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High-fidelity projective readout of a qubit's state in a single experimental repetition is a prerequisite for various quantum protocols of sensing and computing. Achieving single-shot readout is challenging for solid-state qubits. For Nitrogen-Vacancy (NV) centers in diamond, it has been realized using nuclear memories or resonant excitation at cryogenic temperature. All of these existing approaches have stringent experimental demands. In particular, they require a high efficiency of photon collection, such as immersion optics or all-diamond micro-optics. For some of the most relevant applications, such as shallow implanted NV centers in a cryogenic environment, these tools are unavailable. Here we demonstrate an all-optical spin readout scheme that achieves single-shot fidelity even if photon collection is poor (delivering less than 10 3 clicks/second). The scheme is based on spin-dependent resonant excitation at cryogenic temperature combined with spin-to-charge conversion, mapping the fragile electron spin states to the stable charge states. We prove this technique to work on shallow implanted NV centers, as they are required for sensing and scalable NV-based quantum registers.

Global Positioning System (GPS) dissemination of frequency standards is ubiquitous at present, providing the most widespread time and frequency reference for the majority of industrial and research applications worldwide. On the other hand, the ultimate limits of the GPS presently curb further advances in high-precision, scientific and industrial applications relying on this dissemination scheme. Here, we demonstrate that these limits can be reliably overcome even in laboratories without a local atomic clock by replacing the GPS with a 642-km-long optical fiber link to a remote primary caesium frequency standard. Through this configuration we stably address the 1 S 0 - 3 P 0 clock transition in an ultracold gas of 173 Yb, with a precision that exceeds the possibilities of a GPS-based measurement, dismissing the need for a local clock infrastructure to perform beyond-GPS high-precision tasks. We also report an improvement of two orders of magnitude in the accuracy on the transition frequency reported in literature.

Listeria monocytogenes is a notably invasive bacterium associated with life-threatening food-borne disease in humans. Several surface proteins have been shown to be essential in the adhesion of L. monocytogenes, and in the subsequent invasion of phagocytes. Because the control of the invasion of host cells by Listeria could potentially hinder its spread in the infected host, we have examined the effects of a protease treatment on the ability of L. monocytogenes to form biofilms and to invade tissues. We have chosen serratiopeptidase (SPEP), an extracellular metalloprotease produced by Serratia marcescens that is already widely used as an anti-inflammatory agent, and has been shown to modulate adhesin expression and to induce antibiotic sensitivity in other bacteria. Treatment of L. monocytogenes with sublethal concentrations of SPEP reduced their ability to form biofilms and to invade host cells. Zymograms of the treated cells revealed that Ami4b autolysin, internalinB, and ActA were sharply reduced. These cell-surface proteins are known to function as ligands in the interaction between these bacteria and their host cells, and our data suggest that treatment with this natural enzyme may provide a useful tool in the prevention of the initial adhesion of L. monocytogenes to the human gut.

Respiratory viruses, including rhinoviruses, frequently promote bacterial opportunistic infections, through mechanisms that still deserve to be investigated in detail. This work was aimed at understanding how a viral infection mostly affecting the upper respiratory tract, such as the common cold, can repeatedly promote opportunistic infections in the lower airways, a site where viral replication is limited. The adhesivity and invasivity of Staphylococcus aureus were evaluated, in permissive and non-permissive cells, infected with Rhinovirus-1b. The role of inflammatory cytokines, and of ICAM-1 overexpression in the Rhinovirus-S. aureus cooperation was evaluated. Rhinovirus-1b enhanced the efficiency of internalisation of S. aureus irrespective of cellular permissivity, even when very low viral multiplicities of infection were used. Experiments performed with UV inactivated and heat inactivated viral particles suggested that this enhancement does not depend upon viral replication, but requires viral adhesion. Experimental data suggest that Rhinovirus-1b can significantly increase the ability of S. aureus to internalise into pneumocytes with a mechanism that involves the virus induced release of IL-6 and IL-8, and the overexpression of ICAM-1. Overall data disclose a possible mechanism through which rhinoviruses can promote bacterial infections in the lower respiratory tract.

Samples of neoplastic and normal urothelium were obtained from cows originating from areas of southern Italy, a region in which chronic enzootic haematuria is endemic and bracken fern infestation is widespread. Specimens were analysed for bovine papillomavirus type 2 (BPV-2) DNA, BPV-2 E5 expression and telomerase activity. A total of 46 of 60 tumours and 17 of 34 normal bladder mucosa samples harboured BPV-2 DNA. Analysis of a subset of samples showed E5 protein expression and telomerase activity in tumour tissue only. No normal samples positive for BPV DNA showed E5 protein expression or telomerase activity, suggesting the presence of DNA in a latent state. Taken together, these data on naturally occurring bovine bladder tumours corroborate the hypothesis of their virus origin.

Epidermodysplasia verruciformis (EV) can be defined as a genetic disorder that determines a lifelong infection of the skin by human Papillomaviruses (HPV). The benign lesions contain different HPVs; whereas in the tumors HPV types 5, 8, and, much less frequently, types 14, 17, 20 and 47 can be detected. Variants of HPV5 have been recognized on the basis of the genetic heterogeneity of the E6 open reading frame. We report a typical case of EV in which the presence and expression of the HPV type 5 were clearly detectable. Direct sequence analysis demonstrated a perfect homology with the sequence of the HPV5b variant. This variant was first isolated in a Japanese patient and thereafter in a Polish one. Its presence in the Italian patient indicates that the same variant can be detected in different geographic areas and therefore that some cellular genes may exert a selection pressure so strong as to induce the emergence of this stable HPV5b infectious variant.

High DNA viral load in Human Papillomavirus type 16 cervical dysplastic lesions has been proposed as a marker of progression towards malignancy. To evaluate the prognostic value of DNA viral load in infection by HPV type other than HPV-16, a longitudinal study of patients with Anogenital Condylomata (AC) was designed. The study included 40 patients, 25 men and 15 women. Clinical data were collected through standard interview and routine serum analyses. Viral type in lesions was assayed by PCR with general primers (MY09-MY11) and Restriction Fragment Length Polymorphism (RFLP). Viral load was determined by PCR end point determination with type specific primers on serial sample dilutions. The duration of ano-genital lesions upon liquid nitrogen standard cryotherapy or Podofillin chemotherapy was correlated with viral and clinical data. Patients with higher lesional viral load at study entry showed a markedly longer disease clinical course than patients with lower viral loads. The HPV-6 was observed in 31 out of 40 patients (77.5%), much more frequently than HPV-11 (7/40 17.5%), the latter being associated with higher titres of viral load. Two cases (5%) remained undetermined. No HPV-16 or other high-risk type was detected, while HPV-61, not uncommon in Italy, was found once in a double infection carried by an HIV positive patient. No significant association was observed between clinical outcome and demographic and epidemiological data, or risk factors.

A case of urothelial carcinoma in situ of urinary bladder is reported in a 10-year-old cow naturally grazing on bracken-infested land. The cow suffered from enzootic hematuria for more than 5 years. The presence of bovine papillomavirus type 2 (BPV-2) DNA sequences was detected by polymerase chain reaction. The carcinoma in situ was characterized by the presence of anaplastic cells with amphophilic cytoplasm and pleomorphic nuclei containing granular, irregularly dispersed chromatin. Focal areas within the tumor contained large isolated and/or clustered cells. These cells had pale acidophilic cytoplasm, large nuclei with single or multiple nucleoli, and well-defined borders resembling Paget's cells. Immunohistochemically, all malignant cells were negative for vimentin and S-100 and positive for cytokeratins. In addition, normal and neoplastic cells expressed fragile histidine triad (FHIT) protein; surprisingly, some pagetoid cells did not. FHIT, the tumor suppressor gene at 3p14.2, encodes a protein of 147 amino acids (16.8 kd) with diadenosine triphosphate hydrolase activity and is a common target of deletions in human cancers of epithelial origin. Antibody to laminin detected a continuous epithelial basement membrane, thus clearly showing that neoplastic changes were limited to urothelial cells without invading stromal tissue. To our knowledge, this is the first report of an unusual pattern of spread of urothelial carcinoma in situ in a cow.

Human papillomaviruses (HPV) are associated with cervical cancer and interact with growth factors that may enhance malignant transformation of cervical carcinoma cells. Endothelin-1 (ET-1) is released from HPV transfected keratinocytes and induces increased growth response in these cell lines in comparison with normal cells. In the present study several cervical carcinoma cell lines have been analyzed to investigate the expression of ET-1 and its receptors as well as their involvement in tumor growth. All HPV-positive cancer cells secreted ET-1 and expressed mRNA for ET-1 and its receptors, whereas a HPV-negative carcinoma cell line expressed only the ETBR mRNA and didn't secrete ET-1. Binding studies showed that HPV-associated cells expressed an increased number of functional ETAR. ET-1 stimulated a marked dose-dependent increase in [3H]-thymidine incorporation with respect to the normal cells whereas ET-3 and ETBR agonists had no effect. In HPV-positive cancer cells, a specific antagonist of ETAR inhibited the proliferation induced by ET-1 and substantially reduced the basal growth rate of unstimulated cervical tumor cells, whereas the ETBR antagonist had no effect. These results demonstrate that ET-1 participates in the progression of neoplastic growth in HPV-associated carcinoma, in which ETAR are increased and could be targeted for antitumor therapy.

Antibody response to HPV16 E7 oncoprotein may represent a marker of cervical cancer. A HPV16 GST-E7 fusion protein was used in a Western Blot assay to analyse the HPV16 E7 antibody response in 30 patients before and after treatment for cervical carcinoma (stage IIB or IIIB). Patients were treated with three courses of cisplatin/bleomycin therapy followed by surgery, or with surgery alone. Thirteen out of 30 patients had serum antibodies to HPV16 E7 antigen. Three months after chemotherapy little or no change in antibody titre was detected. In contrast, after surgery, a significant decrease in antibody titre was observed in 9/10 patients. In two cases the titre declined to zero 3 and 9 months after treatment, respectively. These results confirm the usefulness of studying anti-HPV16 E7 antibody profile in cervical cancer patients and suggest that the serum response correlates with tumour burden.