The pituitary gland is a small endocrine gland about the size of a pea that rests below the hypothalamus of the brain. When thyrotropin-releasing hormone (TRH) is released from the hypothalamus, TSH will be produced.
Failure of the thyroid gland to respond to the TSH produced by the pituitary gland will result in an excess amount of TSH found in the blood.
To check if the symptoms that you are experiencing such as tiredness, weight gain, constipation, dry skin, frequent menstrual period, or intolerance of cold are caused by hypothyroidism (under-active thyroid gland). To check if the symptoms that you are experiencing such as rapid heart rate, weight loss, diarrhea, nervousness, irregular menstrual period, or intolerance of hot are caused by hyperthyroidism (overactive thyroid gland). For people with hypothyroidism, to keep track if the thyroid replacement therapy is still effective. To keep track if the function of the thyroid gland in people with hyperthyroidism who are undergoing treatment like antithyroid medicine, radiation therapy or surgery. No special preparation will be needed before the test but there are certain medications and circumstances that may affect the quality of the test. In 2003, the American Association of Clinical Endocrinologists (AACE) issued a press release to update changes in the TSH’s normal values. It is for the AACE’s belief that this new range will result in proper diagnosis especially for people with subtle symptoms that are still untreated.
Unfortunately, many traditional doctors are still not aware with this new update and stick with the old range. The pituitary gland is damaged that it is prevented from releasing TSH (secondary hypothyroidism). Too much thyroid medicine was taken during the course of treatment for an underactive thyroid gland. Whether low or high, a TSH result that is out of the normal range should be addressed promptly. For patients who are having a surgery and with myxedema coma, rapid treatment is necessary. There is no known way to prevent hypothyroidism but the screening tests conducted on newborns a day after their delivery can help to detect congenital hypothyroidism. Like hypothyroidism, there is no existing way to prevent hyperthyroidism but quitting smoking may cut your risk.
Science, Technology and Medicine open access publisher.Publish, read and share novel research. Celikel (1995) found a correlation between mean AgNOR and ploidy and proliferation activity of urinary bladder tumor as measured by DNA flow cytometry. 2009aExpression of 53protein in liver and sera of patients with liver fibrosis, cirrhosis or hepatocellular carcinoma associated with chronic HCV infection. 2009bAgNORs count and DNA ploidy in liver biopsies from patients with schistosomal liver cirrhosis and hepatocellular carcinoma.
1999DNA ploidy of liver biopsies from patients with liver cirrhosis and hepatocellular carcinoma: a flow cytometric analysis.
2007Immunochemical identification and detection of serum fibronectin in liver fibrosis patients with chronic hepatitis C. 1994Increased risk of hepatocellular carcinoma development in patients with cirrhosis and with high hepatocellular proliferation. 1998Hepatocyte proliferation rate is a powerful parameter for predicting hepatocellular carcinoma development in liver cirrhosis.
1995DNA Flow cytometric studies and AgNOR counts in patients with transitional cell carcinoma of bladder. 2003Argyrophilic nuclear organizer regions in nasopharyngeal carcinoma and paraneoplastic epithelia.
2007Manganese superoxide dismutase activity and incidence of hepatocellular carcinoma in patients with Child-Pugh class A liver cirrhosis: a 7-year follow-up study. 2011Pre-operative liver biopsy in cirrhotic patients with early hepatocellular carcinoma represents a safe and accurate diagnostic tool for tumour grading assessment. 1987Nucleolar organizer region associated proteins in cutaneous melanocytic lesions: a quantitative study. 2009Predicting recurrence after liver transplantation in patients with hepatocellular carcinoma exceeding the up-to seven criteria.
2011Cytometry of DNA replication and RNA synthesis: Historical perspective and recent advances based on "click chemistry".
The basis for different glands secreting saliva of differing composition can be seen by examining salivary glands histologically. Acini in the parotid glands are almost exclusively of the serous type, while those in the sublingual glands are predominantly mucus cells. In the histologic sections of canine salivary gland shown above, the cells stained pink are serous cells, while the white, foamy cells are mucus-secreting cells. Secretion of saliva is under control of the autonomic nervous system, which controls both the volume and type of saliva secreted. Potent stimuli for increased salivation include the presence of food or irritating substances in the mouth, and thoughts of or the smell of food.
Lubrication and binding: the mucus in saliva is extremely effective in binding masticated food into a slippery bolus that (usually) slides easily through the esophagus without inflicting damage to the mucosa.
Oral hygiene: The oral cavity is almost constantly flushed with saliva, which floats away food debris and keeps the mouth relatively clean. Initiates starch digestion: in most species, the serous acinar cells secrete an alpha-amylase which can begin to digest dietary starch into maltose.
Provides alkaline buffering and fluid: this is of great importance in ruminants, which have non-secretory forestomachs.
Evaporative cooling: clearly of importance in dogs, which have very poorly developed sweat glands - look at a dog panting after a long run and this function will be clear. Diseases of the salivary glands and ducts are not uncommon in animals and man, and excessive salivation is a symptom of almost any lesion in the oral cavity. If you are on birth control pills then light flow is usual, but if you are not taking birth control pills or similar non-hormonal IUD and you still have light flow, there is a chance that you have a hormonal imbalance. If you have unrelated or sudden bleeding to the normal cycle you are having a breakthrough bleeding. If you have intense cramps throughout your periods and think that it is usual, you are making a mistake. 4.If your period is late and there is no chance of pregnancy, you are likely to have hormonal imbalance. If your period is late, and if there is a possibility that you are pregnant, that is great.
5.If your discharge has saturated red color similar to “cranberry juice” than you have a normal secretion. Get Rid Of Those Ugly Dark Stains and Patches On The Inner Thighs, Underarms and Neck in Just 15 Minutes! A child with a thyroid gland that is deficient in making thyroid hormone may later become mentally retarded.
It also has one sole duty, the conversion of your iodine intake into thyroid hormones that help regulate the body’s metabolism, body temperature, growth and development.
TSH level will help determine if it is caused by the damage on the thyroid gland or other factors (like problem in the hypothalamus or pituitary gland). It indicates that signals coming from the pituitary gland go well with the activity of the thyroid gland.
And to investigate further, additional tests will be needed to ascertain the cause of the abnormal value. Beta-blockers like propranolol may be used to manage some symptoms like sweating, rapid heart rate and anxiety. Replacement pill will be taken by a person whose thyroid gland is destroyed by radiation therapy or is surgically removed. Blood lymphocytes (blue) used as external control for DNA ploidy analysis by flow cytometry.
Libbrecht et al., (2001) indicated that the presence of liver cell dysplasia in a needle liver biopsy of patients with the viral induced chronic liver disease is an independent risk factor for the development of HCC.
It is a normal side-effect in case you are on birth control pills but otherwise you should contact a doctor.
However, if the chances of you being pregnant are next to zero, there are a few things you should know.
In effect, the TSH targets our thyroid gland to produce thyroid hormones namely T3 (triiodothyronine) and T4 (thyroxine) that help in controlling the metabolism of our body.
Other tests that your doctor may order are thyroid antibodies (for thyroid disease that is suspected to be autoimmune-related) and T3 test.
The doctors may use other side effects or signals you manifest alongside with your TSH level to determine if other issues are of concern. Figure B shows high percentage of apoptotic cells (indicated by arrows) in comparison with low percentage shown in figure A. Figures A,C from tumor part and B,D from residual part (non tumor part) of HCC liver biospes.

The findings were viewed following staining under the light microscope using 100? oil immersion lens and cedar wood oil.
At least two possibilities can be considered to explain the diploidy in liver cirrhosis biopsies with dysplasia. Adelaide Nardone, from Brown University, light period may indicate problems with the thyroid or pituitary gland. Breakthrough bleeding can be a sign of cervical or uterine polyps, which results from too much estrogen in the body. Daniela Carusi, director of General Gynecology at Brigham and Women’s Hospital in Boston, young women think that they need to endure intense cramps, but that is not the case. Advanced liver fibrosis and HCC have been indicated to develop in about 30% of patients with chronic hepatitis B or C. Therefore, they indicated that the evaluation of the hepatocyte proliferation rate is very important to identify patients requiring stricter follow-up program for early diagnosis of HCC. There is also a probability that you suffer from a Mullerian anomaly, which is a disorder causing a malformed hymen, which can impede with the flow of the menstrual blood. Carusi, you might not have regular ovulations or you could have PCOS, which makes it more difficult for you to know your fertile days.
High AgNOR count in cirrhotic liver as a marker of regenerative capacity has been associated with a significantly increased incidence in development of HCC in chronic liver disease (Derenzini et al., 1993).
Oddly light periods can also be linked to polycystic ovarian syndrome (PCOS) and autoimmune disorders.
In case you suffer from obesity or have a low percent of body fat, you may have a severe hormonal imbalance and it is time you change your lifestyle. HCC typically develops in the cirrhotic liver in about 80% of cases and develops only in the non-cirrhotic liver in less than 20% of cases (Alkofer, 2011). It is difficult to identify a small change, such as a defect in the short arm of a chromosome.
However, in liver cirrhosis, dysplasia had a significant relationship with ploidy and AgNORs. Obese women who have irregular periods need to check to see if they suffer from Type 2 diabetes, since ovaries are directly affected by insulin resistance.
When diagnosed at early stage, HCC remains eligible for potential curative options such as surgical resection, orthotopic liver transplantation or percutaneous destructions. It is difficult to identify a small change, such as a defect short arm of a chromosome or an abnormal or small sized chromosome. So, the incidence of high AgNORs count, dysplasia, and presence of aneuploidy by DNA flow cytometry suggest that liver cirrhosis may be considered as a more serious condition in the evolution of HCC. Furthermore, the aneuploidy was found more commonly in large cell dysplasia (LCD) compared with small cell dysplasia (SCD), but no multiple aneuploid peaks (polypoidy) were found in either LCD or SCD. Recently, we compared the results of DNA flow cytometry and AgNORs with the histopathological data. However, most of HCCs have widespread dissemination within the liver at diagnosis (intermediary stage) or show extrahepatic dissemination within the portal tract, lymph nodes or distant visceral metastasis (Llovet et al., 2008).
As recommended by Llovet and Bruix (2008), new and efficacious therapies are needed, along with new diagnostic biomarkers for early detection of liver cancer. A gradual increase in the AgNOR count per nucleus was noted as disease progressed from normal liver through liver cirrhosis to HCC tumor lesions.
Liver biopsy, since its initial introduction by Klatskin as a clinical tool 100 years ago, soon became the major diagnostic test for liver disease (Afdhal & Manning, 2008). The presence of an aneuploid DNA pattern had been reported by Hoso and Nakanuma (1991), to a precancerous change in cases with atypical adenomatous hyperplasia, of the liver. However, biomarkers alone are not sufficient to allow definitive decisions to be made for a given patient (Halfon et al., 2008).
Yet, the presence of liver cell dysplasia, although not specific as premalignant condition, could be considered as a serious hand mark in cases of liver cirrhosis. However, histological examination of liver biopsy specimens is still the gold standard for evaluating the presence of liver fibrosis, pathogenesis of liver injury and assessment of anti-viral treatment.
However, Thomas et al., (1992) reported a lower incidence (25 %) of DNA aneuploidy in patients with liver cirrhosis. On the other hand, Argyrophilic nucleolar organizer regions (AgNORs) proteins are a set of argyrophilic nucleolar proteins that accumulate in highly proliferating cells, whereas their expression is very low in nonproliferating cells (Farina et al., 2011).
The simple and universally applicable methods for staining fixed and non fixed liver cells will be described. This may be a attributed to poor synthesis of albumin by these dysplastic hepatic cells as a result of change in the nuclear DNA contents of these aneuploid cells. The hepatocellular carcinomas were graded according to the Edmondson-Steiner histological grading system. Methods for staining of cell suspension from liver biopsies, and deconvolution of DNA-content-frequency histograms to estimate the percentage of cells in major phases of the cell cycle and frequency of apoptotic cells with fractional DNA content will be presented. So we can conclude that liver cell dysplasia is associated with aneuploidy, as measured by flow cytometry, may carry the risk for progression to HCC and liver cirrhotic patients with aneuploidy must be followed up.6. In addition, the advantage and disadvantages of the DNA-FCM and AgNORs count in the assessment of cellular kinetics of liver biopsies in liver disease will be investigated.
Unlike cytogenetic analysis, in which 20 to 40 cells are routinely analyzed, cytometric analysis can measure ten thousands of cells or nuclei. So, quantification of AgNOR is considered to have putative relationship with ploidy in HCC. One of the most advantages of cytometric DNA content analysis is that it does not rely on metaphase cells, and cell in all phases of the cell cycle are generally included in the analysis. Our study presents complementary methods to histopathology, which are valuable to pathologists, when they have difficulty in diagnosis of early stages of HCC on the basis of liver biopsy.
The critical limitation of cytometric analysis is the significantly lower resolution compared with karotyping.
Finally, both AgNORs count and DNA ploidy analysis are useful and valuable indicators of cellular kinetics in HCC.8.
A flow cytometer is similar to a microscope, except that, instead of producing an image of the cell, flow cytometry offers "high-throughput" (for a large number of cells) automated quantification of set parameters.
Conclusion remarks and future researchThe assessment of cellular kinetics of liver biopsies is of great importance for understanding the development and diagnosis of liver disease. To analyze tissues, such as liver biopsies, a single-cell suspension must first be prepared using mechanical or enzymatic methods.
The limitation imposed on cytometric analysis even using high precision instruments and the most stiometric DNA binding fluorescence dye; DAPI (4, 6 diamidino 2-phenylindole), prevent cytometric from detecting chromosomal gains or losses involving more than about 5 % of the total DNA. Flow cytometric analyses of ploidy and cell cycle were one of the first applications of flow cytometry and this technique remains the only rapid and efficient means of making such measurement particularly for core liver biopsies.
The idea of flow cytoemtry is based on a beam of light (usually laser light) of a single wavelength is directed onto a hydrodynamically-focused stream of fluid (Cho et al., 2010).
In practical terms, this implies that the smallest change that can be detected theoretically is the gain or loss of equivalent of one large (e.g. AgNOR proteins are a set of argyrophilic nucleolar proteins that accumulate in highly proliferating cells, whereas their expression is very low in nonproliferating cells. A number of detectors are aimed at the point where the stream passes through the light beam: one in line with the light beam (Forward Scatter or FSC) and several perpendicular to it (Side Scatter (SSC) and one or more fluorescent detectors). The AgNOR tissue marker can be used as a routine complementary histopathologic study and DNA ploidy by flow cytometry, since the variations in its number and distribution indicate existence of cell alterations in a given lesion and the use of this technique is easy and inexpensive. Each suspended particle from 0.2 to 150 micrometers passing through the beam scatters the ray, and fluorescent chemicals (such as propidium iodide which used for binding to DNA) found in stained cells or attached to the particle (as conjugated fluoresce antibodies used for immunophenotyping) may be excited into emitting light at a longer wavelength than the light source (de Tute, 2011). Diagnosis of HCC using both DNA ploidy and mean AgNORs are useful in the detection of cellular and structural abnormalities. This combination of scattered and fluorescent light is picked up by the detectors, and, by analyzing fluctuations in brightness at each detector. AcknowledgementsThis work was partially funded through the Science and Technology Development Fund (STDF), Egypt "Grant no. It provides a method for sorting a heterogeneous mixture of biological cells into two or more containers, one cell at a time, based upon the specific light scattering and fluorescent characteristics of each cell. It is a useful scientific instrument, as it provides fast, objective and quantitative recording of fluorescent signals from individual cells as well as physical separation of cells of particular interest. The cell sorted machines have ability to sort cells with highly sterility and purity more than 98%, so these cells can be use for tissue culture applications (Cho et al., 2010). In situ, the expression of AgNOR proteins is measured globally by quantification of the level of silver staining using morphometry and image analysis (Sirri et al., 2000). The distribution of AgNORs and their quantities have been associated with proliferative activity and ploidy in different neoplastic and preneoplastic conditions.
Cell cycle phasesThe cell cycle is commonly divided into “phases” – interphase and mitosis. Staining of nucleolar organizer regionsIn our laboratories, liver biopsy samples were taken from the most representative area of the diseased tissue. The biopsy specimens were then fixed in 10% buffered formalin and subsequently processed to prepare paraffin blocks. G2 was originally defined as the second gap between S and Mitosis, but is now known to function as a time of DNA damage repair, and likely, preparation for entering Mitosis (M phase).

One set stained by the hematoxylin and eosin (H&E) technique for routine histological evaluation and confirmation of diagnosis. Mitosis has been traditionally sub-divided into stages defined by nuclear morphology – prophase, prometaphase, metaphase, anaphase A and B, and telophase. Another set of sections were stained for localization and quantification of AgNORs according to the method described by Lindner et al., (1993). A final phase, division of the cytoplasm that overlaps telophase and is often lumped with mitosis, is cytokinesis (CK). The sections were deparaffinized in xylene, and hydrated through 100% and 95% ethanol to water. The major cell cycle sub-phases, G1, S, G2+M can typically be identified by direct quantitative measurement of the DNA by flow cytometry based on staining of DNA with fluorescence dye such as propidium iodide (Fig. Silver staining solution was prepared by dissolving 2% gelatin in 1% formic acid at room temperature and filtered through syringe filter (Sigma Chemical Co., St. One part of the solution was mixed with two parts of 50% silver nitrate immediately before use. Staining was done in dark at room temperature for 30 min, then sections were immersed in 5% sodium thiosulphate for 5 min. Often is also considered to be a prognostic indicator of tumor progression and outcome of the treatment. Counting of silver stained nucleolar organizer regions (AgNORs) AgNORs appear as brown black dots inside the nucleus using silver stain. Counting of AgNORs was done without any knowledge of the histological or the clinical data. In essence, when DNA measurement is done correctly and accurately, either of these approaches is expected to yield similar estimate of DI of aneuploid cells.
From our experience, we can use normal lymphocytes, including lymphocytes from the same patient, as external standard control of DI = 1.0 (Fig. 3).For comparison with the tumor it is necessary to use normal cells both as external and internal control standards. Figure B shows high percentage of apoptotic cells (indicated by arrows) in comparison with low percentage shown in figure A.laser and detector settings as tumor sample (Shankey and Rabinovitch, 2002). The external lymphocyte control cells should be measured prior to- and also after- measurement of tumor sample. Also, AgNORs count in the hyperplastic foci was significantly higher than that in the controls (Wakasa et al., 1998). Often, normal stromal- or tumor infiltrating cells are already present in the tumor sample and they can be used as an internal control of DNA ploidy.
This is due to the fact that DNA stains ability after formaldehyde fixation and paraffin embedding is markedly altered making external standards useless (Darzynkiewicz et al., 2010). Controls are used to assure proper instrument performance and to validate the DNA staining technique (Robinson et al., 2002). These standards are used to define the amount of DNA related fluorescence that is equivalent to the amount that would be obtained from a diploid human cell.
Figure 4.Blood lymphocytes (blue) used as external control for DNA ploidy analysis by flow cytometry. Another study failed to show a relationship between AgNORs count and Ki-67 and BrdU-labeling indexes (Nyska, 1995), or between diploid and aneuploid tumors (Nagao et al., 1995). This is in a disagreement with a study of mean AgNOR in trophoblastic disease, which indicated that mean AgNOR is a reflection of ploidy and proliferative activities (Maier et al., 1990). Doublet discriminationBecause cells and especially fixated cells tend to stick together, cell aggregates have to be excluded from analysis through a process called doublet discrimination. However, in contrast to immunohistochemical methods merely determining the growth fraction of the cells by Ki-67 and proliferating cell nuclear antigen (PCNA) analysis AgNORs apparently reflect time-related proliferation rates and ploidy.
Thus, AgNOR analysis may provide further insights concerning the biological behavior of the cell and is simple, inexpensive, and reliable method of evaluating the proliferative activity and ploidy of the cell (Meng et al., 1996). Analysis of cellular DNA content by flow cytometryFlow cytometry analysis of ploidy and cell cycle analysis were one of the first applications of flow cytometry and this technique remains the only rapid and efficient means of making such measurements (Lopez-Otero et al., 2010). Different histograms DNA ploidy measured using flow cytometry based on liver biopsied from HCC and liver cirrhosis with different DNA index are shown in Fig. Figures A,C from tumor part and B,D from residual part (non tumor part) of HCC liver biospes.3. Simple and universally applicable method for staining fixed and non fixed liver cells and analysis by flow cytoemtryIn cirrhotic patients with early HCC, a pre-operative liver biopsy should be performed because it has a low complication rate and it provides clinically useful information for the management of these patients (Colecchia et al., 2011). Knowledge of pre-operative tumor grade is crucial in the management of HCC because it can influence recurrence and survival after surgery.
Fine needle aspiration biopsy (FNA) is currently the preferred technique for diagnosing liver masses because of its safety and rapidity. The cytological diagnosis is usually taken as definitive, which makes it critical for patient management (Kocjan G 2010). For this reason, additional studies that can be performed on cytological samples are desirable to aid diagnosis.
DNA analysis has diagnostic and ? or prognostic significance in different human neoplasias, and can be performed on FNA samples. The following is the simple and universally applicable method for staining fixed and non fixed liver cells and analysis by flow cytoemtry. Single-cell suspensions can be prepared by mechanical dissociation of the fresh biopsy specimen in RPMI-1640 medium (Sigma Chemical Co., St. Louis, MO) or phosphate buffer saline (PBS) followed by filtration through a piece of fine nylon mesh (45 ?m pore-size) and centrifugation to remove debris and cell clumps. Core liver biopsy or cell suspension can be preserved in media (sterile DMEM media or phosphate buffer saline) in freezer – 20 °C for more than one month. Flow cytometric analysis can be performed with a any flow cytometer (such as Coulter EPICS profile II Coulter Corp., Hialeah, FL ), configured with a 488 nm argon ion laser. DNA aneuploidy was defined as any population with a distinct additional peak (s) or the presence of a tetraploid population greater than 15%.
The coefficient of variation (CV) was defined as the standard deviation as a percentage of the mean DNA value of the diploid peak. DNA-FCM in the assessment of cellular kinetics of liver biopsies in liver cirrhosis and HCC It is scientifically and clinically important to better understand the intermediate events that predispose to neoplastic progression. Many neoplasms generate DNA-flow cytometry histograms which differ from the normal in having more than one peak with a different DNA content, the term aneuploid being used to describe these populations.
According to the multi-step theory of neoplastic progression, cancer development is associated with the evolution of a clone of cells with an acquired genomic instability and abnormal proliferation, i.e. In our previous study, (Attallah et al., 1999b), the cellular DNA content, known as ploidy state, of cells and expressed as DI revealed that normal liver showed a diploid pattern. The high percentage of aneuploidy in residual liver lesions of the HCC group or the cirrhotic liver group did not agree with the data of Rua et al. In the study of Rua et al., parafin-embedded tissues were used while our samples were tested fresh.
Patients with cirrhosis and high cell proliferation rates are at increased risk of developing cancer as reported by Baker et al. So, the presence of liver cell dysplasia (66%) and a high proliferation rate may explain the high percentage of aneuploidy in the cirrhotic patients group and in residual liver tissues of HCC. The increase of aneuploidy in liver cirrhosis patients and residual liver lesions of HCC demonstrated that liver cirrhosis is a premalignant lesion for HCC and that high proliferating activity and aneuploid cirrhotic patients must be followed up, as reported by Ballardini et al.
We can conclude that, DNA ploidy analysis of core liver biopsy specimens is useful in the investigation of the cell kinetics of liver cirrhosis and HCC. Dysplasia, cell proliferation and aneuploidy in liver cirrhosis may be an indication of a risk factor of liver cirrhosis for development of HCC.
Also quantitative DNA content analysis using small needle liver biopsies is simple and technically convenient for flow cytometric ploidy analysis.5.
DNA ploidy and liver cell dysplasia in liver biopsies from patients with liver cirrhosisThere is controversy among pathologist when assessing the presence or absence of liver cell dysplasia in liver biopsies taken from cirrhotic patients (El-Sayed et al., 2004). Flow cytometry is a rapid cell proliferation (technique and can assess thousands of cells in a few minutes. FCM can therefore be considered as a better ancillary technique in the assessment of liver cell dysplasia because it is objective and not able to misinterpretation or inter-observer discordance (Attallah et al., 2009b). In our previous study (El-Sayed et al., 2004), liver cell dysplasia was found in 60 % of patients with liver cirrhosis, small cell dysplasia in 38 % and large cell dysplasia in 62 %. We used FCM to determine the DNA ploidy, cellular DNA content and proliferation index (PI).
We found that 81.5 % of cases that were histopathologically shown to have liver cell dysplasia also had aneuploidy.

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