Two short films have made it one step closer to an Oscar nomination after picking up a Light in Motion (LIM) award at the Foyle Film Festival’s closing ceremony in Brunswick Moviebowl.

The Transport Department today invited tenders for operating six major licensed ferry services for outlying islands for five years from April 1, 2021.

The six routes are between Central to Cheung Chau, Inter-islands between Peng Chau, Mui Wo, Chai Ma Wan and Cheung Chau, Central to Mui Wo, Central to Peng Chau, Central to Yung Shue Wan and Central to Sok Kwu Wan.

The department said that for the purposes of maintaining financial viability of the six major routes, alleviating the burden of fare increases on passengers, enhancing service quality and promoting a green city development, the Government would continue to provide special measures to the routes.

Such measures would include launching a new Vessel Subsidy Scheme to help selected ferry operators replace the fleets of the six major routes and introduce greener vessels in phases, straddling 10 years from 2021.

Having regard to factors such as passenger demand, overall fleet requirements, flexibility in vessel deployment, operational efficiency, and after consulting the Islands District Council, the six major routes are grouped into two packages for tendering.

The first package covers Central-Cheung Chau, Peng Chau-Mui Wo-Chi Ma Wan-Cheung Chau (Inter Islands) and Central-Mui Wo routes.

The other package covers Central-Peng Chau, Central-Yung Shue Wan and Central-Sok Kwu Wan routes.

Tenderers are required to propose a fare for each fare type of each relevant route and it must not exceed the existing fare level of the corresponding route by more than 5%. Tenders should also include in their submissions fare concession proposals that will be considered in the tender evaluation.

In support of the Government's development of a smart city, tenderers should propose measures including dissemination of real-time arrival/departure time of ferry routes, number of remaining seats via mobile phone apps and opening up such data for the public’s use under data.gov.hk.

The tenders must be sealed in envelopes and placed in the Transport Department Tender Box adjacent to the reception counter of the department on the 10th floor of South Tower, West Kowloon Government Offices, 11 Hoi Ting Road, Yau Ma Tei before noon on June 30.

The cover of the tender submission should be marked with tender reference TD 382/2019 and include the subject of the tender. It should also be addressed to the Commissioner for Transport.

Tender documents will be available for collection at the Transport Department’s Ferry & Paratransit Division on the 14th Floor, South Tower, West Kowloon Government Offices, 11 Hoi Ting Road, Yau Ma Tei from April 27 during office hours.

The Transport Department today announced that its driving test (road test) service will resume on May 4 to align with the resumption of public services in phases.

The department said candidates scheduled to take a road test on or after May 4 should take the test according to the test date, time and the driving test centre specified in their appointment letter.

Candidates affected by the suspension of driving test centres will be notified by post of the rescheduled test.

Driving test (written test Part A) and taxi written test services will resume on May 26.

Candidates affected by the suspension of written test services will be rescheduled to take the tests from May 26 and thereafter and notified by post of the rescheduling.

All candidates participating in the driving, road and written tests, must wear surgical masks inside the test centres and throughout the test.

They must also clean their hands with alcohol sanitisers and undergo body temperature screening.

If a candidate does not wear a surgical mask, refuses body temperature screening or has a body temperature higher than 37.5 degrees Celsius, the candidate will not be allowed to enter the venue and the test will be rescheduled.

To improve air circulation inside the vehicle compartment, driving examiners may open vehicle windows during the test.

The department urged candidates not to attend a driving test if they are unwell.

For applicants who are absent from a driving test on medical grounds, they may submit their postponement applications with the original sick leave or medical certificate to the Driving Test Appointment Office within one month from the test date for rescheduling.

Candidates applying for a postponement, temporary cancellation of driving test appointment or change of driving test region may submit signed application letters, copies of identification document and driving test appointment letter via the drop-in box at the Kowloon Licensing Office or post them to the Driving Test Appointment Office on 2/F, Cheung Sha Wan Government Offices, 303 Cheung Sha Wan Road.

Application of driving test appointment services for fresh candidates and repeaters will continue to be suspended until further notice. Call 2771 7723 for enquiries.

Vehicle Examination Centres will resume full services on May 4. The department will continue to process applications for licensing and related services submitted by applicants with scheduled appointments via the drop-in boxes, by post or online except for the direct issue of a full Hong Kong driving licence.

Applications may be submitted via the drop-in boxes at the offices concerned between 9am and 5pm from Monday to Friday except public holidays.

Those who have made online appointments for renewal of a full driving licence, vehicle licence, and transfer of vehicle ownership or international driving permit can visit the Licensing Office concerned at the scheduled time.

To maintain social distancing to reduce the risk of spreading COVID-19, the offices concerned will continue to suspend walk-in counter services.

Meanwhile, the Highways Department's public services will return to normal from May 4.

The department will implement social distancing and precautionary measures, including the introduction of flexible working and lunch hours, meeting adjustments and stepping up the cleansing of its offices.

Members of the public may call the 24-hour hotline 2926 4111 or 1823 for making suggestions, enquiries or complaints. The department can also be contacted by email or fax.

The Education Bureau and the Agriculture, Fisheries & Conservation Department (AFCD) today announced their latest arrangements on public services starting May 4.

All Regional Education Offices, the Joint Office for Kindergartens & Child Care Centres, the School Places Allocation Section, the Qualifications Framework Secretariat and other service units will resume normal opening hours from May 4.

However, the Hong Kong Teachers' Centre, Special Education Resource Centre, Central Resources Centre and Young Achievers' Gallery located at Kowloon Tong Education Services Centre, as well as the Arts & Technology Education Centre will remain closed until further notice.

The AFCD’s licensing services and reception counters at Cheung Sha Wan Government Offices and the Marine Parks Office at CDW Building in Tsuen Wan will resume normal services Monday to Friday from 8.30am to 12.30pm and 1.30pm to 5.45pm.

The anti-rabies dog inoculation and dog licensing services at the animal management centres will also resume as normal from May 4.

Meanwhile, the Hong Kong Wetland Park (except some indoor facilities and play areas), eight Country Park Visitor/Education Centres, seven Hong Kong Geopark Park Visitor Centres and the Endangered Species Resources Centre will reopen from May 4.

However, school and public programmes will remain suspended until further notice.

The barbecue sites and campsites in country parks will also continue to be closed until May 7.

Click here for the latest public service arrangements.

Secretary for the Civil Service Patrick Nip today visited the Transport Department (TD) and the Social Welfare Department (SWD) to learn more about the preparations made by both departments to resume public services.

The Government announced that the resumption of public services will start from May 4 under a phased approach.

While the TD will resume road tests on that day, written tests will resume on May 26.

The SWD will also gradually resume the services of its Integrated Family Service Centres (IFSC).

Mr Nip first visited the driving test centre in Ho Man Tin where he was briefed by the Commissioner for Transport Mable Chan on arrangements for the resumption of road tests and the implementation of infection control measures at driving test centres.

Such measures will require candidates to wear surgical masks and undergo body temperature screening at entrances.

As for the written tests, Mr Nip said he was pleased to learn that the TD will keep seats apart at appropriate distances and step up cleansing of computers at test centres.

Mr Nip then visited the Causeway Bay Integrated Family Service Centre where he was briefed on the plan to gradually resume services at IFSCs and measures to reduce social contact.

Mr Nip also learned that various infection control measures to safeguard the health of colleagues as well as service users will be put in place. For example, group activities will be conducted for no more than four participants and physical partitioning will be erected in meeting rooms for counselling services.

Mr Nip emphasised that many people hope that the Government can provide more public services when the epidemic situation becomes stable.

“I am pleased to learn that the departments have made all the necessary preparations for providing the services needed by the public while striving to safeguard public health.

“Government departments will continue to maintain a high degree of vigilance and adopt all the necessary precautionary measures. The Government will also closely monitor the situation and determine when to embark on a full resumption of normal business.”

Mr Nip expressed hope that the public will continue to fight the virus together with the Government.

He also thanked civil servants for their commitment and dedicated efforts to serve the public during the epidemic.

To align with the phased resumption of public services, various government departments have announced their latest arrangements.

The Department of Justice’s public services will resume normal from May 4. The reception counters of the department’s Prosecutions Division and Civil Division will be open Monday to Friday from 9am to 1pm and 2pm to 6pm.

The Legal Aid Department’s offices will resume normal office hours on May 4 to provide public services including civil and criminal legal aid application and litigation services.

The Government Logistics Department’s collection office will resume normal operation from May 4 and open Monday to Friday from 8.30am to 1pm and 2pm to 5pm.

The Lands Department's offices will resume public counter services and enquiry telephone lines on May 4, providing services such as payment of government rent and premiums, map sales, and collection of compensation for land resumption.

Except for the surveying services to be conducted outside Hong Kong, all services of the Marine Department will resume starting May 4.

The Rating & Valuation Department will resume normal operation of public services from May 4, with its enquiry counters opening from 8.15am to 6pm from Monday to Friday, except on public holidays.

The Civil Engineering & Development Department said office hours for the Civil Engineering Library, applications for dumping licences and sand removal permits, and the Mines Division’s Permit & Licensing Office will return to normal on May 4.

Public services of the Water Supplies Department, the Planning Department, the Drainage Services Department and the Inland Revenue Department will resume normal on May 4.

Starting from May 4, the Registration & Electoral Office’s enquiry counter and offices, the Buildings Department and the Environmental Protection Department’s public counter services and service counters under the Labour Department will reopen.

The Independent Commission Against Corruption's headquarters and regional offices will also resume normal services from May 4.

The Housing Authority's estate management offices, Public Rental Housing Application Office in the Lok Fu Customer Service Centre, the Clearance Housing Office and the Redevelopment Sub-section Offices will reinstate normal services from that date.

Flat selection sessions of the Territory-wide Overcrowding Relief Exercise, the Living Space Improvement Transfer Scheme and Sale of Home Ownership Scheme Flats 2019 will gradually resume from May 8.

Click here for the latest public service arrangements.

To align with the phased resumption of public services, the Information Services Department, Government Records Service and Civil Aviation Department have announced their latest arrangements.

The Information Services Department will resume sales counter services at its Publications Sales Unit and Photo Library at North Point Government Offices from May 4.

The sales office will be open from 9am to 12.30pm and from 2pm to 6pm from Monday to Friday.

To reduce social contact and avoid people gathering, citizens are encouraged to purchase government publications and photos through the online bookstore and photo store.

The Government Records Service's Public Records Office will provide search room services, including onsite loan and circulation services, from Monday to Friday, 9am to 5.45pm, for people who have made reservations through the online catalogue.

The office will also resume search room services from 9am to 1pm on Saturdays for people to inspect holdings which have been reserved online. 

Requests by up to 15 users will be accepted for search room services for each open day on a first-come, first-served basis. Users will be notified of the reservation results by email.

Visitor and public education programmes will continue to be suspended, while the Exhibition Hall at the Public Records Building will be temporarily closed for preparation of the new thematic exhibition until further notice.

Enquiries can be made to 2195 7700 or via email.

The Civil Aviation Department’s Personnel Licensing Office will continue to provide counter services from next Monday and resume relevant licensing examinations.

The office's opening hours are 8.45am to 12.45pm and 1.45pm to 5pm, Monday to Friday. Enquiries can be made to 2910 6046 or via email.

Click here for the latest public service arrangements.

The 'unicorn letter', sent by some of the best-funded private technology companies in the country, asks the chancellor to form an urgent taskforce to give them access to government-backed lending schemes during the pandemic

The Housing Authority’s Commercial Properties Committee today approved the extension of rent concessions to over 8,300 non-domestic tenants or licensees for six months from April 1 to September 30.

The authority had earlier granted a 50% rent concession to its eligible retail and factory tenants for six months from April 1.

Under the extension, their rent concession will be increased to 75% over the same period with retrospective effect from April 1. The rent concession does not include rates and air-conditioning charges.

The authority said such further measures are to support the Government's new series of measures announced in early April to relieve the financial burden of individuals and businesses.

A total of 2,450 retail and 3,300 factory tenants will benefit from the approved increase in the rent concession.

The 75% rent concession will also be extended to cover tenants and licensees of bus kiosks and most advertising signboards, as well as car park users for the monthly parking of commercial vehicles.

About 40 tenancies for bus kiosks, 80 advertising signboards and about 2,500 car park users stand to benefit from the concession.

Tenants of premises in the authority's properties which are required to be closed under relevant regulations or the Government's directions, may also apply to the authority for a 100% rent concession for the period during which they are required to be closed.

The authority added that the approved measures will be implemented as soon as possible. For rent and licence fees already paid for the months of April and May, arrangements will be made for offsetting in the payment for subsequent months.

The committee has approved three rounds of rent concessions since last September. Together with this round, the total rent and licence fees foregone by the authority is estimated to reach more than $1 billion.

Police today said it is highly concerned about recent criminal offences involving police officers and that a number of these officers have been arrested.

Among them, officers in connection with serious offences have been or will be interdicted.

The force said it is furious and disappointed about the officers who are suspected of having breached the law and that the incidents have impaired public confidence in Police.

Police attach the utmost importance to the discipline and integrity of officers.

All officers, irrespective of their ranks, must abide by the law at all times.

Police management has zero tolerance towards any acts that breach the law or discipline by police officers, it added.

The force also said it attaches great importance to the integrity of its officers.

The Complaints & Internal Investigations Branch formulated the Integrated Integrity Management Framework to promote integrity and honesty among officers as well as to regulate their discipline and conduct.

To avoid similar incidents, Police said its management is reviewing the force’s internal management strategy.

Police reiterated that no bad element in the force can be tolerated.

If an officer is suspected of having breached the law or committed a breach of discipline, Police will conduct an investigation according to the established mechanism and take appropriate actions.

17 individuals in the mathematical sciences are among the 126 new members and foreign associates elected to the National Academy of Sciences (NAS) in 2020.

Members: Ivet Bahar, University of Pittsburgh School of Medicine; Abhijit Banerjee, Massachusetts Institute of Technology; Gerard Ben Arous, Courant Institute of Mathematical Sciences, New York University; Bonnie Berger, Massachusetts Institute of Technology; Laura G. DeMarco, Northwestern University; Ronald Fagin, IBM Almaden Research Center; Katherine Freese, The University of Texas at Austin; Dennis Gaitsgory, Harvard University; Robert L. Griess, University of Michigan, Ann Arbor; Jacob Lurie, Institute for Advanced Study; Terence T. Hwa, University of California, San Diego; Wilfried Schmid, Harvard University; Jeffrey D. Ullman, Stanford University; Lai-Sang Young, Courant Institute of Mathematical Sciences, New York University; and Ofer Zeitouni, Weizmann Institute of Science; Foreign Associates: Yoav Benjamini, Tel Aviv University (Israel) and Jürg Fröhlich, ETH Zurich (Switzerland). Berger, DeMarco, Griess, Schmid, and Zeitouni are members of the AMS and Fellows of the AMS. Fagin is a member of the AMS.

The NAS recognizes achievement in science by election to membership, and—along with the National Academy of Engineering and the National Academy of Medicine—provides science, engineering, and health policy advice to the federal government and other organizations. See the full list of this year's honorees. (Image courtesy of the National Academy of Sciences.)

updated April 1, 2020

In response to current challenges that colleges and universities face as a result of the spread of COVID-19, the American Mathematical Society is offering libraries and institutions additional support, in line with recommendations in the ICOLC Statement on the Global COVID-19 Pandemic and Its Impact on Library Services and Resources.

The AMS is also participating in the Copyright Clearance Center Education Continuity License program, providing access to our content for distance learning and other educational uses at no cost to the user.

We are extending grace access for content hosted on our platforms (including MathSciNet) through the end of May for our existing customers. We will re-evaluate this timing as needed.

As courses transition to online, we can provide instructors with complimentary electronic “reserve” copies of our textbooks for cases in which students do not have access to their print copies.

E-books purchased through the perpetual access model on the AMS platform are always available DRM-free with unlimited simultaneous use. In addition, we are partnering with ProQuest to allow multi-user access through mid-June to all e-books purchased on their platforms. Read ProQuest’s statement.

We are providing remote access to all our content, including MathSciNet. In normal circumstances, this remote access can be set up while on campus or while connected via institution VPN (in order to validate IP-based access). We realize many students, faculty, and researchers did not have an opportunity to initiate this access before leaving campus, so we have given instructions to our library partners on how patrons can connect to our content. Please contact your librarian for assistance.

Libraries: if you have not received instructions to share with your patrons, please email us at cust-serv@ams.org or be in touch about any other of your library’s needs.

Review all AMS Resources & Updates.

(Georgia Institute of Technology) The science behind sticky gecko's feet lets gecko adhesion materials pick up about anything. But cost-effective mass production of the materials was out of reach until now. A new method of making them could usher the spread of gecko-inspired grabbers to assembly lines and homes.

Prostate cancer (PCa) cells heavily rely on an active androgen receptor (AR) pathway for their survival. Enzalutamide (MDV3100) is a second-generation antiandrogenic drug that was approved by the Food and Drug Administration in 2012 to treat patients with castration-resistant prostate cancer (CRPC). However, emergence of resistance against this drug is inevitable, and it has been a major challenge to develop interventions that help manage enzalutamide-resistant CRPC. Erythropoietin-producing human hepatocellular (Eph) receptors are targeted by ephrin protein ligands and have a broad range of functions. Increasing evidence indicates that this signaling pathway plays an important role in tumorigenesis. Overexpression of EPH receptor B4 (EPHB4) has been observed in multiple types of cancer, being closely associated with proliferation, invasion, and metastasis of tumors. Here, using RNA-Seq analyses of clinical and preclinical samples, along with several biochemical and molecular methods, we report that enzalutamide-resistant PCa requires an active EPHB4 pathway that supports drug resistance of this tumor type. Using a small kinase inhibitor and RNAi-based gene silencing to disrupt EPHB4 activity, we found that these disruptions re-sensitize enzalutamide-resistant PCa to the drug both in vitro and in vivo. Mechanistically, we found that EPHB4 stimulates the AR by inducing proto-oncogene c-Myc (c-Myc) expression. Taken together, these results provide critical insight into the mechanism of enzalutamide resistance in PCa, potentially offering a therapeutic avenue for enhancing the efficacy of enzalutamide to better manage this common malignancy.

Prevention of aberrant cutaneous wound repair and appropriate regeneration of an intact and functional integument require the coordinated timing of fibroblast and keratinocyte migration. Here, we identified a mechanism whereby opposing cell-specific motogenic functions of a multifunctional intracellular and extracellular protein, the receptor for hyaluronan-mediated motility (RHAMM), coordinates fibroblast and keratinocyte migration speed and ensures appropriate timing of excisional wound closure. We found that, unlike in WT mice, in Rhamm-null mice, keratinocyte migration initiates prematurely in the excisional wounds, resulting in wounds that have re-surfaced before the formation of normal granulation tissue, leading to a defective epidermal architecture. We also noted aberrant keratinocyte and fibroblast migration in the Rhamm-null mice, indicating that RHAMM suppresses keratinocyte motility but increases fibroblast motility. This cell context–dependent effect resulted from cell-specific regulation of extracellular signal-regulated kinase 1/2 (ERK1/2) activation and expression of a RHAMM target gene encoding matrix metalloprotease 9 (MMP-9). In fibroblasts, RHAMM promoted ERK1/2 activation and MMP-9 expression, whereas in keratinocytes, RHAMM suppressed these activities. In keratinocytes, loss of RHAMM function or expression promoted epidermal growth factor receptor–regulated MMP-9 expression via ERK1/2, which resulted in cleavage of the ectodomain of the RHAMM partner protein CD44 and thereby increased keratinocyte motility. These results identify RHAMM as a key factor that integrates the timing of wound repair by controlling cell migration.

β-1,3-d-Glucan is a ubiquitous glucose polymer produced by plants, bacteria, and most fungi. It has been used as a diagnostic tool in patients with invasive mycoses via a highly-sensitive reagent consisting of the blood coagulation system of horseshoe crab. However, no method is currently available for measuring β-1,6-glucan, another primary β-glucan structure of fungal polysaccharides. Herein, we describe the development of an economical and highly-sensitive and specific assay for β-1,6-glucan using a modified recombinant endo-β-1,6-glucanase having diminished glucan hydrolase activity. The purified β-1,6-glucanase derivative bound to the β-1,6-glucan pustulan with a KD of 16.4 nm. We validated the specificity of this β-1,6-glucan probe by demonstrating its ability to detect cell wall β-1,6-glucan from both yeast and hyphal forms of the opportunistic fungal pathogen Candida albicans, without any detectable binding to glucan lacking the long β-1,6-glucan branch. We developed a sandwich ELISA-like assay with a low limit of quantification for pustulan (1.5 pg/ml), and we successfully employed this assay in the quantification of extracellular β-1,6-glucan released by >250 patient-derived strains of different Candida species (including Candida auris) in culture supernatant in vitro. We also used this assay to measure β-1,6-glucan in vivo in the serum and in several organs in a mouse model of systemic candidiasis. Our work describes a reliable method for β-1,6-glucan detection, which may prove useful for the diagnosis of invasive fungal infections.

Inter-α-inhibitor is a proteoglycan essential for mammalian reproduction and also plays a less well-characterized role in inflammation. It comprises two homologous “heavy chains” (HC1 and HC2) covalently attached to chondroitin sulfate on the bikunin core protein. Before ovulation, HCs are transferred onto the polysaccharide hyaluronan (HA) to form covalent HC·HA complexes, thereby stabilizing an extracellular matrix around the oocyte required for fertilization. Additionally, such complexes form during inflammatory processes and mediate leukocyte adhesion in the synovial fluids of arthritis patients and protect against sepsis. Here using X-ray crystallography, we show that human HC1 has a structure similar to integrin β-chains, with a von Willebrand factor A domain containing a functional metal ion-dependent adhesion site (MIDAS) and an associated hybrid domain. A comparison of the WT protein and a variant with an impaired MIDAS (but otherwise structurally identical) by small-angle X-ray scattering and analytical ultracentrifugation revealed that HC1 self-associates in a cation-dependent manner, providing a mechanism for HC·HA cross-linking and matrix stabilization. Surprisingly, unlike integrins, HC1 interacted with RGD-containing ligands, such as fibronectin, vitronectin, and the latency-associated peptides of transforming growth factor β, in a MIDAS/cation-independent manner. However, HC1 utilizes its MIDAS motif to bind to and inhibit the cleavage of complement C3, and small-angle X-ray scattering–based modeling indicates that this occurs through the inhibition of the alternative pathway C3 convertase. These findings provide detailed structural and functional insights into HC1 as a regulator of innate immunity and further elucidate the role of HC·HA complexes in inflammation and ovulation.

β-Glucocerebrosidase (GBA) hydrolyzes glucosylceramide (GlcCer) to generate ceramide. Previously, we demonstrated that lysosomal GBA1 and nonlysosomal GBA2 possess not only GlcCer hydrolase activity, but also transglucosylation activity to transfer the glucose residue from GlcCer to cholesterol to form β-cholesterylglucoside (β-GlcChol) in vitro. β-GlcChol is a member of sterylglycosides present in diverse species. How GBA1 and GBA2 mediate β-GlcChol metabolism in the brain is unknown. Here, we purified and characterized sterylglycosides from rodent and fish brains. Although glucose is thought to be the sole carbohydrate component of sterylglycosides in vertebrates, structural analysis of rat brain sterylglycosides revealed the presence of galactosylated cholesterol (β-GalChol), in addition to β-GlcChol. Analyses of brain tissues from GBA2-deficient mice and GBA1- and/or GBA2-deficient Japanese rice fish (Oryzias latipes) revealed that GBA1 and GBA2 are responsible for β-GlcChol degradation and formation, respectively, and that both GBA1 and GBA2 are responsible for β-GalChol formation. Liquid chromatography–tandem MS revealed that β-GlcChol and β-GalChol are present throughout development from embryo to adult in the mouse brain. We found that β-GalChol expression depends on galactosylceramide (GalCer), and developmental onset of β-GalChol biosynthesis appeared to be during myelination. We also found that β-GlcChol and β-GalChol are secreted from neurons and glial cells in association with exosomes. In vitro enzyme assays confirmed that GBA1 and GBA2 have transgalactosylation activity to transfer the galactose residue from GalCer to cholesterol to form β-GalChol. This is the first report of the existence of β-GalChol in vertebrates and how β-GlcChol and β-GalChol are formed in the brain.

Lens proteins become increasingly cross-linked through nondisulfide linkages during aging and cataract formation. One mechanism that has been implicated in this cross-linking is glycation through formation of advanced glycation end products (AGEs). Here, we found an age-associated increase in stiffness in human lenses that was directly correlated with levels of protein–cross-linking AGEs. α-Crystallin in the lens binds to other proteins and prevents their denaturation and aggregation through its chaperone-like activity. Using a FRET-based assay, we examined the stability of the αA-crystallin–γD-crystallin complex for up to 12 days and observed that this complex is stable in PBS and upon incubation with human lens–epithelial cell lysate or lens homogenate. Addition of 2 mm ATP to the lysate or homogenate did not decrease the stability of the complex. We also generated complexes of human αA-crystallin or αB-crystallin with alcohol dehydrogenase or citrate synthase by applying thermal stress. Upon glycation under physiological conditions, the chaperone–client complexes underwent greater extents of cross-linking than did uncomplexed protein mixtures. LC-MS/MS analyses revealed that the levels of cross-linking AGEs were significantly higher in the glycated chaperone–client complexes than in glycated but uncomplexed protein mixtures. Mouse lenses subjected to thermal stress followed by glycation lost resilience more extensively than lenses subjected to thermal stress or glycation alone, and this loss was accompanied by higher protein cross-linking and higher cross-linking AGE levels. These results uncover a protein cross-linking mechanism in the lens and suggest that AGE-mediated cross-linking of α-crystallin–client complexes could contribute to lens aging and presbyopia.

Mammalian cytochrome P450 enzymes often metabolize many pharmaceuticals and other xenobiotics, a feature that is valuable in a biotechnology setting. However, extant P450 enzymes are typically relatively unstable, with T50 values of ∼30–40 °C. Reconstructed ancestral cytochrome P450 enzymes tend to have variable substrate selectivity compared with related extant forms, but they also have higher thermostability and therefore may be excellent tools for commercial biosynthesis of important intermediates, final drug molecules, or drug metabolites. The mammalian ancestor of the cytochrome P450 1B subfamily was herein characterized structurally and functionally, revealing differences from the extant human CYP1B1 in ligand binding, metabolism, and potential molecular contributors to its thermostability. Whereas extant human CYP1B1 has one molecule of α-naphthoflavone in a closed active site, we observed that subtle amino acid substitutions outside the active site in the ancestor CYP1B enzyme yielded an open active site with four ligand copies. A structure of the ancestor with 17β-estradiol revealed only one molecule in the active site, which still had the same open conformation. Detailed comparisons between the extant and ancestor forms revealed increases in electrostatic and aromatic interactions between distinct secondary structure elements in the ancestral forms that may contribute to their thermostability. To the best of our knowledge, this represents the first structural evaluation of a reconstructed ancestral cytochrome P450, revealing key features that appear to contribute to its thermostability.

pncRNA-D is an irradiation-induced 602-nt long noncoding RNA transcribed from the promoter region of the cyclin D1 (CCND1) gene. CCND1 expression is predicted to be inhibited through an interplay between pncRNA-D and RNA-binding protein TLS/FUS. Because the pncRNA-D–TLS interaction is essential for pncRNA-D–stimulated CCND1 inhibition, here we studied the possible role of RNA modification in this interaction in HeLa cells. We found that osmotic stress induces pncRNA-D by recruiting RNA polymerase II to its promoter. pncRNA-D was highly m6A-methylated in control cells, but osmotic stress reduced the methylation and also arginine methylation of TLS in the nucleus. Knockdown of the m6A modification enzyme methyltransferase-like 3 (METTL3) prolonged the half-life of pncRNA-D, and among the known m6A recognition proteins, YTH domain-containing 1 (YTHDC1) was responsible for binding m6A of pncRNA-D. Knockdown of METTL3 or YTHDC1 also enhanced the interaction of pncRNA-D with TLS, and results from RNA pulldown assays implicated YTHDC1 in the inhibitory effect on the TLS–pncRNA-D interaction. CRISPR/Cas9-mediated deletion of candidate m6A site decreased the m6A level in pncRNA-D and altered its interaction with the RNA-binding proteins. Of note, a reduction in the m6A modification arrested the cell cycle at the G0/G1 phase, and pncRNA-D knockdown partially reversed this arrest. Moreover, pncRNA-D induction in HeLa cells significantly suppressed cell growth. Collectively, these findings suggest that m6A modification of the long noncoding RNA pncRNA-D plays a role in the regulation of CCND1 gene expression and cell cycle progression.

The dextransucrase DSR-OK from the Gram-positive bacterium Oenococcus kitaharae DSM17330 produces a dextran of the highest molar mass reported to date (∼109 g/mol). In this study, we selected a recombinant form, DSR-OKΔ1, to identify molecular determinants involved in the sugar polymerization mechanism and that confer its ability to produce a very-high-molar-mass polymer. In domain V of DSR-OK, we identified seven putative sugar-binding pockets characteristic of glycoside hydrolase 70 (GH70) glucansucrases that are known to be involved in glucan binding. We investigated their role in polymer synthesis through several approaches, including monitoring of dextran synthesis, affinity assays, sugar binding pocket deletions, site-directed mutagenesis, and construction of chimeric enzymes. Substitution of only two stacking aromatic residues in two consecutive sugar-binding pockets (variant DSR-OKΔ1-Y1162A-F1228A) induced quasi-complete loss of very-high-molar-mass dextran synthesis, resulting in production of only 10–13 kg/mol polymers. Moreover, the double mutation completely switched the semiprocessive mode of DSR-OKΔ1 toward a distributive one, highlighting the strong influence of these pockets on enzyme processivity. Finally, the position of each pocket relative to the active site also appeared to be important for polymer elongation. We propose that sugar-binding pockets spatially closer to the catalytic domain play a major role in the control of processivity. A deep structural characterization, if possible with large-molar-mass sugar ligands, would allow confirming this hypothesis.

Bacterial biofilms are cellular communities that produce an adherent matrix. Exopolysaccharides are key structural components of this matrix and are required for the assembly and architecture of biofilms produced by a wide variety of microorganisms. The human bacterial pathogens Escherichia coli and Salmonella enterica produce a biofilm matrix composed primarily of the exopolysaccharide phosphoethanolamine (pEtN) cellulose. Once thought to be composed of only underivatized cellulose, the pEtN modification present in these matrices has been implicated in the overall architecture and integrity of the biofilm. However, an understanding of the mechanism underlying pEtN derivatization of the cellulose exopolysaccharide remains elusive. The bacterial cellulose synthase subunit G (BcsG) is a predicted inner membrane–localized metalloenzyme that has been proposed to catalyze the transfer of the pEtN group from membrane phospholipids to cellulose. Here we present evidence that the C-terminal domain of BcsG from E. coli (EcBcsGΔN) functions as a phosphoethanolamine transferase in vitro with substrate preference for cellulosic materials. Structural characterization of EcBcsGΔN revealed that it belongs to the alkaline phosphatase superfamily, contains a Zn2+ ion at its active center, and is structurally similar to characterized enzymes that confer colistin resistance in Gram-negative bacteria. Informed by our structural studies, we present a functional complementation experiment in E. coli AR3110, indicating that the activity of the BcsG C-terminal domain is essential for integrity of the pellicular biofilm. Furthermore, our results established a similar but distinct active-site architecture and catalytic mechanism shared between BcsG and the colistin resistance enzymes.

The developing nervous system is remarkably sensitive to environmental signals, including disruptive toxins, such as polybrominated diphenyl ethers (PBDEs). PBDEs are an environmentally pervasive class of brominated flame retardants whose neurodevelopmental toxicity mechanisms remain largely unclear. Using dissociated cortical neurons from embryonic Rattus norvegicus, we found here that chronic exposure to 6-OH–BDE-47, one of the most prevalent hydroxylated PBDE metabolites, suppresses both spontaneous and evoked neuronal electrical activity. On the basis of our previous work on mitogen-activated protein kinase (MAPK)/extracellular signal-related kinase (ERK) (MEK) biology and our observation that 6-OH–BDE-47 is structurally similar to kinase inhibitors, we hypothesized that certain hydroxylated PBDEs mediate neurotoxicity, at least in part, by impairing the MEK–ERK axis of MAPK signal transduction. We tested this hypothesis on three experimental platforms: 1) in silico, where modeling ligand–protein docking suggested that 6-OH–BDE-47 is a promiscuous ATP-competitive kinase inhibitor; 2) in vitro in dissociated neurons, where 6-OH–BDE-47 and another specific hydroxylated BDE metabolite similarly impaired phosphorylation of MEK/ERK1/2 and activity-induced transcription of a neuronal immediate early gene; and 3) in vivo in Drosophila melanogaster, where developmental exposures to 6-OH–BDE-47 and a MAPK inhibitor resulted in offspring displaying similarly increased frequency of mushroom-body β–lobe midline crossing, a metric of axonal guidance. Taken together, our results support that certain ortho-hydroxylated PBDE metabolites are promiscuous kinase inhibitors and can cause disruptions of critical neurodevelopmental processes, including neuronal electrical activity, pre-synaptic functions, MEK–ERK signaling, and axonal guidance.

Extracellular matrix-evoked angiostasis and autophagy within the tumor microenvironment represent two critical, but unconnected, functions of the small leucine-rich proteoglycan, decorin. Acting as a partial agonist of vascular endothelial growth factor 2 (VEGFR2), soluble decorin signals via the energy sensing protein, AMP-activated protein kinase (AMPK), in the autophagic degradation of intracellular vascular endothelial growth factor A (VEGFA). Here, we discovered that soluble decorin evokes intracellular catabolism of endothelial VEGFA that is mechanistically independent of mTOR, but requires an autophagic regulator, paternally expressed gene 3 (PEG3). We found that administration of autophagic inhibitors such as chloroquine or bafilomycin A1, or depletion of autophagy-related 5 (ATG5), results in accumulation of intracellular VEGFA, indicating that VEGFA is a basal autophagic substrate. Mechanistically, decorin increased the VEGFA clearance rate by augmenting autophagic flux, a process that required RAB24 member RAS oncogene family (RAB24), a small GTPase that facilitates the disposal of autophagic compartments. We validated these findings by demonstrating the physiological relevance of this process in vivo. Mice starved for 48 h exhibited a sharp decrease in overall cardiac and aortic VEGFA that could be blocked by systemic chloroquine treatment. Thus, our findings reveal a unified mechanism for the metabolic control of endothelial VEGFA for autophagic clearance in response to decorin and canonical pro-autophagic stimuli. We posit that the VEGFR2/AMPK/PEG3 axis integrates the anti-angiogenic and pro-autophagic bioactivities of decorin as the molecular basis for tumorigenic suppression. These results support future therapeutic use of decorin as a next-generation protein therapy to combat cancer.

Barttin is the accessory subunit of the human ClC-K chloride channels, which are expressed in both the kidney and inner ear. Barttin promotes trafficking of the complex it forms with ClC-K to the plasma membrane and is involved in activating this channel. Barttin undergoes post-translational palmitoylation that is essential for its functions, but the enzyme(s) catalyzing this post-translational modification is unknown. Here, we identified zinc finger DHHC-type containing 7 (DHHC7) protein as an important barttin palmitoyl acyltransferase, whose depletion affected barttin palmitoylation and ClC-K-barttin channel activation. We investigated the functional role of barttin palmitoylation in vivo in Zdhhc7−/− mice. Although palmitoylation of barttin in kidneys of Zdhhc7−/− animals was significantly decreased, it did not pathologically alter kidney structure and functions under physiological conditions. However, when Zdhhc7−/− mice were fed a low-salt diet, they developed hyponatremia and mild metabolic alkalosis, symptoms characteristic of human Bartter syndrome (BS) type IV. Of note, we also observed decreased palmitoylation of the disease-causing R8L barttin variant associated with human BS type IV. Our results indicate that dysregulated DHHC7-mediated barttin palmitoylation appears to play an important role in chloride channel dysfunction in certain BS variants, suggesting that targeting DHHC7 activity may offer a potential therapeutic strategy for reducing hypertension.

(Institute for Operations Research and the Management Sciences) Job performance has long been understood to be the primary equalizing factor affecting promotions for men and women in the workplace, but research shows, women don't gain as much from the same performance improvements as men do. New research in the INFORMS journal Information Systems Research shows training plays an important part in promotions for women in the field of information technology.

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