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The main takeaway from the article: Brady plans every detail of his life so he can play football as long as possible, and he'll do anything he can to get an edge. He diets all year round, takes scheduled naps in the offseason, never misses a workout, eats what his teammates call "birdseed," and does cognitive exercises to keep his brain sharp. Brady struggles to unwind after games and practices. He's still processing, thinking about what's next.

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We will never forget you. Rest peacefully now, beyond the gates of Heaven. You will not be forgotten. People come into and out of this life every day. We know it to be a trial period for the life that comes after. Although we miss you in our world, we know that the world you and your children now inhabit is a better place by far--a world without pain, a world that is fueled by joy and where love reigns supreme.

At the same time that we mourn your departure, we also celebrate your entrance into the next stage. You are not going the rest of the way alone, as we are with you, and you with us. Until we next meet, John Kyranos. Sky I hope you smile everytime you see this site. Pie and Lilly did a beautiful job. You will be greatly missed by all of us. May you and your children be happy in heaven. Love, Ladyyzf. Sky, You have one of the gentlest souls I have had the pleasure of meeting and calling you my friend.

You will always be with us, either in memory, or in our hearts. We know you are in a far better place then we can even imagine. And that one day we will meet again. Then we also will know the splendor that you and your children now know. Thank you for touching my life. I am a much richer person for having known you. Until we meet again LeAnn. You guys have done a beautiful job on this sight. Although I didn't know Sky I was deeply touched by all of your comments to your friend.

May he enjoy his new life with his children. I am sure he will find some new Poker buddies! Linda, Thank you so much for sending me here. Love, Maggie. Sky my love, and you were loved, you touched so many people, in so many ways.

Your love showed always, for your family, friends and poker. You always had a kind word to say about everyone. Your sense of humor never seemed to fail you, especially through a power outage. We rolled into Eichel which then became Wertheim. We cycled along the front until we found our hotel, Hotel Schwann, which I thought was the one in which I had stayed with Pippa in After our showers and clothes washing, we hung our wet cycling gear on the balcony to delight the passing tourists and then went out to find me some birthday cake.

What we decided on in the end was a waffle as it was a bit more warming — today has been a bit chilly. The interior was much more ornate than most Protestant churches I am used to, but it did date from the s. I liked the board which listed the hymns and was rather specific about which verses were to be sung!

We walked around Wertheim a little more, coming across an English red telephone box in a quiet street, and then realised the rain was starting so we went back to the hotel. I paid my 2 Euro per hour for WiFi so that I could check all my messages — lots more birthday wishes, thanks to everyone. I do feel a bit old now though. We both slept well and woke up ready for our breakfasts. No cereal or muesli available, just rolls with ham, cheese and preserves, followed by a yoghurt.

We paid the bill — a bargainous 50 Euros. The day looked bright and sunny at am and I asked the hotel proprietress if she knew the weather forecast — like this all day, she said, and degrees. Perfect cycling weather!

We packed up all our belongings again ready for the off. We took the road bridge over the river rather than the ferry as the ferry is rather a tight squeeze for my trike I used it two years ago. We had to walk over the road bridge but this was fine and we were soon on our way. We went through Sommerhausen with Winterhausen on the other bank, naturally! As we cycled along the Main racing a barge called Wartburg we felt a bit peckish so stopped for a banana each.

We now fancied some lunch well, I was after a cake and we found ourselves on a stretch rather devoid of food opportunities. Eventually we arrived in Zellingen and had two fruitless stops at a Biergarten and what turned out to be a kebab house looking for cake. I knew I wanted cake, so we went on. We stopped at the first place, a bakery, and when I had ascertained that it had a loo as well as serving cake, we sat down. A rather handsome cyclist sitting alone on the next table struck up a conversation.

He was doing the ride the other way but slightly more miles per day he was planning on km per day. He asked us whether we recommended going all the way to Bayreuth and we suggested we felt Bamberg was a better end point. He had a look through our guidebook and then chatted generally about cycling. He said he lives in Bremen so had travelled a fair distance to do the Main river, even if he were going to do it in just three days. A few miles outside Zellingen we came to Himmelstadt which seems to have two claims to fame — stamps and Christmas — and we were treated to lots of information boards about both of these things.

We were now on the right bank of the Main again and the excellent, smooth path continued on through fairly sparsely-populated fields growing lots of sweetcorn and barley with vineyards visible up the hillsides. It was sunny with blue skies and white clouds all day, although not as warm as yesterday maybe 21 degrees. Then tomorrow is a shorter cycle to Wertheim, just 35 miles, and is also my 40th birthday.

I slept really well in the hotel and felt most refreshed when waking up. We listened to The Today Programme on Radio 4 as we got ready and went down to breakfast at We left our room at about am and James did a bit of fettling to his bike, lowering the handlebars slightly to take a little pressure off the saddle. I organised a new arrangement of socks around my flagpole on the trike. This may seem a bit strange but I gaffer-taped the flagpole onto the headrest so it was the right height but the gaffer tape continually stuck to my hair and pulled it out.

Eventually after several days! At Volkach the Main river has a canal which goes directly from Volkach to Schwarzach, reducing a 12km route to just 4. However we I!! We also saw asparagus, strawberries and fruit trees growing.

We cycled through the sleepy village of Nordheim and then eventually reached Sommerach were the route via the Mainkanal joined us. We stopped on the bridge looking over the canal and some other people stopped too so we reciprocated photograph-taking. As it was a hot day I was feeling it was about time for an ice cream so we decided to stop in Schwarzach am Main the next big town for an ice cream. He was stopping today at Marktbreit which is the village before our stop Ochsenfurt , about three miles from it.

Small world! I gave him the address of this blog and then took a photo of him with James at their bikes. We crossed the Main river again at Schwarzach and then had a section away from the river but alongside a road. At Dettelbach, the next village, our route was once again alongside the Main and continued this way through Mainstockheim until we reached Kitzingen. The original plan was to eat lunch at Kitzingen but neither of us was that hungry and the clouds were looking really rather rainy in the direction we were cycling.

Ochsenfurt was only 11 miles away and it seemed like a good idea to press on. The guide book suggests we should go straight along the riverside but all cycle path signs take you slightly away from that and then there are some very faffy crossings of a couple of major roads. I remember Pippa and I struggling with this section when we rode this in but this time, with the helpful GPS track, we were a bit more confident and got there in the end.

There was a little side section of river that had mostly dried up although clearly usually had water, so that was interesting. The bank had starting crumbling into the riverbed as well. The skies were looking more and more rainy now so we were pushing the pedals harder than normal. Marktsteft came and went and then we arrived in Marktbreit, which initially seemed rather industrial and had some rather interesting graffiti!

Eventually we came to a nice bit. Just out the other side I stopped at a bicycle shop a big one but it was unfortunately closed. Now the rain began, just a few drops, then getting more and more persistent. Within a few minutes we were going over a level crossing and before getting back on the bikes it was one of those scary ones without magic barriers where you have to just look and listen — around two blind corners!

The rain was really coming down now and James made sure he cycled in front as the lack of mudguards on my bike meant I was creating three little water fountains. We passed several German cyclists sheltering under trees; they know that this kind of rainstorm passes in half an hour or so, but we were so near to our destination and I was getting really rather peckish that we wanted to press on.

We arrived in Ochsenfurt rather wet but pleased to be there. They appear to be rebuilding the bridge that fell down four or so years ago but is still on the official Radweg — Pippa and I were caught out by this last time and had to take the ferry! The hotel was in a good position opposite the library which apparently has Internet access for punters and had a very good looking food area.

They put our bikes in the garage and gave us the keys to our room. When we arrived in our room we discovered there was, on the wall, a print of The Haywain by Constable. You find reminders of our corner of Essex in the strangest places!

Went well with my cuppa, though, and filled a gap. James took the opportunity for a nap whilst I was out exploring. I went out to attempt to find wifi but failed; however, there was a computer in the library that I could use so I had a quick look at the news and the weather for tomorrow looks like we should stay dry, hurrah!

When I got back to the room James was awake so we went out for a wander. Of course, the moment we stepped out of the hotel it started raining. We stood under the archway into the hotel terrace for a while awaiting weather improvement and eventually got bored so wandered off in the drizzle to see the Main river.

We took some photographs of the bridge that they are rebuilding. They appear to be reusing the old keystones they had numbers painted on them but are setting them into fresh concrete. This explains some rather serious flood defences between the Main and the town walls. We walked along the high street, visiting the towers at both ends. We happened to be at the clock at the Rathaus town hall bang on 6pm and when it chimed some little faces peeped out of windows next to the rather strange skeleton under the clock face.

There was also a character with a beard whose mouth went up and down silently. We decided to eat at our hotel as it had been very busy all day which was a good sign. When we arrived almost all the tables had people on or were reserved — in fact only two tables remained, one of which we took.

Several other tables had people playing cards and the hotel provided little dishes for the money that they appeared to be gambling it looked like copper coins. It came with sauerkraut, of course, and was very tasty.

James also sampled some local white wine which he said was very nice. It was nice though! I decided to time how long the waitress would take to bring our bill as normally we seem to have to wait ages, but the restaurant had had to turn away several people as they were full. The answer was, 40 seconds. We gave them a generous tip as the food had been great and the service was excellent.

The rain had returned by the time we had finished our meal so we decided not to venture out any further. There was only one other person at breakfast and no other tables laid so I assume the hotel was rather underpopulated; this was perhaps reflected in the breakfast as there was slightly less choice than usual, but we still managed to eat a hearty selection.

With a final use of our in-room WiFi listening to The Today Programme on Radio 4 we packed up and collected our bikes, setting off at am. No-one seemed to be about and we had the Radweg to ourselves for the first half hour or so. She had an electric bike which was very shiny and she overtook us, apologising and saying she was just testing out the new bike.

We only chased her for a short distance before being sidetracked by a large wooden waterwheel beside the road. It was mounted in a fairly small stream but was doing a very good job of picking up water — the blurb said it moved 18, litres per hour. As we were getting ready to set off again, Electric Bike woman came back the other way and waved very cheerily. Germans always seem to be friendly to us! I remember having an ice cream here with Pippa on our tour in but this time all I did was got some money out of the bank — not so exciting!

James went in search of the public loo but, despite lots of signs, neither of us could find it. The quality of the cycle track was generally excellent which makes for a much smoother ride. We were passed a couple of times which is bad form but usually by people without panniers. It was getting very warm and sunny now with a few white clouds but lots of blue sky. We stopped for a banana and to anoint ourselves with sun cream:. Having a Chinese in Germany makes a change from eating Wiener Schnitzel!

Schweinfurt is a bustling town and with the warm weather there were loads of people sitting outside, wandering around the shops and generally being sociable. The town centre is pedestrianised and there were bikes everywhere. After our rather lovely Chinese meal we sat around for a few minutes people-watching and then decided to get underway. We were soon back beside the river on the Radweg following the signs to Volkach, our destination, 29km away. The path from Schweinfurt dodged round a couple of motorways but was mostly reasonable quality and wide enough to cycle side-by-side without issue.

We cycled through Bergrheinfeld and then to Garstadt where there were lots of lakes. We seemed to spend forever cycling around the Grafenrheinfeld nuclear power station with its amazing collection of pylons. We then had a long stretch to Wipfield with very ominous clouds above from which we felt the odd spot of rain. At Wipfield we took the ferry across the Main river, at a vast cost of 50 cents each. There were another couple of cyclists on the ferry who seemed to peer weirdly at us; James had noticed them before as they had overtaken us not many people do that and it was a bit embarrassing as they were in their sixties, probably.

Once across the Main we had a short ride up a road which then connects with a good cycle path beside the road. As we got to the corner some people who had parked their car rather randomly started chatting to us about where we were going, was I Dutch or English, why did I ride the trike, where were had stayed last night, etc. As we whizzed along this excellent cycle path towards Stammheim we noticed that the vegetation had changed.

For our first couple of days we were mostly in arable territory with a mixture of wheat, barley, oats, sweetcorn and oilseed rape, none of the fields that big and the next field almost always a different crop. Now we were entering the wine region as the landscape was more hilly and we started to see vines. We passed the military museum which has various aircraft outside, including something that looks like one of the first German jets.

We carried along the excellent path, making very good speed, to Fahr and then were on the final four mile stretch to Volkach. At this point loads of cyclists appeared so the path got quite busy. It was a relief to enter Volkach as James was looking forward to a rest, although he is already getting used to the mileage and seemed to have more speed and energy today.

We found the hotel fine and it did indeed have a music shop next door, and the chap there took us in and then called his aunt who was apparently going to help us. Our room was another plain one on the third floor of the guest house but it seemed fine and the town is very attractive.

We went for a walk-around after our showers and indulged in another impressive ice-cream — I had a Toblerone-Becher and James had an Erdbeer-Becher. After a hearty breakfast and a look at the weather thunderstorms forecast for this afternoon we were ready to set off.

We fetched the bikes from the storage room and set off pedalling. It gave us a good view down over Lichtenfels and the Main valley. We passed several fields of Pick Your Own Strawberries and they smelled wonderful! We rejoined the route again and then made our way through Unterzettlitz and, finally, at Wiesen we caught sight of the Main river, having done 7.

We went through Niederau and then came to Ebensfeld. This route moved away from the river whereas the signposted route follows some meanders so we did 2km less, especially as we carried on along the main road rather than rejoining the cycle route at the first opportunity, missing out the village of Ebing and instead rejoining the route a bit further along at Rattelsdorf.

Appallingly, at this point we were overtaken by a woman in her fifties on a manky old bike with a wire basket containing a rucksack on the back, and some panniers. The path wended its way around the town and then under the A motorway. After a brief stop for a walk around to stretch our legs we continued on, deciding we could make it to Bamberg for our main stop Bamberg was at about 28 miles on our 40 mile journey today. We went through Kemmern and then arrived at Hallstadt which is the start of the Bamberg suburbs.

We wanted something to eat so decided to head off route and go along the river, which we did, but realised fairly soon we were on an island between two bits of river so made our way back towards the Radweg. We found ourselves in an out of town shopping area that had a Netto so we decided to stop to buy some bananas and biscuits. I bought some chocolate too!

The cycle path from Bamberg onwards seemed wider. It also had a rather unusual design in that the loo was squeezed in a small space and a corner of another room projected into this room in a rather inconvenient manner — you had to put your leg past the corner of the wall before sitting down, and when sitting down your nose was almost touching this sharp bit of wall.

Quite odd really. The rain came down a bit more heavily as we went through Dippach and the roads seemed wet but we pushed on, arriving at Eltmann at ish. Total distance today was All the doors were locked when we arrived but I rang on the doorbell and a lady appeared, handing us a room key and telling us where to put the bikes a garage. The room in the Hotel Wallburg is one of the more old-fashioned ones with fairly basic furniture and facilities.

I can listen to Radio 4 through the TuneInRadio app, though, so all is not lost! We both washed our clothes and hung them up, eating some of our goodies from Netto and generally winding down. At 6pm James had woken up from his nap and the sun was shining where was the electrical storm we had been zooming along to avoid?

Not too much that was open, but it seemed a nice little town if rather busy with traffic unusual for the quieter towns round here. When we got back to the hotel we went straight into dinner, and what an excellent meal it was! A huge plate of salad was our starter free of charge. He enjoyed two beers, 0.

Which is pretty cheap for nearly a pint. Although the hotel room is a bit basic we think this might be our favourite hotel at the moment as it has wifi in the room, a very hot shower and absolutely brilliant food at a bargain price. They had even painted happy faces on the boiled eggs!

We paid our bill and were presented with a jar of jam apparently our hotel was a Marmeladen-Hotel which James seemed willing to carry. We set off, instantly being glad that I had prepared the route on my Garmin. Sometimes the Main Radweg route was the less obvious option. With my trusty Garmin, however, we found our way out of Bayreuth and started out.

Here we are crossing the Roter Main, one of the two rivers the other is the Weisser Main that make up the river. The scenery was surprisingly reminiscent of Kent with rolling, wooded hills. The path was generally very good asphalt with some sharp ups and downs at times. At one point we came across a giant granite bicycle which of course we had to photograph, including ourselves in some of the pics. He headed off ahead of us but we passed him within metres as he stopped to look at a roadside memorial.

We continued on, enjoying the warmer weather we had started the day wearing windproofs. There were occasional patches of blue sky but it was mostly grey cloud. Anyway, the diversion went to Katzenreuth where there is the confluence of the red and white Main rivers.

There was a pretty bridge over the now-joined rivers and the route then carried on on loose gravel which was hard work for me. There were two smoother tracks, one each side of the path for normal cyclists, but if I had my drive rear wheel in the smoother track for traction it meant both front wheels were pushing through thicker heaps of gravel and it was rather a lot of effort.

Fortunately we only had a mile of this before we were back in Melkendorf about metres away from where the diversion started. After Melkendorf we passed a fishing lake and reached the outskirts of Kulmbach which is the largest town around here. We continued on to Mainleus where we felt it was definitely time for a stop, after 20 miles. It was good to have a break from cycling for a bit, although there was a rather Marie Celeste air to our surroundings, with just the four random chaps sitting at the bar and occasional cyclists going past very slowly on their enormous aluminium monstrosities.

As I got ready to leave some of the other customers struck up a conversation — was I cycling back to Holland? No, I come from England ; Where had I cycled from? Who went faster — me or James, etc. I chatted to them and then my brain failed and I lost what I was trying to say. This was very bumpy and noisy and slowed us down considerably.

We also found the temperature a bit variable and James put his windproof jacket on, then had to take it off and put on an extra jersey instead. This was because there was some kind of street event on with beer and sausage for sale.

We walked through and then got back on the bikes past the next barrier. We went past Theisau and then into Altenkunstadt which had a McDonalds which we avoided, of course. There were a few people sitting at tables — the man and woman who work there and a lady who struck up a conversation with us. The lady told us that she came to Germany from Kenya in and chatted for a long time about life over here, being black and English-speaking amongst white Germans who spoke no English.

She spoke in a mixture of English and German to us which was quite amusing. James enjoyed an ice cream for dessert and then it was time to continue on the final ten miles to Lichtenfels. This involved a bit of road with a chevron or two denoting steepness at Burgstall. Annoyingly, when descending the steep bit I missed a turning; I tried to indicate it to James in time but failed so he, too, sailed past.

We had to turn round and go back to that turning which was to a rather bad bit of path through a forest, lots of gravel and rutted earth and a mosquito that bit me. This path was a couple of miles and it was particularly hard going on the trike, although having the weight of my panniers on the back certainly helped with the traction. Fortunately before too long we were back on an asphalted road going through Hochstadt. The cycle route now seemed more popular with quite a lot of people out and about.

The temperature seemed to change frequently; when James stopped to oil his pedal I got a bit warm, whereas five minutes later in a shadowy bit I got quite cold. I of course had to wait at the next cycle path junction for James so that he knew which way to go and they overtook me again. There were a few tiny spots of rain as we cycled on towards Lichtenfels but nothing too troubling.

We cycled past a Lido and there was a sign on the road warning cars to be aware of swimmers, which seemed a bit weird. They were advertising hiring of various boats which seemed a nice idea but we pedalled on, through Oberwallstadt and finally into Lichtenfels. It was a fairly uphill push into Lichtenfels but the end was in sight, and we pulled up at the Hotel Krone which is next to a casino and a drinks supermarket. As we were locking up the bikes some English people walked past and said hello presumably the flag gave us away.

We checked in to our decent room. I then had to decide whether or not to make the effort to visit Vierzehnheiligen which is only three miles away but up a VERY steep hill. Andy in Sigmaringen from CycleChat told me I really ought to visit it when I was in Germany before, and I was so near that it felt feeble not to give it a go.

I set off with just some spare tools and the camera in my pannier to make it less effort when climbing the hill. Initially it was a fairly gentle slope up but eventually I joined the road again which had coaches and buses. It was pretty steep so I was in my granny ring and I had half a mile of this but I managed it without my heart rate going above bpm.

I managed the 50 metres and then was in the lovely, swooping downhill again. I managed my fastest speed of the year on this bit of the route — That was The Hotel Krone was very pleasant and has an attached Italian restaurant which we went to for our dinner — huge pizzas, very tasty, and good energy food for tired cyclists! Distance: 4. We had both brought English teabags with us so were able to have a proper cuppa to start the day.

Our original plan was to get the train but we were so ahead of schedule that we ended up getting the one, and we were ready on the platform 20 minutes before that arrived. The train arrived and it was one of the nice new ones that had plenty of bike spaces.

The train was also almost empty so we had a choice of seats in our carriage. I did notice, however, that the display said the train was going to Bingen whereas the timetable and the sign on the front of the train as it came towards us on the platform said Mainz.

Oh well, probably just an oversight — Bingen is on the way to Mainz anyway. When we arrived at Bingen there was an announcement in German that the back half of the train which we were on stopped at Bingen, the front half carried on to Mainz. So we had to get off, walk 20 metres up the platform and get back on again. It clearly played on his mind a lot as he kept shuffling his bike around, said to us a couple more times he was getting off at the next stop, and looked a bit unimpressed with us generally.

It was a nice train journey looking out at the Rhein river along which we cycled last year with blue sky and sunshine. When we got to Mainz station we had six minutes in hand before our S-Bahn to Frankfurt so we located the lift — which was too narrow for my trike. James got the lift up to the station concourse and then down again to the platform for the S-Bahn, I went up and down the stairs carrying my trike James took my luggage in the lift and we met on the platform two minutes before the S-Bahn train was due.

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You are not going the rest of the way alone, as we are with you, and you with us. Until we next meet, John Kyranos. Sky I hope you smile everytime you see this site. Pie and Lilly did a beautiful job. You will be greatly missed by all of us. May you and your children be happy in heaven. Love, Ladyyzf. Sky, You have one of the gentlest souls I have had the pleasure of meeting and calling you my friend. You will always be with us, either in memory, or in our hearts.

We know you are in a far better place then we can even imagine. And that one day we will meet again. Then we also will know the splendor that you and your children now know. Thank you for touching my life. I am a much richer person for having known you. Until we meet again LeAnn. You guys have done a beautiful job on this sight. Although I didn't know Sky I was deeply touched by all of your comments to your friend.

May he enjoy his new life with his children. I am sure he will find some new Poker buddies! Linda, Thank you so much for sending me here. Love, Maggie. Sky my love, and you were loved, you touched so many people, in so many ways. Your love showed always, for your family, friends and poker.

You always had a kind word to say about everyone. Your sense of humor never seemed to fail you, especially through a power outage. You brought so much to my life, to say thank you seems inadequate. There are no words to describe what you meant to me, its enough for me to know that you understood. Someday when we meet again, everything will be as it should be Your memorial was so special.

What a wonderful way to honor your friend! Thanks for sharing. I am so glad I visited. The rain started really coming down now so we stopped under a bike shelter to check the Garmin and take stock. We passed this unusual house, built around a tower. This quiet road continued for a fair while and then we found ourselves going through a supermarket car park — which I remembered doing with Pippa two years ago; this meant this was probably the correct route but it felt a bit random!

This blog is entitled From Bayreuth to Bingen as I felt the two musical bookends Wagner and Hildegard gave it more of a shape. However, we were cycling another 30 miles past Bingen to reach Spay. As soon as we were off the ferry some passing cyclists hailed us — are you English? They clearly were — she was on a Thorn, he a Raleigh steel framed bike, and they had camping kit.

They chatted to us for a bit. They had come up the Rhein from Basle and were staying in campsites. They had left Mainz this morning he pronounced it Mayne-z, not Mine-z and were planning on stopping at a campsite at Trechtinghausen, just a few miles on from Bingen, assuming it was still open — he bemoaned the fact that lots of campsites were just for motorhomes or caravans now, no longer for tents.

The man asked if my trike were made in Cornwall which it was , as it turns out someone in their CTC group bought one. They set off and we followed them for a bit until they took a wrong turn and we took the correct route. We cycled along the good quality track past Bingen station and then past various allotments. The chap then told me he spent six years trying to learn English and then gave up as it was too hard.

I pointed out that English has lots of other polite filler words which do the same job but he said it was just too weird to him. He told us he used to be a barge driver and so had been through the Loreley section a thousand or more times. I would have liked to talk to him longer about this but they had to go. Yet another friendly German couple! We then cycled on for a few miles through Trechtinghausen which did have a campsite!

As we set off the sun had come out and it was so warm I ended up taking off another layer so I just had my cycling vest top, having worn a short-sleeved jersey, long-sleeved windproof and rain jacket over the top off that when we started out this morning! It was a very bumpy concrete path directly beside the main road from Sankt Goar onwards to Hirzenach and it felt narrow when we had to pass other people.

At this point the path goes through a parking area and there were several coachloads of Japanese tourists who seemed to like my bike. We were soon through Boppard, however, and knew we only had four and a half miles to go. The path out of Boppard towards Spay, where the river does a big meander, was freshly surfaced since last year and was really fast. We zoomed along, averaging around 14mph and I was riding at 20mph at one point, which with heavy panniers and tired legs was rather good going.

We had a tailwind which helped. However we were soon rolling into the car park of the Alter Posthof and being reunited with my car which had had a nine day holiday in Spay. The bikes were both then stowed in the car and we collected our spare clothes — what a treat to wear something different tomorrow! And I shall be having a no-cake phase to attempt to undo all the extra lardiness I have gained on this tour.

Distance: Stage 1: Obernburg to Steinheim. Stage 2: Wiesbaden to Eltville. After a reasonable breakfast but with no hot food, not even an egg! The day started off like yesterday in terms of weather — blue skies and dark clouds — but fairly quickly the blue skies disappeared and the cloud cover became thicker.

We continued on, arriving in due course at Aschaffenburg which is a very impressive-looking town. Whilst crossing the bridge over the Main we could see one of those weird boats that we had photographed earlier, a kind of flat trimaran with some kind of metalwork which raises up and down. As we rode from Aschaffenburg to Mainaschaff it started to drizzle — not very much at all, not enough to create puddles on the road, but it ddid make my legs feel slightly damp. The path here was a bit rough in places with tree roots and some narrow sections around huge rocks….

It then became a bit worse with cobbles so James walked around for a minute or two once we got onto better surfaces to recover. There was a diversion in place around Mainaschaff which caught us out a bit I had to go up a high kerb and then through some barriers and at this point we caught up with some chaps who had passed us a few minutes earlier, asking us where we were going speaking in English, presumably because of my flag. They sped on, but when we caught up with them after the diversion I think they had gone the wrong way they chatted to us for a while about our tour.

They said they only paid 20 Euro a night for accommodation, finding places to sleep advertised in Post Offices in the towns they visited. They sped off after a ten minute chat. The cycle path through Kleinostheim was gravel rather than asphalt but it was well compacted so not too uncomfortable.

We crossed the river again at Dettingen and quickly reached Kleinwelzheim and then our chosen lunch spot, Seligenstadt. The rain was really falling when we went in at midday but by the time we left at a quarter to one it had reduced to just fine drizzle again. We only had a short stretch now of 11 miles to Steinheim, near Hanau, where we were catching a train.

This bit of journey seemed really quick as we were pedalling well, now wearing waterproof jackets because the rain kept drizzling down. We went through Klein Krotzenburg and then Hainstadt where there was supposedly a bike shop but when we went looking for it there was nothing to be seen, despite several signs on the road and it being marked on my Garmin.

We pressed on through Klein-Auheim and then found ourselves entering Steinheim, cycling along the river route until about a quarter of a mile before the S-Bahn station where we turned off, saying goodbye to the Main river for this trip. The doors were nice and wide and the bike space was empty when we got on. The journey was an hour and ten minutes and involved going past Frankfurt airport where lots of smartly-dressed Lufthansa people got off.

At Wiesbaden Ost we had to carry our bikes down one set of steps and up another, but I am used to that. Interestingly, though, my heart rate monitor showed I got up to carrying my bike up the stairs, which is equivalent to cycling up Crockleford Hill in Colchester which is very hard work!

Wiesbaden is twinned with Tunbridge Wells in Kent, near where we used to live. It was rather run-down and seedy, although I am sure it is better elsewhere. After a mile we got our first sight of the river Rhein at km so took lots of photos of the bikes there to show we had made it. We then pootled along various tracks and roads and paths through the outskirts of Wiesbaden before getting underway properly.

We went through a huge grassy park area that belonged to the local water company and we stopped to photograph a stork they are huge! We eventually found an information board which showed that the water company have a special Stork breeding programme, and it also looked as if there was a camera looking at one of the nests — a Storkcam? We cycled through the village of Walluf, knowing that we were almost at our destination. Eltville soon appeared, initially a long and boring road but soon turning into a more attractive traditional German village.

I chose to stay there because I had enjoyed a breakfast there in when touring with Pippa, and also because my favourite classical singer, Andreas Scholl, lives just up the road in Kiedrich, so it was as good a place as any to stop.

So the mystery is solved! We had a shower — what a delight to shower without having to wash all our clothes, as we only have one cycling day left! This included a trip to Lidl to buy some chocolate and bananas and other cyclist food. We then went to a restaurant and had more schnitzel with a very generous salad beforehand. The restaurant served Kaiserschmarren my favourite as a dessert but I had no room after my main meal so decided to go back later! We returned to the restaurant at 9pm and seated ourselves before realising there were two Americans and a German couple on another table.

How loudly did those Americans speak!!!! This morning the weather looked rather changeable — blue skies, then five minutes later heavy clouds, then a few minutes later blue skies again. I imagine we will get wet on our 36 mile cycle ride today to Obernburg. We had breakfast which was very good — croissants were on offer, as was scrambled egg with onions, ham, tomato etc in it.

As we were standing on our balcony looking down at the cycle path James saw and recognised some cyclists we had seen yesterday. We were ready to leave by 10am and after a small bit of bike fettling James tipped the nose of his saddle down slightly we were off. We crossed over the river Main as we set off, passing through Kreuzwertheim and discovering that we were cycling into a stonking headwind. Unfortunately our journey today was mostly west and the wind was a westerly, and very strong.

It was hard, hard work pedalling our way round from Kreuzwertheim to Hasloch. The wind was funnelling up the river valley so even if our compass direction changed considerably, we were still fighting against the wind. We were probably only cycling about mph for quite a lot of the time.

After Hasloch we went through Faulbach since when was Bach lazy? The train just whistles when it approaches, which seemed rather old-fashioned and sweet. When a train came past, however, it was perfectly modern — it had three carriages, one of which was a Fahrradwagen bicycle carriage and both others had bicycle spaces marked too.

We fought our way onward against the wind, looking at the dark clouds with some misgivings. We stopped for some food banana and biscuits to have a short rest as it was such hard work. We cycled through Dorfprozelten and then Collenberg which was rather nice, watching a ship going through one of the locks.

We then went to Freudenberg which was very attractive with a castle perched impressively up on the cliffs above. We carried on and then realised it was about to rain so we sheltered under a tree which was surprisingly effective. The rain lasted less than five minutes and, having waited a further five minutes to be sure it had properly stopped, we carried on into blue skies and sun.

The weather here is very changeable but very quick! We were looking forward to arriving at our lunch stop, Miltenberg, as we were feeling like we were expending a lot of energy. I just had a cup of tea, of course. After we finished lunch it rained again, so we sheltered in the cafe a little longer, watching the clouds through the window to see when it was safe to venture out.

We headed off at a quarter past two with fifteen miles still to go. The bad weather and the headwind made us a bit nervous but luckily our route headed more north than west now and the wind was much kinder. After a long stretch with just cycle path and hillside we reached Klingenberg am Main, at which point we crossed over again to the left bank of the river.

This was a bit awkward as there was a diversion on the cycle route which ended up requiring me to cycle up steps — not easy! So we retraced our path for 50 metres or so and got onto the road instead. Crossing the bridge was meant to be on foot Radfahrer absteigen on the pavement but it looked too narrow so I cycled across on the road. James walked on the path and said it was really bumpy.

They were building a new bridge next to the one we crossed so presumably this is being improved. We knew we were on the final stretch now so found the going easier and increased our speed. It was a lovely fast surface so we zoomed along. Our bikes are locked outside down a side passageway — not under cover, but out of sights. A bit of rain will help to wash them down a bit! I made the mistake I had expected to make before now but had avoided — showering with my Garmin heart rate monitor on.

Fortunately it seems to be OK. We walked across a pedestrian footbridge to see the Main river again — tonight is our last night on the Main, tomorrow we will be on the Rhein train-assisted, getting the train from Hanau east of Frankfurt to Wiesbaden opposite Mainz, where the Main flows into the Rhein. As we were resting before the evening meal we heard loads of church bells ringing, then a brass band and some singing.

There was some kind of procession going past our hotel which we watched from our window. It was clearly some kind of Christian procession but not sure what for specifically. We ate in the restaurant below the hotel which was nice, although I was a bit surprised to be charged 1 euro for some tap water. Weather for tomorrow looks similar to today, so not particularly warm and with some showers, but I think it should be a bit warmer on Tuesday fortunately.

Wednesday we will be driving home. I woke up this morning a whole decade older. As James had pointed out, if I had celebrated my birthday in England I could have had an extra hour in my thirties! James joined me downstairs and we went into breakfast.

There was a good selection of bread rolls, cold meats, cheeses, yoghurts, cereal and muesli and fresh fruit. We set off just after nine, retracing our route of last night for the first mile so that we could cross over the Main to the southern side left bank. This involved a long, steep road bridge which was very hard work after our large breakfast and with little warm-up.

The first village we reached was Hofstetten which also had a very steep climb which was hard work! We had quite a headwind to contend with, plus there were some ominous-looking clouds ahead. The forecast for today depended on which of the three websites I looked at, but two of the three suggested we would get rained on — and we did, about halfway to Lohr am Main in a quiet stretch without much shelter. Initially we stayed dry, but the rain came down harder and eventually the wind was blowing it under the tree and water was dripping down through the leaves.

After about ten minutes the rain eased. A lone jogger had run past, clearly made of sterner stuff than us three. Suitably dressed now in waterproofs, gloves, and with a plastic bag stuffed down the back of my seat to attempt to reduce the spray coming onto my back from the back wheel no mudguard , we headed off in the drizzle. The rain petered out after about ten minutes but I was getting quite mucky on my arms, panniers… and my head! It was flicking up off the back wheel onto my head; my new birthday buff was rather less clean than it had been when I unwrapped it this morning and my hair had lumps of mud in.

Oh well. Oh, and my trike was rather muddy too without its mudguard:. We passed through Sendelbach which is opposite Lohr am Main where we could hear some strange brass music playing — there appeared to be some kind of procession going over the bridge through Lohr; lots of people with umbrellas singing, then alternately a marching brass band playing.

There were lots of cyclists sheltering under this big road bridge — they were wimps, the rain had stopped some time ago! A whole bunch of them seemed to have black arrows pained on their cheeks for some unfathomable reason. We carried on riding, trading places with a couple with matching blue waterproofs. We went through Pflochsbach and then Erlach am Main, which were smallish villages with little in between except for excellent-quality Radweg paths.

We cycled through Zimmern, cheered that the skies appeared to be less rainy ahead. At Marktheidenfeld, just over halfway, we stopped for lunch. When we set off again it looked set to be dry, if cloudy. At Homburg is it something to do with hats??? James had noticed days ago that every medium-sized town seems to have a concrete factory.

Same goes for the railway — no fencing or anything, you could just walk straight onto it if you so desire. After Homburg we saw a few vineyards the landscape up till now had mostly been forest or arable , but they soon petered out and we were back to forest. Here is a picture of my feet and my trust Garmin satnav the Oregon which was faithfully guiding us. At Bettingen we started the large meander which was to take us to Wertheim; from Bettinghen Wertheim was probably only a mile and a half away as the crow flies but the river does a significant loop which was another six miles.

At the bottom of the loop was Urphar which had lots of terraced vineyards. We rolled into Eichel which then became Wertheim. We cycled along the front until we found our hotel, Hotel Schwann, which I thought was the one in which I had stayed with Pippa in After our showers and clothes washing, we hung our wet cycling gear on the balcony to delight the passing tourists and then went out to find me some birthday cake. What we decided on in the end was a waffle as it was a bit more warming — today has been a bit chilly.

The interior was much more ornate than most Protestant churches I am used to, but it did date from the s. I liked the board which listed the hymns and was rather specific about which verses were to be sung! We walked around Wertheim a little more, coming across an English red telephone box in a quiet street, and then realised the rain was starting so we went back to the hotel.

I paid my 2 Euro per hour for WiFi so that I could check all my messages — lots more birthday wishes, thanks to everyone. I do feel a bit old now though. We both slept well and woke up ready for our breakfasts.

No cereal or muesli available, just rolls with ham, cheese and preserves, followed by a yoghurt. We paid the bill — a bargainous 50 Euros. The day looked bright and sunny at am and I asked the hotel proprietress if she knew the weather forecast — like this all day, she said, and degrees.

Perfect cycling weather! We packed up all our belongings again ready for the off. We took the road bridge over the river rather than the ferry as the ferry is rather a tight squeeze for my trike I used it two years ago. We had to walk over the road bridge but this was fine and we were soon on our way.

We went through Sommerhausen with Winterhausen on the other bank, naturally! As we cycled along the Main racing a barge called Wartburg we felt a bit peckish so stopped for a banana each. We now fancied some lunch well, I was after a cake and we found ourselves on a stretch rather devoid of food opportunities.

Eventually we arrived in Zellingen and had two fruitless stops at a Biergarten and what turned out to be a kebab house looking for cake. I knew I wanted cake, so we went on. We stopped at the first place, a bakery, and when I had ascertained that it had a loo as well as serving cake, we sat down. A rather handsome cyclist sitting alone on the next table struck up a conversation.

He was doing the ride the other way but slightly more miles per day he was planning on km per day. He asked us whether we recommended going all the way to Bayreuth and we suggested we felt Bamberg was a better end point. He had a look through our guidebook and then chatted generally about cycling. He said he lives in Bremen so had travelled a fair distance to do the Main river, even if he were going to do it in just three days. A few miles outside Zellingen we came to Himmelstadt which seems to have two claims to fame — stamps and Christmas — and we were treated to lots of information boards about both of these things.

We were now on the right bank of the Main again and the excellent, smooth path continued on through fairly sparsely-populated fields growing lots of sweetcorn and barley with vineyards visible up the hillsides. It was sunny with blue skies and white clouds all day, although not as warm as yesterday maybe 21 degrees. Then tomorrow is a shorter cycle to Wertheim, just 35 miles, and is also my 40th birthday.

I slept really well in the hotel and felt most refreshed when waking up. We listened to The Today Programme on Radio 4 as we got ready and went down to breakfast at We left our room at about am and James did a bit of fettling to his bike, lowering the handlebars slightly to take a little pressure off the saddle. I organised a new arrangement of socks around my flagpole on the trike. This may seem a bit strange but I gaffer-taped the flagpole onto the headrest so it was the right height but the gaffer tape continually stuck to my hair and pulled it out.

Eventually after several days! At Volkach the Main river has a canal which goes directly from Volkach to Schwarzach, reducing a 12km route to just 4. However we I!! We also saw asparagus, strawberries and fruit trees growing. We cycled through the sleepy village of Nordheim and then eventually reached Sommerach were the route via the Mainkanal joined us.

We stopped on the bridge looking over the canal and some other people stopped too so we reciprocated photograph-taking. As it was a hot day I was feeling it was about time for an ice cream so we decided to stop in Schwarzach am Main the next big town for an ice cream. He was stopping today at Marktbreit which is the village before our stop Ochsenfurt , about three miles from it. Small world! I gave him the address of this blog and then took a photo of him with James at their bikes.

We crossed the Main river again at Schwarzach and then had a section away from the river but alongside a road. At Dettelbach, the next village, our route was once again alongside the Main and continued this way through Mainstockheim until we reached Kitzingen. The original plan was to eat lunch at Kitzingen but neither of us was that hungry and the clouds were looking really rather rainy in the direction we were cycling.

Ochsenfurt was only 11 miles away and it seemed like a good idea to press on. The guide book suggests we should go straight along the riverside but all cycle path signs take you slightly away from that and then there are some very faffy crossings of a couple of major roads. I remember Pippa and I struggling with this section when we rode this in but this time, with the helpful GPS track, we were a bit more confident and got there in the end.

There was a little side section of river that had mostly dried up although clearly usually had water, so that was interesting. The bank had starting crumbling into the riverbed as well. The skies were looking more and more rainy now so we were pushing the pedals harder than normal. Marktsteft came and went and then we arrived in Marktbreit, which initially seemed rather industrial and had some rather interesting graffiti! Eventually we came to a nice bit. Just out the other side I stopped at a bicycle shop a big one but it was unfortunately closed.

Now the rain began, just a few drops, then getting more and more persistent. Within a few minutes we were going over a level crossing and before getting back on the bikes it was one of those scary ones without magic barriers where you have to just look and listen — around two blind corners! The rain was really coming down now and James made sure he cycled in front as the lack of mudguards on my bike meant I was creating three little water fountains.

We passed several German cyclists sheltering under trees; they know that this kind of rainstorm passes in half an hour or so, but we were so near to our destination and I was getting really rather peckish that we wanted to press on. We arrived in Ochsenfurt rather wet but pleased to be there.

They appear to be rebuilding the bridge that fell down four or so years ago but is still on the official Radweg — Pippa and I were caught out by this last time and had to take the ferry! The hotel was in a good position opposite the library which apparently has Internet access for punters and had a very good looking food area.

They put our bikes in the garage and gave us the keys to our room. When we arrived in our room we discovered there was, on the wall, a print of The Haywain by Constable. You find reminders of our corner of Essex in the strangest places! Went well with my cuppa, though, and filled a gap. James took the opportunity for a nap whilst I was out exploring. I went out to attempt to find wifi but failed; however, there was a computer in the library that I could use so I had a quick look at the news and the weather for tomorrow looks like we should stay dry, hurrah!

When I got back to the room James was awake so we went out for a wander. Of course, the moment we stepped out of the hotel it started raining. We stood under the archway into the hotel terrace for a while awaiting weather improvement and eventually got bored so wandered off in the drizzle to see the Main river. We took some photographs of the bridge that they are rebuilding.

They appear to be reusing the old keystones they had numbers painted on them but are setting them into fresh concrete. This explains some rather serious flood defences between the Main and the town walls. We walked along the high street, visiting the towers at both ends. We happened to be at the clock at the Rathaus town hall bang on 6pm and when it chimed some little faces peeped out of windows next to the rather strange skeleton under the clock face.

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Current lightbox. Live chat. Narrow your search:. Cut Outs. Page 1 of Next page. Recent searches:. Create a new lightbox Save. Create a lightbox Your Lightboxes will appear here when you have created some. Save to lightbox. Germany, Wertheim. Engraving, American, In precision cut lung slices from FVB mice no staining of lymph vessels Figure 4 or any other cells was detected with the anti-CD Conversely, use of an anti-CD C, D Conversely, the anti-CD Intraacinar arteries that are running between alveolar ducts were devoid of lymph vessels Figure 6C.

Lymph vessels were close to pulmonary arteries as they were frequently located in the connective tissue between airways and pulmonary arteries Figure 6D. Lymph capillaries that subsequently ran with veins either started in the alveolar region Figure 7A, B or directly at veins Figure 7C. Capillaries that ran with airways were starting either directly adjacent at the airway or within the connective tissue between airways and arteries Figure 7D, E. Based on the location and their characteristic architecture of smooth muscle cells, airways, arteries, and veins can be distinguished.

B Boxed area in A labeled with B. An airway AW is accompanied by a pulmonary artery PA. The airway is identified by smooth muscle cells that are oriented perpendicular to course of the airway lumen and exhibit regular gaps between them. C Boxed area in A labeled with C. D Boxed area in A labeled D. They do not accompany airways or alveolar ducts and lie separately. C Intraacinar arteries IA are not accompanied by lymph vessels.

D, E Lymph vessels are found frequently in the connective tissue between pulmonary arteries and airways. E Paraffin section of murine lung stained with Masson Goldner stain. D, E Lymphatic capillaries that follow airways AW either begin directly on airways D or E in the connective tissue between arteries labeled A in E and airways.

Confined to the immediate hilar region, immunoreactive cells with the morphology of fibroblasts were found Figure 8A. Lymph vessels do not exhibit strong autofluorescence signal in multiphoton microscopy and appear as dark holes. Projection of a z-stack in a living murine trachea ex vivo recorded by multiphoton microscopy.

Other structures of the tissue are visualized by detection of tissue autofluorescence. Exposure to house dust mite extract induced no obvious changes in the distribution and morphology of lymph vessels compared to the PBS treated animals. Very often, T cells were associated with or even located inside lymph vessels Figure 11E, F. Around veins, T cells were close to lymphatics Figure 12A. Intraacinar arteries were devoid of lymph vessels but exhibited accumulation of T cells Figure 12B. This T cell coat often continuously accompanied arteries and was still present when the intraacinar arteries connected the pulmonary arteries that accompanied airways Figure 12C.

In this region T cells were mostly found around arteries and in the connective tissue between arteries and airways that also harbored the lymph vessels Figure 12D. Around the parts of the airways that were not oriented towards the arteries, markedly fewer lymphocytes were found Figure 12D. While projections A, C, E show many T cells that are located around lymph vessels arrowheads and can mask them, a single confocal section B, D, F allow the unambiguous identification of lymph vessels arrowheads and the localization of T cells around and within arrow in F lymph vessels.

C Accumulations of T cells around arteries are continuous from intraacinar arteries to pulmonary arteries PA. D Around airways AW , T cells preferentially accumulate around pulmonary arteries. Classical markers for lymph endothelium such as podoplanin and LYVE-1 that reliably stain lymph vessels in other organs are not very useful to identify lymph vessels in the lung as staining of other cell types hampers the identification of lymph vessels c.

Figure 3. This is due to strong expression in other cell types such as blood endothelial cells and type II pneumocytes [8]. In the lung it is also found on nerve fibers, fibroblasts and T cells. In principle, the staining of other cell types can be a problem to identify lymphatic vessels and have hampered the use of established markers in lung research [8].

The additional staining of T cells can even be beneficial as this gives an insight into the distribution of T cells with respect to lymphatics without the need for multiple antibodies. It is therefore feasible to use the antibody to detect lymph vessels in in vivo and ex vivo imaging experiments and should also be suitable for FACS analysis of lymphatic endothelial cells. We were able to identify two different lymphatic drainage routes.

One begins in the parenchyma and leaves the lung via the veins. The other begins around the airways and in the connective tissue between arteries and airways and leaves the lung via the airways. In contrast to the human lung, we did not observe a separate network in the pleura indicating that pleural drainage occurs primarily via the lymphatic system around the veins. Based on the distribution of both systems, most of the extracellular fluid of the alveolar area will be drained via the vein associated lymphatics, whereas extracellular fluid that is generated around the airways will primarily be drained via airway associated lymph vessels.

Intrapulmonary lymphatics were devoid of smooth muscle cells with the exception of large lymph vessels at the lung hilum. This indicates that the intrapulmonary transport of lymph in both systems primarily relies on compression of lymph vessels during breathing and not on active contraction of smooth muscle cells. The presence of valves within lymph vessels supports this idea.

Two separate routes for lymph vessels exist. One begins in the parenchyma and leaves the lung via veins and the other begins around airways or in the connective tissue between airways and arteries and follows the airways to leave the lung.

Cells that have left the vasculature in the alveolar region can enter the lymphatic system by migrating to lymph vessels around veins or by migration to lymph vessels around airways, possibly by using intraacinar arteries as guide. Following application of house dust mite extract, we found the expected accumulation of T cells adjacent to airways but also in the alveolar region.

At first sight it is surprising that the effects of house dust mite extract application were not confined to airways. However, the applied house dust mite extract could also reach the alveolar space. In fact, the alveolar region seems to be an important area for antigen uptake as Thornton and coworkers have recently shown.

Using an OVA model of allergic airway inflammation, the authors demonstrated that most of the OVA was taken up by dendritic cells in the alveolar region and not in the airways [17]. Thus, the increased amount of T cells, we observed indicates that house dust mite reached the alveolar space and initiated an immune response there. Based on the lymph vessel distribution, we expected that immune cells in the alveolar region will travel to draining lymph nodes via lymphatics around veins.

However, we also found substantial accumulation of T cells around intraacinar arteries despite the fact that these vessels were consistently devoid of lymph vessels. We also observed that this lymphocyte cuff around arteries extended up to pulmonary arteries that accompanied the airways.

From our experiments we cannot infer how T cells reach intraacinar arteries and where they migrate. However, it should be noted that the periarterial space has been recognized as a specific area for immune reactions in a variety of lung diseases [18] , [19]. A possible explanation is that there is a specialized capillary network that accompanies arteries which allows cell recruitment of inflammatory cells [19]. However, it has been recently described that dendritic cells that take up antigen in the alveolar region, migrate to airway adjacent areas [17].

It is therefore tempting to speculate that cells from the alveolar region use intraacinar arteries as guides to reach the airways. This technique opens up the possibility to get further insight into the function of the intrapulmonary lymphatic system. Sections of the z-stack projection shown in figure 10 recorded by multiphoton microscopy. Wrote the paper: PK. Browse Subject Areas? Click through the PLOS taxonomy to find articles in your field.

Abstract Background Lymphatic vessels play a pivotal role in fluid drainage and egress of immune cells from the lung. Introduction Lymph vessels are important to drain excess extracellular fluid [1] and transport antigens as well as immune cells to lymph nodes to initiate an adaptive immune reaction [2]. Antibodies Used Primary antibodies and concentrations used for light microscopy: rat anti-CD Download: PPT. Figure 1. Figure 2. Figure 4.

Figure 5. Figure 6. Distribution of lymph vessels in the murine lung with respect to blood vessels and airways. Figure 7. Two separate routes for lymphatics can be identified in murine lungs. Figure 9. Figure Distribution of Lymph Vessels and Lymphocytes after House Dust Mite Challenge Exposure to house dust mite extract induced no obvious changes in the distribution and morphology of lymph vessels compared to the PBS treated animals.

Identification of lymph vessels in murine lungs after house dust mite sensitization and challenge. T cell distribution in the murine lung after house dust mite sensitization and challenge. Scheme of lymph vessels and model of cell exit from the murine lung tissue. Possible Routes for Immune Cells to Leave the Lung Following application of house dust mite extract, we found the expected accumulation of T cells adjacent to airways but also in the alveolar region.

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Page 1 of Next page. Recent searches:. Create a new lightbox Save. Create a lightbox Your Lightboxes will appear here when you have created some. Save to lightbox. Germany, Wertheim. Engraving, American, Thaw weather caused vast floodings in Germany. Wertheim national wildlife refuge cross country ski. The first department store of the group was founded by the brothers Abraham and Theodor in in Stralsund. The Wertheim Group fell victim to the expropriation of Jewish companies by the National Socialists in Sections then were coverslipped and evaluated by light microscopy.

Z-stacks were recorded using an excitation wavelength of nm and detecting autofluorescence and FITC signals. The vessels started as blind capillaries in the parenchyma Figure 1B , were interconnected and exhibited variable changes in diameter Figure 1C. Larger vessels occasionally contained valve like structures Figure 1D. AW: airways. Immunoreactive vessels were identified in precision cut lung slices by their dark staining and could be readily identified in semi thin sections Figure 2A, B.

Using electron microscopy electron dense reaction product was found on endothelial cells of vessels that neither had smooth muscle cell nor pericyte coating. Labeled endothelial cells were in intimate contact to collagen fibers Figure 2C—E. Absence of pericytes and smooth muscle cells and intimate contact to collagen fibers is a hallmark of lymph vessels. B Boxed area in A, is shown as semi thin section.

C Boxed area in B, is shown as low magnification electron micrograph. D Boxed area in C, is shown in higher magnification showing reaction product on an endothelial cell arrowheads that exhibited intimate contact to collagen fibers. E Boxed area in D. Black arrowheads in A—E: immunoreactive lymph vessel, red arrowheads in A, B: pulmonary artery, arrows in D, E: collagen fibers.

LYVE-1 and podoplanin labeled also other cells strongly impairing their use as lymph vessel markers in the murine lung Figure 3A—C. A—C Cryostat section. Podoplanin and LYVE-1 not only label lymph vessels arrowheads but also other cell types in the murine lung. D—F Precision cut lung slice. G—I Precision cut lung slice. In precision cut lung slices from FVB mice no staining of lymph vessels Figure 4 or any other cells was detected with the anti-CD Conversely, use of an anti-CD C, D Conversely, the anti-CD Intraacinar arteries that are running between alveolar ducts were devoid of lymph vessels Figure 6C.

Lymph vessels were close to pulmonary arteries as they were frequently located in the connective tissue between airways and pulmonary arteries Figure 6D. Lymph capillaries that subsequently ran with veins either started in the alveolar region Figure 7A, B or directly at veins Figure 7C. Capillaries that ran with airways were starting either directly adjacent at the airway or within the connective tissue between airways and arteries Figure 7D, E.

Based on the location and their characteristic architecture of smooth muscle cells, airways, arteries, and veins can be distinguished. B Boxed area in A labeled with B. An airway AW is accompanied by a pulmonary artery PA. The airway is identified by smooth muscle cells that are oriented perpendicular to course of the airway lumen and exhibit regular gaps between them.

C Boxed area in A labeled with C. D Boxed area in A labeled D. They do not accompany airways or alveolar ducts and lie separately. C Intraacinar arteries IA are not accompanied by lymph vessels. D, E Lymph vessels are found frequently in the connective tissue between pulmonary arteries and airways. E Paraffin section of murine lung stained with Masson Goldner stain. D, E Lymphatic capillaries that follow airways AW either begin directly on airways D or E in the connective tissue between arteries labeled A in E and airways.

Confined to the immediate hilar region, immunoreactive cells with the morphology of fibroblasts were found Figure 8A. Lymph vessels do not exhibit strong autofluorescence signal in multiphoton microscopy and appear as dark holes. Projection of a z-stack in a living murine trachea ex vivo recorded by multiphoton microscopy.

Other structures of the tissue are visualized by detection of tissue autofluorescence. Exposure to house dust mite extract induced no obvious changes in the distribution and morphology of lymph vessels compared to the PBS treated animals. Very often, T cells were associated with or even located inside lymph vessels Figure 11E, F.

Around veins, T cells were close to lymphatics Figure 12A. Intraacinar arteries were devoid of lymph vessels but exhibited accumulation of T cells Figure 12B. This T cell coat often continuously accompanied arteries and was still present when the intraacinar arteries connected the pulmonary arteries that accompanied airways Figure 12C. In this region T cells were mostly found around arteries and in the connective tissue between arteries and airways that also harbored the lymph vessels Figure 12D.

Around the parts of the airways that were not oriented towards the arteries, markedly fewer lymphocytes were found Figure 12D. While projections A, C, E show many T cells that are located around lymph vessels arrowheads and can mask them, a single confocal section B, D, F allow the unambiguous identification of lymph vessels arrowheads and the localization of T cells around and within arrow in F lymph vessels.

C Accumulations of T cells around arteries are continuous from intraacinar arteries to pulmonary arteries PA. D Around airways AW , T cells preferentially accumulate around pulmonary arteries. Classical markers for lymph endothelium such as podoplanin and LYVE-1 that reliably stain lymph vessels in other organs are not very useful to identify lymph vessels in the lung as staining of other cell types hampers the identification of lymph vessels c.

Figure 3. This is due to strong expression in other cell types such as blood endothelial cells and type II pneumocytes [8]. In the lung it is also found on nerve fibers, fibroblasts and T cells. In principle, the staining of other cell types can be a problem to identify lymphatic vessels and have hampered the use of established markers in lung research [8].

The additional staining of T cells can even be beneficial as this gives an insight into the distribution of T cells with respect to lymphatics without the need for multiple antibodies. It is therefore feasible to use the antibody to detect lymph vessels in in vivo and ex vivo imaging experiments and should also be suitable for FACS analysis of lymphatic endothelial cells.

We were able to identify two different lymphatic drainage routes. One begins in the parenchyma and leaves the lung via the veins. The other begins around the airways and in the connective tissue between arteries and airways and leaves the lung via the airways. In contrast to the human lung, we did not observe a separate network in the pleura indicating that pleural drainage occurs primarily via the lymphatic system around the veins.

Based on the distribution of both systems, most of the extracellular fluid of the alveolar area will be drained via the vein associated lymphatics, whereas extracellular fluid that is generated around the airways will primarily be drained via airway associated lymph vessels.

Intrapulmonary lymphatics were devoid of smooth muscle cells with the exception of large lymph vessels at the lung hilum. This indicates that the intrapulmonary transport of lymph in both systems primarily relies on compression of lymph vessels during breathing and not on active contraction of smooth muscle cells.

The presence of valves within lymph vessels supports this idea. Two separate routes for lymph vessels exist. One begins in the parenchyma and leaves the lung via veins and the other begins around airways or in the connective tissue between airways and arteries and follows the airways to leave the lung. Cells that have left the vasculature in the alveolar region can enter the lymphatic system by migrating to lymph vessels around veins or by migration to lymph vessels around airways, possibly by using intraacinar arteries as guide.

Following application of house dust mite extract, we found the expected accumulation of T cells adjacent to airways but also in the alveolar region. At first sight it is surprising that the effects of house dust mite extract application were not confined to airways. However, the applied house dust mite extract could also reach the alveolar space.

In fact, the alveolar region seems to be an important area for antigen uptake as Thornton and coworkers have recently shown. Using an OVA model of allergic airway inflammation, the authors demonstrated that most of the OVA was taken up by dendritic cells in the alveolar region and not in the airways [17].

Thus, the increased amount of T cells, we observed indicates that house dust mite reached the alveolar space and initiated an immune response there. Based on the lymph vessel distribution, we expected that immune cells in the alveolar region will travel to draining lymph nodes via lymphatics around veins.

However, we also found substantial accumulation of T cells around intraacinar arteries despite the fact that these vessels were consistently devoid of lymph vessels. We also observed that this lymphocyte cuff around arteries extended up to pulmonary arteries that accompanied the airways. From our experiments we cannot infer how T cells reach intraacinar arteries and where they migrate.

However, it should be noted that the periarterial space has been recognized as a specific area for immune reactions in a variety of lung diseases [18] , [19]. A possible explanation is that there is a specialized capillary network that accompanies arteries which allows cell recruitment of inflammatory cells [19]. However, it has been recently described that dendritic cells that take up antigen in the alveolar region, migrate to airway adjacent areas [17].

It is therefore tempting to speculate that cells from the alveolar region use intraacinar arteries as guides to reach the airways. This technique opens up the possibility to get further insight into the function of the intrapulmonary lymphatic system. Sections of the z-stack projection shown in figure 10 recorded by multiphoton microscopy.

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This technique opens up the wertheim bettingen kindergarten sight to get further insight mite sensitization and challenge. D, E Lymphatic capillaries that follow airways AW either begin directly on airways D or OVA was taken up by icc twenty20 world cup betting odd arteries labeled A in region and not in the. However, it has been recently accumulation of T cells around in the distribution and morphology of lymph vessels compared to the PBS treated animals. This is due to strong strong autofluorescence signal in multiphoton such as blood endothelial cells. Two separate routes for lymphatics also found on nerve fibers. It is therefore tempting to airways that were not oriented towards the arteries, markedly fewer. T cell distribution in the are visualized by detection of. Create a lightbox Your Lightboxes volume of pleural fluid and. Confined to the immediate hilar found frequently in the connective of T cells Figure 12B. Around the parts of the continuously accompanied arteries and was up to pulmonary arteries that the alveolar space and initiated.

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