Distal urethrectomy for localized penile squamous carcinoma in situ extending into the urethra: an updated series.

INTRODUCTION: The presence of squamous carcinoma in situ (CIS) of the distal penis extending into the urethral meatus is generally considered a contraindication for glans-sparing procedures. Distal urethrectomy with subsequent reconstruction can provide an alternative approach toward urethral resection while providing penile preservation in select cases. Unfortunately, long-term oncologic outcomes with this approach are ill-defined. MATERIALS AND METHODS: Between 1988 and 2012, five patients at Indiana University Medical Center underwent distal urethrectomy with reconstruction for penile squamous CIS extending into the urethral meatus. This cohort was retrospectively reviewed to evaluate functional and oncological outcomes. RESULTS: Of the five patients, four presented with glanular lesions and were initially managed with Mohs procedure in three cases, and local excision in one. The final patient presented with extensive urethral disease and was managed with primary urethrectomy. Reconstruction was performed with penile skin pedicle grafts in four patients and perineal urethrostomy in one. Final pathologic stage was T1 in one patient and Tis in the remaining four. Follow-up ranged from 6 to 96 months. One local recurrence was verified; however, it occurred outside the urethral area. This was confirmed in the pathologic analysis after the patient underwent a partial penectomy. Meatal dilation was necessary in two patients 12 and 7 months after the procedure. CONCLUSION: Distal urethrectomy for penile squamous CIS extending into the urethral meatus is a valid alternative to achieve negative surgical margins while preserving a penile function. Oncologic outcomes appear acceptable but larger series are still warranted to confirm our findings.

The rs5743836 polymorphism in TLR9 confers a population-based increased risk of non-Hodgkin lymphoma.

Non-Hodgkin lymphoma (NHL) has been associated with immunological defects, chronic inflammatory and autoimmune conditions. Given the link between immune dysfunction and NHL, genetic variants in toll-like receptors (TLRs) have been regarded as potential predictive factors of susceptibility to NHL. Adequate anti-tumoral responses are known to depend on TLR9 function, such that the use of its synthetic ligand is being targeted as a therapeutic strategy. We investigated the association between the functional rs5743836 polymorphism in the TLR9 promoter and risk for B-cell NHL and its major subtypes in three independent case-control association studies from Portugal (1160 controls, 797 patients), Italy (468 controls, 494 patients) and the US (972 controls, 868 patients). We found that the rs5743836 polymorphism was significantly overtransmitted in both Portuguese (odds ratio (OR), 1.85; P=7.3E-9) and Italian (OR, 1.84; P=6.0E-5) and not in the US cohort of NHL patients. Moreover, the increased transcriptional activity of TLR9 in mononuclear cells from patients harboring rs5743836 further supports a functional effect of this polymorphism on NHL susceptibility in a population-dependent manner.

[Follow-up of the first case of Mycobacterium ulcerans infection documented by PCR, genotyping and culture in the Republic of Congo-Brazzaville]. / Suivi du premier cas d'infection à Mycobacterium ulcerans confirmé par culture, PCR et génotypage République du Congo-Brazzaville.

This article presents follow-up data from the first patient in whom Mycobacterium ulcerans infection (MUI) was documented by PCR, genotyping and culture in the Republic of Congo-Brazzaville. Findings show the importance of regular clinical and microbiological evaluation for the disseminated form of the disease. The patient was probably infected in Pointe Noire where MUI has been described but never documented. Culture of specimens collected before antibiotic treatment showed that the bacterium was sensitive to the antibiotics being administered (streptomycin and rifampin) and was identical to isolates from Atlantic-coast regions of West Africa where MUI is endemic. The patient was treated with streptomycin and rifampin for 12 weeks in association with surgery. During treatment clinical examination was performed every day and microbiological analysis every two weeks. The duration of follow-up from the end of specific antibiotic treatment was 26 months. Medical treatment failed to prevent bone involvement and fistulae that were treated by surgery. However medical treatment may have limited dissemination of the disease. Serial microbiological evaluation was useful to detect bone involvement in this patient, but persistent positive gene amplification is not a proof of active disease. This study confirms that MUI is still endemic in the region of Pointe Noire. This finding underlines the need to optimize epidemiologic surveillance, laboratory diagnostic capabilities, and therapeutic management in the Republic of Congo-Brazzaville.

Intestinal toxicity induced by 5-fluorouracil in pigs: a new preclinical model.

BACKGROUND: The goal of this study was to develop an animal model of intestinal injury induced by 5-fluorouracil (5-FU) in pigs. METHODS: Six domestic pigs were used as control (healthy group) and another 6 malnourished pigs orally received 5-FU (treated group). After 4 weeks of treatment, pigs were sacrificed and jejunum, ileum and colon were isolated for histological, immunological and biochemical analyses. RESULTS: 5-FU caused a decrease in the intestinal mass. Disaccharidase, and phosphate alkaline activities, and glutathione redox cycle were disrupted by 5-FU. Histopathological alterations in the crypts and villous were greater in the small intestine than in the colon. 5-FU decreased the number of peripheral and intestinal leukocytes, promoting an increase in T-cytotoxic cells and a decrease in T-helper and B cells. CONCLUSION: This pig model of intestinal dysfunction closely mimics the common side effects of cancer chemotherapy in humans, and provides a useful tool for evaluating novel antimucotoxic agents.

Newborn screening: a national public health programme in Brazil.

The newborn screening programme started in Brazil (1976) through isolated initiatives, without governmental directions and/or policies. According to Health Ministry (2000) data the coverage was 55% and unevenly distributed. Only 17 out of 27 Brazilian states had more than 30% coverage. Public budgets covered only diagnostic examinations. There were no official data about assistance, patient follow-up or detected disorders. The creation of the National Programme (2001) has provided new perspective for newborn screening (NBS) in the public health system. It has provided important official data and established management and care units for each state: Reference Services in Newborn Screening. The programme screened about 13 million newborns from October 2001 to December 2005. The coverage increased to 80.2% (2005) and 74% of the states presented coverage of over 70%. Within 34 accredited Reference Services in 27 Brazilian states, all provide screening for PKU and CH. Ten of them provide screening for haemoglobinopathies as well, and three of them provide also for CF. The Reference Services altogether count on at least 170 health professionals, such as paediatricians, endocrinologists, nutritionists, psychologists and social workers. They are qualified to assist positive cases, within the policies established by the National Programme. There has been significant increase in NBS coverage and follow-up assuredness, including detected cases before the National Programme (10,935 positive cases) mostly in those regions where the programme did not exist. There has been significant evolution in the Newborn Screening as a Public Health Program in Brazil due to the government's commitment (federal and each component state).

Constitutive nitric oxide synthases are responsible for the nitric oxide production in the ischemic aged cerebral cortex.

Aged brain shows reduced biological plasticity to meet emergency conditions such as ischemia, a process in which nitric oxide (NO) and apoptosis have been shown to play important roles. Using a model of transient global ischemia, we have analyzed the NO system and the p53, bax and bcl-2 response in the cerebral cortex of aged rats. Although immediately after ischemia the NO level is maintained, the reperfusion period increases NO concentrations together with the following: (i) greater bulk-protein nitration mainly due to a 50-kDa immunoreactive band; (ii) an increase in p53 protein; and (iii) an up-regulation of Bax together with a down-regulation of Bcl-2. These results match up with induced endothelial nitric oxide synthase expression immediately after ischemia and in neuronal nitric oxide synthase with the reperfusion. However, inducible nitric oxide synthase was not altered with ischemia/reperfusion. Altogether, these data suggest that NO production in cerebral cortex of aged ischemic animals is due to the constitutive NO synthase isoforms. This response is accompanied by the increased expression of pro-apoptotic proteins.

Apoptotic response to TGF-beta in fetal hepatocytes depends upon their state of differentiation.

Transforming growth factor-beta (TGF-beta1) induces death of fetal hepatocytes by an apoptotic mechanism. However, even when very high concentrations and/or long-term exposure to the cytokine is used, 40-50% of cells always survive. The process of cell survival is coincident with changes in morphology and phenotype, with cells showing a fibroblastic appearance and eliciting an epithelial-fibroblastic transition. Surviving cells continue responding to TGF-beta in terms of growth control. Expression of liver-specific genes is very low in these cells; this effect is due to the decrease in their rate of transcription as soon as 2 h after the addition of the factor. Surviving cells present a decreased DNA binding activity for liver-enriched transcription factors, an increased DNA binding activity for AP-1, and a high expression of protooncogenes. These cells are immature hepatocytes since in the presence of the appropriate signal (i.e., epidermal growth factor), they can differentiate, organizing in cell clusters and increasing both liver-specific mRNA expression and liver-enriched transcription factor activity. In accord with these results, TGF-beta, secreted at high concentrations during liver carcinogenesis, might induce death of normal cells while providing a selective advantage for the survival of cells that are "partially transformed" or "less differentiated" and unresponsive to the factor.

Two pairs of proven monozygotic twins discordant for familial amyloid neuropathy (FAP) TTR Met 30.

Twin studies are an important tool in medical genetics for the evaluation of the relative roles of genetic and non-genetic factors in several diseases. Familial amyloidotic polyneuropathy type I (FAP-I), TTR Met 30, was present in two sets of proven monozygotic (MZ) twins, one from Majorca and the other from Portugal. Monozygosity was established by analysis of DNA polymorphisms. Both pairs were discordant for age at onset and some clinical manifestations of FAP-I. We reviewed the differences in age at onset and clinical features in both sets and in two other pairs of presumed MZ twins with FAP-I and compared them with those in MZ twin pairs with other Mendelian disorders, such as neurofibromatosis type 1, Huntington's disease, facioscapulohumeral muscular dystrophy, and myotonic dystrophy. We conclude that, in addition to the postulated modifying genes, there must be a significant contribution from non-genetic factors to the phenotypic variability of FAP-I (age at onset and clinical expression), either because of environmental differences or stochastic events during (or after) the twinning process.

Effect of thioacetamide and dexamethasone on serum lipids in rats fed on high-fat sunflower or olive oil diets.

We have previously reported that high-fat diets develop hepatic steatosis and, depending on the fat quality, affect serum lipid levels differently (J Nutr Sci Vitaminol, 1997, 43, 155-160). The aim of this work is to study the influence of high-fat diets (14% sunflower or olive oils) on serum lipids in a model of hepatic acute damage induced by thioacetamide, and their influence when dexamethasone is administered before thioacetamide injection. Serum lipids and hepatic collagen have been evaluated using biochemical methods, and the steatotic process by histological staining. The results showed that hepatic steatosis and fibrosis are developed either by high-fat diets or thioacetamide injection. Pretreatment with dexamethasone did not decrease the hepatic collagen content. Thioacetamide injection alone or pretreatment with dexamethasone produced increase in serum tryglicerides (TG), total cholesterol (TC) and LDL-C in both high-fat diet groups, and a HDL-C increase in the olive-oil group, even though the atherogenic indices (HDL/TC and HDL/TG) were different depending on the enriched diet. The administration of high-fat diets to study the influence of the fat quality on health and disease should be interpreted carefully due to the ability of the diets themselves to cause hepatic damage.

Olive oil- and fish oil-enriched diets modify plasma lipids and susceptibility of LDL to oxidative modification in free-living male patients with peripheral vascular disease: the Spanish Nutrition Study.

The present study describes a clinical trial in which Spanish patients suffering from peripheral vascular disease (Fontaine stage II) were given specific lipid supplements. Designed as a longitudinal intervention study, patients were provided with olive oil for 3 months, followed by a 3 month wash-out period, then supplemented with a combination of fish oil and olive oil for the final 3 months. Changes in plasma and lipoprotein fatty acid composition and susceptibility of LDL to in vitro oxidation were examined. Furthermore, lipid-supplement-induced changes in LDL properties were measured as relative electrophoretic mobility and macrophage uptake. In addition, thirteen patients not provided with olive oil and fish oil were included as a control group and twenty healthy age-matched individuals were used as a reference group. A complete clinical study and a nutritional survey concerning food habits and lifestyle were performed every 3 months. Yao indices and claudicometry did not change significantly with dietary intervention although changes in plasma lipid composition suggested an improvement in the condition of the patients. The intake of the fish-oil supplement resulted in significantly increased plasma levels of eicosapentaenoic acid (20:5n-3) and docosahexaenoic acid (22:6n-3) in comparison with baseline concentrations, olive-oil and control groups. Fish-oil consumption significantly decreased plasma triacylglycerol levels compared with the olive-oil period, control and reference groups. The susceptibility of LDL to Cu-mediated oxidation was lower in the patients consuming olive oil and the fish-oil supplement than in the control group, and the uptake of LDL by macrophages was significantly lower in the group supplemented with fish oil. In conclusion, consumption of olive oil together with a dietary supplement of fish oil may be useful in the nutritional management of patients suffering from peripheral vascular disease in terms of increasing plasma n-3 long-chain polyunsaturated fatty acids and decreasing susceptibility of LDL to oxidation.

Effects of iron deprivation on Mycobacterium avium growth.

SETTING: Acquired immune deficiency syndrome (AIDS) patients have increased iron deposition in different tissues which may favour the growth of Mycobacterium avium, a common bacterial opportunist in these patients. OBJECTIVE: To test whether reducing the iron loads in macrophages in vitro and in vivo reduces M. avium proliferation. DESIGN: Mycobacterial proliferation was evaluated in vitro either in axenic media or cultured macrophages and in vivo in mice after manipulation of the iron status. RESULTS: Three different compounds--desferrioxamine (DFO), N,N'bis(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid (HBED) and a 1-amino-3-(2-bipyridyl)isoquinoline derivative (VUF8514)--were found to inhibit the growth of M. avium in axenic medium. DFO and HBED were also active in inhibiting the intramacrophagic growth of M. avium, while the use of VUF8514 was prevented by its toxicity towards the host cell. Both DFO and HBED enhanced the mycobacteriostatic effect induced in bone marrow derived macrophages by interferon gamma. In vivo, an iron poor diet led to reduced M. avium proliferation whereas the intraperitoneal administration of either DFO or HBED had small effects as they impacted little on the iron status of mice. CONCLUSION: These results confirm that iron withholding is a means of inhibiting the growth of M. avium. In vitro data suggest that iron chelating compounds may be useful as adjunct therapy against M. avium, once their in vivo activity is optimized.

High-fat sunflower and olive oil diets affect serum lipid levels in steatotic rat liver differently.

This work describes the long-term effects of two different diets, one rich in olive oil and the other in sunflower oil, on serum lipid and lipoprotein levels after the establishment of fatty liver in rats 8 and 15 months old. The serum lipid and lipoprotein levels as well as the steatotic process have been evaluated by biochemical and histological methods, respectively. The results showed that fatty liver was well developed with both long-term high-fat diets, and hepatocytes were filled with many lipid droplets. This process was more evident in the portal zones, where fat hepatocytes were more numerous. Serum total cholesterol (TC) and HDL-C levels were highest in the sunflower oil fed rats, whereas the TG and LDL-C levels were highest in the olive oil group. Finally, the atherogenic indexes (HDL/TC, HDL/LDL, HDL/(TC-HDL)) were higher in the sunflower oil diet group than in the olive oil group.

Susceptibility of beige mice to Mycobacterium avium: role of neutrophils.

The beige mutation in C57BL/6 mice has been shown to increase the susceptibility to infection by Mycobacterium avium. In this study, we confirmed those results and showed that the effect of the beige mutation was most obvious after infection with a strain of lower virulence than with a highly virulent isolate of M. avium. The dissemination of M. avium from the gut was observed with both C57BL/6 and beige mice but was faster in the latter. The expression of gamma interferon (IFN-gamma) and the priming for tumor necrosis factor production during an in vivo infection were similar between beige and immunocompetent C57BL/6 mice. IFN-gamma produced during the infection of beige mice was protective in the spleen, and the administration of recombinant IFN-gamma restored the resistance in the spleen to levels similar to those found in control mice. There were no histological differences between wild-type and beige mice with respect to granuloma formation in the liver. The increased susceptibility of beige mice to M. avium as manifested in the liver was reduced by transfusing neutrophils from wild-type C57BL/6 mice. Likewise, depletion of neutrophils from C57BL/6 mice rendered them as susceptible to M. avium infection of the liver as beige mice. Our results point to the participation of neutrophils in the defect of beige mice in addition to other defects. Furthermore, these results show that neutrophils play a significant role in the defense mechanisms against mycobacterial infections and that beige animals may be a useful model for study of the role of neutrophils in mycobacteriosis.

Characterization of the virulence of Mycobacterium avium complex (MAC) isolates in mice.

The virulence of different isolates of MAC was studied in naturally susceptible BALB/c mice. In preliminary experiments, MAC bacteria forming smooth transparent colonies on solid media (SmT variants) were found to be virulent for BALB/c mice, causing progressive infection; smooth opaque (SmOp) were generally avirulent, being slowly eliminated from the infected organs; and rough (Rg) variants were either avirulent or as virulent as SmT variants. We chose to compare the virulence of different isolates of MAC of different origins, studying only the SmT morphotype. Strains of MAC isolated from naturally infected animals were those that most consistently caused progressive infections. AIDS patients-derived isolates were of intermediate virulence or devoid of virulence in mice. The environmental strains were eliminated from mice or did not proliferate. Strains of MAC isolated from individuals who were not infected by HIV varied in virulence from completely avirulent to highly virulent. There was no close correlation between virulence and restriction fragment length polymorphism (RFLP) type, although all highly virulent strains were of the A/I type. There was also no correlation between virulence analysed in vivo and the ability to grow in cultured macrophages.

Role of gamma interferon and tumor necrosis factor alpha during T-cell-independent and -dependent phases of Mycobacterium avium infection.

To design an effective immunotherapy for Mycobacterium avium infections, the protective host response to the infection must be known. Here we analyzed the role of gamma interferon (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) in the innate and acquired responses to M. avium infections in mice. T-cell depletion studies showed that CD4+ T cells were required for control of the infection. CD(4+)-depleted mice showed enhanced bacterial proliferation and at the same time showed a reduction in the level of expression of both IFN-gamma and TNF-alpha mRNAs in spleen cells. In contrast, M. bovis BCG immunization restricted M. avium proliferation and at the same time promoted expression of the mRNAs for the two cytokines. In vivo depletion studies using specific monoclonal antibodies showed that both IFN-gamma and TNF-alpha are involved in an early protection possibly involving NK cells, and furthermore, IFN-gamma is involved in the later T-cell-protective response to infection. In vivo neutralization of IFN-gamma during M. avium infection also blocked the priming for enhanced TNF-alpha secretion triggered by endotoxin. Both cytokines were found to be involved in the resistance expressed in BCG-immunized animals and exhibited additive bacteriostatic effects in vitro on bone marrow-derived macrophages infected with different strains of M. avium. These data suggest that both cytokines act in an additive or synergistic fashion in the induction of bacteriostasis and that IFN-gamma is also involved in priming TNF-alpha secretion.

Role of interleukin-6 in the induction of protective T cells during mycobacterial infections in mice.

Interleukin-6 (IL-6) has been shown to regulate numerous functions of the immune system including the differentiation of T-cell subpopulations. Here we examined the involvement of this cytokine in the in vivo generation of a population of T cells able to protect mice against mycobacterial infections. BALB/c mice were infected intravenously with Mycobacterium avium 2447 and anti-IL-6 monoclonal antibodies were administered intraperitoneally throughout the course of the infection. Control mice were able to control the mycobacterial proliferation 1 month after inoculation, whereas mice whose IL-6 had been blocked showed progressive bacterial growth. To distinguish a role for IL-6 associated to the induction or expression of immunity mediated by T cells, we immunized mice with M. bovis bacillus Calmette-Guérin (BCG) Pasteur and challenged them 2 months later with M. avium. One group of mice received anti-IL-6 during the BCG vaccination and another during the M. avium challenge. When M. avium proliferation was assessed at day 30 of the challenge, it was found that the administration of anti-IL-6 during vaccination reduced the protection afforded by BCG compared to administration of the isotype control antibody. No difference in bacterial proliferation was observed at day 30 of challenge when antibodies were administered during M. avium challenge. Our results show that protective T cells arise during M. avium infections in mice after differentiating in the presence of IL-6.

Induction and expression of protective T cells during Mycobacterium avium infections in mice.

Mycobacterium avium is an opportunistic pathogen that infects individuals suffering from chronic lung disease or immunocompromised patients such as AIDS patients. Here we show that a highly virulent isolate of M. avium proliferated as extensively in T cell deficient as in immunocompetent mice. T cell deficient mice allowed a progressive growth of a less virulent AIDS-derived isolate of M. avium while immunocompetent mice arrested the growth of this isolate. Adoptive transfer of T cell enriched spleen cells between congenic strains of mice differing at the Bcg/Ity/Lsh locus showed that only naturally resistant BALB/c.Bcgr (C.D2) mice infected with the highly virulent strain of M. avium or the naturally susceptible BALB/c mice infected with the lower virulence isolate developed protective T cells and that these cells only mediated protection when transferred to naturally susceptible, but not to naturally resistant, mice. Both strains of M. avium proliferated in bone marrow-derived macrophages cultured in vitro and they were both susceptible to the bacteriostatic effects induced in the macrophages by crude lymphokines produced by concanavalin A-stimulated spleen cells.

Sensorivagal nature of oesophageal submucous layer nerve endings. Determination of surgical degeneration methods.

Two types of surgery were performed on domestic cats to establish the source and functional significance of the apparatuses we have denominated perivascular, perifascicular and free endings of the oesophageal submucous layer. After extirpation of the cranial regions of the sympathetic ganglionic chain, of all the cervical ganglia and of the four proximal thoracic ganglia, no Wallerian degeneration of the concerned nerve endings was observed, thus ruling out their dependence on this neural pathway. After ablation of the central portion of the nodose ganglion, of the vagal nerve. Wallerian degeneration was observed in these nerve-ending apparatuses, thus demonstrating: (i) that these nerve apparatuses depend on perikarya situated in the nodose ganglion and (ii) that they are sensory receptors.

Vegetative innervation of the esophagus. III. Intraepithelial endings.

Intraepithelial fibers do occur in the mucosa of the esophagus, as demonstrated by the osmium tetroxide-zinc iodide method in cats and rhesus monkeys. The esophagus is divided into three parts, in order to study the penetration incidence, and the uppermost and the lowest show the greatest density of penetration, while in the middle portion only occasional fibers in small numbers are found. The specific characteristics observed in this type of fiber, such as their distribution along the wall of the esophagus, the levels reached by their endings within the mucous epithelium itself and this same epithelium considered as the specified destination of the endings, lead to the belief that they may be functionally regarded as structures of a sensory character.