Implantation - Science topic

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[Skin sensitizers are substances that elicit an allergic response, such as allergic contact dermatitis, following contact with the skin. Allergic contact dermatitis causes itching, swelling, and redness of the skin in susceptible individuals (Murphy et al. 2012). Skin sensitization testing identifies the potential for a substance to cause allergic contact dermatitis.]

[As part of an effort to increase confidence in alternative methods and improve the relevance of test outcomes to human health, ICCVAM has developed a strategic roadmap for establishing new approaches for evaluating the safety of chemicals and medical products (NIEHS 2018). The roadmap implementation plan addresses the development and evaluation of alternative approaches for acute systemic toxicity testing, skin and eye irritation testing, and skin sensitization testing. A key element in development of the strategic roadmap and subsequent progress towards incorporation of replacements to animal tests is an understanding of the data needed and how they are used by government regulatory and research agencies.]

[Conclusion

The future of non-animal methods for skin sensitization testing seems promising; however, the next steps toward implementation and acceptance by research and regulatory authorities will require communication and interaction to establish scientific confidence in their ability to protect human health. ICCVAM is leading this effort in the United States by coordinating a strategic roadmap for establishing new approaches to evaluate the safety of chemicals and medical products; the implementation plan for the roadmap specifically addresses the development, evaluation, and acceptance of alternative approaches for skin sensitization testing (NIEHS 2018). As alternative methods are progressively considered and accepted at the national level, global harmonization will be necessary to achieve global implementation. Partnerships with international regulatory authorities and supporting organizations are critical to the widespread evaluation and implementation of new approaches.]

Dear,

Pinopods have features that enable the embryo to attach and feed during the implantation window period. It is a marker for the realization of implantation, but also an indispensable structure. In implantation, the pinopod structure, as the apical surface of the endometrium, plays an important role in the adhesion of the 6-7 days old blastocyst to this region. Its paracrine and morphological structure attracts the embryo. And besides it looks very good under the electron microscope, it can even be selected under the light microscope.

@Taylor Pini

No I have not tried them... thanks for the suggestion

Hello Tom,

a direct ionization is possible, if the electric field E is able to exceed the ionization energy E_I of a atom over the diameter d of an atom, e.g. e*E*d> E_I (bold - vector). In a metal you have bands and E_I must be substituted by the work function W (field emission by an electric field)..

More probably, you ionize by impact ionisation, if a charged carrier is accelerated by an electric field. Here the acceleration length depends on the mean free path l (distance between two collisions of the carrier with phonons, impurities or other excitations). In this case the relation e*E*l > W is important.

With Regard

R. Mitdank

Yes, According to Anderson et al, 67NR expresses ERalfa and responds to Tamoxifen.

Successful bone-anchored hearing aid implantation in a patient with osteogenesis imperfecta

Article in The Journal of Laryngology & Otology 129(11):1-4 · September 2015

DOI: 10.1017/S0022215115002510

It is hard to tell in which phases the Malaysian BIM implementation is. The implementation phases of BIM are normally based on the types of construction project, size of the project, client requirement, developer commitment, as well as the types of developer.

Big developer, such as Sunway, is very keen on BIM implementation. In fact, this company is the pioneer of BIM in the country. Sometimes, BIM is adopted in the design phases or the construction, mainly due to the client's requirement. Sometimes, it is used in operation phase for project that involving sophisticated design such as hospital.

So, it is unable to tell in what phases that BIM is adopted in general. The following are some articles published in the Malaysian Construction Research Journal, featuring on the background of BIM adoption in Malaysia. Hope they are of useful to your study. Thanks.

We use 5 screws, and put them all in at a slight angle (about 15 degrees) so that they are all angled towards the center of the brain so that vertical force won't take the head cap off.

Dear colleague Shantanu Saha,

Have a nice time and good day.

To decide if the semiconductor is still DBG or IBG Tauc plot should be plotted.

Please see the two attached files. the most fitted relation through the attached file represent the transition;direct or indirect.

If you want to calculate or process the optical calculations follow the next procedure:

How to process the calculation to get Tauc plot for direct and indirect transition and finally the optical relations please follow the next:

In case if you measure the mentioned parameters , T , R and/or A herein fined all what you need about optical properties from A to Z. in details including the direct and indirect transition  also  the references are included.

Due to the format of the Equations, it is included in the attached file so check the Eq. No. the see it in the attached file.

First you measure transmittance, T reflectance, R and/or absrobance using any spectrometer. 

If your spectrometer is not calibrated so you need to  calibrate your measured parameters otherwise go to step 2. Herein fined how to calculate the optical gap and constants of the film also the references are mentioned and all you need to overcame the all problems of optical properties [five items]. Due to the format of Equations it is attached in a separate file, just you know the equation number please check it through the attach file.

The reflectance was measured at normal incidence with an aluminum reference mirror. The absolute values of  transmittance, T and reflectance R of the substrates after correcting the  are given by [[**]A. A. Sagade,  R. Sharma, Sensors and Actuators B: Chemical, 133 (2008) 135-143 [**] M. Dongol, M. M. El-Nahass, A. El-Denglawey, A.F. Elhady, A.A. Abuelwafa. Current Applied. Physics 12 (2012) 1178.]

Equation No.……………………………...... (1)

Ift and Iq are the intensities of the light passing through the film-quartz system and the reference quartz, respectively, and Rq is the reflectance of quartz. In addition, if the intensity of light reflected from the sample mirror reaching the detector is Ifr and that reflected from the reflectance reference mirror is Im, then

Equation No.…………………… (2)

Ifr is the intensity of light reflected from the sample reaching the detector and Rm is the mirror reflectance.

T and R spectra were performed on thin film samples with single face. Extinction coefficient, k and refractive index, n were calculated using T, R and thickness d taking into account the experimental error of   the film thickness. T and R.

T, R and d are used to calculate the absorption coefficient, α

according to [[**]A. El-Denglawey, M. Dongol, M.M  El-Nahass, J. Lumin 130 (2010) 801.]:

Equation No.………………………….…….…… (3)

α is given by:

Equation No. …………………………………… (4)

The calculated α is included within high absorption region (α ≥ 104 (cm)-1).  

Extinction coefficient, k of TiO2 film is calculated by using:

 

Equation No. ……………………………………….…. (5)

            An absorption edge of semiconductors corresponds to the threshold of charge transition between the highest nearly filled band and the lowest nearly empty band. According to inter-band absorption theory, the film’s optical gap can be calculated according to [[**]J. Tauc (Ed.). Amorphous and Liquid Semiconductors, Plenum, New York, 1976].

 

Equation No.…………………………. (6)

 A is a parameter of transition probability, it measures the disorder of material 

A = 4πσmin / nc∆E, Where  σmin is the minimum metallic conductivity, n is the refractive index, c is the light-velocity, and  ∆E = ∆Ec - ∆Ev   represents the  band tailing

[[**]M. M. El-Nahass, M. H. Ali, A. El-Denglawey Trans. Nonferrous Met. Soc.     China 22(2012) 3003−3011].  

is the optical gap of the material, hυ is the incident photon energy and r is the transition coefficient. The reported values of r are 2 for the measurement of indirect optical gap and 1/2 for direct optical band gap.

In case you know the value of Eg from the literature you can use:  ln (αhυ) = ln B+ r ln(hυ- Egopt) and find the value of r from the slope of the line for Ex.

 May you have both direct  and indirect band gap

The indirect optical gap, is evaluated by extrapolating the straight line part of (αhν)1/2 curves with energy axes (hυ) i.e (αhυ)1/2 = 0  according to eq:

Equation No.……………………………. (7)

Direct optical gap,  is evaluated by extrapolating the straight line part of (αhν)2 curves with energy axes (hυ) i.e (αhυ)2 = 0  according to eq:

Equation No.……………………………. (8)

Both direct, and indirect, optical gaps of TiO2 films may be found.

OR

to find the value of r the relation between (αhν)^1/r and incident energy (hν) for all values of r should be figured. The value of r which release a straight line represent the value of r and the electronic transition type.

Both R, and k at different λ were used to calculate refractive index, n according to:

[[**] T. S. Moss. Optical Process in semiconductor. Butter Worths, London, 1959.]

Equation No.     ………………………….… (9)

The dispersion of refractive index of the films is analyzed by the concept of the single oscillator and can be expressed by the Wemple–DiDomenico (WDD) relationship[ [**][S. H. Wemple, M. DiDomenico. Phys. Rev. B 3 (1971) 1338-1351] as: 

Equation No.          ……………….(10)

 Eo is the single-oscillator energy and Ed is the dispersion energy which is a measure of the intensity of the inter-band optical transition, it does not depend significantly on the band gap. The oscillator parameters can be determined  by fitting a straight line to the experimental points according to [[**][A. El-Denglawey. J. Non-Cryst. Solids 357 (2011) 1757-1763].

Plotting  vs (hν)2 allows to determine the oscillator parameters by fitting a straight line to the experimental points. By extrapolating the linear part of WDD optical dispersion relationship towards the infrared spectral region (hν = 0), static refractive index n(0), could be defined by the infinite wavelength dielectric constant ε∞ or n(0)2,

The relation between the lattice dielectric constant εL, and the refractive index, n as

[[**] A. El-Denglawey. J. Non-Cryst. Solids 357 (2011) 1757-1763.]:

Equation No. ………………………………….(11)

where εL is the lattice dielectric constant, N/m* is the ratio of the carrier concentration to the effective mass, c is the speed of light, and e is the electronic charge, εo is the permittivity of free space. The linearity of the plots of n2 versus λ2, verifying of Eq. (11). The value of εL is determined from the extrapolation of these plots to λ2 = 0 and N/m* from the slope of the graph.

Please notice that there is a direct calculation of ( hu) by this relation using wave length only.

E= hu =1240/wavelength (nm). 

I wish that is useful and help you.

Don't hesitate to ask about any thing concerning optical, electrical and structural properties.

Good luck

Yours

A. El-Denglawey

Professor, Dr. Dr. Henning Beier of Aachen, i.e., Henning, an old friend of mine, kindly contacted me through ResearchGate, and found that one of his colleagues, Professor, Dr. Andreas Prescher, M.D., has been conserving old journals and these Tables were in his collections. Henning and Professor Prescher gracefully made an electronic copy of these 3 tables (22 Figures) in an outstanding condition, and sent them to me.  I would like to express my sincere gratitude to Professors Beier and Prescher as well as to ResearchGate, for  making this acquisition possible, despite that fact that neither NIH Library nor National Library of Medicine could find these tables. Koji Yoshinaga

On day 6 embryo Blastocyst seems to have 84.4+/- 5.7 cells which doubled on day 7 to around 125.5+/-19.

Happy new year to you too. We don't really see fluid accumulation but found the tumours produced tended to fall into two populations:-

 Those that grew well quickly and produced a well vascularised (deep red) but less solid tumour (the proportion of these tumour was increased by implanting in 50% matrigel)

Those that initially looked like they were tumours but then hardly grew until 10-12 week then grew into more solid but less well vascularised (ie very pale) we avoided using these as some spontaneously regressed. 

The biggest problem was when a cohort was implanted at 10-14 days we would have a good proportion of apparent tumours but maybe a third would be the second type I described and determining which ones were which at this stage when we needed to randomise in to groups was challenging to say the least.

We suspected that it was determined by the amount of VEGF produced by different cell within the implanted population but have never tested this hypothesis.  Hope that helps a little. All the best.

Hello Béatrice，

I checked the poor rat today. Although he moved normally, but the brain problem seemed more severe. I still don't know what happened to him. I did 7 surgeries and 2 of them had this kind of problem. The other's problem was less severe, but I don't know why. I should ask the vet perhaps.

Thank you John. I deduce from your answer that for you, the EAD measured by the method of Olthoff today represents the primary endpoint, best suited to the evaluation of machines perfusion.

The transcriptome is not the proteome. So you look at quite different things. Microarrays give you a quite complete picture of the transcriptome as a whole, allowing you to investigate regulatory transcription networks. Given the array experiments and the data analysis is done "state of  the art", array results are very reliable. But, to repeat myself: they are about the transcriptome, not about the proteome.

thank you very much Marcia H. Ratner 

UK TAVI database

Clearly thrombophilia brings repeat abortions and  constitute a major cause of infertility by promoting hypercoagulability of placental vessels.  Whether it is genetic and acquired its preventive treatment consists of administering aspirin or heparin in early pregnancy, maintaining treatment until delivery. Current is forced to study mutations in coagulation factors (at least II, V, XII) and MTHFR. Safdarian et al)(Iran J Reprod Med. 2014 Jul;12(7):467-70) found that mutations for factor V and homozygosity for MTHFR are risk factors for IVF faillure . Detachment of placenta, preeclampsia, hypertension or intrauterine growth retardation are other possible complications of pregnancy induced by thrombophilia.

Genetikon Blastomer Fiksasyonu (Modifiye)

2.2.BLASTOMERE FIXATION

2.2.1.MATERIALS:

Microscope slides

Falcon Tubes

Glass Pasteur Pipettes

Mouth Pipette

Elmas uçlu cam çizer

Inverted Microscope (Olympus CK40)

Steriomicrocope (Olympus SZX12)

2.2.2 Solüsyonlar

HCl (1 M)

Methanol

Glacial Acetic Acid

dH2O

KCL

Phosfat Buffer Solution (PBS)

Hipotonik Solüsyonu: 100 ml dH2O + 0,559 gram KCl / 37 oC (0.075 M KCl)

: Alternatif Hipotonik solusyonu: (6mg/ml BSA-Bovin Serum Albümin,

%1 sodium citrate-tribasic:dihydrate)

Fiksatif: 3 birim Metanol + 1 birim Asetik asit (altın kural: fiksatif taze hazırlanacak 15-20 dakika tazelikte)

2.2.3 METOD-Fiksasyon için lamlarının hazırlanması

-Fiksasyon yapılacak alan “cam çizer” ile daire şeklinde çizilir.

-Her blastomer için 1 adet olmak üzere lamlar 100 ml’lik şale’nin içine dizilir.

-Şale’de 100 ml dH2O + 1 ml HCl mevcuttur. Lamlar 30 dakika bekletirilir ve şaleden çıkarılıp havada silkelendikten sonra kuruması için kurutma kağıdının üstüne dik olarak yaslanır, oda sıcaklığında bırakılır.

2.2.4 METOD-Fiksasyon

-Temiz bir pedri yada lam üzerine; 2 ayrı PBS damlacığı, 2 ayrı Hipotonik damlacığı konur.

-Biyopsisi yapılmış blastomer; “içine PBS çekilmiş ağız pipeti” ile 1. PBS damlacığının içine taşınır. 5 kez pipetaj yapılır (yağ ve artıklarından temizlemek amacı ile) sonra 2. PBS damlacığının içine bırakılır.

- Boş ağız pipetine 1. Hipotonik damlacığından hipotonik çekilir. 2. PBS damlacığı içindeki blastomer hipotonik çekilmiş ağız pipetine alınır ve 1. Hipotonik damlacığına taşınarak 5 kez pipetaj yapılıp, 2. Hipotonik damlacığına taşınır.

-Blastomer 2. Hipotonik damlacığında 1 (bir) dakika bekletilir. Bu süre aşılmaz.

-İnvert mikroskop altına önceden dairesi çizilmiş ve temiz lam konur. Hipotonik solusyonu içinde bekleyen blastomer mümkün olan en az sıvı ile lamda önceden çizilmiş dairenin içine bırakılır.

-Mikroskopta blastomer görülür.

- Mikroskop ile hipotonik kurumadan hemen önce ayrı bir pipet (ağız pipeti veya mikropipet veya cam pastör pipeti) ile 2 mikrolitre taze hazırlanmış fiksatif, blastomerin üzerine damlatılır.

-Blastomerin; 2. Hipotonik damlacığı içinden minimum sıvı ile fiksasyon lamına taşınması ve üzerine miktar olarak 2 mikrolitre fiksatif damlatılması, “blastomer kaçağını” önlemesi açısından önemlidir. Fiksatif hipotonik içindeki blastomere damlatıldığında, hipotonik solusyonu ve fiksatif karışırken blastomer damlacık içinde hızla hareket edebilir. Bir miktar beklenirse blastomer hareketini durduracak ve lam üstüne fiske olacaktır.

- Fiksatif damlatılırken blastomer sürekli gözlenmelidir. Kuruma aşamasında ikinci, gerekirse üçüncü damla fiksasyon (2 mikrolitre’lik damlalar halinde) damlatılır ve nukleusun apaçık ortaya çıktığı gözlenir. Lam tamamen kurutulur. Lamlar FISH hibridizasyonuna kadar oda sıcaklığında, lam taşıma kutularında, kapağı kapalı temiz şekilde saklanır.

-Lamın etiketlenmesi çok önemlidir. Blastomer numarası vb yazılmalıdır.

Sly, D J, M J McKinley, and B J Oldfield. 2001. “Activation of Kidney-Directed Neurons in the Lamina Terminalis by Alterations in Body Fluid Balance..” American Journal of Physiology. Regulatory, Integrative and Comparative Physiology 281 (5): R1637–46.

I would go for a single arm and no electronics attached. Those are not really helpful to correct your coordinates but just help in doing the surgery more visual and sometimes fast (in case of the display showing axis movements). I would ask for quotes to Stoelting or Kopf (I don't know WPI frames but they may be good as well). I used both of them. I recommend to check also the pricing infos for add ons for any of the frames you are considering, in advance so you know how much it would cost for you to upgrade the instrument. I was using a kopf for cannula implantation. You eventually need a cannula-holder to place the cannulas on the skull. Which could be an additional 250 euros, or something similar (if you are in Portugal).

Stoelting has the same tool and WPI as well... In my opinion KOPF looks more well made, robust and stable... But I don't want to make bad advertising...it could be just due to different lab usage.

Regarding the add-ons, If you don't need to use strange angles but you will do always orthogonal injections - as for cannula implantation - you can even think of buying just rat earbars (they can be used for both rats and mice), so it would be cheaper. If you use mice, you will need a mouse jaw holder (if not included), which is different from the one made for rats of course (is basically the steel support to keep the jaw closed). If you need to move around (also changing angles) maybe the "small animal holder" (to understand, this would be the black steel support on the stoelting website, it comes with smaller earbars and smaller jaw holder) is more useful because it has finer regulation for the height of earbars and jaw holder. In addition, due to the more advanced position in respect to the holder/cannula/pipette support, it allows a wider range of angles and movements (but of course is more expensive). So it depends what you plan to do.

I would avoid electronic assisted frames simply because:

1) if there is an issue you have to call the company and the instrument can't be used (with the basic steel frame this rarely occurs, unless it drops on the floor)

2) if you misplace the skull the system doesn't correct for that so eventually is not really helping in precision. Plus, if you know where to inject, you probably have already a brain atlas you have consulted before doing surgery (so you don't need a live update with computer screen and software for that, it is just costly).

3) the only electronic device (if you have the money) I would reccommend is the digital display that shows you the movement update on each axis (this saves you time looking for the right coordinates on the metric scales on the frame arms and allows precision and consistency.

so my suggestion would be

from this catalog http://www.bilaney.com/kopf/Kopf%20Catalog%202013.pdf

frame #900 single manipulator, 100 micron (page6)

animal adaptor (page 16) model # 926

cannula holder 1776 P1 (page20) if fits with your cannulas

earbars #857 for rats (adaptable on mice) (page 19)

but you can compare the pricing from Stoelting and decide based on the difference

Dear Anne, apologize if I again am guessing....

but IF you use the following method for staining, please let me know...

citing ammonium sulfide (only) as reagent for staining implantation sites was found @ http://www.ncbi.nlm.nih.gov/pubmed/9408976 :

Teratology. 1997 Oct;56(4):252-61.

Critical period of carbon tetrachloride-induced pregnancy loss in Fischer-344 rats, with insights into the detection of resorption sites by ammonium sulfide staining.

Narotsky MG, Brownie CF, Kavlock RJ.

Source: U.S. Environmental Protection Agency, Reproductive Toxicology Division, National Health and Environmental Effects Research Laboratory, Research Triangle Park, North Carolina 27711, USA. narotsky.michael@epamail.epa.gov

=citation=:"Non-gravid uteri were examined for resorption sites, placed in 10% ammonium sulfide, and re-examined for stained resorption sites approximately 1 and 4.5 hr later. ..... Ammonium sulfide staining clearly yielded a more accurate account of the incidence of FLR. The technique was most effective when the staining period was extended to 4.5 hr, as two cases of FLR were revealed that had been undetected after 1 hr of staining. " [nofollowing histo- technique or images of histology sections or staining] =End of citation=

Another article would be:

Jikken Dobutsu. Experimental Animals [1985, jan,34(1):17-22]

[Studies on implantation traces in rats. I. Size, observation period and staining].

[Article in Japanese]

Yamada T, Hara M, Ohba Y, Inoue T, Ohno H.

=Citation=: "All kinds of implantation traces were stained distinctly with 10% ammonium sulfide, 0.2% sodium hydroxide and 2% potassium ferrocyanide. In this staining method, sodium hydroxide has an excellent effect on the staining of implantation traces. Specimens washed in water after being stained with sodium hydroxide and fixed in formalin can be preserved without discoloration for a long period of time." =End of citation=.

also found:

Yamada T, Hara M, Ohba Y, Inoue T, Ohno H: [Studies on implantation traces in rats. II. Staining of cleared uteri, formation and distribution of implantation traces]. Jikken Dobutsu; 1985 Jul;34(3):249-60

=citation:= This staining method was convenient for observation of the implantation traces.

Though the implantation traces in rats which had been delivered four times were arranged like beads, they were recognizable by NaOH stain as either old or new traces, the former appearing smaller without agglomerate cells and a little more brown in comparison with the latter. =end of citation=

another source:

"staining the uterus with ammonium sulfide or sodium hydroxide (Salewski, 1964 * ; Yamada et al. 1988 **)

- *SALEWSKI VE (1964): Faerbemethode zum makroskopischen Nachweis von Implantationstellen am Uterus der Ratte, Nauyn-Schmiedebergs Arch. Exp.Pathol. und Pharmak. 247, p 367ff.

- ** Yamada T, Ohsawa K, & Ohno H (1988): The usefulness of alkaline solutions for cle...