Positronium Laser Cooling via the 1

We report on laser cooling of a large fraction of positronium (Ps) in free flight by strongly saturating the 1^{3}S-2^{3}P transition with a broadband, long-pulsed 243 nm alexandrite laser. The ground state Ps cloud is produced in a magnetic and electric field-free environment. We observe two different laser-induced effects. The first effect is an increase in the number of atoms in the ground state after the time Ps has spent in the long-lived 2^{3}P states. The second effect is one-dimensional Doppler cooling of Ps, reducing the cloud's temperature from 380(20) to 170(20) K. We demonstrate a 58(9)% increase in the fraction of Ps atoms with v_{1D}<3.7×10^{4} ms^{-1}.

SAR user guide to the rectal MR synoptic report (primary staging).

Rectal MR is the key diagnostic exam at initial presentation for rectal cancer patients. It is the primary determinant in establishing clinical stage for the patient and greatly impacts the clinical decision-making process. Consequently, structured reporting for MR is critically important to ensure that all required information is provided to the clinical care team. The SAR initial staging reporting template has been constructed to address these important items, including locoregional extent and factors impacting the surgical approach and management of the patient. Potential outputs to each item are defined, requiring the radiologist to commit to a result. This provides essential information to the surgeon or oncologist to make specific treatment deisions for the patient. The SAR Initial Staging MR reporting template has now been officially adopted by the NAPRC (National Accreditation Program for Rectal Cancer) under the American College of Surgery. With the recent revisions to the reporting template, this user guide has been revamped to improve its practicality and support to the radiologist to complete the structured report. Each line item of the report is supplemented with clinical perspectives, images, and illustrations to help the radiologist understand the potential implications for a given finding. Common errors and pitfalls to avoid are highlighted. Ideally, rectal MR interpretation should not occur in a vacuum but in the context of a multi-disciplinary tumor board to ensure that healthcare providers use common terminology and share a solid understanding of the strengths and weaknesses of MR.

Lyman-α source for laser cooling antihydrogen.

We present a Lyman-α laser developed for cooling trapped antihydrogen. The system is based on a pulsed Ti:sapphire laser operating at 729 nm that is frequency doubled using an LBO crystal and then frequency tripled in a Kr/Ar gas cell. After frequency conversion, this system produces up to 5.7 µW of average power at the Lyman-α wavelength. This laser is part of the ATRAP experiment at the antiproton decelerator in CERN.

Spin Dependence of η Meson Production in Proton-Proton Collisions Close to Threshold.

Taking advantage of the high acceptance and axial symmetry of the WASA-at-COSY detector, and the high polarization degree of the proton beam of COSY, the reaction p[over →]p→ppη has been measured close to threshold to explore the analyzing power A_{y}. The angular distribution of A_{y} is determined with the precision improved by more than 1 order of magnitude with respect to previous results, allowing a first accurate comparison with theoretical predictions. The determined analyzing power is consistent with zero for an excess energy of Q=15 MeV, signaling s-wave production with no evidence for higher partial waves. At Q=72 MeV the data reveal strong interference of Ps and Pp partial waves and cancellation of (Pp)^{2} and Ss^{*}Sd contributions. These results rule out the presently available theoretical predictions for the production mechanism of the η meson.

The functional significance of microRNA-145 in prostate cancer.

BACKGROUND: MicroRNAs (miRNAs) are small noncoding RNAs that have important roles in numerous cellular processes. Recent studies have shown aberrant expression of miRNAs in prostate cancer tissues and cell lines. On the basis of miRNA microarray data, we found that miR-145 is significantly downregulated in prostate cancer. METHODS AND RESULTS: We investigated the expression and functional significance of miR-145 in prostate cancer. The expression of miR-145 was low in all the prostate cell lines tested (PC3, LNCaP and DU145) compared with the normal cell line, PWR-1E, and in cancerous regions of human prostate tissue when compared with the matched adjacent normal. Overexpression of miR-145 in PC3-transfected cells resulted in increased apoptosis and an increase in cells in the G2/M phase, as detected by flow cytometry. Investigation of the mechanisms of inactivation of miR-145 through epigenetic pathways revealed significant DNA methylation of the miR-145 promoter region in prostate cancer cell lines. Microarray analyses of miR-145-overexpressing PC3 cells showed upregulation of the pro-apoptotic gene TNFSF10, which was confirmed by real-time PCR and western analysis. CONCLUSION: One of the genes significantly upregulated by miR-145 overexpression is the proapoptotic gene TNFSF10. Therefore, modulation of miR-145 may be an important therapeutic approach for the management of prostate cancer.

A survey of tuberculosis hospitals in India.

SETTING: Hospitals with beds for tuberculosis (TB) in India. OBJECTIVES: To assess diagnostic and treatment practices at institutions offering secondary or tertiary level care for TB patients, and to determine the resources being used at these institutions. DESIGN: Countrywide cross-sectional survey of TB hospitals using a mailed semi-structured questionnaire sent to all 105 hospitals with 100 or more beds and to all State Directorate Health Services. RESULTS: The 94 hospitals that returned the questionnaire had 15773 TB beds, one third of the total TB beds in the country. Nearly 1 million patients sought treatment in the TB hospitals and one third were diagnosed with TB; the ratio of smear-positive to smear-negative patients was 1:2.7. Sixty-four per cent of hospitals prescribed unobserved rifampicin in the continuation phase, and 56% of sputum smear-positive patients were hospitalised. The annual expenditure for the TB hospitals was more than the total annual budget for the TB control programme of the country. CONCLUSIONS: In view of the high number of patients seen and the suboptimal practices observed, urgent steps should be taken to ensure implementation of correct diagnostic and treatment policies in hospitals with TB beds.

Molecular screening of unculturable bacteria present in men with reproductive failure.

Objective: To determine the presence of unculturable bacteria using polymerase chain reaction (PCR) in infertile men with pyosperrmia. Design: Perspective clinical study. Setting: The study took place at the Department of Reproductive Medicine, Owaisi Hospital and Research Center; In vitro Fertilization Unit, Mahavir Hospital and Research Center; Center for Cellular and Molecular Biology; and Bharat Biotech Foundation, Hyderabad, India. Patients: A total of 68 infertile men and 15 donors, all with no symptoms of genito-urinary tract infections and sterile semen cultures were included in the study. Interventions: None. Main outcome measures: Screening bacteria using routine bacterial cultures and PCR based screening with universal eubacterial primers. Results: The statistical analysis of all the semen parameters in asthenazoospermic, azoospermic, ceyptazoospermic, severe oligospermic and mild oligospermic patients were found to be significant compared with the controls. All the groups were found to be significant compared with the controls (P < 0.05) except for volume and pus cells in the cryptozoospermia group. The Student's t-test also was significant for the seminal parameters before and after treatment of 68 selected individuals with pyospermia and sterile cultures. A total of 44.11% (30/68) samples were collected from the negative culture of pyospermic infertile men have shown the presence of bacteria on amplification using PCR with universal eubacterial primers. The DNA was purified and sequenced. The sequences were checked for homology using DNASTAR and Ribosomal DataBase Project II. A total of 90% of the samples have shown the nearest evolutionary relation to Pantoea P102 (AF394539) and 10% of samples have shown close relation with Burkholderia cepacia (AF042161). Conclusion: The routine bacteriological cultures were unable to detect certain bacterial species particularly with members of enterobacteriaceae family (Pantoea species). Polymerase chain reaction, when used for screening bacteria, can detect the unculturable form of bacteria in infertile men. No amplification for bacterial DNA was obtained in control samples (fertile men with sterile semen cultures.) (Reprod Med Biol 2004; 3: 77- 84).

Impact of national consultants on successful expansion of effective tuberculosis control in India.

SETTING: India, during a period of rapid expansion of DOTS services. DOTS expansion has been slow in many countries. OBJECTIVE: To document use of consultants to expand DOTS effectively. DESIGN: Staff were contracted to monitor DOTS expansion and implementation. To estimate the impact of these staff, we compared areas with and without consultants, and individual areas before and after consultants were assigned. Consultants were preferentially assigned to the more difficult areas; the temporary absence of consultants reflected non-availability of candidates. RESULTS: Areas with consultants met pre-defined criteria and began DOTS service delivery faster (median 9 vs. 18 months of preparation) than areas without consultants. Rates of sputum conversion (87% vs. 83%, P < 0.001) and treatment success (83% vs. 78%, P < 0.001) were significantly higher in areas with consultants present. CONCLUSION: Assignment of consultants resulted in much more rapid implementation of the DOTS strategy, and better quality performance. Continued effective performance in these areas will rely on many factors, but the need for consultants appears to be decreasing, suggesting that they have provided sustainable improvements. The effectiveness of local consultants may have important implications for efforts to scale up public health interventions for tuberculosis, malaria, AIDS and other diseases in developing countries.

Controlling tuberculosis in India.

BACKGROUND: Tuberculosis kills nearly 500,000 people in India each year. Until recently, less than half of patients with tuberculosis received an accurate diagnosis, and less than half of those received effective treatment. METHODS: We analyzed the effects of new policies introduced in 1993 that have resulted in increased resources, improved laboratory-based diagnosis, direct observation of treatment, and the use of standardized antituberculosis regimens and reporting methods. RESULTS: By September 2001, more than 200,000 health workers had been trained, and 436 million people (more than 40 percent of the entire population) had access to services. About 3.4 million patients had been evaluated for tuberculosis, and nearly 800,000 had received treatment, with a success rate greater than 80 percent. More than half of all those treated in the past 8 years were treated in the past 12 months. CONCLUSIONS: India's tuberculosis-control program has been successful in improving access to care, the quality of diagnosis, and the likelihood of successful treatment. We estimate that the improved program has prevented 200,000 deaths, with indirect savings of more than $400 million--more than eight times the cost of implementation. It will be a substantial challenge to sustain and expand the program, given the country's level of economic development, limited primary health care system, and large and mostly unregulated private health care system, as well as the dual threats of the human immunodeficiency virus and multidrug-resistant tuberculosis.

Rapid DOTS expansion in India.

Since late 1998 the coverage of the DOTS strategy in India has been expanded rapidly. In both 2000 and 2001 the country probably accounted for more than half the global increase in the number of patients treated under DOTS and by early 2002 more than a million patients were being treated in this way in India. As a result, nearly 200 000 lives were saved. The lessons learnt relate to the importance of the following elements of the programme: (1) getting the science right and ensuring technical excellence; (2) building commitment and ensuring the provision of funds and flexibility in their utilization; (3) maintaining focus and priorities; (4) systematically appraising each area before starting service delivery; (5) ensuring an uninterrupted drug supply; (6) strengthening the established infrastructure and providing support for staff; (7) supporting the infrastructure required in urban areas; (8) ensuring full-time independent technical support and supervision, particularly during the initial phases of implementation; (9) monitoring intensively and giving timely feedback; and (10) continuous supervision. Tuberculosis (TB) control still faces major challenges in India. To reach its potential, the control programme needs to: continue to expand so as to cover the remaining half of the country, much of which has a weaker health infrastructure than the areas already covered; increase its reach in the areas already covered so that a greater proportion of patients is treated; ensure sustainability; improve the patient-friendliness of services; confront TB associated with human immunodeficiency virus (HIV) infection. It is expected that HIV will increase the number of TB cases by at least 10% and by a considerably higher percentage if HIV becomes much more widespread. India's experience shows that DOTS can achieve high case-detection and cure rates even with imperfect technology and often with an inadequate public health infrastructure. However, this can only happen if the delivery programme is appropriately designed and effectively managed.

Effect of bone biopsy in guiding antimicrobial therapy for osteomyelitis complicating open wounds.

BACKGROUND: Osteomyelitis associated with infected overlying wounds represents a difficult diagnostic and therapeutic problem; bone biopsies can be done during debridement of the overlying wounds, but it is unclear how often the results of these bone cultures actually affect subsequent antibiotic decisions. The present study was undertaken to evaluate the usefulness of bone biopsies in guiding antibiotic therapy for this type of osteomyelitis. METHODS: Culture results of 44 bone biopsies taken during surgical debridement in 41 patients over the period from June 1994 to August 1998 were compared with those from the overlying wounds to determine whether the data affected the subsequent choice of antibiotics. The study design was that of a retrospective chart review in which the standard operative and microbiological procedures in place at the Milwaukee Veterans Affairs Medical Center were used. RESULTS: Sixty-one wound and 55 bone isolates were obtained during this study. Thirty-one isolates were found in bone, but not the overlying wound; diphtheroids were the most common organism obtained in this fashion. Correlation between wound and bone isolates was generally poor. Antibiotics were subsequently changed in 20 of the 44 cases after results of the bone biopsy became known, with the bone isolates already being covered in 10 cases and the bone biopsy results ignored in 14 cases. CONCLUSION: Because bone biopsy results seem to aid in tailoring antibiotic therapy in almost half the cases when bone is sampled during wound debridement surgery, this technique may be very helpful in certain cases and should be regularly undertaken when these procedures are carried out.

The status and prospects of tuberculosis control in India.

SETTING: India, where much of the global strategy for tuberculosis control was established, but where, every year, there are an estimated 2 million cases of tuberculosis. OBJECTIVE: To describe the policies, initial results, and lessons learned from implementation of a Revised National Tuberculosis Control Programme using the principles of DOTS (Directly Observed Treatment, Short-course). DESIGN: A Revised National Tuberculosis Control Programme (RNTCP) was designed and implemented starting in 1993. With funding from the Government of India, State Governments, the World Bank and bilateral donors, regular supply of drugs and logistics was ensured. Persons with chest symptoms who attend health facilities are referred to microscopy centres for diagnosis. Diagnosed cases are categorized as per World Health Organization guidelines, and treatment is given by direct observation. Systematic recording and cohort reporting is done. RESULTS: From October 1993 through mid-1999, 146,012 patients were put on treatment in the programme. The quality of diagnosis was improved, with the ratio of smear-positive to smear-negative patients being maintained at 1:1. Case detection rates varied greatly between project sites and correlated with the percentage of patients who were smear-positive among those examined for diagnosis, suggesting heterogeneous disease rates. Treatment success was achieved in 81% of new smear-positive patients, 82% of new smear-negative patients, 89% of patients with extra-pulmonary tuberculosis, and 70% of re-treatment patients. CONCLUSION: The RNTCP has successfully treated approximately 80% of patients in 20 districts of 15 states of India. Treatment success rates are more than double and death rates are less than a seventh those of the previous programme. Starting in late 1998, the programme began to scale up and now covers more than 130 million people. Maintaining the quality of implementation during the expansion phase is the next challenge.

Mechanism of recruitment of DnaB helicase to the replication origin of the plasmid pSC101.

Although many bacterial chromosomes require only one replication initiator protein, e.g., DnaA, most plasmid replicons depend on dual initiators: host-encoded DnaA and plasmid-encoded Rep initiator protein for replication initiation. Using the plasmid pSC101 as a model system, this work investigates the biological rationale for the requirement for dual initiators and shows that the plasmid-encoded RepA specifically interacts with the replicative helicase DnaB. Mutations in DnaB or RepA that disrupt RepA-DnaB interaction cause failure to load DnaB to the plasmid ori in vitro and to replicate the plasmid in vivo. Although, interaction of DnaA with DnaB could not substitute for RepA-DnaB interaction for helicase loading, DnaA along with integration host factor, DnaC, and RepA was essential for helicase loading. Therefore, DnaA is indirectly needed for helicase loading. Instead of a common surface of interaction with initiator proteins, interestingly, DnaB helicase appears to have at least a limited number of nonoverlapping surfaces, each of which interacts specifically with a different initiator protein.

National tuberculosis control programme.

PIP: In 1962, the government of India launched a National Tuberculosis Control Program to detect as many tuberculosis cases as possible, provide effective treatment, establish district tuberculosis centers, extend short-course chemotherapy, and strengthen existing state tuberculosis training and demonstration centers. To date, district tuberculosis centers have been established in 454 of India's 496 districts and a total of 330 tuberculosis clinics are in operation. The tuberculosis mortality rate dropped from 80/100,000 population in 1970 to 53/100,000 in 1993. However, a 1992 review of the national program revealed inadequate budgetary outlays and drug shortages, an overemphasis on clinical and radiologic diagnosis, insufficient utilization of sputum microscopy facilities, an emphasis on case detection rather than cure, and a lack of consensus on treatment regimens. On the basis of these findings, the national strategy has been revised to achieve an 85% cure rate through administration of short-course (6-8 months) chemotherapy and to detect 70% of estimated cases. Strategies for achieving these objectives include use of sputum testing as the primary diagnostic method among self-referred cases, a standardized treatment regimen, an uninterrupted supply of drugs at all levels of the health system, increased budgetary outlays, creation of a sub-district supervisory unit, and greater emphasis on training and operations research. A pilot project testing this approach in five areas in 1993 resulted in significant improvements in the pulmonary smear positive to negative ratios and cure rates. As a result, the government of India will expand the project strategy to 17 sites covering a population of 15.83 million.^ieng

Effects of Mg2+ and denaturants on the unfolding pattern of DNA-T--a replication protein of E. coli.

Escherichia coli-DNA-T protein is a key component of a multiprotein complex called the primosome which is involved in the initiation of DNA replication. The thermal and urea induced unfolding transition of this protein in the presence and absence of Mg2+ was studied using circular dichroism (CD) and fluorescence spectroscopy as probes. Quenching of the intrinsic fluorescence of DNA-T was observed in the thermal unfolding while formation of a hyperfluorescent form of the protein was found in the urea induced unfolding process. The CD studies showed a monophasic transition curve for thermal unfolding in the presence and absence of Mg2+. Biphasic curves indicative of the formation of intermediates was observed in the urea induced unfolding. The results suggest that the pathways of unfolding of thermal- and urea-induced transitions are different. MgCl2, which affects the conformation of the protein and stabilises the secondary structure, also affects the unfolding pattern.

Overexpression of Escherichia coli single stranded DNA binding protein under bacteriophage T 7 promoter.

A recombinant vector for overproduction of the E. coli single stranded DNA binding protein (E. coli SSBP) has been constructed. An E. coli strain carrying this plasmid produces up to 150 mg pure SSBP per litre of bacterial culture in a laboratory shake flask. Electron microscopy of the single stranded DNA complexed with SSBP shows characteristic "beaded string"-like appearance. Strong clustering of protein molecules on ssDNA is indicative of a highly cooperative binding.

Sequence-specific and polarized replication termination in vitro: complementation of extracts of tus- Escherichia coli by purified Ter protein and analysis of termination intermediates.

We have developed an in vitro replication system in which purified replication termination protein (Ter) elicits specific and polarized termination of DNA replication at terminator sites (tau) in a cell extract of tus- Escherichia coli that does not encode Ter protein. Using this system and two-dimensional agarose gel electrophoresis, we have identified intermediates with stalled replication forks. The replication bubbles contain both arrested leading strands and lagging strands that were initiated at the ColE1 origin of replication and that had progressed unidirectionally until arrested at tau. To dissect the system further, we have analyzed the kinetics of the formation of the termination intermediates and have discovered that the earliest termination intermediate had properties consistent with an arrested D loop. The D loop contained an arrested leading strand. Thus, in this test system, there appears to be a transient uncoupling of leading- and lagging-strand synthesis during termination of replication at tau sites.