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Ischemic heart disease is the leading cause of death worldwide, and interventions to reduce myocardial infarction (MI) complications are widely researched. Photobiomodulation therapy (PBMT) has altered multiple biological processes in tissues and organs, including the heart.This study aimed to assess the temporal effects of PBMT on cardiac fibrosis activation after MI in rats. In this proof-of-concept study, we monitored the change in expression patterns over time of genes and microRNAs (miRNAs) involved in the formation of cardiac fibrosis post-MI submitted to PBMT.Experimental MI was induced, and PBMT was applied shortly after coronary artery ligation (laser light of wavelength 660 nm, 15 mW of power, energy density 22.5 J/cmPBMT, when applied after ischemia, reversed the changes in mRNA expression of myocardial extracellular matrix genes induced by MI. Surprisingly, PBMT modified cardiac miRNAs expression related to fibrosis replacement in the myocardium. Expression correlations between myocardial mRNAs were assessed. The correlation coefficient between miRNAs and target mRNAs was also determined. A positive correlation was detected among miR-21 and transforming growth factor beta-1 mRNA. The miR-29a expression negatively correlated to Col1a1, Col3a1, and MMP-2 mRNA expressions. In addition, we observed that miR-133 and Col1a1 mRNA were negatively correlated.The results suggest that PBMT, through the modulation of gene transcription and miRNA expressions, can interfere in cardiac fibrosis activation after MI, mainly reversing the signaling pathway of profibrotic genes.

Background Current sudden cardiac death (SCD) risk stratification relies heavily on the assessment of left ventricular ejection fraction (LVEF), but markers that could refine risk assessment are needed. Total fibrosis mass (TFM) and “gray zone” of myocardial fibrosis (GZF) on late gadolinium enhancement cardiac magnetic resonance (LGE-CMR) have been proposed as potential arrhythmogenic substrates. The aim of our study was to determine whether TFM and GZF can predict the occurrence of ventricular arrhythmias in patients with previous myocardial infarction. Methods We performed a single centre retrospective study enrolling all consecutive patients with previous myocardial infarction undergoing LGE-CMR before implantable cardioverter-defibrillator (ICD) implantation for primary or secondary prevention. TFM and GZF were defined as myocardial tissue with signal-intensities >6 SD and 2–6 SD above the mean of reference myocardium, respectively. The primary endpoint was a composite of sudden arrhythmic death, appropriate ICD shock, ventricular fibrillation (VF), or sustained ventricular tachycardia (VT) as detected by the device. Results A total of 55 patients (mean age 62±12 years, 87% male, mean LVEF 30% ± 8%) were included. During a mean follow-up period of 34±15 months, 10 patients reached the primary endpoint (8 appropriate ICD shock, 2 sustained VT or VF). Patients who attained the primary endpoint had similar TFM (28.6g ± 14.5 vs. 23.1g ± 14.5; P=0.283) but larger GZF (25.3g ± 11.0 vs 15.6g ± 7.3; P=0.001). After adjustment for LVEF, GZF remained independently associated with the composite arrhythmic endpoint (adjusted hazard ratio [aHR]: 1.10; 95% CI: 1.03–1.17; P=0.005), whereas TFM did not (aHR: 1.02; 95% CI: 0.98–1.06; P=0.394). Decision tree analysis identified 16.4g of GZF as the best cut-off to predict life-threatening arrhythmic events. The primary endpoint occurred in 9 out of the 22 patients (41%) with GZF >16.4g, but in only 1 of the 33 patients (3%) with GZF ≤16.4g – Figure. Conclusions The extent of GZF seems to be a better predictor of ventricular arrhythmias than TFM. This LGE-CMR parameter may be useful to identify a subgroup of patients with previous myocardial infarction at an increased risk of life-threatening arrhythmic events. Funding Acknowledgement Type of funding sources: None.

Background and aim Prophylactic implantation of an implantable cardioverter-defibrillator (ICD) is class 1 recommendation for heart failure (HF) patients with reduced ejection fraction (HFrEF) even though its proven advantage is weaker among nonischemic aetiology. In fact, in an era where both optimal medical therapy (OMT) and cardiac resynchronization therapy (CRT) significantly reduce sudden cardiac death (SCD), it is questionable whether ICD still have additional value. The aim of this study was to assess the current benefit of ICDs in preventing sudden cardiac death through resuscitated cardiac arrest (RCA), appropriate therapy for sustained ventricular tachycardia (VT) or fibrillation (VF) in a contemporary population of idiopathic HFrEF patients. Methods Single-centre retrospective study of consecutive symptomatic (NYHA class II to IV) idiopathic HFrEF patients with an ICD (either alone or in association with CRT), and remote monitoring with the corresponding software (MerlinTM, LatitudeTM, CarelinkTM, MicroPortTM or BiotronikTM) to assure appropriate event supervising. Idiopathic aetiology was assumed after excluding other probable causes. Coronary angiogram was required to exclude ischemic aetiology. Only those with prophylactic ICD implantation were included. RCA was defined as collapse with clinical signs of cardiac arrest and VF or VT appropriately terminated by ICD. In order to be sustained, VT episode had to have last at least 30 seconds. Results From 781 remote monitoring controlled patients, a total of 187 consecutive symptomatic idiopathic HFrEF patients with an ICD (125 men, mean age 64±18 years) were enrolled. Patients were on optimal medical therapy (ACEi/ARB: n=168, 90%; BB: n=154, 82%; mineralocorticoid antagonists: n=91, 49%; CRT: n=130, 70%; see Table). After a median follow-up of 99 months (IQR 62.2), RCA occurred in 10.7% (n=20) and 36.9% (n=69) had appropriately terminated VT. Both left ventricular ejection fraction (LVEF) improvement and CRT implantation did not independently reduce the incidence of RCA and VT requiring ICD therapy (OR, 1.02; 95% CI, 0.99–1.05; P=0.146 and OR, 0.85; 95% CI, 0.34–2.13; P=0.728; respectively). All cause mortality was 20 (10.7%). Inappropriate therapy was given as shocks to 41 patients (21.9%) and as antitachycardia pacing (ATP) to 30 (16%), opposing with appropriately given therapy to 43 (23%) and 63 (33.7%) patients, respectively (see Figure), contributing to a net clinical benefit (NCB) of 18.8%, favouring ICD implantation. Conclusion In this contemporaneous real-world population of symptomatic idiopathic HFrEF patients, episodes of impending cardiac death were frequent. Prophylactic ICD implantation seems to have added further benefit reducing SCD on top of optimal medical therapy, LVEF improvement and coexisting CRT. Funding Acknowledgement Type of funding source: None

The establishment of a precise and accurate method for the determination of the analyte is the first step in the development of certified reference materials (CRMs), which play a key role in metrological traceability. To assist the production of a glucose CRM in human serum for clinical chemistry routine tests, we developed a gas chromatography-isotope dilution mass spectrometry method using trimethylsilylation and performed two calibration techniques: exact matching approach and calibration curve. Performance of the method and calibration techniques were evaluated for their suitability for the characterization of a CRM candidate carrying out studies to assign the property value and to assure homogeneity as well as short- and long-term stability. Both calibration approaches were equivalent in terms of accuracy and precision and presented bias lower than 1.0 % with relative standard deviations lower than 3.0 %. Relative measurement uncertainties obtained with the exact matching approach calibration method were between 1.0 % and 2.5 % and were lower than the ones obtained with calibration curve method. This finding justifies the use of the time-consuming exact matching method for the assignment of the property value, while the calibration curve procedure suits the other certification studies.

Depending on the presence of mutations that determine isoniazid (INH) susceptibility (katG and inhA), Mycobacterium tuberculosis may be susceptible to high doses of INH or ethionamide (ETH).To describe the INH resistance profile and association of katG mutation with previous INH treatment and level of drug resistance based on rapid molecular drug susceptibility testing (DST) in southern Brazil and central Mozambique.Descriptive study of 311 isolates from Ribeirão Preto, São Paulo, Brazil (2011-2014) and 155 isolates from Beira, Mozambique (2014-2015). Drug resistance patterns and specific gene mutations were determined using GenoType(®) MTBDRplus.katG gene mutations were detected in 12/22 (54.5%) Brazilian and 32/38 (84.2%) Mozambican isolates. inhA mutations were observed in 9/22 (40.9%) isolates in Brazil and in 4/38 (10.5%) in Mozambique. Both katG and inhA mutations were detected in respectively 1/22 (5%) and 2/38 (5.2%). The difference in the frequency of katG mutations in Brazil and Mozambique was statistically significant (P = 0.04). katG mutations were present in 68.8% (33/48) of patients previously treated with INH and 31.2% (15/48) of patients without previous INH. This difference was not statistically significant (P = 0.223).INH mutations varied geographically; molecular DST can be used to guide and accelerate decision making in the use of ETH or high doses of INH.

p21 is a potent inhibitor of cyclin-dependent kinases capable of arresting cell cycle progression. p21 is primarily regulated at the transcriptional level by several transcription factors, including p53. Previously, we reported that certain members of the E2F family of transcription factors may activate p21 transcription via a p53-independent mechanism. To further elucidate the consequences of E2F-1-regulated induction of p21, we developed cell lines with a tamoxifen-dependent form of E2F-1. We confirmed direct interaction of E2F-1 with the proximal region of the p21 promoter. Interestingly, elevated E2F-1 activity was sufficient to arrest a substantial subset of cells in S phase and this effect was correlated to and dependent on the induction of p21 protein. Since E2F proteins control genes required for cell cycle progression and are activated by various oncogenic events, we believe that the p21-dependent arrest described in this report represents an additional mechanism that guards against unrestricted cell proliferation.

The tumor suppressor p53 is the most frequently mutated gene in human tumors. In response to DNA damage, aberrant growth signals, or chemotherapeutic drugs, p53 is stabilized and induces apoptosis and/or cell cycle arrest. While the mechanisms of p53-dependent apoptosis are not well understood, p53-dependent cycle arrest is primary mediated by the CDK inhibitor p21. p53 is a transcriptional activator and it is not surprising that a majority of p53 mutations occur in the core DNA binding domain and affect DNA binding and transactivation of p53 targets in tumors. We used the capability of p53 to activate transcription for developing a new assay that permits rapid determination of the status of p53 in cancer cell lines of different origin. Our strategy involved using a retrovirus containing a p53-regulated lacZ reporter gene that was introduced into colon and breast tumor cell lines to determine p53 status. Simple staining for beta-galactosidase allowed us to confirm that the colon cancer cell lines LIM1215 and HCT116, as well as the breast cancer cell line MCF7. have wild-type p53, and the colon cancer cell line Caco-2 as well as breast cancer cell lines MDA-MB-435 and MDA-MB-231 have mutant p53. This method may be applied to novel cell lines of any origin with unknown status of p53.

Activation of Akt, or protein kinase B, is frequently observed in human cancers. Here we report that Akt activation via overexpression of a constitutively active form or via the loss of PTEN can overcome a G(2)/M cell cycle checkpoint that is induced by DNA damage. Activated Akt also alleviates the reduction in CDC2 activity and mitotic index upon exposure to DNA damage. In addition, we found that PTEN null embryonic stem (ES) cells transit faster from the G(2)/M to the G(1) phase of the cell cycle when compared to wild-type ES cells and that inhibition of phosphoinositol-3-kinase (PI3K) in HEK293 cells elicits G(2) arrest that is alleviated by activated Akt. Furthermore, the transition from the G(2)/M to the G(1) phase of the cell cycle in Akt1 null mouse embryo fibroblasts (MEFs) is attenuated when compared to that of wild-type MEFs. These results indicate that the PI3K/PTEN/Akt pathway plays a role in the regulation of G(2)/M transition. Thus, cells expressing activated Akt continue to divide, without being eliminated by apoptosis, in the presence of continuous exposure to mutagen and accumulate mutations, as measured by inactivation of an exogenously expressed herpes simplex virus thymidine kinase (HSV-tk) gene. This phenotype is independent of p53 status and cannot be reproduced by overexpression of Bcl-2 or Myc and Bcl-2 but seems to counteract a cell cycle checkpoint mediated by DNA mismatch repair (MMR). Accordingly, restoration of the G(2)/M cell cycle checkpoint and apoptosis in MMR-deficient cells, through reintroduction of the missing component of MMR, is alleviated by activated Akt. We suggest that this new activity of Akt in conjunction with its antiapoptotic activity may contribute to genetic instability and could explain its frequent activation in human cancers.

The aim of the work presented was to study the precipitation of the dipeptide derivative AcPheLeuNH2 in order to improve its yield and productivity during enzymatic synthesis catalysed by α-chymotrypsin in tetradecyltrimethylammonium bromide (TTAB)/heptane/octanol reversed micelles. AcPheLeuNH2 solubility in reversed micellar solutions was examined, revealing that a decrease in TTAB concentration, water to surfactant ratio (Wao) and octanol concentration in the organic mixture led to a decrease in solubility. In contrast, no significant influence of buffer molarity and pH was detected. These results were explained by suggesting that the main environment for the solubilization of the dipeptide molecules is the micellar interface rather than the bulk organic phase or the micellar water pool. The synthesis of AcPheLeuNH2 was then carried out using those process conditions that led to the lower solubility values. The activity and stability of α-chymotrypsin, dipeptide yield, selectivity, dipeptide recovery an...

The "bystander effect," produced by ganciclovir-mediated killing of cells transduced with a herpes simplex virus thymidine kinase (HSVtk) gene, defines the cooperative killing of non-HSVtk-transduced cells. In vitro, a major contributor to this phenomenon is metabolic cooperation involving transfer of cytotoxic small molecules between cells via gap junctions. In this study, the bystander effect was assessed in vivo using cells of oral squamous cell carcinoma origin. Mixtures of HSVtk+ and HSVtk- tumor cells were implanted subcutaneously in the left flank of nude mice, and naive HSVtk- cells were implanted subcutaneously in the right flank. When tumors attained a size of 0.5 to 1 cm, the animals were treated with ganciclovir on a daily basis. The tumors comprised of mixed cells in the left flank resolved, consistent with a predicted bystander effect. The naive tumors in the right flank either resolved or became cytostatic showing little further growth compared to controls. Similar results were obtained when naive tumors were grown in both flanks and the tumor in the left flank received intratumoral injection of HSVtk retroviral producer cells or PA317 (HSVtk+) packaging cells, but not parental NIH 3T3 cells. Concomitant treatment with dexamethasone impaired the antitumor effect on the contralateral side. When these experiments were performed in SCID-Beige mice, there was a reduced antitumor effect on the ipsilateral flank and no antitumor response in the contralateral flank. Together with histology of regressing tumors, which showed an infiltration of lymphoid cells, these results are suggestive of an immune-related antitumor response that could account for the distant bystander effect.

In this work, the authors have used technics of multivariate analysis to determine the structure-activity relationships of 46 podophyllotoxin derivatives and analogs studied for their antineoplastic and antiviral activities. The obtained results allow to envisage the possible synthesis of more specific molecules by making modifications in the structure of the model molecular archetype.

In theory, sunshine exposure is sufficient to maintain normal vitamin D concentrations for the optimal growth of newborn infants. To determine whether season of birth, latitude (north v. south) and increasing dosages of vitamin D supplements would influence the growth rate for the first 6 months of life, 255 healthy fall-and spring-born infants from two northern and two southern cities in China were randomly assigned to receive either 100, 200, or 400 IU of vitamin D a day. The study showed that season of birth and dose of vitamin D did not affect the growth rate of infants born in the same latitude, but a significant difference was found in the gain in length over the 6-month period between infants from the north and infants from the south (P = 0.0001). Regional differences among the Chinese people, other than sunshine exposure, may have influenced the difference in length gain.

7527 Background: Initial treatment strategies in FL include observation (obs), rituximab (R), chemotherapy ± R, and XRT. A recent study suggests initial therapy may impact FL survival (JCO 23:8447). NLCS is the first prospective observational study in the US designed to assess FL presentation, prognosis, treatment, and clinical outcomes. We describe initial therapeutic strategy in NLCS FL pts. Methods: FL pts diagnosed within 6 months with no prior lymphomas were recruited in this ongoing study. Data collected includes histology, stage, therapy, response, relapse and death. There is no specified treatment regimen. Results: From 3/04 to 11/05 1493 pts enrolled at 237 sites in the United States. Demographics have been initially reported at ASH 2005 (Blood 106:293a) and are comparable to SEER. Initial therapeutic strategy was: obs, 19%; R-monotherapy, 13%; chemo+R, 51%; XRT, 5%; chemo only, 4%. Chemo+R regimens were: R-CHOP, 59%; R-CVP, 19%; R-fludarabine based,11%; other, 11%. Choice to initiate therapy was associated with FLIPI, stage, and grade (p < 0.0001). Significant regional differences (p < 0.0001) were noted: obs was used in 13% of pts in Southeast and 31% in Northeast (NE); fludarabine-based R-chemo was used in 18% of pts in Southwest and only 3% in NE. Academic sites were more likely than community sites to treat pts on clinical trials (12% vs 4%). In pts treated with R or chemo+R, a higher FLIPI was not associated with decision to utilize chemotherapy. 26% of initially observed pts have switched to active therapy after a median of 2.8 months on obs since diagnosis; this was associated with baseline grade (III>II>I), but not stage or FLIPI. Conclusions: Diverse regimens are used for initial management of FL in the United States. Few pts are treated on clinical trials. Significant differences among regions and between center types suggest physician preference may drive initial therapy. Studies such as NLCS are needed to better understand the impact of initial therapy on short- and long-term outcomes. [Table: see text]

L'A. retrace les changements de statut social et juridique des femmes philippines en examinant l'evolution de l'heritage juridique et colonial philippin a partir de l'immigration des Malays musulmans au XIV e siecle jusqu'a nos jours. L'A. demontre que les femmes beneficiaient d'un rang social eleve, dans ce qui est desormais la Republique des Philippines, jusqu'a la colonisation par l'Espagne au XVII e siecle, date a laquelle les coutumes, la religion et le droit espagnols ont impose de nombreuses restrictions sur les femmes. Il faut attendre l'instauration de la Republique en 1946 pour que des efforts commencent a renverser ces politiques restrictives. L'A. insiste particulierement sur la Constitution de 1986 et les changements operes dans le Code de la famille, qui ont mis en valeur l'egalite juridique des femmes au sein de la famille. L'A. appuie sur le fait qu'un travail politique doit etre soutenu pour assurer une application des lois existantes et poursuivre les changements juridiques en faveur des femmes

The Philippines, an archipelago of 7,107 islands, (about 2,000 of which are inhabited) with a land area of 115,600 square miles, has a population of 42 million. Some 87 major dialects are spoken all over the islands. English and Filipino are the official languages with English as the medium of instruction in higher education. According to the latest census, the literacy rate is 83.4 per cent. Agriculture constitutes the largest single sector of the economy. The Philippines has a total labor force of 14.2 million.Filipino culture is a blend of East and West, although foundationally and originally Malay. Its Eastern heritage – the result of centuries of interaction with neighboring countries of Asia – is a synthesis of three mainstreams: the Malay, Chinese and Hindu. Its Western cultural traits are the result of her colonization by two Western powers: Spain and the United States.

Studies on international human rights had amorphous beginnings before 1945, which gradually evolved into an intricate but “untrodden area of systematic research.” This is largely attributed to the adoption of the United Nations Charter which sets forth the international protection of human rights as a basic purpose. The proclamation of the Universal Declaration of Human Rights by the United Nations General Assembly in December 10, 1948, provided the impetus for the development of new rules of international law defining in specific terms, individual rights and freedoms. The result is not only a substantial and rapidly developing body of law, both substantive and procedural, that has called for a systematic scholarly analysis but a bibliographic output on the subject which has reached alarming proportions to what is now being referred to as “the human rights documentation explosion.”

Legal research traditionally has been concerned with the development or elaboration of legal doctrines, and the raw materials of such research have been statutes, administrative regulations and rulings, and court decisions. This type of research is termed as doctrinal research. Characteristically, a legal scholar undertaking doctrinal research takes one or more legal propositions as a starting point and focus of his study. Research then takes place in the law library, where the scholar tries to locate all relevant statutes, cases and all discussions of his proposition found in encyclopedias, textbooks, treatises, and legal periodicals. If the legal concept is taken from a statute, his sources would necessarily include the statute, its legislative history and, if possible, comparable statutes in other jurisdictions. He then analyzes his readings, formulates his conclusions and writes up his study in the form of a memorandum, a brief, a periodical article or a treatise.

Bacterial beta-galactosidase, encoded by the lacZ gene, serves as a sensitive cytochemical marker in eukaryotic cells and tissues. In transient expression experiments, human and simian cells stain blue 48 hr after transfection with a plasmid containing a lacZ gene, whose expression is directed by a simian virus 40 promoter containing a synthetic lactose operator sequence. Transfection efficiency was about 0.6%. Incorporation of an operator sequence within the promoter permits regulation of beta-galactosidase gene expression by the lacI gene product, the lac repressor. When cells were cotransfected with the lacZ plasmid and a second plasmid containing the lacI gene, beta-galactosidase activity was extinguished. Its activity could be reestablished to original levels upon application of isopropyl beta-D-thiogalactoside to transfected cells. A cell line that stably carries both the lacI and lacZ genes was efficiently induced to synthesize beta-galactosidase after isopropyl beta-D-thiogalactoside administration. In transient expression experiments and in stably transfected lines, repression and induction of beta-galactosidase activity were predominantly at the transcriptional level.

After underlining the serious psychological consequences of radical mastectomy for carcinoma and the importance of breast reconstruction for the patient's complete recovery, the paper briefly reviews the main reconstruction techniques employed in restoring breast volume which is essentially achieved by inserting an artificial prosthesis below a myocutaneous skin flap often taken from the great dorsal. Some mention is also made of recently developed alternative techniques.

The biological and synthetic materials employed over the years for filling breasts after mastectomy or for congenital malformations are reviewed. At the start of the century, breast shape and volume were changed by adipose tissue grafts. In the 50s and 60s dermo-fibro-adipose grafts and later auto-grafts and grafts of the large epiploon were used. Synthetic materials have also developed, from early century attempts using paraffin injections to the discovery for industrial use of plastics. Following the first polyvinyl prostheses, Cronin developed silicon prostheses for medical purposes and in later years these went through numerous changes in form, volume and design up to the recent skin expanders. Polyurethane has also been used in attempts to reduce capsular retraction.

After pointing out the need for reconstruction of the nipple-areola complex and the restoration of symmetry between the two breasts after radical mastectomy for cancer, the paper briefly examines the techniques currently available for the achievement of these "finishing touches" that usually take longer then the primary and fundamental breast reconstruction.

A careful review of the literature stimulated these comments on current developments in conservative surgery as an alternative to radical mastectomy in stage I and II breast cancer with particular reference to their implications for cosmetic rehabilitation. It is emphasised that the data so far available indicate comparable results in terms of locoregional recurrences, metastases and survival. It is also pointed out that the efficacy of the conservative techniques reflects new concepts of the natural history of breast cancer. The hope is expressed that these encouraging data will be confirmed by more extensive and systematic clinical experiment in order to minimise the indications to more radical surgery which notoriously has such devastating physical and psychological effects that it takes extremely complex and demanding plastic surgery to reshape the patient to psychological sexual and social health.

The use of external fixators in the treatment of complex mandibular fractures is described, as are the advantages offered by the technique which permits accurate, controlled reduction of the bone fragment without immobilising the temporomandibular joint.

A review of the literature on complications in septorhinoplasty is presented. Though rare, complications may arise in the surgical treatment of such a delicate anatomical area as the nasal septum and both theoretical and practical knowledge of the problem is necessary in order to prevent and/or handle such unexpected events. Complications are grouped according to aetiopathogenesis into infectious, haemorrhagic, traumatic, systemic and miscellaneous.

The entire literature on abdominoplasty is reviewed with a description of the various techniques developed since the introduction of the operation.

The recent computerisation of operating theatre and forensic files facilitated a retrospective study of 136 plastic surgery operations performed on 89 patients presenting traumatic or surgical scalp tissue loss in the San Camillo Hospital, Rome between 1/1/1980 and 31/12/1987. The study provided some useful data for the planning of the surgical approach to this delicate anatomical area. Where the dimensions of the defect and the trophic condition of the local tissues permitted, the best functional and cosmetic results were obtained by using flaps of contiguous skin. In this field, new prospects are offered by the introduction of skin expanders whose gradual and controlled distension of the skin permit excellent functional and cosmetic results.