Lots of work to do

It's been very hectic this week. If it hadn't been for the book post I'd prepared earlier, you wouldn't have heard anything.

After the end of exams and a curry in Kegworth with Dipti on Friday night, I had Saturday morning off and then headed to London for some wild times with Lola II. I watched while she tidied up (I think this is in my job description as Lola the Elder) then we went out for sushi - one of the prerequisites for my visit.

On Sunday, we received a visit from friends who helped Lola II specify and buy a new television. In the event, she also received some useful advice about glazing, damp and rendering. The visit ended on a sad note, with a viewing of the end of The Railway Children on the new TV. "Daddy, my daddy!" (That will make Lola II cry again. She's so soft about The Railway Children.)

This week at university has seen the start of five new modules and the continuation of a sixth. As well as all that, I've been catching up with all the jobs that were put off by a month of revision and two weeks of exams, including three months' backlog of Mr A's accounts. That's not a job I relish, and I expect he dislikes it too, given the amount of nagging and sniping he gets from me for the duration. Why can't he just print out the email invoices as they arrive? Or at least file them somewhere so it's easy for me to find them? Anyway, it's nearly done now. The house is still a filthy tip, though.

On Monday morning we started a module on Food Composition, in which I resolved to weigh all my food in future so that I can estimate the amount of protein and energy in it. I have not done this.

On Monday afternoon we started a new Human Nutrition module, where we covered some previous ground on fat, and I resolved to look up the notes and remind myself of what we'd done before. I have not done this either.

On Tuesday we started a module on Medicine and Pathology, looking at the types of disease we're likely to come across in dietetic practice, like diabetes and renal disease. I need to get a textbook out of the library to support this module. I have not done this yet.

On Wednesday, we had the first session of a module about Communication Skills and Educational Methods, which includes the requirement to choose a recent published article in a professional journal, and write an article aimed at a popular magazine readership. It would be a good idea for me to choose my journal article soon, but I have not done this.

Thursday was the first of Computing Techniques, in which we sat through two hours of tedium on what the abbreviations FTP and HTML stand for, before moving into the computer rooms to create a basic web page (part of our first coursework assignment). I need to get started on that because the deadline is only a week away, but guess what, I haven't done it yet.

This morning we continued with a module about nutrition and metabolic disease, and we heard about phenylketonuria. The last piece of coursework for this module was set last term, and is all about metabolic syndrome. You may not be surprised to hear that I haven't started it yet.

I really need to get started on some of this work, but there's one more session needed on Mr A's accounts first...

What I've been reading

Jonathan Strange & Mr Norrell
by Susanna Clarke

narrated by Simon Prebble

"English magicians were once the wonder of the known world, with fairy servants at their beck and call; they could command winds, mountains, and woods. But by the early 1800s they have long since lost the ability to perform magic. They can only write long, dull papers about it, while fairy servants are nothing but a fading memory."

This is a long, long, book, clocking in at 32 hours audio, but so beautifully written that it was worth every minute. One of the reviews I read compares it to Harry Potter, but apart from incorporating magic into reality there's really no comparison in the tone or quality of writing. Not that HP is badly written, but this is just so elegant, and not aimed at children (although older children might enjoy it).



The Food We Eat
by Joanna Blythman

"The average shopper often buys food on trust with little idea of its true quality. This work by an award-winning writer examines all the foods we routinely buy, asking how they match up to the standards that modern food consumers are increasingly demanding."

It's an old, old book now - this edition came out in 1996 - so it's difficult to know whether information is still current. Do they still allow 'smoked' bacon to be smoke-flavoured? I found it interesting, if dated, until I came to the part where she says that table salt has iodine added "to give the salt a healthy image." That ruined the whole thing - if she hasn't done this basic research, then all her other facts are suspect.



Trawler
by Redmond O'Hanlon

"Redmond O'Hanlon describes his extraordinary three-week trip on an Orkney trawler as it journeys far into the north Atlantic in search of its catch. Young skipper Jason Schofield has a 2 million pound overdraft on his boat, the Norlantean, which is why he has to go out in a Category One Force 12 hurricane when the rest of the Scottish fleet has run for shelter."

Long ago I read some amusing books by Mr O'Hanlon, and I mostly enjoyed this one too. He has written it in a style that tries to convey the chaos aboard this trawler - the motion, the noise, the discomfort, the sleep deprivation, the trawlermen, the machinery, the fish, the fatigue. It probably has the most italics and exclamation marks and ellipses in any book I've ever read, and was actually tiring to read, like being incoherently shouted at the whole time. I shall pass it on - Bookmooch, anyone?

Answer Me This!

I'd like to highlight a little spot of stardom achieved by my co-blogger and supporting actor, Lola II. We both listen to the Answer Me This podcast, which Lola II found first and then pestered me until I agreed to listen to it, and now I can't stop. It's a fairly simple premise: people send in questions by email, text, phone or Skype, and Helen and Olly answer the questions at the same time as slapping down Martin the Sound Man when he tries to put forward any opinion. It's an 'explicit' rated podcast so there's usually some fruity language, on the website as well as the podcast.

Lola II has been ringing up with questions on a regular basis for a while, without success. Now she has achieved the glory she deserves, in episode 81, right at the end (after about 28 minutes). I really can't be bothered to work out how to embed a player into this page (it was tough enough doing the YouTube clip), so you'll have to use the link and listen to it there. The comments on that page are quite fun too, and will make sense after you've listened. So head over there now!

The end in sight

While the public face of the blog has been getting all technical about Hepatitis B and celebrating 200 posts, I've actually been sitting at my desk stuffing my brain full of Facts. The heart develops from the mesoderm germline. The cardiogenic mesoderm induces the liver to form from an adjacent stretch of the primitive gut. The kidney develops from a process of reciprocal induction between the metanephric mesenchyme and the ureteric bud sprouting from the Wolffian duct.

The first exam was Molecular Pharming, and took place, as all the lectures had, on the main campus rather than on the Biosciences satellite campus. This is about 20 minutes further from home for me, and I'd never been to the building where the exam was, and on the few occasions when there had been trouble on the motorway, the journey takes about three hours instead of one. The exam was scheduled to start at 9 a.m. I left home at 6.30 - which would probably still have made me late if there had been any trouble on the motorway. The first version of Golden Rice substituted an enzyme from daffodil but didn't produce enough vitamin A. The second version was more successful, and an enzyme from maize was chosen from several based on the strength of the yellow hue in culture. Regulatory, organizational and political factors are still holding up widespread field trials of vitamin A-enhanced Golden Rice.

Immunology was next, and I actually went to see the lecturer to check out a few things that I couldn't make sense of from the lectures or the books. In the event, I found the exam too easy. What I mean by that is: I aim high in exams, putting a lot of effort into revision. And I mean a lot of effort. If the exam is too easy, then people who haven't done much work can get similar marks, and I resent that. I want lazy people to fail. That doesn't make me a bad person. Mast cells are stimulated by antigen-IgE complexes and by C5a complement. The role of complement in the blood is to enhance phagocytosis by opsonisation, create a membrane attack complex to lyse bacterial pathogens, and to help activate B lymphocytes to produce antibodies.

Yesterday was biochemistry, and satisfyingly difficult. I wrote quite a lot about the filaments that form the cellular cytoskeleton, which enable cells to move about, change shape, transport chemicals about and pull chromatids apart when cells divide. It does occur to me that this is an area of knowledge that has little practical use, but I do find it so interesting and it all adds up to a wide-ranging education. Focal contacts allow signals from the extracellular matrix to be sensed by a transmembrane integrin dimer, resulting (if favourable) in the formation of a protein complex recruiting actin microfilaments that are able to do work, pushing the leading edge of the cell forward.

The last exam is Endocrinology and Metabolism tomorrow, and I was feeling a bit aggrieved that it runs from 4.30 to 6.30 p.m. until I found out that some people have exams on Saturday. The parathyroid gland secretes parathyroid hormone in response to low blood calcium, which acts on bone to release calcium and phosphate into the circulation, and on the kidney to retain more calcium but excrete the excess phosphate in the urine. The kidney also produces more active vitamin D, which enhances calcium absorption in the gut.

I'm going out for a curry afterwards, then off to visit Lola II for the weekend. With hardly time to take a breath, the new semester starts first thing Monday with the first lecture of a Food Composition module.

200th post: Accentuate the Positive

In this 200th post I would like to celebrate all those who have made this blog possible.

• Mr A, even though he doesn't read it
• Lola II, who provides endless bloggable material
• Mum and dad, who are my biggest fans.

A few real people that I know have told me that they read my stuff occasionally (or regularly), and this makes me a) happy, and b) slightly embarrassed.

It also wedges my editorial hat firmly in place, because I don't want to write anything that makes anyone feel uncomfortable or reveals secrets or might be vaguely controversial. This is no place to have a sweary rant, or slag off people who have made me cross, or point out imperfections in people who can't help it. Or show Lola II's real hair before her recent new cut. I even feel slightly uncomfortable when I mildly criticise people who can't possibly know about the blog, but if they did read it would recognise themselves as the target. So I have tried to confine myself to the advice of the great Bing Crosby (or rather, Johnny Mercer and Harold Arlen, you see it's always credit where credit's due with me):

You've got to accentuate the positive
Eliminate the negative
Latch on to the affirmative
Don't mess with Mister In-Between

[The YouTube clip I found to illustrate this comes from 'The Singing Detective', which I remember being broadcast to critical acclaim in the 80's, but I didn't watch it. Michael Gambon stars, and I think he's a splendid actor and quite a character, so I've put it on my LoveFilm list for future viewing.]

Anyway, I'm quite surprised that I've kept going this long, and have turned into a blogging addict. But I still think about having a more anonymous place to write tales that are slightly less salubrious, more angry, or less fitting for a tea-time audience. Until I do, this blog has my full attention.

So my 200th post is nearly over, and I've thanked people I know, so it only remains to say thanks to other readers that I don't 'know' in the real world, but who regularly visit and sometimes leave comments. I get an average of perhaps 12 visitors a day, which is lovely. Do leave a comment, especially if you haven't done it before - no need to write loads if you don't want, just say Hi so I know you're there. Or just leave me your favourite word: today, mine is 'plump'.

An edible vaccine for Hepatitis B - part 3

Part 3 - 2008 and into the future

If you're keen you could go back to read part 1 and part 2. They describe progress on vaccines against Hepatitis B virus from genetic modification of yeast to the production of vaccine in edible plants like potato, lettuce and tomato.

The technique for genetic modification I've described so far produces 'stably transformed' plants. This is enabled by the wonderful ability of the plant kingdom to grow a whole new plant out of a mature cell of an adult plant instead of doing the pollen-egg-seed sexual fertilisation thing.

The fragment of virus DNA (the antigen) plus its promoters and targeting sequences that you've constructed are packaged up and fired into plant cells. You can then pick out just those plant cells that have successfully taken up the new DNA and integrated it into their own, and grow new plants that incorporate your gene for virus antigen in every cell, including the pollen and egg cells. So your transformed plants produce a new generation of transformed plants of their own, however you choose to propagate, by cuttings or transferring pollen or whatever.

There's another way to get your carefully constructed DNA into a plant, and that's by using plant viruses. The way that viruses work is that they hijack a host's cells in order to reproduce and spread, so if you put your Hepatitis antigen DNA into a plant virus and then infect a plant with it, the plant produces your desired protein as well as propagating the virus. It's called 'transient transformation' because unless the virus gets to every cell in the plant (which would probably kill it), the viral DNA plus your new DNA isn't incorporated into the whole plant and isn't passed on to the next generation.

The advantage of this transient transformation technique using plant viruses is that you get lots of protein very quickly. To start with, they found that the levels dropped off very rapidly, as if the plant had worked out that something was going on and mobilised defences against it. This leads us to an area of genetic research called 'gene silencing' and a whole new topic that I won't go into here (and in the end, I left it out of my critique too). A way to combat this gene silencing effect was found, and let's leave it at that.

Most of this viral transformation work has been done with non-food plants, like our friend tobacco. One study, though, used a virus that can infect edible plants like cucumber, and perhaps we'll be able to eat the virus-infected plant to get immunised (the study hasn't reached that conclusion yet). Another used a fancy new viral transfection technique, admittedly on tobacco leaves again, but managed to produce extraordinary high levels of Hepatitis B antigen in only 14 days.

The interesting thing about this last study is that it was done by the same team that published the original paper I was reviewing, when they were expressing the antigen in potato and feeding it to mice. They are still working on an edible vaccine, but I think the concentrations they are getting are so low and it takes so long to grow the different plants that this parallel line of research is quite attractive, Perhaps producing Hepatitis B antigen by this viral transformation might form a bridge between the yeast-derived vaccine and a future stably transformed plant supplying an edible vaccine.

Then there are one or two other issues that will need to be sorted out for an edible vaccine. In the paper describing this success with viral transformation, they write: "Although our original research goal was to provide inexpensive, easily administered vaccines in the form of edible transgenic plant material, it is realized that plant-derived vaccines, like any other drug, will be subject to the strict regulations of the US Food and Drug Administration or other similar national agencies. Requirements, including dose standardization, must be met before a plant-derived vaccine is approved. To ensure dosing consistency, vaccine antigen-expressing plant materials must be subjected to downstream processing to some extent, including steps ranging from simple freeze-drying to chromatography purification."

For my coursework I had to put all of this into 2000 words. Obviously I left out some of the explanation, but I had to put in quite a lot more technical detail. I read more than 30 scientific papers, and referenced 20 of them in the final version. I have absolutely no idea whether I will get high, middling or low marks for this bit of work, because it will depend on whether I've interpreted the assignment correctly and written about what they expected me to write about with the correct amount of detail and accurate referencing. I've found it thoroughly fascinating, though, and it all stands me in good stead for the exam.

The last lecture in this module was delivered by one of the team who was responsible for Dolly the cloned sheep. It's been the only lecture that touched on genetic modification of animals; all the rest has been about plants. It's been the best module of the six I've been doing this semester, and isn't even part of the core syllabus. Psychology has been very interesting too, and I've blogged a little bit about that. The modules about the effects of nutrition, exercise and hormones on metabolism have also been good.

The other two modules have only been OK. Immunology is an interesting subject but was taught very badly, and what was described as Mammalian Biochemistry turned out to be about embryology and early organ development. If you think that viral vectors, transcription factors and manipulating DNA are complicated, it's nothing compared to the complexity of complement cascades in the immune system, or the factors that affect the early development of your heart, liver, kidneys and the rest of your innards, starting with one single cell - the fertilised egg. Miraculous.

An edible vaccine for Hepatitis B - part 2

Part 2: from 2000 to the present.

You might like to read Part 1 of this story first, but no matter if you aren't that keen. I left you with an experiment to try and produce an edible Hepatitis B virus vaccine in potatoes. The principle was proved successful, but there wasn't enough antigen in the potatoes to make it a serious contender at this stage.

The first option that might strike you as sensible is to try a different plant. Not just because it might work better, but because raw potato isn't generally considered an appetising food. Apart from the lettuce experiment, different teams have done similar trials in tomatillo (a Mexican fruit), banana, rice and tomato, although I'm not sure what the rice team were thinking, given that cooking the rice would destroy most of the protein. None of them tried feeding their plants to humans, but the team that wrote 'my' paper had a go with potato.

Most, but not all of the human volunteers who ate the transgenic potato had the desired immune response, and they didn't even bother with the adjuvant this time (if you remember, the adjuvant is something to make sure the immune system doesn't ignore your vaccine). Still, none of these foods made enough antigen yet to make a viable edible vaccine.

There were two other paths still being explored, both of them involving tinkering with the transgene (the virus DNA fragment that is put into the plant) by a) choosing a different bit of the virus as the antigen, in case the bit they first chose isn't the best bit - if they could get a better immune response from another fragment, then they wouldn't need as much protein in the vaccine, or b) putting different promoters and targeting constructs in it.

The promoter is the bit of DNA that tells the plant cell machinery to "start transcribing DNA into mRNA from here". You may know that there's an awful lot of random DNA that we don't think is being used - 'junk' DNA. One way the cell knows which bits to use and which bits to leave alone because of certain sequences called promoter regions. Sometimes these promoters are permanently blocked so that the DNA is never transcribed and the protein is never made. For example, every cell in the body contains the DNA that could make the milk protein casein or the blood protein haemoglobin, but we only really want them to be made in mammary cells or bone marrow cells respectively, so it's permanently blocked everywhere else.

Of course, I'm writing as if the cell 'knows' what to do. That's one of the miracles of biology - of course it knows nothing. At molecular levels, which is where this is all going on, it's all about electrical charges and attractions between individual atoms based on where their electrons are. Molecules float about in cells, and randomly bump into other molecules, which give them a push in one direction or other in the same way that comets float about the universe getting pushed about by stars' gravitational fields. Billions of years of evolution (and magic) means that this random motion is directed into interactions that make life possible. We're all about chemistry inside our cells, and chemistry is all about physics.

Where was I? Oh yes, promoters and targeting constructs. If promoters are at the beginning of a gene saying 'start making your mRNA here', then targeting constructs are at the end of the gene, and tell the cell where to make the protein, or what to do with it after it's made. This is where you try to direct the vaccine protein to the fruit or tuber rather than the leaves or flowers, or where you could try and get it secreted, or stored in chloroplasts or the vacuole or other internal areas of the cells. This bit is quite technical, but they've tried a few different ones with variable results. The trouble is that storing a lot of foreign protein carries the risk of poisoning the plant, so there's an amazingly clever technique called inducible expression.

I've mentioned that there are promoters that are permanently blocked in some cells so that their associated gene DNA can't be transcribed. Transcription factors are molecules or complexes that do this blocking, by attaching themselves to the promoter region of DNA. It's possible to treat the plant with something that changes the configuration of the transcription factor so it stops being attached to the promoter. That means that a gene that previously was not transcribed becomes unblocked, and can start making mRNA and then protein. The thing that can change the transcription factor could be as simple as cold, or ethanol, or 'wounding'.

So you could grow your normal potato or tomato or whatever, harvest it normally, then chill it, expose it to ethanol fumes or pound it to a pulp. All the transcription factors would detach from the promoters in every cell, and the DNA would get busy and make your Hepatitis B antigen protein. Brilliant. This has been tried out in tobacco leaves, but not in an edible plant yet.

So we are now at the stage when we've proved the principle of the edible vaccine, but we’re struggling to get the concentration high enough in the edible part of the plant without killing it in the process. There's one other alternative, but for that you'll have to wait for Part 3.

The picture of the potatoes came from www.bbc.co.uk

An edible vaccine for Hepatitis B - part 1

[I first posted this and the following two segments in December, and then decided to take them down because some students hadn't yet reached the deadline for handing in their work.]

I finished the Molecular Pharming coursework at 8.30 p.m. on Thursday, for handing in on Friday. That was a tough assignment, but so very interesting. It also brought together most of the topics that we've covered in this module, and emphasised how very cutting edge this course has been.

Picture of potato plants from http://www.veggieharvest.com/

The research paper I chose to write about was published in 2000, and covered a study on producing a vaccine for Hepatitis B in potato tubers.* We had to produce a critique of the paper, discussing the findings and its context, including what has happened since that time.

To create a vaccine, you have to find something (an 'antigen') that is so similar to the disease that the body responds by creating antibodies and immune memory cells specific to that antigen. That way, if the real thing turns up later, the immune system is ready to fight straight away and the disease doesn't have a chance to gain a foothold. In the most famous case, a cowpox antigen was similar enough to prime the immune system against smallpox. Sometimes they use dead or weakened bacteria, and in the case of Hepatitis B they found a bit of the virus 'envelope', its outer coat, that did the trick.

The next step came about through the new science of genetic manipulation. Normally DNA makes mRNA which makes protein. In this case, they took the bit of virus coat protein and reverse engineered a bit of DNA, then inserted it into the genome of a species of yeast. Through what can only be described as magic, the yeast produced a bit of protein that was identical to this bit of the virus. If the yeast is allowed to get on with living in a huge vat, eventually they can extract the protein made by the yeast (Hepatitis B virus antigen), purify it, process it, and create a vaccine by mixing it with an 'adjuvant': something to wave a flag at the immune system to say "Oi - look over here - I'm a disease!"

I think (without looking at my notes) that a commercial vaccine was first produced in this way in 1981 or 1982. [Aside: I find it fascinating that this was well within my lifetime, but before nearly all of my fellow students were born.] It's very effective as a vaccine, but there are disadvantages: it's expensive to make, needs to be kept sterile and refrigerated, and must be administered by injection on three separate occasions. In the places where Hepatitis B is most prevalent, it's not affordable, sterile refrigerated conditions are rare and getting patients to conform to the three injection regime is difficult.

What I've learned this term about genetic modification, and the ability to manipulate bits of DNA (genes) is staggering. The lecturers talk blithely about creating desired sequences, knocking out genes, detecting single nucleotide substitutions, and even targeting the translation of mRNA into specific locations as if it were the most normal thing in the world. You want to make a protein in fruit but not in leaves? Easy, stick a fruit-specific stretch of DNA on the front of the gene. You want your protein secreted out of the cell once it's made? No problem, stick a secretory signal peptide on the end of the DNA. Want it? You got it. Unbelievable.

Using 'simple' organisms like yeast and bacteria to make useful proteins for us seems pretty routine. Insulin which used to be extracted from pigs can now be made in human-identical form without any input from the animal kingdom. To me, that would seem to reduce animal-based research (unless you consider bacteria or yeast to be animals), but it's genetic modification, because you've modified the genes of the bacteria and the yeast. And some people are simply against genetic modification. If I've learned anything, I've learned that every case is different. Some GM techniques benefit humanity. Some GM techniques benefit large global corporations and their shareholders. Some GM techniques take us close to ethical boundaries, if not across them. But it all needs to be paid for, and nobody can afford to spend a lot of time at the cutting edge without drawing a salary of some kind.

Anyway, I digress. I left you with an effective Hepatitis B vaccine produced by yeast, albeit with drawbacks. Meanwhile, things had been moving in the world of plant science. If a yeast could be genetically modified, why not a plant? Turns out, it can, and what's more, the antigen produced by the plant is better than the yeast, because it needs much less processing afterwards - plants assemble it into a form that resembles a virus much more closely than the yeast can manage. They started doing experiments with tobacco plants (don't ask me why tobacco), and extracted enough protein from leaves to show that an immune response could be established by injecting it into mice.

Obviously there's still a processing overhead when you have to extract your product from tobacco leaves. The obvious next step was to try this out in edible leaves. In 1999 there was a small scale trial where three people ate transgenic lettuce together with the adjuvant (the stuff that makes sure the immune system is paying attention), and bingo! their immune systems recognised the virus. Success. Well, actually, only two out of three showed a response, but still.

Meanwhile, the team that wrote 'my' paper were hard at work making a potato to do the job. First they replicated the mouse experiment and showed that their transgenic potato provoked an immune response when the mice simply ate the tuber with adjuvant - no purification from leaves or injection needed. The main disadvantage of eating the vaccine is that you need a lot more of the antigen than you do when it's injected in a pure form, partly because you digest some of it, and partly because the immune cells in your gut are a bit less sensitive, otherwise they might respond wrongly to everything you eat. And plants only make teeny tiny amounts. You'd have to eat a whole lot of raw potato (or lettuce).

So they tinkered with the DNA to try and make the potato express more protein, and did quite well, increasing the level about forty-fold. As always, there's a catch - potato plants don't naturally produce a viral envelope protein, so when they are asked to make quite a lot of it, they get a bit sick. This is where the paper in 2000 ends: edible potato producing vaccine protein, but still not enough of it. For the next eight years' work, you'll have to wait for Part 2.

* Richter, LJ, Thanavala, Y, Arntzen, CJ & Mason, HS 2000, 'Production of hepatitis B surface antigen in transgenic plants for oral immunization', Nature Biotechnology, vol. 18, no. 11, pp. 1167-71.

Whose hair?

(Click on each picture to enlarge.)





I know whose I'd choose. Feel free to leave your vote.

Exams, here we come

At last, the exams are about to start. This year, revision has been so much harder than last year, mostly reflecting the complexity of the subject areas. And I think that last year the focus was on just covering the basics, making sure that everybody was up to speed, including those who hadn't previously studied biology. Lucky for those whose degrees take just three years - my course is four years, and last year doesn't count at all towards the final grade. Which is a pity, since there's no way I can match last year's marks.

In previous revision periods, I've written little blog summaries of what I've been learning as I've gone along. I'm told by my blog statistics that 'Digestion: the facts' is one of the most popular pages in the blog. That pretty much hasn't been possible this time, partly because I've felt much less confident that I'd get everything done in time, and partly because immunology, endocrinology and mammalian development are so complicated that simple blog summaries aren't possible. I might have done it with the molecular pharming module, but there hasn't been time. Perhaps I'll do some after the pressure's off - I'd really like to write about Golden Rice. And I'll be re-posting the three segments about the edible Hepatitis B vaccine very soon.

The other difference is that last year, the the exams were scheduled one week earlier. Given that I've put in a similar number of hours revision per day this year, I would have been in a bad way without that extra week. As it is, I'm just about ready for this first one, although my brain is fried, and holding a conversation is quite beyond me this evening.

Holiday #3: We all go home

It was a great holiday, even though I didn't do enough walking outdoors stuff. Unlike our home, it was warm enough inside to study comfortably despite the subzero temperatures outside, although there wasn't a convenient table in a quiet corner. The crowd, as always, was kind, thoughtful, zany, and fun to be with. We did the usual amount of reminiscing about when we were at university and previous New Year holidays with various personnel changes over the years, and talked about setting up a Facebook group. How times have changed.

All of us have certainly aged, as you would expect, but for most of the time we don't notice. On occasional events such as this, the changes seem to show up with more clarity. Earlier bedtimes, much less alcohol consumption, fewer snacks, and we used up the leftovers from previous meals rather than starting afresh each day and having to throw away a lot of old food at the end of the week. I made a couple of cracking soups simply by sweeping the contents of the fridge into a saucepan and adding water. I was particularly pleased with the potato, Brussels sprouts, peas, greens and Stilton soup. One of the later meals consisted of soup, salad, a baked Camembert, fruit, malt loaf, chocolate and apricot pudding, and cheese and biscuits.

Another characteristic of aging we found is a tendency to talk about the ailments we are currently nursing. Mr A was onto a winner with his high liver enzyme, although he stuck to his alcohol-free regime with surprising ease. Lola II obviously had potential tetanus and a virulent hair disease, Nick had his gate-related injury, I was using my exercise ball seat because of back/leg trouble (it's working a treat, hardly hurts at all now), and Pippa recently had an emergency appendectomy. When we got talking about blood pressure, everyone had a story to tell - Julia's on medication, and Jim wasn't allowed to join a squash club because of his.

In years gone by, in the wild carefree days of our youth, would we ever have imagined sitting around an enormous table on New Year's Eve passing around a blood pressure monitor, comparing readings between individuals and before and after dinner?

Numbers dwindled towards the end of the week. Mr A needed to get back to the office, Joseph had trials for a football team, Mark and Nick were off to visit Mark's mum, the Wells-Bells were also visiting family. By the last night there were only four of us left. After a movie, more cheese and biscuits and a game of Pictionary, we all turned in, and that was the end of our holiday.

Holiday #2: New Year's Eve

After our terrific morning walk and lunch in a pub, everyone but Mr A, Lola II and me went and did another afternoon walk. We headed off to buy a few last minute provisions for the evening's cooking, and have a wander about the town. It turned out to be far too cold to be wandering about, so instead we installed ourselves in a tea shop with the newspaper and waited for the others.

After dinner, we settled in the sitting room. With Lola II in the party, games in the evenings were compulsory, not an optional extra. The first few nights we found ourselves feeling unexpectedly elderly, heading for bed at around 10 p.m. With a bit of endurance training, we managed to extend the evenings nicely, so that by New Year's Eve we all managed to make it to midnight (except those under the age of 10). So there was enough scope for Lola II's imagination to run riot.

Our main game this year was Balderdash, which is similar to the TV quiz game 'Call My Bluff'. For our special NYE game, however, we were treated to a version of 'Mr & Mrs' compered by our very own Derek Lola Batey II.

[For the purposes of research, I had a quick look for any pertinent websites, and I found this. Not only is there already a Mr & Mrs game in a box, but there's also a DVD, you can rent Derek Batey's own house in Florida for a holiday, he is a "talented, witty and amusing after dinner speaker" and you can even hire the man himself to "write and present special ‘Mr. & Mrs.’ or ‘Partners’ competitions at Corporate events, Company functions or Conferences." Well I never.]

I really should have known that Mr A hates golf more than he hates aerobics, but I definitely did know that 'Whole Lotta Rosie' was the tune to get him out onto the dance floor, and that if the doorbell rang at 7 a.m. on a Saturday, he'd be the one to get out of bed and answer the door. Nevertheless, we trailed the field throughout and came last. The deserving winners (who got 9 out of 10) aren't even married. Pah.

Rainer treated us all to the traditional German punch that is Feuerzangenbowle as midnight came around, the extra leap second ticked over, and it was 2009.

Lola II's Lockjaw Update

Hello, Lola II here.

I've got some good news and some bad news.

The good news is that it's unlikely that I am going to develop lockjaw (from my serious injury on holiday, sustained when some barbed wire callously attacked me when I wasn't looking - see Lola I's previous posting).

The bad news is that it's unlikely that I am going to develop lockjaw. Apologies to my family. You'll have to come up with something else to lessen my enthusiasm for communication.

This morning I went to my doctor to talk Tetanus. The nice smiley man took a good look at my immensely serious and life-threatening, though now surprisingly hardly visible, hand injury, and we talked inoculations.

Due to responsible parenting and superb documentation by those same parents (and some tidy recording of my own), I can tell you that I have had a total of five Tetanus shots over the years. It seems that Lockjaw is not scheduled to happen in my lifetime. I'll have to look for some other challenge in 2009.

The doctor man was very nice and laughed at all my jokes. While we were at it he took my blood pressure, but seemed understandably confused when I pointed out that it was very unlikely I was going to be able to roll my sleeve up sufficiently. I had to explain that I was wearing several layers of clothing, including two thermal vests (though as Paris fashion dictates, one with sleeves, one without). We both struggled for a moment but he very soon saw sense and gave up. You can never wear too many thermal vests, that's my motto. Although I'm considering changing my motto in the summer, we'll see how it goes.

Just before my departure I showed MrMedicalMan all the fruit I was taking to work. He seemed VERY impressed. I thought that maybe I'd get some patient points for healthy eating. Instead I think he probably got some patience points...

Holiday #1: The Walk

As usual I should be revising, but if I don't take some time out to write about the holiday then I'll have forgotten the best bits, which were mostly on New Year's Eve. That's because I spent most of the rest of the holiday working, and nearly everybody went home before the end of the week. Quitters.

I did go out with everyone for a walk on New Year's Eve. It was a great day, and a fabulous walk, and I'd like to put it on record that the book said to generally follow the fence, and Lola II didn't get it wrong. It was the word 'generally' that meant we walked through a bog and climbed over two fences, to get to the bridge that we could have found just by following the fence more generally and a little less specifically. Rainer took over the map reading after that - it helped that he also had a map.

It was probably the coldest week of the year - most days the temperature didn't rise above freezing, and there was frost forming on top of previous days' frost, which made for some lovely effects on leaves and spiders' webs and sheep's wool caught on fences. There was quite a heavy mist at breakfast time and the forecast said it would persist, so we decided not to do the long walk along the top of the hill. The forecast turned out to be less than accurate - there were some lovely sunny patches, but quite a lot of fog too.

Lola II sustained the worst injury of the day when she speared herself on some barbed wire. Luckily, she worked out a full plan of action in case she were to succumb to lockjaw, involving lines of communication, transport plans to hospital, and the use of Jim's iPhone to find the number for Interflora (these things can get overlooked in the excitement, she said). Some of us were hoping that lockjaw would set in sooner rather than later... The wound has almost healed up now, although when I spoke to her yesterday she'd looked up tetanus and the symptoms may not appear for a week or more, so there's still a chance.

The second worst injury of the day was sustained by Nick. Rather than opening a gate between fields, he and others decided to climb over. Rainer, however, thought he would open the gate instead, and after opening it wide, he let it go. I managed to take a photo at the exact moment when it slammed shut against the gatepost.

I've got to get back to work now, but over the next few days you may get further instalments. I've still got plenty of good pictures to post.

The New Year begins

We're back! The holiday is over, and I'm back at my desk, about to start swotting again. Of course I took all the books with me to the holiday and did some revision most days, except for New Year's Eve.

Unfortunately, my lovely new laptop did not survive the holiday. One morning when I switched it on, it simply wouldn't boot. Yesterday we made all kinds of recovery attempts using the supplied CDs, but nothing worked. Very frustrating, but at least I've still got the option of the old PC for the time being, and nothing incredibly critical is lost.

I'll have to find time to write about the holiday, which was great fun despite the revision. I'm also gradually catching up with the regular blogs I read - I have 73 subscriptions in Google Reader, but luckily not all are active. Still, it's taking a while even without leaving comments.

Other more urgent things include upgrading my phone contract so as to replace my broken phone, which gives a sad, muted 'beep' instead of ringing properly, and sorting out the house insurance, which is due next week. By then I'll have forgotten what a holiday feels like.