bluuebunny/crossref_metadata_2025 huggingface dataset:
Botulism: Cause, Effects, Diagnosis, Clinical and Laboratory Identification, and Treatment Modalities
Authors: Dembek Zygmunt F. , Smith Leonard A. , Rusnak Janice M.
Published: October 2008

Botulism is a neuroparalytic disease caused by neurotoxins produced by the bacteriaClostridium botulinum. Botulinum neurotoxins (BoNTs) are among the most potent naturally occurring toxins and are a category A biological threat agent. The 7 toxin serotypes of BoNTs (serotypes A–G) have different toxicities, act through 3 different intracellular protein targets, and exhibit different durations of effect. Botulism may follow ingestion of food contaminated with BoNT, from toxin production ofC botulinumpresent in the intestine or wounds, or from inhalation of aerosolized toxin. Intoxication classically presents as an acute, symmetrical, descending flaccid paralysis. Early diagnosis is important because antitoxin therapy is most effective when administered early. Confirmatory testing of botulism with BoNT assays orC botulinumcultures is time-consuming, and may be insensitive in the diagnosis of inhalational botulism and in as many as 32% of food-borne botulism cases. Therefore, the decision to initiate botulinum antitoxin therapy is primarily based on symptoms and physical examination findings that are consistent with botulism, with support of epidemiological history and electrophysiological testing. Modern clinical practice and antitoxin treatment has reduced botulism mortality rates from ∼60% to ≤10%. The pentavalent botulinum toxoid is an investigational product and has been used for more than 45 years in at-risk laboratory workers to protect against toxin serotypes A to E. Due to declining immunogenicity and potency of the pentavalent botulinum toxoid, novel vaccine candidates are being developed. (Disaster Med Public Health Preparedness. 2007;1:122–134)
Regional Health System Response to the 2007 Greensburg, Kansas, EF5 Tornado
Authors: Ablah Elizabeth , Tinius Annie M. , Konda Kurt , Synovitz Carolyn , Subbarao Italo
Published: October 2008

Background:On May 4, 2007 an EF5 tornado hit the rural community of Greensburg, KS, destroying 95% of the town and resulting in 12 fatalities.

Methods:Data was requested from the emergency medical services units that initially responded and the regional hospitals that received people injured in the tornado within 24 hours following the tornado. Requested data included patient age and sex, and injury severity score orICD-9codes. Critical mortality, or the number of deaths of critically injured patients, was also calculated.

Results:The extensive damage caused by the tornado effectively destroyed the infrastructure of the community and created enormous challenges for emergency medical services responders, who were unable to record any triage data. Area hospitals treated 90 patients, who had an average injury severity score of 6.4. Age was found to be related to injury severity, but no relationship between sex and injury severity was found. Critical mortality was found to be 18% for this event.

Conclusions:Injury severity score has seldom been used to analyze natural disasters, especially tornadoes, although such analysis is helpful for understanding the magnitude of the disaster, comparing to other disasters, and preparing for future incidents. Advanced warning and personal preparedness are important factors in reducing tornado-related injuries and deaths. (Disaster Med Public Health Preparedness. 2007;1:90–95)
Prediction Modeling to Determine the Adequacy of Medical Response to Urban Nuclear Attack
Authors: Dallas Cham E. , Bell William C.
Published: October 2008

Background: Government reports have persistently indicated the intent of terrorists and hostile nations to acquire and “weaponize” nuclear materials for deliberate attack on a major US metropolitan city.

Methods: A modeling analysis of the effects of 20- and 550-kiloton nuclear detonations on the 2 major metropolitan centers of Los Angeles and Houston is presented with a focus on thermal casualties. Brode’s work as modified by Binninger was used to calculate thermal fluence, using thermal fractions. The EM-1 and WE programs were used to calculate blast effects. Fallout radiation was calculated using the Defense Threat Reduction Agency’s Hazard Prediction and Assessment Capability V404SP4 with “urban effects” turned on. The ESRI ArcView program calculated affected populations from 2000 US Census block-level data for areas affected by thermal effects.

Results: The population affected by a 550-kiloton nuclear weapon detonated in Los Angeles and Houston is staggering: surviving thermal casualties are estimated at 185,000 and 59,000, respectively. Even the 20-kiloton detonations in Los Angeles and Houston are significant: the numbers of surviving thermal casualties requiring care exceed 28,000 and 10,000, respectively.

Conclusions: The surviving health care community postdetonation would be faced with an unprecedented burden of care for thermal casualties. A great expansion of personnel involved in emergency burn care response is critical. Bold, new approaches such as regionalization and predetermined medical air transport need to be considered. (Disaster Med Public Health Preparedness. 2007;1:80–89)
The New International Health Regulations: Considerations for Global Public Health Surveillance
Authors: Sturtevant Jessica L. , Anema Aranka , Brownstein John S.
Published: October 2008

Global public health surveillance is critical for the identification and prevention of emerging and reemerging infectious diseases. The World Health Organization recently released revised International Health Regulations (IHR) that serve as global legislation and provide guidelines for surveillance systems. The IHR aim to identify and prevent spread of these infectious diseases; however, there are some practical challenges that limit the usability of these regulations. IHR requires Member States to build necessary infrastructure for global surveillance, which may not be possible in underdeveloped countries. A large degree of freedom is given to each individual government and therefore different levels of reporting are common, with substantial emphasis on passive reporting. The IHR need to be enforceable and enforced without impinging on government autonomy or human rights. Unstable governments and developing countries require increased assistance in setting up and maintaining surveillance systems. This article addresses some challenges and potential solutions to the ability of national governments to adhere to the global health surveillance requirements detailed in the IHR. The authors review some practical challenges such as inadequate surveillance and reporting infrastructure, and legal enforcement and maintenance of individual human rights. (Disaster Med Public Health Preparedness. 2007;1:117–121)
Unexplained Deaths in Connecticut, 2002–2003:Failure to Consider Category A Bioterrorism Agents in Differential Diagnoses
Authors: Palumbo John P. , Meek James I. , Fazio Darcy M. , Turner Susan B. , Hadler James L. , Sofair André N.
Published: October 2008

Background: Recognition of bioterrorism-related infections by hospital and emergency department clinicians may be the first line of defense in a bioterrorist attack.

Methods: We identified unexplained infectious deaths consistent with the clinical presentation of anthrax, tularemia, smallpox, and botulism using Connecticut death certificates and hospital chart information. Minimum work-up criteria were established to assess the completeness of diagnostic testing.

Results: Of 4558 unexplained infectious deaths, 133 were consistent with anthrax (2.9%) and 6 (0.13%) with tularemia. None were consistent with smallpox or botulism. No deaths had anthrax or tularemia listed in the differential diagnosis or had disease-specific serology performed. Minimum work-up criteria were met for only 53% of cases.

Conclusions: Except for anthrax, few unexplained deaths in Connecticut could possibly be the result of the bioterrorism agents studied. In 47% of deaths from illnesses that could be anthrax, the diagnosis would likely have been missed. As of 2004, Connecticut physicians were not well prepared to intentionally or incidentally diagnose initial cases of anthrax or tularemia. More effective clinician education and surveillance strategies are needed to minimize the potential to miss initial cases in a bioterrorism attack. (Disaster Med Public Health Preparedness. 2008;2:87–94)
Chronic Disease and Related Conditions at Emergency Treatment Facilities in the New Orleans Area After Hurricane Katrina
Authors: Sharma Andrea J. , Weiss Edward C. , Young Stacy L. , Stephens Kevin , Ratard Raoult , Straif-Bourgeois Susanne , Sokol Theresa M. , Vranken Peter , Rubin Carol H.
Published: October 2008

Background:Disaster preparations usually focus on preventing injury and infectious disease. However, people with chronic disease and related conditions (CDRCs), including obstetric/gynecological conditions, may be vulnerable to disruptions caused by disasters.

Methods:We used surveillance data collected after Hurricane Katrina to characterize the burden of visits for CDRCs at emergency treatment facilities (eg, hospitals, disaster medical assistance teams, military aid stations). In 6 parishes in and around New Orleans, health care providers at 29 emergency treatment facilities completed a standardized questionnaire for injury and illness surveillance from September 8 through October 22, 2005.

Results:Of 21,673 health care visits, 58.0% were for illness (24.3% CDRCs, 75.7% non-CDRCs), 29.1% for injury, 7.2% for medication refills, and 5.7% for routine or follow-up care. The proportion of visits for CDRCs increased with age. Among men presenting with CDRCs, the most common illnesses were cardiovascular disease (36.8%), chronic lower-respiratory disease (12.3%), and diabetes/glucose abnormalities (7.7%). Among women presenting with CDRCs, the most common were cardiovascular disease (29.2%), obstetric/gynecological conditions (18.2%), and chronic lower-respiratory disease (12.0%). Subsequent hospitalization occurred among 28.7% of people presenting with CDRCs versus 10.9% of those with non-CDRCs and 3.8% of those with injury.

Conclusions:Our data illustrate the importance of including CDRCs as a part of emergency response planning. (Disaster Med Public Health Preparedness. 2008;2:27–32)
SWiFT: A Rapid Triage Tool for Vulnerable Older Adults in Disaster Situations
Authors: Dyer Carmel B. , Regev Mor , Burnett Jason , Festa Nicolo , Cloyd Beth
Published: October 2008

Background: In 2005, Hurricane Katrina caused extensive damage to parts of Mississippi, Louisiana, and Alabama, causing many people, including vulnerable older adults, to evacuate to safe surroundings. Approximately 23,000 evacuees—many of them 65 years old or older, frail, and lacking family to advocate for their care—arrived at the Reliant Astrodome Complex in Houston, Texas. There was no method for assessing the immediate and long-term needs of this vulnerable population.

Methods: A 13-item rapid needs assessment tool was piloted on 228 evacuees 65 years old and older by the Seniors Without Families Team (SWiFT), to test the feasibility of triaging vulnerable older adults with medical and mental health needs, financial needs, and/or social needs.

Results: The average age of the individuals triaged was 66.1 ± 12.72 (mean ± standard deviation [SD]) years. Of these, 68% were triaged for medical and or mental health needs, 18% were triaged for financial assistance, and 4% were triaged for social assistance. More than half of the SWiFT-triaged older adults reported having hypertension.

Conclusions: The SWiFT tool is a feasible approach for triaging vulnerable older adults and provides a rapid determination of the level of need or assistance necessary for vulnerable older people during disasters. The tool was only piloted, thus further testing to determine reliability and validity is necessary. Potentially important implications for using such a tool and suggestions for preparing for and responding to disaster situations in which vulnerable older adults are involved are provided. (Disaster Med Public Health Preparedness. 2008;2(Suppl 1):S45–S50)
Trial of Prophylactic Inhaled Steroids to Prevent or Reduce Pulmonary Function Decline, Pulmonary Symptoms, and Airway Hyperreactivity in Firefighters at the World Trade Center Site
Authors: Banauch Gisela I. , Izbicki Gabriel , Christodoulou Vasilios , Weiden Michael D. , Webber Mayris P. , Cohen Hillel , Gustave Jackson , Chavko Robert , Aldrich Thomas K. , Kelly Kerry J. , Prezant David J.
Published: October 2008

Background: Inhaled corticosteroids (ICS) are the most effective anti-inflammatory treatment for asthmatics. This trial evaluated the effects of prophylactic ICS in firefighters exposed to the World Trade Center disaster.

Methods: Inhaled budesonide via a dry powder inhaler (Pulmicort Turbuhaler, AstraZeneca, Wilmington, DE) was offered on-site to New York City firefighters between September 18 and 25, 2001. One to 2 years later, firefighters (n = 64) who completed 4 weeks of daily ICS treatment were evaluated and compared with an age- and exposure-matched comparison group (n = 72) who did not use ICS.

Results: When spirometry results at the final visit were compared with those from the weeks following the 9/11 disaster, the treatment group had a greater increase in forced vital capacity (P = .009) and possibly a slower decline in forced expiratory volume at 1 second (P = .11), as well as a greater improvement in perceived well-being as assessed by the St George's Respiratory Questionnaire (P < .01). There was no difference in airway hyperreactivity and no evidence of adverse effects from ICS.

Conclusions: Because the potential for hazardous exposures is great at many disasters, disease prevention programs based on environmental controls and respiratory protection are warranted immediately. Our results suggest that, pending further study with a larger sample, prophylactic ICS should be considered, along with respiratory protection, to minimize possible lung insult. (Disaster Med Public Health Preparedness. 2008;2:33–39)
Implementing the Cities Readiness Initiative: Lessons Learned From Boston
Authors: Koh Howard K. , Elqura Loris J. , Judge Christine M. , Jacob John P. , Williams Amy E. , Crowther M. Suzanne , Serino Richard A. , Auerbach John M.
Published: October 2008

The federally funded Cities Readiness Initiative (CRI) requires seamless federal, state, and local public health coordination to provide antibiotics to an entire city population within 48 hours of an aerosolized release of anthrax. We document practical lessons learned from the development and implementation of the Boston CRI plan. Key themes center on heightened emphasis on security, a new mass protection model of dispensing, neighborhood-centric clinic site selection, online training of Medical Reserve Corps volunteers, and the testing of operations through drills and exercises. Sharing such lessons can build national preparedness. (Disaster Med Public Health Preparedness. 2008;2:40–49)
Analyzing Postdisaster Surveillance Data: The Effect of the Statistical Method
Authors: DiMaggio Charles , Galea Sandro , Abramson David
Published: October 2008

Data from existing administrative databases and ongoing surveys or surveillance methods may prove indispensable after mass traumas as a way of providing information that may be useful to emergency planners and practitioners. The analytic approach, however, may affect exposure prevalence estimates and measures of association. We compare Bayesian hierarchical modeling methods to standard survey analytic techniques for survey data collected in the aftermath of a terrorist attack. Estimates for the prevalence of exposure to the terrorist attacks of September 11, 2001, varied by the method chosen. Bayesian hierarchical modeling returned the lowest estimate for exposure prevalence with a credible interval spanning nearly 3 times the range of the confidence intervals (CIs) associated with both unadjusted and survey procedures. Bayesian hierarchical modeling also returned a smaller point estimate for measures of association, although in this instance the credible interval was tighter than that obtained through survey procedures. Bayesian approaches allow a consideration of preexisting assumptions about survey data, and may offer potential advantages, particularly in the uncertain environment of postterrorism and disaster settings. Additional comparative analyses of existing data are necessary to guide our ability to use these techniques in future incidents. (Disaster Med Public Health Preparedness. 2008;2:119–126)
Cash Grants in Humanitarian Assistance: A Nongovernmental Organization Experience in Aceh, Indonesia, Following the 2004 Indian Ocean Tsunami
Authors: Doocy Shannon , Johnson Diane , Robinson Courtland
Published: October 2008

Background: Historically, cash interventions, as opposed to material or in-kind aid, have been relatively uncommon in the humanitarian response to emergencies. The widespread implementation of cash-based programs following the 2004 Indian Ocean tsunami provided an opportunity to examine cash distributions following disasters. The Mercy Corps cash grant program in Aceh, Indonesia, was a short-term intervention intended to assist in recompensing losses from the December 2004 tsunami.

Methods: An evaluation of the Mercy Corps cash grant program was conducted for the 12-month period following the tsunami using program monitoring data and a systematic survey of cash grant beneficiaries.

Results: in 2005, the cash grant program disbursed more than US$3.3 million to more than 53,000 beneficiaries; the average cash grant award was US$6390, which was shared by an average of 108 beneficiaries. In a beneficiary survey, more than 95% of respondents reported the grant allocation processes were fair and transparent and that grant funds were received.

Conclusions: The Mercy Corps experience with cash programs suggests that cash interventions in the emergency context, when properly administered, can have an immediate impact and serve as an efficient mechanism for providing assistance. Organizations involved in humanitarian relief, particularly donors and nongovernmental organizations, should consider incorporating cash-based interventions as an element of their response in future emergencies. (Disaster Med Public Health Preparedness. 2008;2:95–103)
Emergency Medical Services Response to a Major Freeway Bridge Collapse
Authors: Hick John L. , Ho Jeffrey D. , Heegaard William G. , Brunette Douglas D. , Lapine Anne , Ward Tom , Clinton Joseph E.
Published: October 2008

Background: The Interstate 35W Bridge in Minneapolis collapsed into the Mississippi River on August 1, 2007, killing 13 people and injuring 127.

Methods: This article describes the emergency medical services response to this incident.

Results/Discussion: Complexities of the event included difficult patient access, multiple sectors of operation, and multiple mutual-aid agencies. Patient evacuation and transportation was rapid, with the collapse zone cleared of victims 95 minutes after the initial 9-1-1 call. A common regional emergency medical service incident management plan that was exercised was critical to the success of the response.

Conclusions: Communication and patient tracking difficulties could be improved in future responses. (Disaster Med Public Health Preparedness. 2008;2(Suppl 1):S17–S24)
Surge Capacity Concepts for Health Care Facilities: The CO-S-TR Model for Initial Incident Assessment
Authors: Hick John L. , Koenig Kristi L. , Barbisch Donna , Bey Tareg A.
Published: October 2008

Facility-based health care personnel often lack emergency management training and experience, making it a challenge to efficiently assess evolving incidents and rapidly mobilize appropriate resources. We propose the CO-S-TR model, a simple conceptual tool for hospital incident command personnel to prioritize initial incident actions to adequately address key components of surge capacity. There are 3 major categories in the tool, each with 4 subelements. “CO” stands for command, control, communications, and coordination and ensures that an incident management structure is implemented. “S” considers the logistical requirements for staff, stuff, space, and special (event-specific) considerations. “TR” comprises tracking, triage, treatment, and transportation: basic patient care and patient movement functions. This comprehensive yet simple approach is designed to be implemented in the immediate aftermath of an incident, and complements the incident command system by aiding effective incident assessment and surge capacity responses at the health care facility level. (Disaster Med Public Health Preparedness. 2008;2(Suppl 1):S51–S57)
Mass Casualty Triage: An Evaluation of the Data and Development of a Proposed National Guideline
Authors: Lerner E. Brooke , Schwartz Richard B. , Coule Phillip L. , Weinstein Eric S. , Cone David C. , Hunt Richard C. , Sasser Scott M. , Liu J Marc , Nudell Nikiah G. , Wedmore Ian S. , Hammond Jeffrey , Bulger Eileen M. , Salomone Jeffrey P. , Sanddal Teri L. , Lord Graydon C. , Markenson David , O'Connor Robert E.
Published: October 2008

Mass casualty triage is a critical skill. Although many systems exist to guide providers in making triage decisions, there is little scientific evidence available to demonstrate that any of the available systems have been validated. Furthermore, in the United States there is little consistency from one jurisdiction to the next in the application of mass casualty triage methodology. There are no nationally agreed upon categories or color designations. This review reports on a consensus committee process used to evaluate and compare commonly used triage systems, and to develop a proposed national mass casualty triage guideline. The proposed guideline, entitled SALT (sort, assess, life-saving interventions, treatment and/or transport) triage, was developed based on the best available science and consensus opinion. It incorporates aspects from all of the existing triage systems to create a single overarching guide for unifying the mass casualty triage process across the United States. (Disaster Med Public Health Preparedness. 2008;2(Suppl 1):S25–S34)
Disaster Triage Systems for Large-scale Catastrophic Events
Authors: Bostick Nathan A. , Subbarao Italo , Burkle Frederick M. , Hsu Edbert B. , Armstrong John H. , James James J.
Published: October 2008

Large-scale catastrophic events typically result in a scarcity of essential medical resources and accordingly necessitate the implementation of triage management policies to minimize preventable morbidity and mortality. Accomplishing this goal requires a reconceptualization of triage as a population-based systemic process that integrates care at all points of interaction between patients and the health care system. This system identifies at minimum 4 orders of contact: first order, the community; second order, prehospital; third order, facility; and fourth order, regional level. Adopting this approach will ensure that disaster response activities will occur in a comprehensive fashion that minimizes the patient care burden at each subsequent order of intervention and reduces the overall need to ration care. The seamless integration of all orders of intervention within this systems-based model of disaster-specific triage, coordinated through health emergency operations centers, can ensure that disaster response measures are undertaken in a manner that is effective, just, and equitable. (Disaster Med Public Health Preparedness. 2008;2(Suppl 1):S35–S39)
Participatory Action Research Methodology in Disaster Research: Results From the World Trade Center Evacuation Study
Authors: Gershon Robyn R.M. , Rubin Marcie S. , Qureshi Kristine A. , Canton Allison N. , Matzner Frederick J.
Published: October 2008

Objective: Participatory action research (PAR) methodology is an effective tool in identifying and implementing risk-reduction interventions. It has been used extensively in occupational health research, but not, to our knowledge, in disaster research. A PAR framework was incorporated into the World Trade Center evacuation study, which was designed to identify the individual, organizational, and structural (environmental) factors that affected evacuation from the World Trade Center Towers 1 and 2 on September 11, 2001. PAR teams—comprising World Trade Center evacuees, study investigators, and expert consultants—worked collaboratively to develop a set of recommendations designed to facilitate evacuation from high-rise office buildings and reduce risk of injury among evacuees.

Methods: Two PAR teams worked first separately and then collectively to identify data-driven strategies for improvement of high-rise building evacuation.

Results: The teams identified interventions targeting individual, organizational, and structural (environmental) barriers to safe and rapid evacuation.

Conclusions: PAR teams were effective in identifying numerous feasible and cost-effective strategies for improvement of high-rise emergency preparedness and evacuation. This approach may have utility in other workplace disaster prevention planning and response programs. (Disaster Med Public Health Preparedness. 2008;2:142–149)
Nestedness in riverine mussel communities: patterns across sites and fish hosts
Authors: Rashleigh Brenda
Published: October 2008

The pattern of nestedness, where species present in depauperate locations are subsets of species present in locations with higher species diversity, is often found in ecological communities. Mussel communities examined in four rivers in the upper Tennessee River basin appeared significantly nested. Mussel species distributions were mostly unrelated to differences in immigration and only weakly related to downstream direction, giving some indication of structuring by differences in extinction. Mussel species distributions were not related to the number of fish species used as hosts for mussel larvae. Mussel species were more likely to overlap on common fish hosts; however, the host‐use matrix was not nested – groups of mussel species used different sets of host fish species in a pattern that appeared phylogenetically related. Sites with high fish host abundance may support high mussel diversity by promoting the survival of mussel species that are less able to attract and infect hosts. Thus, nestedness in freshwater mussel communities may be driven by the array of host fish resources, combined with differences in species’ abilities to use fish hosts. An understanding of the nested pattern in this region can aid conservation of this imperiled fauna.
Impact of simulated moose densities on abundance and richness of vegetation, herbivorous and predatory arthropods along a productivity gradient
Authors: Suominen Otso , Persson Inga‐Lill , Danell Kjell , Bergström Roger , Pastor John
Published: October 2008

Large herbivores can affect vegetation structure and species composition as well as material and energy flows in the ecosystem through their selective feeding, defecation, urination and trampling. These changes have a large potential to indirectly affect other trophic levels, but the mechanisms are poorly known. We studied the impacts of moose Alces alces browsing along a gradient of site productivity by experimentally simulating four different moose densities. Here we show that moose can affect the richness and abundance of three trophic levels in Swedish boreal forests through complex direct and indirect impacts, but in qualitatively different ways depending on how the physical habitat or food resources of a trophic level are affected. Vegetation richness had a hump‐shaped (unimodal) response to increased moose density. Leaf litter production decreased when browsing increased, which in turn depressed the abundance of flying prey for spiders. Consequently, spider abundance and richness declined monotonically. The responses of spider richness to moose density were further conditioned by site productivity: the response was positive at productive and negative at unproductive sites. In contrast, herbivorous Hemiptera were not affected by moose, most likely because the abundance of their food plants was not affected. The highest simulated moose density had an impact on all variables responding to moose even after a few years of treatment and can be considered as overabundance. We also show that the impacts of low or moderate moose density can be positive to some of the organisms negatively affected by high density. The level of herbivore population density that leads to substantial community impacts also depends on site factors, such as productivity.
Lineage specific composition of cyclin D–CDK4/CDK6–p27 complexes reveals distinct functions of CDK4, CDK6 and individual D‐type cyclins in differentiating cells of embryonic origin
Authors: Bryja V. , Pacherník J. , Vondráček J. , Souček K. , Čajánek L. , Horvath V. , Holubcová Z. , Dvořák P. , Hampl A.
Published: October 2008

Abstract.  Objectives: This article is to study the role of G1/S regulators in differentiation of pluripotent embryonic cells. Materials and methods: We established a P19 embryonal carcinoma cell‐based experimental system, which profits from two similar differentiation protocols producing endodermal or neuroectodermal lineages. The levels, mutual interactions, activities, and localization of G1/S regulators were analysed with respect to growth and differentiation parameters of the cells. Results and Conclusions: We demonstrate that proliferation parameters of differentiating cells correlate with the activity and structure of cyclin A/E–CDK2 but not of cyclin D–CDK4/6–p27 complexes. In an exponentially growing P19 cell population, the cyclin D1–CDK4 complex is detected, which is replaced by cyclin D2/3–CDK4/6–p27 complex following density arrest. During endodermal differentiation kinase‐inactive cyclin D2/D3–CDK4–p27 complexes are formed. Neural differentiation specifically induces cyclin D1 at the expense of cyclin D3 and results in predominant formation of cyclin D1/D2–CDK4–p27 complexes. Differentiation is accompanied by cytoplasmic accumulation of cyclin Ds and CDK4/6, which in neural cells are associated with neural outgrowths. Most phenomena found here can be reproduced in mouse embryonic stem cells. In summary, our data demonstrate (i) that individual cyclin D isoforms are utilized in cells lineage specifically, (ii) that fundamental difference in the function of CDK4 and CDK6 exists, and (iii) that cyclin D–CDK4/6 complexes function in the cytoplasm of differentiated cells. Our study unravels another level of complexity in G1/S transition‐regulating machinery in early embryonic cells.
The metalloproteinase ADAM‐12 regulates bronchial epithelial cell proliferation and apoptosis
Authors: Rocks N. , Estrella C. , Paulissen G. , Quesada‐Calvo F. , Gilles C. , Guéders M. M. , Crahay C. , Foidart J.‐M. , Gosset P. , Noel A. , Cataldo D. D.
Published: October 2008

Abstract.  Objectives: The ADAMs (a disintegrin and metalloproteinase) enzymes compose a family of membrane‐bound proteins characterized by their multi‐domain structure and ADAM‐12 expression is elevated in human non‐small cell lung cancers. The aim of this study was to investigate the roles played by ADAM‐12 in critical steps of bronchial cell transformation during carcinogenesis. Materials and methods: To assess the role of ADAM‐12 in tumorigenicity, BEAS‐2B cells were transfected with a plasmid encoding human full‐length ADAM‐12 cDNA, and then the effects of ADAM‐12 overexpression on cell behaviour were explored. Treatment of clones with heparin‐binding epidermal growth factor (EGF)‐like growth factor (HB‐EGF) neutralizing antibodies as well as an EGFR inhibitor allowed the dissection of mechanisms regulating cell proliferation and apoptosis. Results: Overexpression of ADAM‐12 in BEAS‐2B cells promoted cell proliferation. ADAM‐12 overexpressing clones produced higher quantities of HB‐EGF in their culture medium which may rely on membrane‐bound HB‐EGF shedding by ADAM‐12. Targeting HB‐EGF activity with a neutralizing antibody abrogated enhanced cell proliferation in the ADAM‐12 overexpressing clones. In sharp contrast, targeting of amphiregulin, EGF or transforming growth factor‐α failed to influence cell proliferation; moreover, ADAM‐12 transfectants were resistant to etoposide‐induced apoptosis and the use of a neutralizing antibody against HB‐EGF activity restored rates of apoptosis to be similar to controls.Conclusions: ADAM‐12 contributes to enhancing HB‐EGF shedding from plasma membranes leading to increased cell proliferation and reduced apoptosis in this bronchial epithelial cell line.
Regulation of apoptosis by type III interferons
Authors: Li W. , Lewis‐Antes A. , Huang J. , Balan M. , Kotenko S. V.
Published: October 2008

Abstract.  Objective: Two types of interferons (IFNs), type I (IFN‐α/β) and type III (IFN‐λs), utilize distinct receptor complexes to induce similar signalling and biological activities, including recently demonstrated for IFN‐λs antitumour activity. However, ability of type III IFNs to regulate cell population growth remains largely uncharacterized.Materials and methods: Intact and modified human colorectal adenocarcinoma HT29 cells were used to study regulation of apoptosis by IFN‐λs. Results and Conclusions: We report that the IFN‐λR1 chain of the type III IFN receptor complex possesses an intrinsic ability to trigger apoptosis in cells. Signalling induced through the intracellular domain of IFN‐λR1 resulted in G1/G0 phase cell cycle arrest, phosphatidylserine surfacing and chromosomal DNA fragmentation. Caspase‐3, caspase‐8 and caspase‐9 were activated; however, pancaspase inhibitor Z‐VAD‐FMK did not prevent apoptosis. In addition, the extent of apoptosis correlated with the level of receptor expression and was associated with prolonged IFN‐λ signalling. We also demonstrated that the ability to trigger apoptosis is a unique intrinsic function of all IFN receptors. However, more robust apoptosis was induced by signalling through type III IFN receptor than through type I or type II (IFN‐γ) receptors, suggesting higher cytotoxic potential of type III IFNs. In addition, we observed that IFN‐γ treatment sensitized HT29 cells to IFN‐λ‐mediated apoptosis. These results provide evidence that type III IFNs, alone or in combination with other stimuli, have the potential to induce apoptosis.
Leishmania donovaniinfection down-regulates TLR2-stimulated IL-12p40 and activates IL-10 in cells of macrophage/monocytic lineage by modulating MAPK pathways through a contact-dependent mechanism
Authors: Chandra Dinesh , Naik Sita
Published: October 2008

The failure of Leishmania, an intracellular pathogen, to stimulate a pro-inflammatory response following entry into macrophages has been well reported. This occurs in spite of the fact that ligands for the toll-like receptors (TLR) have been recently shown on the parasite surface and their role in disease protection well documented. The outcome of infection in leishmaniasis is determined by the Th1 versus Th2 nature of the effector response and the generation of IL-12 and IL-10 by the infected macrophages is important for this decision. We evaluated the effect of L. donovani infection of monocytes (cell line THP-1, and monocytes derived from human peripheral blood) on Pam3cys (TLR2 ligand) and lipopolysaccharide (TLR4 ligand) stimulated production of IL-12p40 and IL-10. L. donovani infection caused suppression of TLR2 and TLR4-stimulated IL-12p40, with an increase in IL-10 production. Parasites also modulated the TLR2-stimulated mitogen-activated protein kinase (MAPK) pathway by suppressing MAPK P38 phosphorylation and activating extracellular regulated kinase (ERK)1/2 phosphorylation. These effects could be reversed either by using a MAPK P38 activator, anisomycin, or ERK1/2 inhibitor, U0126. L. donovani caused modulation of TLR2-stimulated MAPK pathways in a contact-dependent mechanism. In addition parasite structural integrity but not viability was required for suppression of TLR2-stimulated IL-12p40 and activation of IL-10. These observations suggest that L. donovani has evolved survival strategies that subvert the pro-inflammatory response generated through TLRs.
Impaired immune responses in streptozotocin-induced type I diabetes in mice. Involvement of high glucose
Authors: Rubinstein R , Genaro A M , Motta A , Cremaschi G , Wald M R
Published: October 2008

Diabetes is widely believed to predispose to serious infections. However, the mechanisms linking diabetes and immunosuppression are not well defined. One potential mediator of the altered defence mechanisms is hyperglycaemia. It has been identified as the main factor contributing to the development of diseases associated with diabetes mellitus. In this study we analyse the immune response in diabetes and the direct effect of hyperglycaemia on T and B lymphocyte reactivity. Diabetes induced an early decrease in IgG levels in the secondary response. However, both primary responses against a T-cell-dependent or independent antigen were affected after 6 months of diabetes induction. T- and B- cell proliferation was only decreased at this time. To gain insight into the potential mechanisms involved, we evaluated the influence of hyperglycaemia over the immune response. Pre-incubation of lymph node and spleen cells in a high glucose (HG) containing medium led to a significant time- and dose-dependent decrease in T- and B-cell proliferation. This effect was associated with the presence of HG-derived supernatants. Still viable cells after HG exposition were able to improve their proliferative response when cultured with the mitogen in a fresh standard medium. HG diminished cell viability, increased apoptosis and induced oxidative stress in lymphocytes. These results indicate that HG concentrations can directly affect lymphoid cell growth. An increase in oxidative stress would be implicated in this deleterious effect. The possibility that prolonged exposure to pathologically HG concentrations would result in the immunosuppressive state observed in diabetes is also discussed.
A RNA transcript (Heg) in mononuclear cells is negatively correlated with CD14 mRNA and TSH receptor autoantibodies
Authors: Christensen N J , Habekost G , Bratholm P
Published: October 2008

During a study of gene expression of foxp3 in blood mononuclear cells we observed a DNA product of an unknown RNA fragment. The area of this peak correlated with CD14 mRNA in a small group of subjects. The sequence was localized to chromosome 1. We tested the hypothesis that gene expression of the poly A(−) transcript (designated Heg) in mononuclear cells was correlated with CD14 mRNA in normal subjects and with CD14 mRNA and TSH receptor autoantibodies in patients with acute and untreated Graves' disease. mRNA was expressed in amol/µg DNA. The main study groups were: (i) normal subjects; (ii) patients with early and untreated Graves' disease; and (iii) patients with Graves' disease studied after treatment. In 18 normal subjects and in 20 patients with treated Graves' disease CD14 mRNA was negatively correlated with Heg (P < 0·001). In 17 untreated patients with Graves' disease Heg and thyroid receptor autoantibodies were negatively correlated (P < 0·009). Incubation studies with mononuclear cells showed that the addition of a fragment of the central part of Heg (949 bases) to mononuclear cells decreased CD14 mRNA markedly to zero or nearly zero (P < 0·001). This response was not specific in the sense that siRNA and lipopolysaccharide also decreased CD14 mRNA, probably due to activation of the CD14/Toll-like receptor complex. Single-stranded RNA is likely to increase interferon production. Due to the anti-inflammatory effect Heg may also inhibit the early phase of TSH receptor autoantibody production.
Expression of the CXCR6 on polymorphonuclear neutrophils in pancreatic carcinoma and in acute, localized bacterial infections
Authors: Gaida M M , Günther F , Wagner C , Friess H , Giese N A , Schmidt J , Hänsch G M , Wente M N
Published: October 2008

The chemokine receptor CXCR6 has been described on lymphoid cells and is thought to participate in the homing of activated T-cells to non-lymphoid tissue. We now provide evidence that the chemokine receptor CXCR6 is also expressed by activated polymorphonuclear neutrophils (PMN) in vivo: Examination of biopsies derived from patients with pancreatic carcinoma by confocal laser scan microscopy revealed a massive infiltration of PMN that expressed CXCR6, while PMN of the peripheral blood of these patients did not. To answer the question whether CXCR6 expression is a property of infiltrated and activated PMN, leucocytes were collected from patients with localized soft tissue infections in the course of the wound debridement. By cytofluorometry, the majority of these cells were identified as PMN. Up to 50% of these PMN were also positive for CXCR6. Again, PMN from the peripheral blood of these patients were nearly negative for CXCR6, as were PMN of healthy donors. In a series of in vitro experiments, up-regulation of CXCR6 on PMN of healthy donors by a variety of cytokines was tested. So far, a minor, although reproducible, effect of tumour necrosis factor (TNFα) was seen: brief exposure with low-dose TNFα induced expression of CXCR6 on the surface of PMN. Furthermore, we could show an increased migration of PMN induced by the axis CXCL16 and CXCR6. In summary, our data provide evidence that CXCR6 is not constitutively expressed on PMN, but is up-regulated under inflammatory conditions and mediates migration of CXCR6-positive PMN.
Anti-oxLDL antibody isotype levels, as potential markers for progressive atherosclerosis in APOE−/− and APOE−/−CD40L−/− mice
Authors: Smook M L F , van Leeuwen M , Heeringa P , Damoiseaux J G M C , Theunissen R , Daemen M J A P , Lutgens E , Cohen Tervaert J W
Published: October 2008

In humans and animal models of atherosclerosis, antibodies against oxidized LDL have been associated with atherosclerotic lesion development. It has been suggested that IgM anti-oxLDL antibodies are anti-atherogenic, whereas IgG anti-oxLDL antibodies are pro-atherogenic. In this study, we examined the relation between IgM and IgG antibody levels and atherosclerosis severity in APOE−/−CD40L−/− mice, which are deficient for IgG and develop moderate advanced atherosclerosis, and compared results with mice developing severe (APOE−/−) or no atherosclerosis (C57Bl/6). Mice were followed in time for anti-oxLDL antibodies while on high-fat diet or normal chow. Anti-oxLDL antibody levels were determined by ELISA. Results revealed that 24-week-old APOE−/−CD40L−/− mice had enhanced IgM anti-oxLDL antibody levels when compared with wild-type mice, but similar levels to those of APOE−/− mice. As expected, IgG anti-oxLDL antibody levels were almost absent in APOE−/−CD40L−/− mice. The transition from early to advanced lesions in APOE−/− mice was reflected by elevated IgM anti-oxLDL antibody levels. IgM anti-oxLDL levels did not further increase during progression to more advanced lesions. No relation was found between IgG anti-oxLDL levels and atherosclerosis severity. In conclusion, the severity of advanced atherosclerosis in mice is not reflected by IgM and/or IgG anti-oxLDL antibody levels. Furthermore, less advanced atherosclerotic lesion development in APOE−/−CD40L−/− mice does not seem to be the result of higher levels of protective IgM anti-oxLDL antibodies. Therefore, our study does not support the idea that the previously observed inconsistency in the relation between anti-oxLDL and atherosclerosis severity is due to differences in antibody isotypes.
B cells are involved in the modulation of pathogenic gut immune response in food-allergic enteropathy
Authors: Cardoso C R , Provinciatto P R , Godoi D F , Vieira T S , Ferreira B R , Teixeira G , Rossi M A , Cunha F Q , Silva J S
Published: October 2008

Food enteropathies involve uncontrolled or hypersensitivity reactions to ingested nutrients and may result in IgE and T-helper type 2 (Th2) responses as in food allergy. However, the precise role of B cells in the development of food enteropathies remains uncertain. In this work, we used B cell-deficient mice (B KO) and a model of peanut sensitization to examine the involvement of B lymphocytes in the pathogenesis of food allergy. Results showed that priming of wild-type (WT) mice with peanut proteins induced specific IgG1 and IgE responses in serum, with edema, tissue destruction, epithelial exulceration and inflammatory infiltrate in the gut of sensitized and challenged (S + Peanut) WT animals. In contrast, there was no sera immunoglobulin detection and absence of tissue destruction in the gut of B KO mice, which presented moderate inflammatory infiltrate and villous enlargement after peanut challenge. These animals presented marked decrease in IL-4 and TNF-α and high levels of IL-10, TGF-β, IL-12p40 and IFN-γ mRNA in the gut. Moreover, the expression of CCL5, CCL11 and CXCL1 was reduced in the gut of B KO mice, in contrast to elevated messages of CCL2 or similar detection of Th1-related chemokines in S + Peanut WT mice. Finally, we provided evidence that B cells are necessary to the development of food-related enteropathies and induction of gut inflammation during allergic reactions to food.
Non-obese diabetic–recombination activating gene-1 (NOD–Rag 1 null) interleukin (IL)-2 receptor common gamma chain (IL 2 rγnull) null mice: a radioresistant model for human lymphohaematopoietic engraftment
Authors: Pearson T , Shultz L D , Miller D , King M , Laning J , Fodor W , Cuthbert A , Burzenski L , Gott B , Lyons B , Foreman O , Rossini A A , Greiner D L
Published: October 2008

Immunodeficient hosts engrafted with human lymphohaematopoietic cells hold great promise as a preclinical bridge for understanding human haematopoiesis and immunity. We now describe a new immunodeficient radioresistant non-obese diabetic mice (NOD) stock based on targeted mutations in the recombination activating gene-1 (Rag1null) and interleukin (IL)-2 receptor common gamma chain (IL2rγnull), and compare its ability to support lymphohaematopoietic cell engraftment with that achieved in radiosensitive NOD.CB17–Prkdcscid (NOD–Prkdcscid) IL2rγnull mice. We observed that immunodeficient NOD–Rag1null IL2rγnull mice tolerated much higher levels of irradiation conditioning than did NOD–Prkdcscid IL2rγnull mice. High levels of human cord blood stem cell engraftment were observed in both stocks of irradiation-conditioned adult mice, leading to multi-lineage haematopoietic cell populations and a complete repertoire of human immune cells, including human T cells. Human peripheral blood mononuclear cells also engrafted at high levels in unconditioned adult mice of each stock. These data document that Rag1null and scid stocks of immunodeficient NOD mice harbouring the IL2rγnull mutation support similar levels of human lymphohaematopoietic cell engraftment. NOD–Rag1null IL2rγnull mice will be an important new model for human lymphohaematopoietic cell engraftment studies that require radioresistant hosts.