Hazara nairovirus elicits differential induction of apoptosis and nucleocapsid protein cleavage in mammalian and tick cells.

The Nairoviridae family within the Bunyavirales order comprise tick-borne segmented negative-sense RNA viruses that cause serious disease in a broad range of mammals, yet cause a latent and lifelong infection in tick hosts. An important member of this family is Crimean-Congo haemorrhagic fever virus (CCHFV), which is responsible for serious human disease that results in case fatality rates of up to 30 %, and which exhibits the most geographically broad distribution of any tick-borne virus. Here, we explored differences in the cellular response of both mammalian and tick cells to nairovirus infection using Hazara virus (HAZV), which is a close relative of CCHFV within the CCHFV serogroup. We show that HAZV infection of human-derived SW13 cells led to induction of apoptosis, evidenced by activation of cellular caspases 3, 7 and 9. This was followed by cleavage of the classical apoptosis marker poly ADP-ribose polymerase, as well as cellular genome fragmentation. In addition, we show that the HAZV nucleocapsid (N) protein was abundantly cleaved by caspase 3 in these mammalian cells at a conserved DQVD motif exposed at the tip of its arm domain, and that cleaved HAZV-N was subsequently packaged into nascent virions. However, in stark contrast, we show for the first time that nairovirus infection of cells of the tick vector failed to induce apoptosis, as evidenced by undetectable levels of cleaved caspases and lack of cleaved HAZV-N. Our findings reveal that nairoviruses elicit diametrically opposed cellular responses in mammalian and tick cells, which may influence the infection outcome in the respective hosts.

Two challenges to the double effect doctrine: euthanasia and abortion.

The validity of the double effect doctrine is examined in euthanasia and abortion. In these two situations killing is a method of treatment. It is argued that the doctrine cannot apply to the care of the dying. Firstly, doctors are obliged to harm patients in order to do good to them. Secondly, patients should make their own value judgments about being mutilated or killed. Thirdly, there is little intuitive moral difference between direct and indirect killing. Nor can the doctrine apply to abortion. Doctors kill fetuses as a means of treating the mother. They also kill them as an inevitable side effect of other treatment. Drawing a moral distinction between the direct and the indirect killing gives counterintuitive results. It is suggested that pragmatic rules, not ethics, govern practices around euthanasia and cause it to be more restricted than abortion.

Depressive illness delayed Hamlet's revenge.

If Hamlet had not delayed his revenge there would have been no play. Many explanations of the delay have been offered in the last four centuries. None is convincing. The interpretation which best fits the evidence best is that Hamlet was suffering from an acute depressive illness, with some obsessional features. He could not make a firm resolve to act. In Shakespeare's time there was no concept of acute depressive illness, although melancholy was well known. Melancholy, however, would have been seen as a character defect. In the tragic model the hero brings himself and others to ruin because of a character defect. Thus, at the time, the play conformed to the tragic model. With today's knowledge, it does not. This analysis adds to, but does not replace, other insights into the play.

Long-term preservation of strains of Burkholderia cepacia, Pseudomonas spp. and Stenotrophomonas maltophilia isolated from patients with cystic fibrosis.

Long-term preservation methods are important in the maintenance of bacteria for downstream research applications. Most clinical laboratories have only limited resources for archiving isolates and therefore require cost-effective and simple methods. An effective and cheap storage method using debrinated blood and maintenance at -80 degrees C is described.

Intuitions, principles and consequences.

Some approaches to the assessment of moral intuitions are discussed. The controlled ethical trial isolates a moral issue from confounding factors and thereby clarifies what a person's intuition actually is. Casuistic reasoning from situations, where intuitions are clear, suggests or modifies principles, which can then help to make decisions in situations where intuitions are unclear. When intuitions are defended by a supporting principle, that principle can be tested by finding extreme cases, in which it is counterintuitive to follow the principle. An approach to the resolution of conflict between valid moral principles, specifically the utilitarian and justice principles, is considered. It is argued that even those who justify intuitions by a priori principles are often obliged to modify or support their principles by resort to the consideration of consequences.

Age as a basis for healthcare rationing. Support for agist policies.

It is inevitable that publicly funded, technically sophisticated medical treatments will be increasingly rationed. Other things being equal, there is a greater duty to use them to prolong the lives of younger than older people. Age is therefore an ethical rationing criterion. Denial of this may actually harm older people.

Non-therapeutic (elective) ventilation of potential organ donors: the ethical basis for changing the law.

Non-therapeutic ventilation of potential organ donors would increase the supply of kidneys for transplantation. There are no major ethical objections to it. The means of permitting it are forbidden by laws with an ethical basis. A law permitting it would need an ethical basis. Introducing a third legal method of diagnosing death would be unethical. Expanding the power of the advance directive to permit procedures involving minimal harm would be ethical but not helpful. Extending the power of proxies to permit specific non-therapeutic procedures which caused or risked minimal harm to incompetent patients is the best way forward.

Ensuring equity and quality of care for elderly people: a critique of the College report.

Rationing methods which discriminate against patients on account of their age can be ethical. Sometimes the young and sometimes the old should be favoured. It may depend on who benefits most and whether the rationing is by waiting list or by the exclusion of individuals. Equality is not equity.

Acts of commission, omission, and demission or pulling the plug.

The difference between withholding and withdrawing treatment and actively killing is analysed. The intention of the physician and the effect on the patient are similar but the wider effect on others is different. The harmful effects of withdrawing life-preserving treatment which is no longer beneficial can be reduced by clarification of its purpose. This is to prolong life with value to the patient not just existence.

In defence of ageism.

Health care should be preferentially allocated to younger patients. This is just and is seen as just. Age is an objective factor in rationing decisions. The arguments against 'ageism' are answered. The effects of age on current methods of rationing are illustrated, and the practical applications of an age-related criterion are discussed. Ageist policies are in current use and open discussion of them is advocated.

Quantification of proteinuria: a re-evaluation of the protein/creatinine ratio for elderly subjects.

The difficulties of 24-hour urine collection are well recognized, especially for elderly people for whom a means of estimating protein excretion from a single casual unit sample would be preferable. We compared measurement of total urinary protein estimated by the protein/creatinine ratio with 24-h urine collection and examined the role of the ratio in discriminating between clinically important levels of proteinuria. Although the protein/creatinine ratio appeared to have an excellent correlation with the 24-h collection, the absolute agreement between the two methods was poor. However, the clinical role of the protein/creatinine ratio as a semi-quantitative test of proteinuria remained, provided adjustment for expected creatinine excretion was made.

Assessment of protein intake in early progressive renal disease.

Mean protein intake in a 4-day dietary survey of 34 patients with early renal disease was 1.2 +/- 0.27 g/kg ideal body weight daily. Nine subjects were taking over twice the recommended minimum intake. An increased protein intake was associated with a greater phosphate and sodium intake and an increased excretion of urea, phosphate, and sodium. Daily intake fluctuated markedly during the study (coefficient of variance 22.5 +/- 10.9%). Protein catabolic rate calculated from urea excretion correlated with protein intake, r = 0.60. Mean difference between the two estimates was 0.04 +/- 0.28 g/kg per day. It is proposed that if urea excretion exceeds 4.5 mmol/kg per day in a single collection (protein catabolic rate 0.98 g/kg per day), serial measurements should be made to determine protein intake more accurately and give dietary advice.

Which urine sample for detection of proteinuria?

The standard practice of screening for proteinuria in the concentrated early morning urine was examined. Protein concentration and dipstick readings were determined in early morning and daytime urines of 56 normal subjects and 110 patients with proteinuria attending a renal clinic. In normal subjects the protein concentration was similar in both urine samples and dipsticks gave the same frequency of false positive results, usually only trace positive. The protein concentration was higher in the daytime urine in 76% of patients. Dipstick readings were similar in 64.5% but higher in the daytime urine in 31% and in the early morning urine in only 4.5%. It is recommended that daytime urine should be screened for proteinuria in patients presenting in out-patient clinics.

Biological equivalence of natural bovine and recombinant human alpha-endothelial cell growth factors.

The cDNA encoding human alpha-endothelial cell growth factor (alpha-ECGF) has been engineered for high-level expression in Escherichia coli. Induction of bacterial cultures harboring the recombinant plasmid pMJ26 results in the appearance of a prominent 16-kDa polypeptide. This protein has been purified from bacterial lysates using a rapid, 2-step procedure employing heparin-Sepharose affinity based chromatography and reversed-phase high pressure liquid chromatography. Recombinant human alpha-ECGF was compared to bovine brain-derived alpha-ECGF in three biological assays: receptor binding on murine lung capillary endothelial cells (LE-II cells), stimulation of [3H]thymidine incorporation in LE-II cells, and stimulation of human umbilical vein endothelial cell proliferation. The results demonstrate that the recombinant human mitogen has the same biological potency as the bovine brain-derived material. Fluorescence spectroscopy was used to study the interaction between recombinant ECGF and heparin. Heparin-binding resulted in a 40% reduction in the intrinsic fluorescence of ECGF, consistent with a heparin-induced conformational change. The intrinsic fluorescence of ECGF also varied as a function of pH.