Exposure to NO<inf>2</inf> in occupationalbuilt environmnets in urban centre in Lahore

Increased economic growth, urbanisation and substantial rise in automobile vehicles has contributed towards the elevated levels of air pollution in major cities in Pakistan. Aone week study was conducted by using passive samplers to assess NO2 concentration in occupational built environments at two most congested and populated sites of Lahore. Both sites were locatedon the busy roads of Lahore. At Site-I the highest concentration was in outdoors followed by corridor and indoor. While at Site II all the sampling location wereindoors and level were comparable to that of outdoor levelsat Site I. The results suggest the likely contribution of ambient sources in exposure to indoor NO2 in educational and other occupational built environments in urban centres

SH3 Domain-Mediated Recruitment of Host Cell Amphiphysins by Alphavirus nsP3 Promotes Viral RNA Replication

Among the four non-structural proteins of alphaviruses the function of nsP3 is the least well understood. NsP3 is a component of the viral replication complex, and composed of a conserved aminoterminal macro domain implicated in viral RNA synthesis, and a poorly conserved carboxyterminal region. Despite the lack of overall homology we noted a carboxyterminal proline-rich sequence motif shared by many alphaviral nsP3 proteins, and found it to serve as a preferred target site for the Src-homology 3 (SH3) domains of amphiphysin-1 and -2. Nsp3 proteins of Semliki Forest (SFV), Sindbis (SINV), and Chikungunya viruses all showed avid and SH3-dependent binding to amphiphysins. Upon alphavirus infection the intracellular distribution of amphiphysin was dramatically altered and colocalized with nsP3. Mutations in nsP3 disrupting the amphiphysin SH3 binding motif as well as RNAi-mediated silencing of amphiphysin-2 expression resulted in impaired viral RNA replication in HeLa cells infected with SINV or SFV. Infection of Balb/c mice with SFV carrying an SH3 binding-defective nsP3 was associated with significantly decreased mortality. These data establish SH3 domain-mediated binding of nsP3 with amphiphysin as an important host cell interaction promoting alphavirus replication

Vacuolar ATPase Regulates Surfactant Secretion in Rat Alveolar Type II Cells by Modulating Lamellar Body Calcium

Lung surfactant reduces surface tension and maintains the stability of alveoli. How surfactant is released from alveolar epithelial type II cells is not fully understood. Vacuolar ATPase (V-ATPase) is the enzyme responsible for pumping H+ into lamellar bodies and is required for the processing of surfactant proteins and the packaging of surfactant lipids. However, its role in lung surfactant secretion is unknown. Proteomic analysis revealed that vacuolar ATPase (V-ATPase) dominated the alveolar type II cell lipid raft proteome. Western blotting confirmed the association of V-ATPase a1 and B1/2 subunits with lipid rafts and their enrichment in lamellar bodies. The dissipation of lamellar body pH gradient by Bafilomycin A1 (Baf A1), an inhibitor of V-ATPase, increased surfactant secretion. Baf A1-stimulated secretion was blocked by the intracellular Ca2+ chelator, BAPTA-AM, the protein kinase C (PKC) inhibitor, staurosporine, and the Ca2+/calmodulin-dependent protein kinase II (CaMKII), KN-62. Baf A1 induced Ca2+ release from isolated lamellar bodies. Thapsigargin reduced the Baf A1-induced secretion, indicating cross-talk between lamellar body and endoplasmic reticulum Ca2+ pools. Stimulation of type II cells with surfactant secretagogues dissipated the pH gradient across lamellar bodies and disassembled the V-ATPase complex, indicating the physiological relevance of the V-ATPase-mediated surfactant secretion. Finally, silencing of V-ATPase a1 and B2 subunits decreased stimulated surfactant secretion, indicating that these subunits were crucial for surfactant secretion. We conclude that V-ATPase regulates surfactant secretion via an increased Ca2+ mobilization from lamellar bodies and endoplasmic reticulum, and the activation of PKC and CaMKII. Our finding revealed a previously unrealized role of V-ATPase in surfactant secretion

WSES guidelines for management of Clostridium difficile infection in surgical patients

In the last two decades there have been dramatic changes in the epidemiology of Clostridium difficile infection (CDI), with increases in incidence and severity of disease in many countries worldwide. The incidence of CDI has also increased in surgical patients. Optimization of management of C difficile, has therefore become increasingly urgent. An international multidisciplinary panel of experts prepared evidenced-based World Society of Emergency Surgery (WSES) guidelines for management of CDI in surgical patients.Peer reviewe

Algorithm-based online software for patients' self-referral to breast clinic as an alternative to general practitioner referral pathway.

Introduction The study aimed to assess the accuracy of online software in the use of self-referral to breast surgery clinics for patients with new signs and symptoms. The study also evaluated the appropriateness of GP referrals to breast clinics and evaluated patients' perceptions of an online self-referral portal to the breast clinic for the assessment of breast signs and symptoms.  Design and methods The pilot study was divided into two phases. In the first phase, prospective questionnaire-based data was collected from patients who were referred by a GP and presented to the regional breast unit with new signs and symptoms for breast conditions, Princess Alexandra Hospital NHS Trust (May - October 2018). The questionnaire assessed the time at each stage required by the patient to have a visit at the breast unit. It also asked the patient's opinion about an online self-referral portal to the surgical clinic. They were given hypothetical scenarios to evaluate their understanding of breast conditions. In the second phase, the patients presenting to symptomatic breast clinics were provided with the iPad to fill in their medical information in the online software. The data was collected between July and October 2019. The software algorithm was based on the National Institute of Clinical Health and Excellence (NICE) guidelines for breast conditions (2015). Breast surgeons' recommendations acted as a standard to evaluate the accuracy of GPs' referrals and software outcome for each patient.  Results There were 80 patients (mean age 49.1 [SD: 17.7], all females) included in the first phase of the study. The most common clinical presentation was a breast lump (47.6%), followed by breast pain (26.9%) and nipple changes (7.9%). Breast surgeons considered appropriate 75.6% of the referrals made by the GP. Seventy-two percent of the patients got an urgent appointment to see their GP, and 94.8% of the patients were urgently referred by their GP to see the breast surgeon. Only 37.8% of the urgent referrals were correctly referred as urgent. Having a direct online referral system for breast conditions will be beneficial for patients was agreed by 78.4%. The majority (98.1%) of the participants answered correctly for the hypothetical questions requiring breast surgeon review. In the second phase, there were a total of 86 patients with a mean age of 43.9 (SD: 13.3). The most common presentation was breast lump (n=68, 79.1%) and other presentations included breast pain, nipple changes, and discharge. The GPs' accuracy of correct referral was 69.1%. One third (30.9%) of the referrals could have been managed in the community or as a routine review by the breast surgeon. In comparison, the online software's accuracy was 85.1% accurate (p=0.001). The accuracy for detecting patients who needed urgent breast clinic review was 100% for online software.  Conclusion A large proportion of referrals could have been dealt with in the community or referred routinely. Patients would prefer a direct online referral system to the breast clinic. They understand red flag signs and symptoms. Online software has the potential to streamline patients for symptomatic breast clinics. It can reduce the burden on the GPs who are constantly under pressure to diagnose patients accurately and refer to the correct specialty appropriately within a short time