REVENGE OF THE TWO DERELICTS

Derolicts1
“Don’t worry, Buck, she’ll get over it.  Remember when I tried to run in the house with that big dead rat in my mouth?  … The  hysterical screaming,     …the door banging in my face?  We had to stay outside forever, until Dad came home and calmed her down.  Unfortunately, he got my rat away from me, but at least Mom stopped acting silly.”

derolicts2 “Anyway, I told you that dropping the dead possum next to the grill when she was cooking the steak was not a good move. You should have listened to me, but now it’s too late.  OK, she could have cooked it, but apparently she doesn’t like possum.   “Now, we’ll be here for a while, I’m afraid.”

“I guess it doesn’t  matter that the damn possum was playin’ possum and then walked away …”

Never a dull moment, folks.  Never a dull moment.  And I wonder why my hair gets a little more gray each day. 😉

HALF A MILLION PAGE VIEWS! AND IT’S GIVEAWAY TIME…

Can you see me doing a happy dance?  Can you at least imagine me doing a very happy dance?  Today the Bewitching Kitchen hit a special milestone, reaching 500,000 page views, and I’m in heaven!  It seems like only yesterday the site registered the first 100 views, and had the first comment coming from someone other than close friends and family.  😉  Half a million page views!  Great feeling, folks!
Fivehundred

To celebrate, I am giving away a very special book called Jerusalem: A Cookbook, written by Yotam Ottolenghi and Sami Tamimi. The story behind these authors is fascinating! You can read a little bit about it here. Jerusalem is more than a cookbook, it describes the complexities of different cultures co-existing, and the way they influence each other in ways that reach way beyond cooking.  The recipes have that exotic feel that make your mind travel to far away places, imagining each smell and taste.  So, if you would like to get this book as a gift,  leave a comment in this post, and I will draw the lucky winner’s name on December 14th, which happens to be Ottolenghi’s birthday!

Thank you so much for carrying me through the milestone of half a million page views!

HAPPY THANKSGIVING!

Growing up, I knew nothing about Thanksgiving.   I had no idea one day I would be living here and calling the US my new home.  Of all American holidays, Thanksgiving is my favorite.  It is a time to just be together with family and friends, being grateful for the good things we have.  On that note, I wish Black Fridays did not exist. Or that they could be placed somewhere far apart in time from this special holiday.   Still, what matters the most is that we all have things to be grateful for, and I hope your day will focus on that.

For our first Thanksgiving in the Little Apple, we will have a joint party, two labs together, celebrating with all the graduate students who stayed here working hard during the break from classes.  It will be a fun evening, I am sure!  Phil will be in charge of the turkey, and I’ll be his sous chef.   All I have to do is mashed potatoes… I got the easy job this  year.

And here is a shot taken a couple of days ago in our street.  One would imagine that with Thanksgiving right at the corner, those guys would be a little more careful!  😉

HAPPY THANKSGIVING!

YOU ARE ALL INVITED!

Cookies and coffee will be available at 3:45pm…

THE PUZZLES OF FERRIC ENTEROBACTIN TRANSPORT THROUGH FepA

When Gram-negative bacteria acquire iron, the metal crosses both the outer membrane (OM) and the inner membrane (IM). But, existing radioisotopic uptake assays only measure iron passage into the cell as the accumulation of the radionuclide in the cytoplasm. We devised a novel methodology that exclusively observes the OM transport reaction of ferric enterobactin (FeEnt) by Escherichia coli FepA. This technique, called postuptake binding, revealed previously unknown aspects of TonB-dependent ferric siderophore transport reactions. The experiments showed, for the first time, that despite the discrepancy in cell envelope concentrations of FepA and TonB (approximately 35:1), all FepA proteins were active and equivalent in FeEnt uptake, with a maximum turnover number of approximately 5/min.   The accumulation of FeEnt in the periplasm required the binding protein and inner membrane permease components of its overall transport system; postuptake binding assays on strains devoid of FepB, FepD, or FepG did not show uptake of FeEnt through the OM. However, fluorescence labeling data implied that FepA was active in the fepB-minus strain, suggesting that FeEnt entered the periplasm but then leaked out. Further experiments confirmed this futile cycle; cells without FepB transported FeEnt across the OM, but it immediately escaped through TolC.  These ferric siderophore acquisition systems are crucial to the pathogenesis of Gram-negative bacteria, and our results show that cathecolate siderophores, which are transported by OM receptors such as FepA, CirA, FecA and Fiu, play a defining role in colonization of the gut by E.coli.

Anyone who falls asleep will hurt my feelings!   😉

THE LAB MOVE AND NEW BEGINNINGS

Almost three months have gone by since we moved our lab from OU to KSU.  I  wish I had been able to compose a post about it sooner, but let’s just say that life has been tremendously busy ever since.   Those who follow my blog might remember that our home move was a saga of epic proportions.  As a consequence, I lost many nights of sleep worrying about the lab move because it’s a lot more complex, and too many things can go wrong.  One of the major differences between a home move and a lab move, is a higher sense of urgency in the latter. When you move your home, you can always leave boxes hanging around, and set up just the basics:  a place to sleep, a place to cook, a place to sit down and collapse at the end of the day.   But, a lab is a full entity of sorts. The whole thing needs to be put in place as quickly as possible because: no lab, no work.  No work, no progress.  We anticipated a lag of a couple of weeks, and that’s more or less what happened.   A big thanks goes to a company called  TLM (Transportation Logistic Management), that moved all our stuff without breaking or losing anything!  Plus, their whole crew was a pleasure to interact with, very nice group of men and women working hard and in great spirits.  If you can believe it, they went through the trouble of wrapping up our large test tubes individually.  And we have more than 500 of these!
One of the most stressful things about a lab move is that quick decisions need to be made on the spot.  For instance, this “little” piece of equipment, called a French press, weighs a ton (well, almost a ton) and obviously once it sits on a spot, it better be its permanent home!   😉

Of all things that were moved, perhaps the most important was our – 70 C freezer.  It holds all our bacterial strains, as well as  very sensitive reagents.  TLM had a nice strategy to deal with it: they packed the whole lab over a couple of days, and closed the truck. Next morning, at 5am they came to get the freezer, loading it just before starting the 312-mile drive. Six hours later, the freezer would be unloaded first and quickly plugged.  If you don’t open the door, the temperature will stay cold enough to preserve its precious contents.  Well, to add a little excitement to the adventure, once the freezer was unloaded, we realized that the plug and the outlet did not match!  Frantic calls were made to electricians on campus, and they came to the rescue. Thanks to their efficiency, our freezer was plugged back when the temperature had risen only to – 44 C.   Big sigh of relief!
Another very tricky piece of equipment to move was our double water-distiller.  All parts are made of glass, connections are delicate and convoluted.  I bet our grad student was wondering if it would ever be back in working condition…;-)

But it had a happy ending!

The huge baby shown below is a cell culture hood.  We use it to work with mammalian cells, much more fragile and prone to contamination than bacteria. This machine has a special laminar flow of air that isolates the environment inside the hood.  Plus, it also holds a germicidal UV lamp to sterilize that environment.  It weighs a lot more than a ton, and it required a special lift to have its legs installed in place.

But once it is all said and done,  setting up a new lab is a great experience!

Drawers are labeled… and stuff is stored inside….

New, clean bottles are labeled, solutions prepared, sterilized and stored, ready for use…

Chemicals are organized, benches get ready for experiments….

And we are finally back, cranking up some data!  Here is a shot of our SLM Fluorometer 8100, a Rolls Royce of a machine,  that needs to be operated in a dark room. It got a nice upgrade before our move, and it will be a major player in our projects in the near and not so near future.

I hope you enjoyed the glimpse on what a lab move involves.  I can tell that we intend to retire here (not anytime soon, mind you!), because another move would just about kill us!  😉  Seriously, though, we are thrilled to join KSU and feeling energized and ready for this next phase in our professional life.

If you want to know a little more about our research, visit our webpage at KSU by clicking here….

ONE YEAR AGO: Honey-Oat Pain de Mie

TWO YEARS AGO: Carrot and Leek Soup

THREE YEARS AGO: Chicken Parmiggiana 101