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The photochemistry of Fe(III) coordinated to natural uronate-containing polysaccharides has been investigated quantitatively in aqueous solution. It is demonstrated that the photoreduction of the coordinated Fe(III) to Fe(II) and oxidative decarboxylation occurs in a variety of uronate-containing polysaccharides. The photochemistry of the Fe(III)-polyuronic acid system generated a radical species during the reaction which was studied using the spin trapping technique. The identity of the radical species from this reaction was confirmed as CO2•- indicating that both bond cleavage of the carboxylate and oxidative decarboxylation after ligand to metal charge transfer radical reactions may be taking place upon irradiation. Degradation of the polyuronic acid chain was investigated with dynamic light scattering, showing a decrease in the hydrodynamic radius of the polymer assemblies in solution after light irradiation that correlates with the Fe(II) generation. A decrease in viscosity of Fe(IIII)-alginate after light irradiation was also observed. Additionally, the photochemical reaction was investigated in plant root tissue (parsnip) demonstrating that Fe(III) coordination in these natural materials leads to photoreactivity that degrades the pectin component. These results highlight that this Fe(III)-polyuronic acid can occur in many natural systems and may play a role in biogeochemical cycling of iron and ferrous iron generation in plants with significant polyuronic acid content.

Infrared (IR) thermography renders invisible infrared radiation with intuitive coloration in images and videos taken of objects, reactions, and processes. Educators can take advantage of this technology to extend students’ sensory perception of chemical reactions or processes that absorb or release heat in rich detail. In theory, IR thermography can be applied essentially universally for such analysis given that any change in thermal energy must result in, or from, the change of potential energy due to the interactions among atoms, molecules, and photons. Through the use of IR thermography, students can visualize otherwise invisible evidence of what is occurring on the molecular level in a variety of chemical process such as evaporative cooling, phase change, dissolution, titration, and enzymatic reactions. While not new, IR cameras are rapidly becoming affordable with models that connect easily with smartphones and tablets. The price decrease has opened the door for large-scale implementation in the chem...

The PEOPLE trial aimed to identify new immune biomarkers in negative and low programmed death-ligand 1 (PD-L1) (0%-49%) advanced non-small-cell lung cancer (aNSCLC) patients treated with first-line pembrolizumab. Here we report the main outcomes and the circulating immune biomarkers analysis.The primary endpoint of this phase II trial was the identification of immune biomarkers associated with progression-free survival (PFS). Overall survival (OS), objective response rate (ORR), disease control rate (DCR), duration of response (DoR) and safety were secondary endpoints. Absolute cell counts for 36 subsets belonging to innate and adaptive immunity were determined by multiparametric flow cytometry in peripheral blood at baseline and at first radiologic evaluation. An orthoblique principal components-based clustering approach and multivariable Cox regression model adjusted for clinical variables were used to analyze immune variables and their correlation with clinical endpoints.From May 2018 to October 2020, 65 patients were enrolled. After a median follow-up of 26.4 months, the median PFS was 2.9 months [95% confidence interval (CI) 1.8-5.6 months] and median OS was 12.1 months (95% CI 8.7-17.1 months). The ORR was 21.5%, DCR was 47.7% and median DoR was 14.5 months (95% CI 6.4-24.9 months). Drug-related grade 3-4 adverse events were 9.2%. Higher T cell and natural killer (NK) cell count at baseline and at the first radiologic evaluation were associated with improved PFS, DCR and OS. On the contrary, higher myeloid cell count at baseline or at the first radiologic evaluation was significantly associated with worse OS and DCR.Circulating immune biomarkers can contribute to predict outcomes in negative and low PD-L1 aNSCLC patients treated with first-line single-agent pembrolizumab.

The photochemistry of Fe(III) coordinated to natural uronate-containing polysaccharides has been investigated quantitatively in aqueous solution. It is demonstrated that the photoreduction of the coordinated Fe(III) to Fe(II) and oxidative decarboxylation occurs in a variety of uronate-containing polysaccharides. The photochemistry of the Fe(III)-polyuronic acid system generated a radical species during the reaction which was studied using the spin trapping technique. The identity of the radical species from this reaction was confirmed as CO

Toxicological effects of butylparaben (BuP) and ethylparaben (EtP) on zebrafish (Danio rerio) early-life stages are not well established. The present study evaluated, using zebrafish embryos and larvae, the toxicity of BuP and EtP through benchmark dose (BMD) approach. BuP was more toxic than EtP to zebrafish larvae. In fact, Lethal Concentration 50 (LC50) values at 96 h post-fertilization (hpf) for BuP and EtP were 2.34 mg/L and 20.86 mg/L, respectively. Indeed, BMD confidence interval (lower bound (BMDL) - upper bound (BMDU) was 0.91–1.92 mg/L for BuP and 10.8–17.4 mg/L for EtP. Zebrafish embryos exposed to 1 mg/L, 2.5 mg/L of BuP and 5 mg/L, 10 mg/L, 20 mg/L, 30 mg/L of EtP showed several developmental abnormalities and teratological effects compared to negative control. Exposed zebrafish developed reduced heartbeat, reduction in blood circulation, blood stasis, pericardial edema, deformed notochord and misshaped yolk sac. Embryos exposed to the highest concentrations of the chemicals (2.5 mg/L of BuP, 10 mg/L, 20 mg/L and 30 mg/L of EtP) showed the developmental abnormalities at 48 hpf while those treated with 1 mg/L of BuP and 10 mg/L of EtP reported behavioral changes at 72 hpf, including trembling of head, pectoral fins and spinal cord. This research identified the lethal and sublethal effects of BuP and EtP in zebrafish early-life stages and could be helpful to elucidate the developmental pathways of toxicity of parabens.

A quick freeze shows an enzyme's secretsOrganic radicals are chemically useful in enzymatic reactions but are often hard to observe, owing to their short lifetimes. Donget al.used rapid freeze-quench methods to trap two intermediates formed by a noncanonical radical S-adenosylmethionine (SAM) enzyme: a fragmented SAM molecule bound to the iron-sulfur cluster through an iron-carbon bond and a product-like radical. The structure of the SAM-bound enzyme reveals a noncolinear arrangement of carbon, sulfur, and iron atoms. The arrangement of bonds suggests that the organometallic intermediate may be created through a two-electron nucleophilic mechanism. A subsequent radical intermediate is formed on the protein substrate and resolves by oxidation to form the amino acid product diphthamide.Science, this issue p.1247

IscU, the central scaffold protein in the bacterial ISC iron–sulfur (Fe–S) cluster biosynthesis system, has long been recognized to bind a Zn2+ ion at its active site. While initially regarded as an artifact, Zn2+ binding has been shown to induce stabilization of the IscU structure that may mimic a state biologically relevant to IscU’s role in Fe–S cluster biosynthesis. More recent studies have revealed that SufU, a homologous protein involved in Fe–S cluster biosynthesis in Gram-positive bacteria, also binds a Zn2+ ion with structural implications. Given the widespread occurrence of the “IscU-like” protein fold, particularly among Fe–S cluster biosynthesis systems, an interesting question arises as to whether Zn2+ ion binding and the resulting structural alterations are common properties in IscU-like proteins. Interactions between IscU and specific metal ions are investigated and compared side-by-side with those of SufU from a representative Gram-positive bacterium in the phylum Firmicutes. These studies...

IscU, the central scaffold protein in the bacterial ISC iron-sulfur (Fe-S) cluster biosynthesis system, has long been recognized to bind a Zn

Non-expert scientists are frequently involved in research requiring data acquisition over large geographic areas. Despite mutual benefits for such "citizen science", barriers also exist, including 1) difficulty maintaining user engagement with timely feedback, and 2) the challenge of providing non-experts with the means to generate reliable data. We have developed a system that addresses these barriers. Our technologies, KnoWare and InSpector, allow users to: collect reliable scientific measurements, map geo-tagged data, and intuitively visualize the results in real-time. KnoWare comprises a web portal and an iOS app with two core functions. First, users can generate scientific 'queries' that entail a call for information posed to a crowd with customized options for participant responses and viewing data. Second, users can respond to queries with their GPS-enabled mobile device, which results in their geo- and time-stamped responses populating a web-accessible map in real time. KnoWare can also interface with additional applications to diversify the types of data that can be reported. We demonstrate this capability with a second iOS app called InSpector that performs quantitative water quality measurements. When used in combination, these technologies create a workflow to facilitate the collection, sharing and interpretation of scientific data by non-expert scientists. Keywords: Citizen Science, Cell-phone Spectrometer, Water Contaminant Analysis, Web and Mobile Development, Visualization Introduction The prevalence of smartphones represents an opportunity to gather, create, and consume data from individuals on a scale that has not previously been available. Even the 'average' mobile devices in widespread use have significant computing power and are typically GPS-enabled. Scientific research that requires widespread sampling over geographic areas can benefit greatly by mobilizing members of the population equipped with these devices to conduct scientifically useful measurements or activities. Data generated in this way can complement existing data collection methods. One key advantage of these 'citizen science' initiatives is that sourcing large groups of individuals is an efficient strategy to collect large-scale data sets because contributors typically volunteer their time in exchange for meaningful participation. However, in spite of the benefits, multiple barriers work against increased prevalence of citizen science initiatives, including 1) the difficulty maintaining prolonged user engagement with meaningful feedback, and 2) the challenge of providing non-experts with the means to collect reliable data. We have developed technologies to directly address these barriers by providing 'citizen scientists' with 1) an easy-to-use mechanism to participate in scientific investigations with real time visual feedback, and 2) software and a device that interface with iPhones to collect reliable scientific measurements from color-based test kits in widespread use. Our technologies currently focus on allowing users to record water quality measurements since such data collection requires large, repeated measurement, and there are many existing citizen science initiatives already focused on water quality that may benefit. Our technology platform, called KnoWare, comprises a paired web portal and iOS application for data visualization and mapping. In this report, we use KnoWare in combination with an additional iOS application we created (InSpector) to process images taken with the iPhone camera using an add-on device. The InSpector app allows users to obtain reliable readings of colorimetric water quality tests that currently depend on subjective color-comparisons. These technologies work together to provide an engaging workflow for the collection and interpretation of environmental water quality data by non-professional, citizen scientists. Literature Review The technologies we have developed facilitate citizen science based data collection efforts with the following motivations: increasing exposure and 'literacy' in STEM areas, the potential impact of involving the broader public in scientific efforts to advance research, and opportunities to engage students at our own institution in the development of software and devices with 'real-world' applications. …

We analysed our experience with computed tomography (CT)-guided percutaneous cryoablation (PCA) in patients who were not surgical candidates or refused surgery for small to medium-sized renal masses. Two freezing cycles were applied and separated by a passive warming cycle using 1.7- and 2.4-mm cryoprobes under either general anaesthesia or sedation based on patient positioning and respiratory status. Postoperative monitoring included haematological and biochemistry evaluation and CT scan 24 h after PCA. Follow-up consisted of a multislice CT scan at 1 month and every 3 months in the first year then every 6 months thereafter. Thirty-seven patients (38 lesions) underwent 40 PCA procedures; 5/37 (13.5%) had a solitary kidney. Median mass size was 35 (range 12–70) mm. No complications occurred during the procedure. Clavien grade ≥2 anaemia occurred in two patients (5.4 %): one patient required 1 U of packed red blood cells; the other required an arterial embolisation. Serum creatinine did not increase in any case. Two patients showed persisting or recurrent disease at 1 and 9 months, respectively, and both could be re-treated with PCA. All other patients showed a hypodense mass 3 months after PCA, with no contrast enhancement. Subsequent examinations showed that lesion sizes decreased and CT densitometry remained stable or increased minimally, also with no contrast enhancement. PCA proved relatively easy and safe and could be considered an effective alternative for patients who are not surgical candidates or refuse surgery, as well as in patients with medium-sized lesions.

Archaeal and eukaryotic translation elongation factor 2 contain a unique post-translationally modified histidine residue called diphthamide, which is the target of diphtheria toxin. The biosynthesis of diphthamide was proposed to involve three steps, with the first being the formation of a C–C bond between the histidine residue and the 3-amino-3-carboxypropyl group of S-adenosyl-l-methionine (SAM). However, further details of the biosynthesis remain unknown. Here we present structural and biochemical evidence showing that the first step of diphthamide biosynthesis in the archaeon Pyrococcus horikoshii uses a novel iron–sulphur-cluster enzyme, Dph2. Dph2 is a homodimer and each of its monomers can bind a [4Fe–4S] cluster. Biochemical data suggest that unlike the enzymes in the radical SAM superfamily, Dph2 does not form the canonical 5′-deoxyadenosyl radical. Instead, it breaks the Cγ,Met–S bond of SAM and generates a 3-amino-3-carboxypropyl radical. Our results suggest that P. horikoshii Dph2 represents a previously unknown, SAM-dependent, [4Fe–4S]-containing enzyme that catalyses unprecedented chemistry. Diphthamide, the target of diphtheria toxin, is a post-translationally modified histidine residue in archaeal and eukaryotic translation elongation factor 2 (EF2). It has been studied for more than three decades, but some aspects of its mechanism of biosynthesis remain elusive. The process is thought to involve three steps, the first being the formation of a C–C bond between the histidine residue and the 3-amino-3-carboxypropyl group of S-adenosyl-l-methionine (SAM). A study of the archaeon Pyrococcus horikoshii now shows that in this organism at least, the first step in diphthamide biosynthesis requires an unusual iron–sulphur cluster enzyme, Dph2. Biochemical data suggest that — in contrast to the mechanism in other radical SAM enzymes — Dph2 breaks the CγMet–S bond of the SAM cofactor. The enzyme then transfers the 3-amino-3-carboxylpropyl group to EF2, possibly via a radical mechanism. Translation elongation factor 2 (EF2) from archaea and eukaryotes contains a unique, post-translationally modified histidine residue called diphthamide, which is the target of diphtheria toxin. The biosynthesis of diphthamide involves three steps; here it is shown that the first step in the archaeon Pyrococcus horikoshii requires an unusual iron–sulphur-cluster enzyme, Dph2. It catalyses unprecedented chemistry.

Germ-cell tumors of the testis (GCTT) are rare, but have a high social impact. In fact they represent no more than 1% of male tumors (about 700 new cases per year in Italy), but electively occur in young patients, 20 to 40 years old, during their fully mature social and working life. More than 80% of patients are cured and return to a normal social, sexual, and working life. Improvements achieved both in diagnosis, with the use of scans (CT, MRI, US and recently PET) and of serum tumor markers alpha-fetoprotein (AFP), beta-fraction of human chorionic gonadotropin (β-HCG) and lactate dehydrogenase (LDH), and mainly in treatment, through the amelioration of radiotherapy and surgical techniques and, especially, with the introduction of Cisplatin, Etoposide and Ifosfamide in Chemotherapic regimens, have made germ-cell tumor a model of “curable disease”. Retroperitoneal lymph node dissection (RPLND) has indications in patients with clinical stage I (CS1) as well as in advanced disease, where it is integrated in the multimodality treatment. Anatomical studies, as well as a long-term experience, have gradually but consistently modified the surgical techniques of RPLND. Currently, “nerve sparing” RPLND represents a safe management of CS1 nonseminomatous germ cell testicular tumor with minimal morbidity and excellent outcomes. Nonetheless, surveillance and adjuvant chemotherapy are as effective as RPLND, but, in our opinion, associated with some discomforts for the patients. Laparoscopic retroperitoneal lymph node dissection (Lap-RPLND) is gaining popularity as a minimally invasive staging procedure for clinical stage I nonseminomatous testicular carcinoma, but its therapeutic role is still under investigation.

Many different, intersecting strategies are available for managing germ-cell cancers, particularly in early-stage disease. Which is ‘right’ remains a matter of debate, and requires balancing efficacy against late effects, bearing in mind the complexity of treatment strategies and the available expertise.

Riboswitches are RNA elements that control gene expression through metabolite binding. The preQ1 riboswitch exhibits the smallest known ligand-binding domain and is of interest for its economical organization and high affinity interactions with guanine-derived metabolites required to confer tRNA wobbling. Here we present the crystal structure of a preQ1 aptamer domain in complex with its precursor metabolite preQ0. The structure is highly compact with a core that features a stem capped by a well organized decaloop. The metabolite is recognized within a deep pocket via Watson-Crick pairing with C15. Additional hydrogen bonds are made to invariant bases U6 and A29. The ligand-bound state confers continuous helical stacking throughout the core fold, thus providing a platform to promote Watson-Crick base pairing between C9 of the decaloop and the first base of the ribosome-binding site, G33. The structure offers insight into the mode of ribosome-binding site sequestration by a minimal RNA fold stabilized by metabolite binding and has implications for understanding the molecular basis by which bacterial genes are regulated.

The potential for water to participate in RNA catalyzed reactions has been the topic of several recent studies. Here, we report crystals of a minimal, hinged hairpin ribozyme in complex with the transition-state analog vanadate at 2.05 Å resolution. Waters are present in the active site and are discussed in light of existing views of catalytic strategies employed by the hairpin ribozyme. A second structure harboring a 2′,5′-phosphodiester linkage at the site of cleavage was also solved at 2.35 Å resolution and corroborates the assignment of active site waters in the structure containing vanadate. A comparison of the two structures reveals that the 2′,5′ structure adopts a conformation that resembles the reaction intermediate in terms of (1) the positioning of its nonbridging oxygens and (2) the covalent attachment of the 2′-O nucleophile with the scissile G+1 phosphorus. The 2′,5′-linked structure was then overlaid with scissile bonds of other small ribozymes including the glmS metabolite-sensing riboswitch and the hammerhead ribozyme, and suggests the potential of the 2′,5′ linkage to elicit a reaction-intermediate conformation without the need to form metalloenzyme complexes. The hairpin ribozyme structures presented here also suggest how water molecules bound at each of the nonbridging oxygens of G+1 may electrostatically stabilize the transition state in a manner that supplements nucleobase functional groups. Such coordination has not been reported for small ribozymes, but is consistent with the structures of protein enzymes. Overall, this work establishes significant parallels between the RNA and protein enzyme worlds.

Activation-induced deaminase (AID) uses base deamination for class-switch recombination and somatic hypermutation and is related to the mammalian RNA-editing enzyme apolipoprotein B editing catalytic subunit 1 (APOBEC-1). CDD1 is a yeast ortholog of APOBEC-1 that exhibits cytidine deaminase and RNA-editing activity. Here, we present the crystal structure of CDD1 at 2.0-Å resolution and its use in comparative modeling of APOBEC-1 and AID. The models explain dimerization and the need for trans-acting loops that contribute to active site formation. Substrate selectivity appears to be regulated by a central active site “flap” whose size and flexibility accommodate large substrates in contrast to deaminases of pyrimidine metabolism that bind only small nucleosides or free bases. Most importantly, the results suggested both AID and APOBEC-1 are equally likely to bind single-stranded DNA or RNA, which has implications for the identification of natural AID targets.

Many different, intersecting strategies are available for managing germ-cell cancers,particularly in early-stage disease. Which is 'right' remains a matter of debate, and requires balancing efficacy against late effects, bearing in mind the complexity of treatment strategies and the available expertise./font/font/i

Diphthamide, the target of diphtheria toxin, is a unique posttranslational modification on eukaryotic and archaeal translation elongation factor 2 (EF2). The proposed biosynthesis of diphthamide involves three steps and we have recently found that in Pyrococcus horikoshii (P. horikoshii), the first step uses an S-adenosyl-L-methionine (SAM)-dependent [4Fe–4S] enzyme, PhDph2, to catalyze the formation of a C–C bond. Crystal structure shows that PhDph2 is a homodimer and each monomer contains three conserved cysteine residues that can bind a [4Fe–4S] cluster. In the reduced state, the [4Fe–4S] cluster can provide one electron to reductively cleave the bound SAM molecule. However, different from classical radical SAM family of enzymes, biochemical evidence suggest that a 3-amino-3-carboxypropyl radical is generated in PhDph2. Here we present evidence supporting that the 3-amino-3-carboxypropyl radical does not undergo hydrogen abstraction reaction, which is observed for the deoxyadenosyl radical in classical radical SAM enzymes. Instead, the 3-amino-3-carboxypropyl radical is added to the imidazole ring in the pathway towards the formation of the product. Furthermore, our data suggest that the chemistry requires only one [4Fe–4S] cluster to be present in the PhDph2 dimer.

Germ-cell tumors of the testis (GCTT) are rare, but have a high social impact. In fact they represent no more than 1% of male tumors (about 700 new cases per year in Italy), but electively occur in young patients, 20 to 40 years old, during their fully mature social and working life. More than 80% of patients are cured and return to a normal social, sexual, and working life. Improvements achieved both in diagnosis, with the use of scans (CT, MRI, US and recently PET) and of serum tumor markers alpha-fetoprotein (AFP), beta-fraction of human chorionic gonadotropin (b-HCG) and lactate dehydrogenase (LDH), and mainly in treatment, through the amelioration of radiotherapy and surgical techniques and, especially, with the introduction of Cisplatin, Etoposide and Ifosfamide in chemotherapic regimens, have made germ-cell tumor a model of "curable disease". Retroperitoneal lymph node dissection (RPLND) has indications in patients with clinical stage I (CS1) as well as in advanced disease, where it is integrated in the multimodality treatment. Anatomical studies, as well as a long-term experience, have gradually but consistently modified the surgical techniques of RPLND. Currently, "nerve sparing" RPLND represents a safe management of CS1 nonseminomatous germ cell testicular tumor with minimal morbidity and excellent outcomes. Nonetheless, surveillance and adjuvant chemotherapy are as effective as RPLND, but, in our opinion, associated with some discomforts for the patients. Laparoscopic retroperitoneal lymph node dissection (Lap-RPLND) is gaining popularity as a minimally invasive staging procedure for clinical stage I nonseminomatous testicular carcinoma, but its therapeutic role is still under investigation.

Reaction-intermediate analogs have been used to understand how phosphoryl transfer enzymes promote catalysis. Herein we report the first structure of a small ribozyme crystallized with a 3'-OH, 2',5'-linkage in lieu of the normal phosphodiester substrate. The new structure shares features of the reaction coordinate exhibited in prior ribozyme structures including a vanadate complex that mimicked the oxyphosphorane transition state. As such, the structure exhibits reaction-intermediate traits that allow direct comparison of stabilizing interactions to the 3'-OH, 2',5'-linkage contributed by the RNA enzyme and its protein counterpart, ribonuclease. Clear similarities are observed between the respective structures including hydrogen bonds to the non-bridging oxygens of the scissile phosphate. Other commonalities include carefully poised water molecules that may alleviate charge build-up in the transition state and placement of a positive charge near the leaving group. The advantages of 2',5'-linkages to investigate phosphoryl-transfer reactions are discussed, and argue for their expanded use in structural studies.

In mammals, activation-induced deaminase (AID) initiates somatic hypermutation (SHM) and class switch recombination (CSR) of Ig genes. SHM and CSR activities require separate regions within AID. A chromosome region maintenance 1 (CRM1)-dependent nuclear export signal (NES) at the AID C terminus is necessary for CSR, and has been suggested to associate with CSR-specific cofactors. CSR appeared late in AID evolution, during the emergence of land vertebrates from bony fish, which only display SHM. Here, we show that AID from African clawed frog (Xenopus laevis), but not pufferfish (Takifugu rubripes), can induce CSR in AID-deficient mouse B cells, although both are catalytically active in bacteria and mammalian cell systems, albeit at decreased level. Like mammalian AID, Takifugu AID is actively exported from the cell nucleus by CRM1, and the Takifugu NES can substitute for the equivalent region in human AID, indicating that all the CSR-essential NES motif functions evolutionarily predated CSR activity. We also show that fusion of the Takifugu AID catalytic domain to the entire human noncatalytic domain restores activity in mammalian cells, suggesting that AID features mapping within the noncatalytic domain, but outside the NES, influence its function.

The ability of peptides to form stable complexes with MHC class II molecules expressed in the host determines their ability to recruit CD4 T cells during an immune response. In this study, we sought to define the features of the antigenic peptides that control their kinetic stability with I-A d because of the diversity of peptides that this molecule is known to present. Peptide dissociation assays indicated that each pocket of I-A d displays exquisite sensitivity to side chain structure, size, and charge. Most surprising were results related to the P1 pocket, which has been difficult to define by conventional competition assays. Our studies revealed a considerable degree of specificity in the P1 pocket but also an unexpected degree of structural flexibility. Amino acids with neutral side chains such as Met and the alternatively negatively charged Glu are both highly favored at P1. Interestingly, these two options at the P1 pocket in I-A d display dramatically different pH-dependent interactions with the class II molecule. These findings are discussed in the context of a structural model to explain these data and in light of the immunological implications of pH-dependent behavior of class II-peptide complexes in acidic endosomal compartments, where DM-catalyzed loading of class II molecules takes place, and at the neutral pH of the APC cell surface, where class II-peptide complexes promote activation of CD4 T cells.

A comparative study between modified Camey II and Studer ileal orthotopic neobladder was performed. The Camey II was modified as follows: 1) The ureters were implanted, using wallace technique, in an undetubularized ileal loop, 15-18 cm. long, to prevent vesico-ureteral reflux; 2) The neobladder was made using staplers. In such a way, time is saved (about one hour) and results are quite similar, with a low rate of ureteral stenosis in both groups.

Of 19 patients with advanced transitional bladder cancer (T2-T4, N0-N+, M0) who received two or three cycles of pre-emptive MVC (Methotrexate, Vinblastine, Cisplatin), pathological partial (PR) and complete (CR) remissions were observed in 67% (50% and 17% respectively). The toxicity of chemotherapy was generally acceptable but 5 patients required hospitalization for neutropenia and thrombopenia . In one of them chemotherapy was stopped for severe sepsis. No death was observed. In 11 patients follow-up is greater than 12 months. In this group, 10 patients are actually alive and disease-free, while the other one was dead owing to brain metastasis, after eight months from surgery.

Benign prostatic hypertrophy and inguinal hernia are frequently associated. 59 patients underwent transvesical prostatic adenomectomy and simultaneous pre-peritoneal hernioplasty by Nyhus' technique. Results lead the Authors to consider this technique effective, quick to perform, with a recurrence rate similar to that after inguinal hernioplasties.

The authors report preliminary results of a study performed on six patients with by advanced, hormone-refractory prostate cancer. These patients were treated by estramustine phosphate (10 mg/kg) and methotrexate (60 mg/mq). Low toxicity allows the use of this therapy in outpatient regimens. Moreover, the high rate of partial responses (five patients) shows the remarkable efficacy of this therapy, even if follow-up and number of patients are not sufficient to reach definitive conclusions.