August 26

Today was our second and last day of trapping and also the day of recaps. At Long Tin we had two cotton mice (Peromyscus gossypinus) and an opossum (Didelphis virginianus) and at Dry Bay we had one deer mouse (Peromyscus maniculatus). None had ticks and all were recaps. At Four Mile we caught an anolis (Anolis carolinensis) and a narrow mouth toad (Gastrophryne carolinensis) and at Dry Bay we caught a woodhouse toad (Bufo woodhousei).
All of this data will be used to figure out the life-cycle and tendencies deer ticks in the south, which we don’t know a lot of. Let’s hope our data shows us some useful trends. And I guess that’s it for the summer, I hope you enjoyed my blog and learned a lot.

August 25

So first night of trapping brought in three recaptured cotton mice (Peromyscus gossypinus), one from each site and none had ticks. The female from Long Tin was pregnant and very agitated that she was trapped. We also caught two lizards. The 5-lined skink (Plestiodon fasciatus) at Dry Bay was caught under one of our metal cover boards. He was the first lizard caught under any of our cover boards. At Four Mile we caught an anolis (Anolis carolinensis). Neither had ticks. The rest of our morning was spent doing a vegetation survey. This survey is being done at all of the other sites and will be used to compared ground cover (any plants below 20 cm). Ground cover is important because ticks are often found here.

August 24

Hey everyone back again! Well like always day one doesn’t really involve many animals. We opened the pitfalls and flagged at Dry Bay and Long Tin in the morning and opened and bated the mammal traps in the afternoon. It’s still too hot for ticks I think because we didn’t get anything from flagging. Maybe this is the reason why Lyme disease has such a low prevalence in the south, it’s even too hot for the ticks to be out. We also decided to shorten our trapping session to two nights instead of three because we have pretty much captured the entire population, we know this because almost every animal we catch is a recap.

August 11

Last day of the week and we had two lizards, both from Dry Bay, one was an anolis (Anolis carolinensis) and the other was a ground skink (Scincella lateralis). Neither had ticks. We had a few frogs, southern leopard (Rana utricularia), pickerel (Rana palustris) and a southern toad (Bufo terrestris). Last but not least the mammals. We did not catch anything at Four Mile but we did get 2 recaptured cotton mice (Peromyscus gossypinus) and 1 deer mouse (Peromyscus maniculatus) at Long Tin and 2 cotton mice and an opossum at Dry Bay. This was the first medium mammal I processed myself. It was really cool to actually handle it. He had 3 dog ticks on his ears, but no deer ticks.

Picture 1: Pickerel Frog

August 10

Today started at Long Tin Site where we had one recaptured cotton mouse and a young male opossum. He was a lot smaller than the female we caught yesterday. He also had no ticks of any kind. At Dry Bay we checked the cover boards and burlap and didn’t find anything. But we did catch two cotton mice however both were recaps. At Four Mile we didn’t catch anything except two narrow mouth toads in the pitfalls.

August 9

This morning started at Long Tin Site where we had 3 mice (1 was a recap and 2 were new) and all were cotton mice (Peromyscus gossypinus). There were no ticks on any of the mice. Then off to Four Mile where we had our huge catch. There was one cotton mouse that was a recap, one raccoon (Procyon lotor) and one opossum (Didephis virginians). The opossum was a young female and had 3 adult engorged dog ticks. The raccoon was a male and had 7 adult engorged dog ticks and 2 larvae. After this we headed over to Dry Bay where we had 2 cotton mice and 1 deer mouse (Peromyscus maniculatus). Deer mouse had a lot of mites on the back edges of its ears. Today was a big trapping day for us, hopefully this trend continues!
Picture 1. Raccoon in a Tomahawk trap

August 8

First Day of this week and like usual we lifted cover boards at Long Tin Tin. Unfortunately we didn’t catch anything, I think it still is too hot too early in the morning for lizards to be under the boards. Although it was only about 95° today, which is way better the 100. We then opened the pitfalls and were off to Home Depot to get some dowels to make shades for the buckets. The dowels were cut into four pieces and zip tied to a piece of burlap. This afternoon these shades are put in the ground directly over the buckets. Hopefully these help cool the buckets off enough so no more frogs or other animals that fall in die of heat and dehydration. After this we open the rest of the traps. The only thing we did different was in the Tomahawk traps we added a sardine to the cat food.

The pictures are of the pitfall tents at Long Tin.

Catheterization Lab and Cardiothoracic Surgery!

Two unbelievable experiences for me this summer were spending two weeks in the Catheterization Lab (Cath Lab) and a week in Cardiothoracic surgery (CT surgery) at LIJ Medical Center. It was significant for me to understand the procedures completed in the Cath lab first, before observing in CT surgery.

I learned that these two departments relate to each other in the fact that they can both treat a patient with a coronary artery blockage. The Interventionalists in the Cath lab can diagnose a lesion within the Left Main Coronary artery or the Right Coronary artery. They can also determine if there is a lesion in the Left Anterior Descending (LAD) artery and the Circumflex, the branches of the Left Main. By injecting a dye through a catheter, with access through either the groin or radial artery, the lesions can be seen on images called angiograms from x-ray machines. Depending on the anatomy, location of the lesion and risk factors, such as diabetes, a patient with a significant blockage of 70% or higher will either be treated in the Cath lab or will be transferred over to CT surgery.

In the Cath lab there are a few possibilities as to how to treat the patient. A balloon pump can be wired into the artery on a catheter and expanded to try and open up the vessel. Also a stent can be deployed in the vessel to open it up an push the clot/plaques against the artery wall. There is always the risk of in-stent stenosis when this is done, so the patient is normally medically treated with anti-coagulants to prevent clots from forming. Newer stents called drug-eluting stents decrease this risk greatly due to the coat of chemothraputic-like medicine on the stent. This prevents the overgrowth of the vessel wall covering the stent.

In cases when the lesion is in the Left Main Coronary artery, or the patient is at high risk for a stent, or the anatomy makes it difficult for a stent to be placed, they are transferred to CT surgery. This is when a Coronary Artery Bypass Grafting (CABG) is completed. Internal Mammary arteries, or Internal Thoracic arteries, located behind the rib-cage are often used to bypass the LAD artery which supplies blood to the left ventricle of the heart. Vein grafts from the legs and arteries from the arm can be taken to bypass other vessels in the heart to supply the ischemic tissue with blood.

I was fortunate enough to be able to "scrub-in" on these procedures and stand right behind the surgeons and intervenionalists. Seeing a fully exposed heart beat in a patients chest was one of the most unforgettable experiences I have ever had.

July 25

Today was our last morning to check traps and caught a few small mammals. We had 2 cotton mice (Peromyscus gossypinus) at Four Mile and Long Tin and one cotton mouse and one cotton rat (Sigmodon hispidus) at Dry Bay. None of them had any ticks and only one was a new capture.
The exciting news is that our motion sensory cameras have been picking up a lot of animals. We seen nine-banded armadillos (Dasypus novemcintus), wild pigs, adults and babies (Sus scrofa), a bobcat (Lynx rufus), squirells, raccoons in packs (Procyon lotor), oppossums (Didelphis virginianus), and deer of all ages (Odocoilesu virginianus).

Picture 1: A wild pig
Picture 2: Four Raccoons
Picture 3: Bobcat

July 24

Today was an eventful day. We had mice at all of our sites this morning. Long Tin had three! Yes three mice and one was a recap. At Dry Bay we three more mice and a cotton rat (Sigmodon hispidus ). At Four Mile we had one mouse, which was also a recap. The mice did not have any ticks on them but the rat had a lot of mites at the base of its ear, which shows that the ticks are out there. We had no lizards or medium mammals today but that’s ok we still have one more trapping night. We also flagged at Four Mile today and I got our first nymph of the trapping session, and it appears to be Iscap. Tomorrow is supposed to be slightly cooler (still in the 90’s). Let’s hope we continue to get more animals!!

July 23

One of our cotton mice.
We caught our first mammal at Long Tin site this morning. It was a cute cotton mouse (Peromyscus gossypinus), and like most of the other mice we have found he did not have any ticks. At Four mile we had a male scelop (Sceloporus undulatus ). They are so beautiful. He was a dark grey/charcoal color and had brilliant blue scales on his stomach. He had a lot of mites by his armpits and ears but no nymphs or adults. This is a good sign because it means he was walking around where ticks are (it just may be too hot for the other life stages to be out). We also had a recap of a mouse at Dry Bay but he got free before we could finish processing him.
This afternoon we flagged at Dry Bay and found only one adult dog tick (not what we want). In one of the pitfalls at Long Tin we caught an adult female broadheaded skink (Scincella laticeps). She was huge, over 100 mm long. Like the scelop she had a lot of mites but nothing else. Hopefully we get some more animals tomorrow but if it’s the heat that’s keeping them away we may not get so lucky, forecast says its going to 97° tomorrow.

July 22

This is a female fence lizard.

Hey guys first day back in South Carolina and it started at Long Tin tin. We lifted all of the metal coverboards and found a juvenile 5-lined skink (Scincella fasciatus), which was so tiny it was the length of my pinky finger. We also caught a female fence lizard (Sceloporus undulatus), she was chunky and has a wicked bite. Kaety had a her hand on top of her but couldn’t get a grab so I picked her up and she turn and bit my finger. She didn’t break the skin but it’s now bruised. Neither had ticks but the scelop had a re-grown tail.
After this we opened the pitfalls at all three sites and flagged at Long Tin site. We only caught one larval tick. More research needs to be on when is the best time to flag. Later today we went back and opened and bated the mammal traps. Fingers crossed we catch a lot of animals tomorrow!

July 7 and 8

I think the lack of rain and excessive heat are keeping the animals from coming out. During our last two trapping sessions of the week we only caught 3 mice each day at Dry Bay and all were recaps from previous weeks. We caught no medium mammals and only a few lizards (Anolis sagrei and Scincella lateralis), which were caught while they were foraging on the ground. This made our mornings really short and hopefully while we are away there will be more rain.

July 6

The team working hard setting up a pitfall at Long Tin.

Day two of digging and it felt like the longest day ever. First we had to check the traps at Four Mile and Dry Bay. At Four Mile we caught a shrew in one of the pitfalls and like all the others we found tick larvae between the toes and on the feet. We also caught our first lizard under the burlap that’s tied around a tree. It was a beautiful Sceloporus undulatus, aka eastern fence lizard. She did not have any ticks but she was only days away from laying eggs. At Dry Bay we caught another lizard under the burlap this time it was a male five-lined skink (Plestiodon fasciatus) with no ticks. We also had two mice in the Sherman traps, both cotton mice (Peromyscus gossypinus) and both were recaps with no mice.
After checking the other two sites we went to Long Tin Site and checked the traps that were out (the one pitfall and the Sherman and Tomahawk traps) but there wasn’t anything. While the others started digging I did my shift of flagging for ticks but after two transects I didn’t get any. This could be because they are not out or because it is too hot and they are hidden under better cover. After this I joined in on the digging of the last two pitfall arrays. These were unlike the other sites in that we had to dig in clay, not sand or soil. So basically these were significantly harder to dig. By the time we were done we had a few hours to relax until we went back out to open the traps.
We didn’t find anything in the pitfalls at Dry Bay or Long Tin Site but we did find another shrew at Four Mile in one of the pitfalls. It also had tick larvae between its toes. This is really odd that we keep finding them between the toes because the shrew should easily be able to remove the ticks while it’s cleaning itself. Who knows maybe the ticks are not a nuisance and the shrew doesn’t even realize they are there.

July 5

This is a male eastern fence lizard (notice the blue stomach, females only have color under their neck).

Our last site was finally approved!! This site is called Long Tin, it’s right next the original Long Tin site (which is just a bunch of long pieces of tin so we couldn’t use this area because there wasn’t enough room). Today and tomorrow are long days. We checked the tins at the original Long Tin, which is now called Long Tin Tin and found a fence lizard (Sceloporus undulates). Then we went off to Long Tin Site and carried all the supplies in (40 coverboards, 49 Sherman traps, 4 Tomahawk traps, 20 buckets, stakes, shovels, and on and on, basically a lot of stuff). While the others were plotting out the arrays I set up the Sherman traps and coverboards and organized the other materials so it would be easier to setup once everything was mapped out. After a short break we dug the first set of pitfalls (5 buckets and 4 pieces of tin sheets). This took about an hour to do and we intended to dig more but by the time we finished everyone decided the rest could be finished tomorrow, no need to run ourselves down on the first day. We took a few hours break and headed back out later when the temp went down a little.
We opened all the traps at Four Mile, Dry Bay and Long Tin Site. All the mammal traps are good to go at Long Tin so only the pitfalls and weather station need to be done tomorrow. It may not sound like a lot but its going to be a long day because we need to check all the traps at the other two sites, check coverboards, process all the animals we catch and finish up Long Tin Site.
The two exciting things that happened today was we saw a bobcat on one of our motion cameras at Four Mile and then while driving to Four Mile we passed two adult feral pigs with about 7 babies pigs running through the woods.

June 20

Today was our last day of trapping and in my opinion we caught our coolest animal yet. At Dry Bay we trapped a female opossum (Didelphis sp.) with her newborn babies in her pouch. We sedated her as the protocol says and her babies were unharmed they were still moving and feeding. You could actually see the milk in their stomachs because their skin was still slightly see through and hairless. She did not have any Iscap on her. Once we were done we moved her off the array and kept a close eye her to make sure her and her babies were ok. We also caught our tailless rat, Cotton rat (Sigmodon hispiidus) again and for the first time I held a rat. We had her in a bag so we quickly checked her for ticks and then I took her out to release her, but I needed to work on my hold so we went through the basic procedure of checking her for ticks while I held her. We didn’t have to do anything else because we had all her data from two days before, it would just be unnecessary to put her under excess stress for redundant data.
At Four Mile we caught our Norway rat (Rattus norvegicus) again and like our Cotton rat we checked it for ticks and let him go. We also caught two deer mice (Peromyscus maniculatus). Like all of our other mammals we haven’t found any ticks on them so the debate still continues why we haven’t any ticks. Until next time…

June 19

Today was the day of recaptures, at Dry Bay we caught three mice, two of which were recaps. The third mouse (one of the recaps) has a developed an infection or tumor of some sort by its hind leg and was extremely high strung. We didn’t even try to get anymore data on him because we were afraid he may die from the stress. At Four Mile the only mammal we got was the rat we caught yesterday. Other than this we had some leopard frogs and southern toads and a few other amphibians but that was it. It was a really quick morning. It still is really odd that we have not found any ticks on the mammals. Who knows maybe the larvae are on them and we can’t see them or maybe the adults are out yet.

June 17

Checking the Anolis for ticks.

We got a lot more done today than we did yesterday. Today start at Dry Bay where we checked all of the cover boards, we saw a blue tailed lizard (which could be Plestiodon fasciatus, Plestiodon laticeps, or Plestiodon inexpectatus, all three of these lizards are the same coloring as juveniles) but we weren’t fast enough to catch it. We then opened all the pitfall traps. We did the same thing at Four Mile. This didn’t take too long and we had several hours until we could meet with Bess to determine our GPS coordinates at Long Tin, so we decided to search for a drift fence at a set aside called Sand Hill. This drift fence has been up for decades and the people who set it up are a long time gone and since it’s not used anymore not many people know where to find it. But this site has a high population of Racerunners (Aspidoscelis sexlineata), which we haven’t found yet, at our sites. After almost an hour of wandering we found the fence. It was in a really weird set up but we thinking about asking permission to dig buckets at the ends and turn them into pitfalls. I don’t know if it will be approved but it’s worth the shot to find out. After this we had some down time until our meeting.
Turns out our plot we found yesterday was really easy to find the coordinates for so we were done in about an hour. We now have a 130m by 130 m square that is now being looked over for approval. Fingers crossed this will finally work out so we can get all three of these sites up and running together.
Back to Dry Bay and Four Mile to check the pitfall traps (we found an Anolis at Dry Bay in a pitfall and it had no ticks, supporting the belief that Anolis do not carry ticks) and open the mammal traps. At Four Mile we found a juvenile shorttail shrew (Blarina brevicauda) in a pitfall. It was adorable, it only weighed 6 grams and was so soft. This tiny shrew had 10 tick larvae attached to him. All of them were either on the hind feet, between the toes or on the tail. We don’t know what species the tick was but DNA testing will be able to answer that question.

June 18

This is a picture of a short tailed shrew.

Last night was a very successful trapping night. At Dry Bay we caught a mouse, a rat and raccoon. The rat was actually the rat from last trip that lost the skin on its tail. Turns out the tail fell off completely and was healing very well. Just like last time she had no ticks. The raccoon was really cool to see, he played dead for the most part until I put the squish plate into the cage. The plate is just a wood board with a handle that we push into the trap so the animal is forced to the back where we can safely give him a shot of telazol (a mild sedative). He had no ticks or even fleas, he was a very clean raccoon. Unfortunately our mouse died while we were trying to get an ear punch. This is a new part of the procedure. We use a special tool to cut a tiny hole out of its ear. The tissue is used to determine if it has Lyme Disease. None of us really like the idea of cutting part of the ear, especially after he died. Now we are taking tiny snips from the edge of the ear instead which is much faster and less stressful.
After all this we headed to Four Mile and found two shrews in one of the pitfalls. Like the shrew from yesterday they both had a lot of ticks (especially between the toes). A Sherman trap caught a Norway rat (Rattus norvegicus). Once we let him go he ran straight up a vine and disappeared into the canopy. That was the first time I’ve ever seen a rat do that. We also caught an Anolis foraging and surprisingly it had an engorged larvae tick and two mites on it. This is the first one we found to have ticks. Other than that we caught a few frogs and toads like we always do.

Cardiology Blog #3: Cardiothoracic Surgery (CT)

This week I was in the Cardiothoracic Surgery (CT Surgery) unit and had the opportunity to watch multiple open-heart procedures. One interesting thing I have learned at the hospital is that all the different areas of Cardiology connect because in order to evaluate patients the doctors must unite to treat patients’ problems to the best of their ability. For example, the CT surgeons use angiograms to determine how severe the patients condition is, and where they will need to use a CABG (Coronary Artery Bypass Grafts ) to supply the heart with blood flow. Echocardiograms are also used in order for the surgeons to determine how the heart is pumping the blood and whether the valves are functioning properly.
I started off the week by attending a morning conference and learned about the anatomy of the thoracic cavity. Then I followed the cardiothoracic surgeons on their rounds for patients that were post-operative. The doctors examined the patients to evaluate the healing of their surgery incision and checked the overall appearance and lifestyle of the patients two to three weeks post-operation. During the patient evaluations I was able to meet patients that had received valve replacements, Coronary Artery Bypass Grafts (CABG) and people that had Pulmonary Emboli and blood clots removed.
The first surgery I was able to observe was an Aortic Valve Replacement (AVR). This surgery was five hours long and was incredible. The surgeon first ‘cracked open’ the patient’s sternum and helped control bleeding in order to enter the heart with minimal disturbances. A hypothermia protocol and bypass was done in order to preserve brain function and provide the rest of the body with blood flow during the procedure. It was amazing to me that the mechanical valve looked very similar to the congenital valve that everyone is born with. It was also interesting how quickly patients recover after such an intense surgery.
The next two days I was able to watch a double CABG, and a quadruple CABG. These procedures were unbelievable because I was able to watch the heart beat inside of the patient’s chest, and overlooked the surgery standing on a stool in the operating room. During this procedure they remove a vein from the leg and graft it from the aorta to various coronary arteries to supply areas of the heart that aren’t receiving enough blood supply due to severe coronary artery disease.
Seeing the open-heart surgeries in CT surgery was an incredible experience that I will never forget. I have learned so much and really been able to put all the knowledge I’ve gained the past 6 weeks into practical use.

Cardiology Fellowship...Stress Lab & Echocardiograms

The next two weeks of my internship were really fun and informative. I shadowed various nurses, techs, and fellows throughout the Echocardiogram and Stress Labs. Some of the interesting things I observed in the echo rotation included a Trans-esophageal Echocardiogram (TEE), Echo Stress tests, and Trans-thoracic Echocardiograms (TTE). For the echo stress test, the patient runs on the treadmill at increasing levels of difficulty until they reach their target or maximum heart rate, and meanwhile are hooked up to ECG leads; once the patient reaches the target heart rate they are rushed to a table where the tech and doctor performs a Trans-thoracic echocardiogram on their heart. The echocardiogram (TTE) takes pictures of the heart from various angles to determine abnormalities of the heart post exercise and compares the function to the rest pictures taken pre-exercise.
The Trans-esophageal Echocardiogram (TEE) test was also intriguing because the doctor would place a long probe down the patient’s esophagus and take pictures of various parts of the heart from different angles. The TEE monitors valve function, and can show if there is an irritation in the muscle tissue. It gives a 3D picture of the inside of the heart and valves. Trans-esophageal Echocardiograms are used before surgeries to check for blood clots and Pulmonary Emboli’s ; if these things existed the procedure may not be able to be performed because it could put the patient at high risk for a complication.
In the stress lab I learned about Exercise Stress tests and the Medicated Stress Tests, which are the two different kinds of stress tests used. The stress tests are very helpful tests used on patients that have previously had a stent implanted in their coronary arteries; they are useful because if the stress test is positive then this could be an indication that the artery is possibly being blocked or that another artery needs a stent. Stress tests can also be used on patients that are experiencing chest pain after surgery. For the exercise stress test, the patient runs on the treadmill at increasing levels of difficulty until they reach their target or maximum heart rate, which is 220 minus the patient’s age. Then they are given a nuclear substance that is pumped through the heart in order for nuclear pictures to be taken of the heart at both rest and during stress or exercise. The medicated stress test is done on patients who cannot walk on the treadmill long enough for their heart rate to reach the target value. For these patients, adenosine or other drugs are injected into the patient to imitate the effect of exercise or stress on the heart, and a nuclear material is also given so that nuclear images can be taken of the heart. The pictures are taken by a nuclear imaging machine that takes images for three to five minutes of all areas of the heart to check that it is beating in the proper fashion; the stress tests also show if there are areas of the heart where the muscles aren’t contracting properly. If the muscles aren’t contracting properly then this could be an indication that a heart attack may have occurred, causing ischemic damage to that area of the heart.

Jennalee Trombley

*The Final Steps...*

The week of July 26, I continued to assist in the Forest Restoration Project with Dr. Aronson. This project was very successful and it helped me significantly with my plant identification. I also began to work on the analysis of my bulbil dispersal experiment. I worked closely with Nick, one of the students from South Side high school, on the preliminary data and made sure that he understood the process of analyzing data in a research project.

The week of August 3, I did some more field work and began to work on the methods section of my paper. I worked with Dr. Sanford on understanding how to use statistical programs and analyzing the data for my bulbil dispersal experiment. We performed a regression analysis on bulbils that had buoyancies below 0 and the ones that had buoyancies above 0. The results showed that there was no relation between the bulbil’s buoyancy and time to sink. However, my research did show that bulbils can travel 0.002 m/sec.

I also performed some more statistical tests on my diversity sampling data with the help of Dr. Aronson. There was a significant difference in species richness between invaded and non-invaded plots at Muttontown Preserve. The average invaded mean was 5.6 and the average non-invaded mean was 7.3. There was no significant difference in diversity between invaded and non-invaded plots at Muttontown Preserve. The invaded mean was 1.31 and the non-invaded mean was 1.04. At Prospect Park, there was no significant difference in species richness between invaded and non-invaded plots. The invaded mean was 1.8 and the non-invaded mean was 1.6. There was not a significant difference between diversity in invaded and non-invaded plots at Prospect Park as well. The invaded mean was 0.46 and the average non-invaded mean was 0.26. The number of plots was limited due to the small size of the invasion as well as environmental conditions that consisted of Norway Maples casting deep shade. Therefore, the skewed results reflect the conditions of the site.

The Greenhouse portion of my experiment was not as successful as I intended it to be. I have not observed any growth, except for a few weeds, and it has been about a month. Therefore, Dr. Aronson and I have decided to remove the flats from the high school and bring them back to Hofstra. We will keep them in a growing chamber, where the lighting conditions would better support the plants. We believe that the seeds have not been stratified long enough. As a result, we will perform a new seed addition and start the process over right away.

Week 6

WT1

Dr. Clendening and I had decided late last week that we would start a new project on Monday. We wanted to investigate the protein WT1. WT1 seemed like a good candidate for two reasons. First of all, we knew that WT1 worked synergistically with SF1 to promote the downstream expression of SOX9 and AMH, eventually leading to testes differentiation. Secondly, we knew that WT1 had two different splice sites, resulting in four different isoforms. These isoforms, we predicted, might be regulated according to temperature.

Dr. Clendening and I spent the beginning of the week isolating total RNA samples from both the adrenal/kidney and gonadal tissues. I was not the easiest task, because we had to make sure that we were working in a clean environment. Everything we used had to be treated with RNAse Zap, a chemical that destroys ribonucleases. Luckily, the samples that we extracted were good. An agarose gel gave us the results we were looking for.

DAX1 and SF1 (return to the agarose gel)

Now I know that I had posted a comment in an earlier blog stating that Dr. Clendening and I wouldn't be using agarose gels, but...things weren't working well with the IEF gel. After doing some research, we found a protocol for native blotting of proteins using agarose. However, this protocol called for a different type of agarose, a high resolution blend (HRB). We had a bottle of the HRB in the lab, so we decided to go ahead and try it. I prepared samples, and ran a gel during the remainder of the week. The western provided the following results.

Week 5

Western

Monday morning I came into the lab and began developing last weeks membranes for western analysis. I used DAX1 as my primary antibody, since the gel contained a lane of isolated DAX1. Unfortunately, the IEF gel markers were the only bands to appear. At this point, neither Dr. Clendening or I had any idea as to why the transfers were unsuccessful.
Phone Call

Dr. Clendening and I decided it would be best to call up the company that provided us with our IEF gel and try to troubleshoot. The woman that I spoke with recommended using the acetic acid transfer, but letting it go overnight. She said that she would send us a gel right away.

IEF Again

When the IEF gel came in, we decided to run last week's experiment again. Samples of the nuclear fraction, nuclear fraction with formaldehyde/glycine treatment, and isolated DAX1 were loaded into the gel. Experimental conditions were kept constant, except we ran the transfers overnight. I developed the westerns the following day, but we got the same result as before. DAX1 was not detected in any of the samples.

*Greenhouse Greatness!*

Last week, I began the greenhouse portion of my research project. I was extremely happy to start this experiment. On Monday, I made sure that my methods were precise and exact. Also, I prepared all of the materials that we needed to start the experiment. I conducted the experiment on Tuesday at Southside High school in Rockville Centre, Long Island. There were about 4 students present and they were really involved. As a result everything went smoothly. We first mixed soil with activated carbon and placed it into the appropriate treatments of the five species of plants. We did the same for non-invaded soil to avoid potential confounding effects of soil disturbance and mixing. We then added 6 seeds of five different treatments into 40 pots each, totaling 200 pots. We then placed all of the pots randomly in the greenhouse. The plants were watered to saturation with 50 ml of tap water. They will be watered every other day with the same amount and I will check on them periodically throughout the upcoming weeks. When they germinate, I will have all of the students return and assist me in measuring the germination rates.


I also began the revisions to the methods of the bulbil dispersal experiment to make sure that there was no room for error. I created all new data sheets, used all new bulbils and set up stations to ensure that everything would run smoothly. I had two assistants that helped me and we began the project last week. This week, I completed the bulbil dispersal experiment and began to enter all of the new data. I will begin to analyze all of the data next week with the help of Dr. Sanford and Dr. Aronson.



On Tuesday, Dr. Aronson and I went to Alley Pond in New York City to help with a Forest Restoration project . Although, I am always looking forward to being out in the field, we were all eaten alive by malicious mosquitoes, literally! On Thursday, we went to a different site at Roy Wilkins Park in Queens to assist on the same project. This site was way more pleasant and we got a lot accomplished. We have some upcoming sites that I am looking forward to. This is such a great partnership! On Friday, Dr. Aronson and I went over to the high school to check on my plants. Everything looked pretty good and no plants have germinated as yet, except for two weeds. I will return next week to check on them again.

Cardiology Fellowship... EP lab and Consult SVC

Throughout the summer I am interning at LIJ hospital in the Cardiology Department for 10 weeks. I am lucky enough to have received the opportunity to observe a different area of Cardiology each week and shadow various Doctors, Fellows, and Nurse Practitioners throughout the day to learn about what the typical day is like for the Cardiology team. During my first two weeks of the internship I shadowed in the ElectroPhysiology Lab (EP lab) and the Cardiology Consult Service.
In the beginning of each week the Cardiology team meets for a morning conference to discuss how the weekend went, and makes plans for the week to come. On the first day of my internship I started out meeting everyone at the morning conference then jumped right into a bunny suit, which is a sterilized outfit that allows me to observe procedures without possibly harming the patient, and watched my first generator change in the EP lab. It really fascinated me to see how small such an important device can be and that the procedure is done quite quickly. Throughout the rest of the week I observed a few pacemaker implantations and a cardio version as well. For those of you that don’t know what a cardio version is, it occurs when a patient has an irregular heart rate, and this procedure is done to shock the heart in an attempt to reset their heart rate to a safe rhythm. I also followed a Nurse Practitioner to observe the post procedure and learned how defibrillators and pacemakers are controlled.
The next service I followed was the Cardiology Consult service. In the cardiology consult service I learned about the many reasons people are placed under Cardiologists care, and the different tests done in order to determine the best source of treatment for the patient. One interesting procedure I learned about was hypothermia treatment for patients that have a cardiac episode; the hypothermia treatment is performed by cooling the body after stabilizing the patient’s heart in an attempt to preserve brain function, and has been shown to be quite effective. Throughout this week I also was able to listen to a patient’s heart that had a heart murmur, and feel their unique pulse.
These first couple weeks I have had a great time and have learned so much. I can’t wait to experience the next 8 weeks of the internship!

Jennalee Trombley

Week 4

Test Run

The beginning of the week was devoted to running an actual version of my experiment. I incubated samples of DAX1 and SF1 at male producing temperature (MPT), 26 degrees celsius, and female producing temperature (FPT), 31 degrees celsius. Half of the samples received a formaldehyde/glycine treatment to theoretically "fix" the proteins and all of the samples were loaded into a 4-20% polyacrylamide gel. The gel ran for an hour at 100 volts. Afterwards, the proteins were transferred to nitrocellulose membranes through electroblotting (I electroblotted twice to get two, presumably, identical membranes).

The next day I developed western blots, but the results were not satisfactory. Half of the samples did not show up (left side of photos). The lanes that did appear reacted with both DAX1 and SF1 antibodies, but these lanes should have reacted with only one or the other. It seemed as though there might have been some non-specific binding of the antibodies, so Dr. Clendening suggested that I lower the concentration of antibody in the future. We had no explanation for the lanes that did not show up. Perhaps the proteins got degraded?

I spent the middle of the week researching the protein Wilm's Tumor Suppressor (WT1). I had run out of gels, so I had to find something to do in the meantime. It turns out that WT1 plays a role in TSD. It has many different isoforms, which may or may not be regulated by temperature. WT1 would serve as a prime candidate for future experimental applications.

IEF Test Run

By the end of the week the IEF gels had arrived and I was able to return to the DAX1/SF1 experiment. I ran an IEF gel on Friday with samples of nuclear fraction, nuclear fraction with formaldehyde/glycine treatment, and DAX1 isolated. I tried three different methods for transferring: electroblot with acetic acid buffer, electroblot with tris-glycine buffer, and a pressure blot with tris-HCl buffer. The membranes were left to dry over the weekend.

June 16

Hey everyone were back in South Carolina again. We decided to come down two days earlier than we had planned because we got word that our third site was almost approved, all we had to do was get the GPS coordinates and hand them in, then we could start digging. We got up at 5 to get an early start to our day. We went to the Long Tin site and looked under all of the cover boards. Usually this will take us about 2 hours today it was maybe 30 minutes. We did not see one single lizard and we don’t know why. Then we had to meet up with Bess who is with SREL so we could get our GPS coordinates. The plan was to go to the back of Long Tin (which is a piece of land set aside for research). When we got to the back we found out that the Forestry Service had cut down a lot of trees, everywhere. There was not one acre (which is all we need) that was not close to the disturbance by the Forestry service. After walking around we went to the lab to find out what was going on, we were also thinking that we were not going to be able to work there at all. After a few hours we found out that Forestry was done working in that area and said it was all ours. Apparently they had a special permit to go into the area to cut trees down. Lucky for us they were done.
Later that day when the sun started to go down a bit we went back out to try and pick out exactly where we want our plot. We decided on a fairly dense patch of wood that was barely disrupted by Forestry. Tomorrow we will get the coordinates and hopefully get started.

*Revelations in my Research!*

Last week, I continued to work on the project looking at bulbil dispersal. I completed measuring all of the volumes for the bulbils that I am studying. I confirmed my measurements with an assistant in my lab that also took measurements, to make sure that I had repeatable measurements. However, I had an obstacle when it came to the dimensions that I was measuring the bulbils. Dr. Aronson and I were not sure that I was measuring correctly so we researched the process of calculating the volume of an ellipsoid and we realized that I wasn’t quite doing it right. I understood that this was all a part of the research process, even though I was about half way through and it was frustrating to start over.

Ultimately, I calculated the volume, density and buoyancy of these bulbils. I met with Dr. Sanford to discuss what was next in terms of the project and I got started on it right away. I am testing whether my prediction of buoyancy is an accurate measure of the metric in water. I want to simulate how bulbils will sink in a stream, so I used a 10 gallon tank and filled it half way with water. I made sure that there were no currents in the water and got a precise measure of the depth of water. I was very careful to avoid the meniscus effect. I had an assistant with me to calculate the time it took to sink from the surface of water to the bottom of the tank. I was very careful not to touch the water too much by using a long pair of forceps so that the level of water remained the same.

I predicted that the bulbils would have a negative buoyancy which means that they would sink. Most of them did but a lot of them did not. I left them over night to see if anything would happen but unfortunately, I returned to the same results the following morning. The most logical reasoning for the bulbils not sinking was the fact that they began to lose water in the zip lock bags that they were enclosed in, in the freezer. I noticed that when I took the weights of the bulbils after I retrieved them from the water, they actually lost about half of their original mass. Therefore, Dr. Aronson and I decided that we would start the whole experiment over with fresh bulbils that we had stored. This time we agreed that we would do all of the measurements and timing of sinking at one time to prevent the bulbils from drying out between measurements. This is also all a part of the research process. Sometimes it means perfecting an experiment as many times as you need to, in order to ensure that it is successful. I am grateful for all of these experiences because it is helping me to become a better research student.

Myself along with Dr. Aronson, Mr. Weiss, and our Locust Valley High school student all went out to Prospect Park last week Friday to add another site to my diversity sampling of invaded and non-invaded areas. However, there was not much to sample from at this site due to the small size of the invasion and environmental conditions (Norway Maples casting deep shade).

*Getting Down at Muttontown!*

On the week of June 28th, Dr. Aronson and I went out to Muttontown Preserve (http://www.nassaucountyny.gov/agencies/Parks/wheretogo/preserves/north_shore_preserve/Muttontown_Pres.html) to check on our flagged non-invaded areas. We identified some species and flagged the invaded areas of where we would set up our plots. The forest looked absolutely beautiful and the flagged area that we are going to conduct the experiment was perfect.

We went out again later that week to complete our diversity sampling. We teamed up with the research teacher, Mr. Weiss and one student from Southside High school, Nick, as well as another high school student from Locust Valley High school and one of Dr. Aronson’s field assistants from Hofstra. We completed ten plots from the invaded area and ten plots from the non-invaded area. There was a lot of diversity among the forest especially in the non-invaded areas. It really broadened my knowledge to many different species of natives (such as violet, jewel weed, red maple, spice bush, Canada mayflower, may apple, etc.) and non-natives (such as English ivy, garlic mustard, Japanese honeysuckle, etc.) . There was also a lot of poison ivy but we tried our best to avoid it!

English ivy
Jewel weed

After the sampling was completed, we collected soil from the non-invaded area as well as the invaded area. The soil will be used in the greenhouse experiment along with our activated carbon to test for allelopathic chemicals being left behind by this invasive species. I also decided on exactly which company I would order the activated carbon from by using scientific literature, searching on the web and contacting companies. We should have it in time for the greenhouse experiment.

We could not identify some of the species in the plots so we carried back samples from the field site. We could only identify one out of the six species that we were not sure about. We identified it by looking at identification books. This was the only species with flowers. However, we pressed the other species in the lab to save for later identification and we plan to return to the field site once these unknowns are flowering. It is very difficult to identify herbaceous plants that have not flowered yet.