Multi-omic lineage tracing predicts the transcriptional, epigenetic and genetic determinants of cancer evolution.

Cancer is a highly heterogeneous disease, where phenotypically distinct subpopulations coexist and can be primed to different fates. Both genetic and epigenetic factors may drive cancer evolution, however little is known about whether and how such a process is pre-encoded in cancer clones. Using single-cell multi-omic lineage tracing and phenotypic assays, we investigate the predictive features of either tumour initiation or drug tolerance within the same cancer population. Clones primed to tumour initiation in vivo display two distinct transcriptional states at baseline. Remarkably, these states share a distinctive DNA accessibility profile, highlighting an epigenetic basis for tumour initiation. The drug tolerant niche is also largely pre-encoded, but only partially overlaps the tumour-initiating one and evolves following two genetically and transcriptionally distinct trajectories. Our study highlights coexisting genetic, epigenetic and transcriptional determinants of cancer evolution, unravelling the molecular complexity of pre-encoded tumour phenotypes.

[Analysis of medication prescription in an Argentinian geriatric hospital]. / Análisis de la prescripción de medicamentos en una comunidad geriátrica Argentina.

BACKGROUND: Although polypharmacy may be justified in elderly patients with multiple diseases, it may be dangerous, especially when it includes potentially inappropriate medications (PIM). AIM: To identify inappropriate medication and factors associated with the most relevant prescriptions among older people. MATERIAL AND METHODS: Cross-sectional observational analysis of drugs prescribed during the first trimester of 2010 to 179 older adults aged 77 ± 8 years (98 women), living in a geriatric reference hospital in Argentina. The use of potentially inappropriate medications (PIM) in elderly patients was analyzed using Beers Criteria updated to 2012. RESULTS: The mean number of drugs prescribed per individual was 6.1 ± 2.7. The most commonly used drugs were anti-ulcer agents (58.1%), agents acting on the renin-angiotensin system (54.2%), antithrombotic medications (50.8%) and benzodiazepines (50.8 %). The use of antacids and anti-ulcer agents, psychotropic drugs and PIM was significantly higher among patients using six or more drugs daily, compared to the less medicated group (odds ratio (OR) = 6.8, 95% confidence intervals (CI) 3.5-13.2; OR=15.0, 95%CI 5.9-38.4; OR=5.0; 95%IC 2.6-9.8, respectively). Thirty one percent of participants using non-steroidal anti-inflammatory drugs, were not receiving medications for gastric protection. One to four drugs included in the Beers list were prescribed to 66% of participants. CONCLUSIONS: Despite the high prevalence of use of antacids and anti-ulcer agents, these drugs were not prescribed to a significant proportion of patients using non-steroidal anti-inflammatory drugs. Strategies to optimize pharmacotherapy in the elderly population are urgently required.

Análisis de la prescripción de medicamentos en una comunidad geriátrica Argentina / Analysis of medication prescription in an Argentinian geriatric hospital

Background: Although polypharmacy may bejustified in elderly patients with multiple diseases, it may be dangerous, especially when it includes potentially inappropriate medications (PIM). Aim: To identify inappropriate medication and factors associated with the most relevant prescriptions among older people. Material and Methods: Cross-sectional observational analysis of drugs prescribed during the first trimester of 2010 to 179 older adults aged 77 ± 8 years (98 women), living in a geriatric reference hospital in Argentina. The use of potentially inappropriate medications (PIM) in elderly patients was analyzed using Beers Criteria updated to 2012. Results: The mean number of drugs prescribed per individual was 6.1 ± 2.7. The most commonly used drugs were anti-ulcer agents (58.1%), agents acting on the renin-angiotensin system (54.2%), antithrombotic medications (50.8%) and benzodiazepines (50.8 %). The use of antacids and anti-ulcer agents, psychotropic drugs and PIM was significantly higher among patients using six or more drugs daily, compared to the less medicated group (odds ratio (OR) = 6.8, 95% confidence intervals (CI) 3.5-13.2; OR=15.0, 95%CI 5.9-38.4; OR=5.0; 95%IC 2.6-9.8, respectively). Thirty one percent of participants using non-steroidal anti-inflammatory drugs, were not receiving medications for gastric protection. One to four drugs included in the Beers list were prescribed to 66% of participants. Conclusions: Despite the high prevalence of use of antacids and anti-ulcer agents, these drugs were not prescribed to a significant proportion of patients using non-steroidal anti-inflammatory drugs. Strategies to optimize pharmacotherapy in the elderly population are urgently required.

Dento-alveolar features and diet in an Etruscan population (6th-3rd c. B.C.) from northeast Italy.

OBJECTIVES: The purpose of this study was to assess the prevalence of some dento-alveolar features (caries, dental calculus, tooth wear, enamel hypoplasia, abscesses, retraction of the alveolar bone, chipping, and ante mortem tooth loss) on an Iron Age sample from the Etruscan necropolis of Spina (North-Eastern Italy) and to make a relation with dietary evidence found in the archaeological records. These dental features were used to evaluate the oral health status and dietary habits with the aim of shedding light on the lifestyle and living conditions of this population. MATERIALS AND METHODS: The sample analysed consisted of 680 permanent teeth from 80 burials. RESULTS: Overall, individuals at Spina exhibited a low incidence of caries, abscesses and ante mortem tooth loss, high calculus rates and severe attrition, suggesting a subsistence pattern based on a mixture of fishing, pastoralism and agriculture. The low incidence of hypoplasia was probably related to their good nutritional conditions and weak metabolic stresses during early childhood. CONCLUSIONS: These findings, quite similar to those of Etruscans from other parts of Italy, are very different from those of other populations living in the same territory during the same time. This pattern is indicative of a general good health status in this Iron Age population and may be related, at least in part, to the Etruscan diet.

[Drug prescriptions and demographic indicators in a diabetic population in Argentina]. / Patrones de prescripción de medicamentos e indicadores demográficos en una población diabética de Argentina.

OBJECTIVE: To analyse prescription patterns for antihyperglycemic drugs and co-medication in a group of diabetic patients regarding demographic variables. METHOD: Observational, cross-sectional study performed from the prescriptions of diabetic patients who attended a public health service in Argentina during 2008-2009. The programme Epidat 3.1 was used for statistical analysis. RESULTS: We included 813 patients, among which 455 (56%) were women. There was a greater prevalence of women under 40 years old (OR: 1.64; 95% CI: 1.05-2.56). Sixty-six point seven percent (n=542) of the population used at least one drug acting on the cardiovascular system, finding a positive statistical association between frequency of use and age (P<.00001). CONCLUSIONS: We found that there was a higher prevalence of women than men with early onset of type 2 diabetes mellitus. There were no differences between the sexes regarding frequency of use of drugs for the cardiovascular system.

The solitary fibrous malignant tumour of the kidney: clinical and pathological considerations on a case revisiting the literature.

The solitary fibrous tumours (SFT) are rare spindle cell neoplasms which generally originate from the pleura; also described are cases of SFT in other locations, included the genital-urinary tract. Described in the ambit the kidney are 19 cases of SFT and such rarity of localisation makes rather unknown the histogenesis and the prognosis of the lesion. We report the case of a 72 year old lady who attended our Unit for a mass which was clinically palpable at the level of the left hemiabdomen. Following an abdominal ultrasound scan a neoformation was highlighted which a successive tomodensitographic test indicated as being of likely pertinence of the middle third of the left kidney; the mass had a diameter of approximately 19 cm. A radical nephrectomy has been conducted. The histological examen highlighted a solitary fibrous tumour: the presence of hypercellularity, of cellular pleiomorphism and of a high number of mitosis has led to a histopathological diagnosis of malignancy of the neoplasm under examination. Departing from this case a review of the literature is carried out. The SFT of the kidney can have an aggressive character and more the present has hystopathological characters and clinical results are still rather unknown.

[Solitary fibrous tumor of the kidney. A propos of a case].

Solitary Fibrous Tumors (SFT) are rare spindle cell neoplasm that typically originate from the pleura. However, cases of the SFT are described with origin in other organs, including the urinary and genital apparatus. Within the kidney, except from the renal pelvis, only 19 cases of SFT are described and such rarity of localization makes the histogenesis and the prognosis of the tumor rather unknown. We report the case of a 72-year-old lady who attended our Unit for a mass that was clinically palpable at the level of the left hemiabdomen. The tomodensitographic test indicated a 19cm-diameter mass of likely pertinence of the middle bystender of the left kidney. She had undergone left radical nephrectomy. The histological examination highlighted a solitary fibrous tumor (SFT): the presence of hypercellularity, of cellular pleiomorphism and of a high number of mitosis has led to a histological diagnosis of malignancy for the neoplasm analyzed. The SFT are of rare clinical comparison: this does not allow for a deep knowledge of the lesion histogenesis and prognosis; moreover, the clinical behavior should be more precisely defined.

[Scrotal masses: considerations on two cases of epididymal tumors].

The scrotum is a fibromuscular sac that contains the testis, epididymis, spermatic cord and the associated fibrous coatings. All these components can be affected by different variety of pathologic phenomena, including congenital, inflammatory and neoplastic events. When a scrotal mass is observed, there are two basic questions to be answered, i.e. if the mass is intratesticular or extratesticular, and if it is of cystic or solid nature. Apart from a few rare exceptions, intratesticular solid masses should be considered malignant, while extratesticular masses with liquid content are generally benign. CASE REPORTS. Two cases of tumor are hereby presented: they originated from the epididymis, and their clinical presentations did not allow making a differential diagnosis between benign and malignant tumor during the preoperative examinations. After presenting the diagnostic doubts to patients, and once obtained their informed consent, surgeries were performed allowing for a precise histological diagnosis, and at the same time, proving to be also valid therapeutic tools.

IgG antibodies against phospholipase A2 from Crotalus durissus terrificus: cross-reaction with venoms from Bothrops species from Argentina

We examined the ability of IgG anti-crotalic PLA2 to cross-react with Bothrops spp. venoms, from snakes found in the northeast of Argentina. Immunoblotting and ELISA tests showed that IgG anti-crotalic PLA2 recognize antigens of bothropic venoms. Indirect hemolytic activity tests showed that the quantity of antibodies that neutralized 50% of Crotalus durissus terrificus venom (ED50: 2.1 mg IgG anti-crotalic PLA2/100 µg of venom) were also able to neutralize venom from other snakes in the following proportion: 34% of B. alternatus, 18% of B. diporus and 12% of B. jararacussu. Likewise, direct PLA2 activity neutralization tests showed a similar cross-neutralization pattern including 56% of B. alternatus, 29% of B. diporus and 30% of B. jararacussu. In addition, in a myotoxic activity neutralization test, measured by plasma activity of creatine kinase, 35% of B. alternatus venom and 26% of B. diporus venom were neutralized, while no neutralization was detected with B. jararacussu venom. This study presents original data concerning cross-reactions between bothropic venoms from Argentina and IgG anti-crotalic PLA2. Our results suggest that anti-crotalic PLA2 antibodies should not be used to neutralize PLA2 activity of B. alternatus, B. diporus and especially B. jararacussu venoms; nor to enrich commercial antivenoms against these Bothrops species.(AU)

[A case of particular diffusion of prostate cancer].

Neoplastic diffusion can occur due to dissemination, continuity, through lymphatic or haematic vessels, or, more rarely originate from surgical instruments. We report a particular case of prostate cancer spread. A 64-year-old man was diagnosed with undifferentiated prostate cancer through prostate biopsy. The patient was treated with a total androgenic block allowing a decrease in PSA blood level. The patient, wishing to regain his sexual activity compromised by hormonal therapy, interrupted the treatment spontaneously and unchecked. 19 months later he contacted us again: we had to hospitalize him due to a 12-hour anuresis. A urgent right transcutaneous nephrostomy was carried out, yielding an improvement in the patient's condition. A descending pyelography carried out by means of nephrostomy revealed a completely reduced urethral lumen. During the following surgery for transcutaneous urinary derivation we observed the two ureters entangled in whitish tissue, spreading bilaterally up to the renal pelvis. The histological examination of tissue samples showed the presence of neoplastic metastasis of prostatic origin. We assume that this neoplastic diffusion has occurred due to permeability through the lymphatic vessels of the urethral wall, producing a subsequent neoplasm growth: this has been limited by the urethral connective sheath, thus preventing its wide diffusion to the surrounding tissues, but fostering its spreading upwards along the ureter pathway.

[The clinical and pathological comments from a case of sarcomatoid renal carcinoma]. / Considerazioni cliniche e patologiche da un caso di carcinoma sarcomatoide del rene

Sarcomatoid renal carcinoma represents 1-5% of primitive tumours of the kidney. MATERIALS AND METHODS. A young man aged 34 presented with left side colic pain, preceded two days earlier by hematuria without pain; no previous complaints. Echotomography was performed revealing a tumour lesion of the upper left renal pole, which was subsequently confirmed by computerized tomography scanning, with a maximum diameter of approximately 6cm, and evidence of multiple lymph nodes with increased volume. A left-side nephroadrenalectomy was performed with locoregional lymphadenectomy; a sarcomatoid renal carcinoma with massive metastases in 4/19 excised lymph nodes was shown through histological examination. IL-2 therapy was commenced but interrupted due to intolerance. After only three months the disease resumed locally with diffuse lymph nodes metastases and multiple encephalic metastases. A carboplatinum and vinorelbine therapy was started; the possibility of an allogeneic transplant with non-myeloablative conditioning ("miniallogeneic") was suggested, though being impossible to be performed due to the further rapid disease progression. The patient died after approximately 10 months. CONCLUSIONS. Sarcomatoid renal carcinoma is characterised histologically by closely interconnected epithelial and connective tissue elements; sarcomatous cells seem to originate from a phenotype conversion to carcinomatous cells, as they both belong to the same DNA clone. Their behaviour is very aggressive and the prognosis is unfavourable: 6-month average survival after diagnosis. Radical nephrectomy is still the main therapeutic approach, although it is has no significant influence on prognosis and survival rate. This case showed an approximately 10-month patient's survival, with some different chemotherapeutic approaches being followed (first IL-2 and then carboplatinum) after surgery.

Partial characterization of enterocin MR99 from a corn silage isolate of Enterococcus faecalis.

AIMS: To assess the inhibitory activity on Gram-positive and Gram-negative bacteria of several species of enterococci recovered from a natural corn silage. METHODS AND RESULTS: The inhibitory activity of strains of Enterococcus faecalis (58), Enterococcus faecium (35), Enterococcus gallinarum (3) and Enterococcus casseliflavus (4) were studied employing indicator strains from various sources (clinical, food and ATCC). Enterococcus faecalis MR99, the only strain with inhibitory activity, inhibited other enterococci, Listeria spp., Staphylococcus aureus, Clostridium spp., Bacillus spp., Escherichia coli, Shigella sonnei and Shigella flexneri. The bacterium contained only one conjugative pheromone-responsive plasmid. The partially chromatography-purified MR99 enterocin (PPE) had a molecular weight of approx. 5000 Da and a pI of 6.2, was sensitive to proteolytic enzymes and could be extracted in benzene and butanol. It appeared stable to adjustment of pH 4.0, 5.0, 6.0, 7.0 and 8.0 and was resistant to heat. Inactivation was at 15 min at 121 degrees C. Enterocin MR99 was bactericidal on strains of Listeria monocytogenes, Staph. aureus, and bovine mastitis agents, it was bacteriostatic on E. coli. Although enterocins MR99 and AS48 have inhibitory activity on Gram-negative bacilli, PCR studies demonstrated a lack of relationship between them. CONCLUSIONS: The active component had a protein nature, was resistant to heat and presented a wide inhibitory spectrum. SIGNIFICANCE AND IMPACT OF THE STUDY: The biological properties of Ent. faecalis MR99 suggest that this strain merits further investigations so it can be applied in human and veterinary health programmes.

Clinical status and parasitic infection in a Wichí Aboriginal community in Salta, Argentina.

In a study, carried out in 2000, of the clinical and parasitological status of a Wichi Aboriginal community living in the suburbs of Tartagal, northern Salta, Argentina, 154 individuals were screened for parasitic infections. Ninety-five faecal samples were also obtained from the same population. Ninety-three percent of the subjects were positive for 1 or more of the parasites investigated by direct test and 70.5% of them had parasitic superinfection. The most frequent helminths were Strongyloides stercoralis (50.5%) and hookworm (47.4%). We found low reinfection rates and a long reinfection period after treatment and provision of safe water and sanitation. Serum reactivity of these patients was analysed by enzyme-linked immunosorbent assay and indirect immunofluorescent assay and 22.1% of them had anti-Toxocara antibodies, 16.2% were positive for a complex antigen of Leishmania braziliensis, 29.9% were positive for a complex Trypanosoma cruzi antigen, and 17.5% were positive for a specific Trypanosoma cruzi antigen, Ag 163B6/cruzipain.

[When and why to use penile prostheses: seven years of experience]. / Quand et pourquoi la prothèse pénienne: sept ans d'expérience.

During the last years important changes in the management of the patients with erectile dysfunction have been verified. Psychologist, neuro-physiologist and bioengineers have given a valuable help in this field. Implant surgery represents, to our notice, the curative more acceptable choice for patient and the partner, since the results are excellent and the materials used extremely manageable. In each case the degree of satisfaction of the patient remains the only legitimate indicator currently for establish the effectiveness of the therapy.

[Erectile dysfunction in Peyronie's disease. Experience in 132 patients]. / Disfunzioni dell'erezione nella malattia di La Peyronie. Esperienza su 132 pazienti.

BACKGROUND: The exact percentage of sexual dysfunctions resulting from Peyronie's disease remains very difficult to evaluate even today. The purpose of the present paper is to examine the series observed in our Department in order to make an assessment of the erectile function of patients found to be suffering from Peyronie's disease. METHODS: Over the course of 9 years, 132 patients have come to our observation; they were originally examined for sexual function disturbances or for other pathologies of a uro-andrological nature. RESULTS: In all, a total of 208 lesions were observed. Only 21 patients (22.5%) were found to be completely asymptomatic at the time of diagnosis. The most frequent symptom was shaft curvature (alone or in association with pain: 47%). The most commonly observed site for plaques was the back of the corpus cavernosum (43%). Impotentia erigendi was observed in 29 patients (22%) whereas impotentia coeundi was found in 50 (38). The functionally most significant lesions were those affecting the septum or in some way involved with the septum: 19 of 29 patients with impotentia erigendi presented such lesions. CONCLUSIONS: Considering the generally progressive course of the disease, the therapeutic approach taken by our Department is to fit a prosthesis to patients who have been suffering from Peyronie's disease for at least six months; conservative therapy is only undertaken in the rare cases of patients who have a perfectly preserved erection but who suffer from impotentia coeundi (as a result of curvature or pain).

Novel 7-oxyiminomethyl derivatives of camptothecin with potent in vitro and in vivo antitumor activity.

In an attempt to synthesize potential anticancer agents acting by inhibition of topoisomerase I (Topo I) a new series of oxyiminomethyl derivatives in position 7 of camptothecin (CPT) was prepared. The synthesis relied on the condensation of 20S-CPT-7-aldehyde or 20S-CPT-7-ketones with alkyl, aryl, heteroaryl, arylalkyl, and heteroarylalkyl O-substituted hydroxylamines. The compounds were tested for their cytotoxic activity in vitro against H460 non-small lung carcinoma cell line, the activity being for 24 out of 37 compounds in the 0.01-0.3 microM range. A QSAR analysis indicated that lipophilicity is the main parameter correlated with cytotoxicity. Investigation of the DNA-Topo I-drug cleavable complex showed a rough parallelism between cytotoxicity and inhibition of Topo I. Persistence of the DNA cleavage after NaCl-mediated disruption of the ternary complex suggests that for the most potent compounds, e.g., 15, the cytotoxicity was at least in part related to stabilization of the complex, as also supported by the persistence of the DNA-enzyme complex in drug-treated cells. The in vivo antitumor efficacy of the most potent analogue (15) was evaluated in direct comparison with topotecan using human lung tumor xenograft models. In the range of optimal doses (2-3 mg/kg), the improved efficacy of 15 was documented in terms of inhibition of tumor growth and rate of complete response.

Reversible carnitine palmitoyltransferase inhibitors with broad chemical diversity as potential antidiabetic agents.

A series of carnitine related compounds of general formula XCH(2)CHZRCH(2)Y were evaluated as CPT I inhibitors in intact rat liver (L-CPT I) and heart mitochondria (M-CPT I). Derivative 27 (ZR = -HNSO(2)R, R = C(12), X = trimethylammonium, Y = carboxylate, (R) form) showed the highest activity (IC(50) = 0.7 microM) along with a good selectivity (M-CPT I/L-CPTI IC(50) ratio = 4.86). Diabetic db/db mice treated orally with 27 showed a significant reduction of serum glucose levels.

Methicillin resistance study in clinical isolates of coagulase-negative staphylococci and determination of their susceptibility to alternative antimicrobial agents.

AIMS: To achieve reliable detection of methicillin resistance in clinical isolates of coagulase-negative staphylococci. METHODS AND RESULTS: Strains (105) were evaluated by normatized antimicrobial susceptibility methods, and for the presence of the methicillin resistance-determining mecA gene, using the polymerase chain reaction. Correlation between phenotypic and genotypic methods was obtained in 87.6% of the samples. Six strains, classified as methicillin-susceptible by phenotypic assays, revealed the presence of the mecA gene, indicating that methicillin resistance expression was probably repressed. Another seven isolates failed to show mecA amplification after displaying methicillin resistance in phenotypic evaluations. The susceptibility of the methicillin-resistant isolates to other antimicrobial agents was variable. CONCLUSION: Genotypic determination of the mecA gene proved to be the most reliable method for detection of methicillin resistance. SIGNIFICANCE AND IMPACT OF THE STUDY: Correct assessment of methicillin resistance, such as that attained through genotyping, is essential for defining therapeutic strategies, particularly when treating severely compromised patients.

[Importance of the bacterium-host interaction in coagulase-negative staphylococci isolated from neonates]. / Importancia de la relación huésped-parásito en estafilococos coagulasa negativa aislados de neonatos.

OBJECTIVE: Neonates represent a high risk population for infections by coagulase-negative staphylococci (CNS). To have a better understanding of these process our purpose was to compare the expected result of the bacterium-host interaction given by the neonates' risks factors and the micro-organisms' virulence factors with the condition of infecting or colonising strain that emerge from the diagnosis on the basis of the clinical symptoms. METHODS: We studied 24 neonates who were submitted to an epidemiological control establishing as risk factors: catheters, vesicle sounds, previous surgery and immunodepressed conditions. In the CNS recovered from clinical samples we determined the following virulence factors: synergistic hemolysis, slime production, adherence to Teflon catheters and hydrophobicity. RESULTS: We found correlation between the clinical diagnosis and the expected result of the bacterium-host interaction in 21 patients (87.5%). Among them, in 8 patients infection didn't occurred in spite of having the micro-organisms 3 from 4 virulence factors since the patients didn't have risk factors. CONCLUSIONS: A microbiological study based entirely on identification and treatment can alter the biological context. It is necessary to understand the bacterium-host interaction for an appropriate comprehension of the bacterial diseases.

Ex vivo and in vitro effect of human immunodeficiency virus protease inhibitors on neutrophil apoptosis.

Polymorphonuclear leukocytes (PMNL) from human immunodeficiency virus (HIV)-infected patients exhibit accelerated apoptosis and impaired functional activity. HIV protease inhibitor-based therapy produces improvements in both acquired and innate immune responses. Ex vivo and in vitro effects of HIV protease inhibitors on apoptosis and chemotaxis of PMNL were evaluated. After therapy, there was a rapid and significant decrease of PMNL apoptosis, which correlated with increased chemotactic function. These findings were found both in patients with immunological and virological response and in control subjects who showed an increase in CD4(+) T cell counts but no concomitant decline in HIV load. After in vitro treatment with ritonavir or indinavir, apoptosis of both HIV-infected and -uninfected PMNL markedly decreased and correlated with significant enhancement of chemotaxis. These results suggest that HIV protease inhibitors may improve the PMNL function by reducing the apoptosis rate and that this effect may, at least in part, be independent of their antiviral activity.