo, Stefano M M Basso, Franco Lumachi, Alessandro Camporese

  In vivo (Athens, Greece). 26(3):469-72.

  The potential role of active methicillin-resistant Staphylococcus aureus (MRSA) surveillance in the intensive care unit (ICU), has been recently proposed as a guide for antibiotic treatment inThe potential role of active methicillin-resistant Staphylococcus aureus (MRSA) surveillance in the intensive care unit (ICU), has been recently proposed as a guide for antibiotic treatment in patients suspected of being infected with MRSA by using an antibiotic care bundle (ACB) approach. A group of 376 consecutive ICU patients were prospectively screened for nasal carriage of MRSA using a real-time polymerase chain reaction test. The study group consisted of 244 (64.9%) males and (35.1%) females, with a median age of 64 (range 17-95 years). Overall, 26 (6.9%) patients were positive for MRSA, while 350 (93.1%) were MRSA-negative. No difference was observed in gender and age between groups. During ICU stay, 9 (2.4%) patients developed generalized MRSA infection, of whom 8 out of 26 (30.8%) were MRSA-carriers and one out of the 350 (0.3%) was MRSA-negative. Thus, a strong relationship between MRSA infection and MRSA carriage (relative risk=107.7, 95% confidence interval=14.0-828.5, p<0.0001) was found. Subsequently, in our ICU, we developed and introduced a new ACB approach based on rapid nasal screening results for improving the management of critically ill patients. The use of anti-MRSA agents should be re-evaluated daily on the basis of clinical and laboratory features, with positive cultures from sterile site or signs of active infection supporting prolongation of empirical treatment. On the contrary, MRSA-negative clinical cultures support a de-escalation strategy. In conclusion, the early identification of MRSA-carriers using a rapid molecular screening is safe and accurate, allowing MRSA-positive patients, who will more likely develop MRSA infections, to be detected.

• Influence of gender on mortality and need for extracorporeal membrane oxygenation in neonates with congenital diaphragmatic hernia.

  Authors: Sebastian Berlit, Amadeus Hornemann, Thomas Schaible, Wolfgang Neff, Regine Schaffelder, Jörn Siemer, Marc Sütterlin, Sven Kehl

  In vivo (Athens, Greece). 26(3):481-6.

  Aim: To evaluate the influence of gender in neonates with congenital diaphragmatic hernia (CDH) on survival and to assess the necessity of extracorporeal membrane oxygenation (ECMO) therapy. AllAim: To evaluate the influence of gender in neonates with congenital diaphragmatic hernia (CDH) on survival and to assess the necessity of extracorporeal membrane oxygenation (ECMO) therapy. All parturients with newborns suffering from CDH were included. A total of 425 infants with CDH were analyzed. The primary outcome parameters evaluated were the necessity of ECMO and the survival. Secondary outcome parameters were the mode of delivery, the arterial umbilical cord pH value, the Apgar score, and the postpartum day of death. Cases with incomplete data were excluded. An analysis of the gender distribution of neonates in our study revealed that more males (n=248) than females (n=177) suffered from CDH. This represented a male versus female gender ratio of 58.4% to 41.6%, a significantly different ratio from that for neonates without CDH (p=0.003). Comparing both groups, statistical analysis showed no significant differences in mode of delivery, arterial umbilical cord blood pH, Apgar score, or CDH-relevant parameters of postpartum survival, including the need for ECMO therapy. CDH occurred significantly more frequently in male newborns. However, there were no significant differences in postnatal survival nor in the necessity of ECMO therapy.

• Order-of-magnitude estimates of latency (time to appearance) and refill time of a cancer from a single cancer 'stem' cell compared by an exponential and a logistic equation.

  Authors: Ken M Anderson, Marvin Rubenstein, Patrick Guinan, Minu Patel

  In vivo (Athens, Greece). 26(2):183-9.

  The time required before a mass of cancer cells considered to have originated from a single malignantly transformed cancer 'stem' cell reaches a certain number has not been studied. ApplicationsThe time required before a mass of cancer cells considered to have originated from a single malignantly transformed cancer 'stem' cell reaches a certain number has not been studied. Applications might include determination of the time the cell mass reaches a size that can be detected by X-rays or physical examination or modeling growth rates in vitro in order to compare with other models or established data. We employed a simple logarithmic equation and a common logistic equation incorporating 'feedback' for unknown variables of cell birth, growth, division, and death that can be used to model cell proliferation. It can be used in association with free or commercial statistical software. Results with these two equations, varying the proliferation rate, nominally reduced by generational cell loss, are presented in two tables. The resulting equation, instructions, examples, and necessary mathematical software are available in the online appendix, where several parameters of interest can be modified by the reader www.uic.edu/nursing/publicationsupplements/tobillion_Anderson_Rubenstein_Guinan_Patel1.pdf. Reducing the proliferation rate by whatever alterations employed, markedly increases the time to reach 10(9) cells originating from an initial progenitor. In thinking about multistep oncogenesis, it is useful to consider the profound effect that variations in the effective proliferation rate may have during cancer development. This can be approached with the proposed equation, which is easy to use and available to further peer fine-tuning to be used in future modeling of cell growth.

• Black tea polyphenols inhibit tumor proteasome activity.

  Authors: Taskeen Mujtaba, Q Ping Dou

  In vivo (Athens, Greece). 26(2):197-202.

  Tea is a widely consumed beverage and its constituent polyphenols have been associated with potential health benefits. Although black tea polyphenols have been reported to possess potent anticancerTea is a widely consumed beverage and its constituent polyphenols have been associated with potential health benefits. Although black tea polyphenols have been reported to possess potent anticancer activities, the effect of its polyphenols, theaflavins on the tumor's cellular proteasome function, an important biological target in cancer prevention, has not been carefully studied. Here black tea extract (T5550) enriched in theaflavins inhibited the chymotrypsin-like (CT) activity of the proteasome and proliferation of human multiple myeloma cells in a dose-dependent manner. Also an isolated theaflavin (TF-1) can bind to, and inhibit the purified 20S proteasome, accompanied by suppression of tumor cell proliferation, suggesting that the tumor proteasome is an important target whose inhibition is at least partially responsible for the anticancer effects of black tea.

• 3,3'-Diindolylmethane increases serum interferon-γ levels in the K14-HPV16 transgenic mouse model for cervical cancer.

  Authors: Daniel W Sepkovic, Laura Raucci, Johann Stein, Antoine D Carlisle, Karen Auborn, H Barbara Ksieski, Themba Nyirenda, H Leon Bradlow

  In vivo (Athens, Greece). 26(2):207-11.

  While cervical cancer incidence and mortality rates have declined in the United States, this cancer represents a worldwide threat. Human papilloma viral infection causes cervical neoplasia (CIN).While cervical cancer incidence and mortality rates have declined in the United States, this cancer represents a worldwide threat. Human papilloma viral infection causes cervical neoplasia (CIN). 3,3'-Diindolylmethane (DIM) prevents or inhibits the progression from cervical dysplasia to cancer. The aim of this study is to determine the most effective dose of DIM given continuously in food, that significantly increases serum interferon gamma levels (IFN-γ) in the K14-HPV16 transgenic mouse model for cervical cancer. Five doses of DIM in food were administered to the mouse model for 20 weeks. Serum Interferon gamma (IFN-γ) levels and estrogen metabolite levels were quantified. At 1000 ppm DIM, serum IFN-γ concentrations were significantly increased (p<0.0396). The estrogen metabolites were unchanged. IFN-γ concentrations in CIN free mice and the percentage of CIN free transgenic mice were well correlated (r=0.88). Significant increases in IFN-γ serum concentrations that correlate with the percentage of CIN free mice in each group indicate that 1000 ppm of DIM in food may be the most effective dose for future studies. These results may eventually lead to new and effective vaccination strategies in women already infected with the human papilloma virus.

• The activity of 16 new hydantoin compounds on the intrinsic and overexpressed efflux pump system of Staphylococcus aureus.

  Authors: Anna Dymek, Ana Armada, Jadwiga Handzlik, Miguel Viveiros, Gabriella Spengler, Joseph Molnar, Katarzyna Kiec-Kononowicz, Leonard Amaral

  In vivo (Athens, Greece). 26(2):223-9.

  To evaluate a new series of 16 hydantoin derivatives for activity against the intrinsic and overexpressed efflux pumps of the ATTC 25923 Staphylococcus aureus and the clinical Staphylococcus aureusTo evaluate a new series of 16 hydantoin derivatives for activity against the intrinsic and overexpressed efflux pumps of the ATTC 25923 Staphylococcus aureus and the clinical Staphylococcus aureus HPV-107 strain, respectively. The hydantoin compounds were evaluated for activity against the efflux pumps of the ATTC 25923 S. aureus and the clinical S. aureus HPV-107 strains by the aid of the automated ethidium bromide method. Compounds that inhibited the efflux pumps of either strain were evaluated for ability to reduce or reverse resistance of these strains to oxacillin. Although most of the hydantoins inhibited the efflux pumps of the ATTC strain, none reduced the resistance of this strain to oxacillin. In contrast, the inhibition of the Qac efflux pump present in HPV-107 was inhibited to some degree, by much higher concentrations of the hydantoin compounds than that needed for similar activity against the ATTC strain; only hydantoin PI8a significantly reduced the minimum inhibitory concentration of oxacillin against the HPV-107 strain. Hydantoin compound PI8a may have potential for therapy of a methicillin-resistant S. aureus infection whose multidrug-resistant phenotype is due to overexpression of an efflux pump.

• Comparative transcriptional profiling in HIV-infected patients using human stress arrays: clues to metabolic syndrome.

  Authors: Vincenzo Esposito, Lucrezia Manente, Rosaria Viglietti, Giovanni Parrella, Roberto Parrella, Miriam Gargiulo, Vincenzo Sangiovanni, Angelica Perna, Alfonso Baldi, Antonio De Luca, Antonio Chirianni

  In vivo (Athens, Greece). 26(2):237-42.

  Highly active antiretroviral therapy (HAART therapy) for HIV-1 infection has significantly increased the survival and quality of life of patients with this disease. However, in severalHighly active antiretroviral therapy (HAART therapy) for HIV-1 infection has significantly increased the survival and quality of life of patients with this disease. However, in several epidemiological studies the onset of metabolic syndrome is a phenomenon reported to be extremely frequent. In the present study, genes involved in the molecular cascade responsible for the alteration of fat tissue and of lipid and glucose metabolism in patients with HIV-1 infection treated with antiretroviral therapy were identified. Towards this goal, hybridization using Atlas cDNA Expression Arrays allowed simultaneous monitoring of the expression levels of approximately 250 genes and identification of a panel of changes in relation to different therapeutic groups and in the presence of metabolic syndrome, with some genes being up-regulated, while others are down-regulated in the different subgroups of patients. The results of this analysis have shown a panel of transcriptional changes associated with oxidative stress mechanisms that provide a basis for further studies on understanding of mechanisms that, in vivo, are the foundation the metabolic disorders in patients with HIV infection.

• Cartilage defect regeneration by ex vivo engineered autologous microtissue--preliminary results.

  Authors: Ulrich Meyer, Hans-Peter Wiesmann, Jeanette Libera, Rita Depprich, Christian Naujoks, Jörg Handschel

  In vivo (Athens, Greece). 26(2):251-7.

  We analysed the improvement of cartilage defect regeneration by the use of microspheres of autologous chondrocytes. Autologous chondrocytes from minipigs were cultured using the microsphereWe analysed the improvement of cartilage defect regeneration by the use of microspheres of autologous chondrocytes. Autologous chondrocytes from minipigs were cultured using the microsphere technology. Cartilage defects were surgically introduced to tibia-femoral joints. Tissue constructs were then implanted into defect sites. Histological, immunohistological and transmission electron microscopic analyses were performed. Histological and ultrastructural investigations of chondrospheres revealed a cartilage-like tissue formation, indicated by phenotypically differentiated chondrocyte-like cells surrounded by de novo synthesised chondrogenic extracellular matrix. Clinical inspection of defects demonstrated nearly complete cartilage regeneration in the microtissue treated defect sites, whereas controls exhibited irregular fibrous tissue formation. In vitro-expanded articular chondrocytes are able to form a microtissue capable of repairing cartilage defects in vivo, improving regeneration of cartilage defects.

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