Histological Quality Research Articles

Comparison of standard laminectomy with an optimized ejection method for the removal of spinal cords from rats and mice

For researchers seeking to collect spinal cord samples from mice and rats while avoiding acid decalcification, few options are available. Laminectomy is the standard method which yields high quality samples, yet is time consuming and technically difficult. Ejection of the cord from the vertebral column is another technique commonly used; however, the literature suggests that this method can damage the spinal tissues and is typically avoided when histology of samples is the desired endpoint. Here, we describe an optimized method for ejection of spinal cords from rats and mice, and compare histological quality of these samples with those collected via laminectomy. Our results show that ejection can yield high quality spinal cord samples and may be suitable for use as an alternative to laminectomy.

Amount of Torque and Duration of Stretching Affects Correction of Knee Contracture in a Rat Model of Spinal Cord Injury

Joint contractures are a common complication of many neurologic conditions, and stretching often is advocated to prevent and treat these contractures. However, the magnitude and duration of the stretching done in practice usually are guided by subjective clinical impressions. Using an established T8 spinal cord injury rat model of knee contracture, we sought to determine what combination of static or intermittent stretching, varied by magnitude (high or low) and duration (long or short), leads to the best (1) improvement in the limitation in ROM; (2) restoration of the muscular and articular factors leading to contractures; and (3) prevention and treatment of contracture-associated histologic alterations of joint capsule and articular cartilage. Using a rat animal model, the spinal cord was transected completely at the level of T8. The rats were randomly assigned to seven treatment groups (n = 4 per group), which were composed of static or intermittent stretching in combination with different amounts of applied torque magnitude and duration. We assessed the effect of stretching by measuring the ROM and evaluating the histologic alteration of the capsule and cartilage. Contractures improved in all treated groups except for the low-torque and short-duration static stretching conditions. High-torque stretching was effective against shortening of the synovial membrane and adhesions in the posterosuperior regions. Collagen Type II and VEGF in the cartilage were increased by stretching. High-torque and long-duration static stretching led to greater restoration of ROM than the other torque and duration treatment groups. Stretching was more effective in improving articular components of contractures compared with the muscular components. Stretching in this rat model prevented shortening and adhesion of the joint capsule, and affected biochemical composition, but did not change morphologic features of the cartilage. This animal study tends to support the ideas that static stretching can influence joint ROM and histologic qualities of joint tissues, and that the way stretching is performed influences its efficacy. However, further studies are warranted to determine if our findings are clinically applicable.

How can we safely climb the ALPPS?

R0 resection with zero mortality is the ultimate goal for hepatobiliary surgeons, especially for the treatment of extensive hepatobiliary malignancies. The safety of liver resection is dependent on the function of the future liver remnant (FLR), and an inadequate FLR volume is related to a significant increase in postoperative mortality and morbidity. Therefore, various criteria for the FLR volume have been proposed to secure the safety of major hepatectomies according to the extent of underlying injury in the liver [1–5]. However, these criteria for FLR volume often cause clinical dilemmas for surgeons in determining the surgical indications for patients with small FLR volumes because the safety of surgery and oncological radicality are, by nature, conflicting factors. In the history of hepatobiliary surgery, there have been two outstanding approaches for the safe management of patients with very small FLR volumes. The first was the development of techniques that manipulate the portal blood flow to induce hypertrophy of the FLR. Initially achieved using portal vein ligation (PVL) [6–8], these techniques have evolved toward percutaneous portal vein embolization (PVE) [9–12]. Increasing evidence has suggested that hypertrophy of the FLR induced by portal flow modulation is associated with an improved safety of major hepatectomies [3, 10, 13]. In addition, dynamic volume parameters, such as the degree of hypertrophy [13] or the kinetic growth rate [14], are also very informative for estimating the histologic quality and functional reserve of the underlying liver. The second noteworthy approach was the ‘‘twostage surgery’’ for the resection of multiple bilobar hepatic lesions. This sequential procedure was initially proposed by surgeons at the Hopital Paul Brousse in Paris, France, with the expectation of allowing interim liver regeneration between the two sequential hepatic resections [15]. An oncological advantage of the two-stage approach has been reported in patients with extensive colorectal liver metastases [16], and this procedure used in conjunction with or without PVE has been adopted by numerous hepatobiliary centers. These two evolutional approaches have expanded the indications for surgery, and many patients with extensive liver tumors have benefited from surgery using these approaches. However, a remaining issue is that these approaches require at least several weeks to complete the entire clearance of the tumor burden within the liver. Some authors have suggested a risk of tumor progression during the waiting time after PVE [17, 18]. Therefore, the time lag between the preoperative intervention and resection can be critical, especially for the treatment of patients with borderline resectable tumors and/or oncologically highly aggressive tumors. A recent notable paper in the field of hepatobiliary surgery was a case series introducing a new surgical procedure known as ‘‘ALPPS’’ (Associating Liver Partition with Portal vein ligation for Staged hepatectomy) that enables the rapid growth of the FLR [19]. The first stage of this procedure includes a right PVL and the in situ splitting of the liver along the umbilical fissure or the main portal fissure. Schnitzbauer et al. [19] reported that a 74 % volume increase was observed in the FLR at a median of 9 days after the first procedure. Immediately after its publication, this article triggered a large number of reactions from all over the world. Although the clinical outcomes demonstrated in this paper were very impressive and all the patients were able to proceed to a right N. Kokudo (&) J. Shindoh Hepato-Biliary-Pancreatic Surgery Division, Department of Surgery, Graduate School of Medicine, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8655, Japan e-mail: KOKUDO-2SU@h.u-tokyo.ac.jp

1039 Mortality From Non Variceal Upper Gastrointestinal Bleeding in Patients With Liver Cirrhosis: an Individual Patient Data Meta-Analysis

EUS-FNA of solid pancreatic mass lesions. Methods: In this multi-center trial, patients with solid pancreatic masses were randomized to undergo EUS-FNA using a 19G or 25G needle. The Standard 19G needle was used for trans-gastric FNA and the Flexible 19G needle for trans-duodenal FNA. A pathologist blinded to randomization sequence evaluated specimens for onsite diagnostic sufficiency and later for presence of histological core tissue. Main outcome measures: Compare the median number of passes required to establish onsite diagnostic sufficiency, ability to procure histological samples, specimen quality (bloodiness graded as 33%, 34-66%, 67%), technical failures and complications using the 19 vs. 25-gauge FNA needles. Sample size: Estimation was based on median number of passes required to establish onsite diagnostic sufficiency. Results: Of the 72 randomized patients, there was no difference in patient demographics or disease characteristics between both groups. Pancreatic masses were located in the head/uncinate in 38 (58.7%) and in the body/tail in 34 (41.3%) patients with no difference in location between cohorts (p 0.77). The final diagnosis was adenocarcinoma in 49, neuroendocrine tumors in 6, chronic pancreatitis in 11, lymphoma in 2, metastatic or other neoplasm in 2 and indeterminate in 2. One patient randomized to 19G needle had intra-procedural bleeding and postprocedure fever that was managed conservatively. SUMMARY: While there was no difference in the median number of passes required to establish on-site diagnostic sufficiency between the 19 and 25G needles, the 19G needles were significantly better for core tissue procurement. Although more bloody, the current platform of 19G needles can procure samples from any location within the pancreas. Conclusions: As both the 19 and 25G needles perform equally well, the choice of an FNA needle for sampling pancreatic masses should be based on the need for core tissue.

Abstract 2373: The LCBRN: a biospecimen resource for lung cancer biomarker and discovery science.

Abstract The Lung Cancer Biospecimen Resource Network (LCBRN) is an open access biorepository that obtains specimens from lung cancer patients (tissue, blood, urine, bronchial lavage fluid and saliva) and makes them available to biomedical researchers. Specimen collection is supported at 3 collection sites (Medical University of South Carolina, The University of Virginia, and Washington University at St. Louis) in compliance with uniform standard operating procedures. The specimens are annotated with clinical, radiologic and pathologic data. Histologic and molecular quality control procedures are in place for tissue samples. Lung cancer and non-neoplastic lung tissue aliquots are processed into DNA, RNA and protein fractions, which are also available. Fluid biosamples are available from subjects prior to, and after surgery to remove the cancer. A cohort of subjects are followed for recurrence with serial biofluid sampling. Specimens are currently available from over 300 research subjects. The application form for biospecimens and additional information is available at http://LCBRN.org. Citation Format: Christopher A. Moskaluk. The LCBRN: a biospecimen resource for lung cancer biomarker and discovery science. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2373. doi:10.1158/1538-7445.AM2013-2373

Quality assessment of tissue specimens for studies of diabetic foot ulcers

Diabetic foot ulcers (DFUs) represent an important clinical problem resulting in significant morbidity and mortality. Ongoing translational research studies strive to better understand molecular/cellular basis of DFU pathology that may lead to identification of novel treatment protocols. Tissue at the non-healing wound edge has been identified as one of major contributors to the DFU pathophysiology that provides important tool for translational and clinical investigations. To evaluate quality of tissue specimens and their potential use, we obtained 81 DFU specimens from 25 patients and performed histological analyses, immunohistochemistry and RNA quality assessments. We found that depth of the collected specimen is important determinant of research utility, and only specimens containing a full-thickness epidermis could be utilized for immunohistochemistry and RNA isolation. We showed that only two-thirds of collected specimens could be utilized in translational studies. This attrition rate is important for designs of future studies involving tissue specimen collection from DFU.

Megassessões de unidades foliculares e fatores de crescimento plaquetário

Queda de cabelo, seja parcial ou completa, e causa de preocupacao significativa para homens e mulheres, que a veem como um sinal inestetico e visivel de envelhecimento. Avancos e refinamentos das tecnicas culminaram na introducao de megassessoes de microenxertos e minienxertos. Essa tecnica se tornou amplamente aceita como um procedimento simples e seguro, que recria as linhas randomicas naturais do cabelo. Os foliculos pilosos foram retirados da area cervical posterior, onde 500 a 1.500 unidades foliculares podem ser obtidas. A area calva foi implantada atraves de incisoes de lâmina no 11. Apos o procedimento, gaze umedecida em solucao salina foi aplicada sobre a area implantada por 24 horas. Os pontos foram removidos no 7o dia de pos-operatorio. O resultado final foi obtido 8 meses a 12 meses apos o procedimento, em homens, e 12 meses a 14 meses, em mulheres. Nos pacientes em que um segundo procedimento foi necessario, este foi realizado 1 ano apos o transplante inicial. A qualidade e a forca do cabelo transplantado permanecem em alguns pacientes por tempo indeterminado, em decorrencia de caracteristicas particulares, como alta qualidade histologica da area doadora, hereditariedade, hormonios e envelhecimento. A cirurgia de transplante capilar demonstra que o uso de fatores de crescimento plaquetario autologo pode melhorar a densidade capilar. Esse processo oferece uma nova perspectiva ao transplante capilar, representando uma contribuicao importante para a cirurgia de implante com megassessoes de unidades foliculares.

Evaluation of two different vacuum-assisted breast biopsy systems: Mammotome® system 11G/8G vs. ATEC® system 12G/9G

Breast cancer screening programs have been established worldwide and early detection of breast cancer has increased steadily. The most common way to confirm dignity of non-palpable and sonographically-occult suspicious findings on mammography is the stereotactically-guided vacuum-assisted breast biopsy To compare two stereotactically guided vacuum-assisted breast biopsy systems measuring time effectiveness and quality of harvested material in clinical practice. One hundred and forty-six patients presenting with suspicious microcalcifications on mammography were included in the study. Biopsies were carried out with either the Mammotome(®) system (11-gauge and 8-gauge) or the ATEC(®) system (12-gauge and 9-gauge). Lesions with a diameter <15 mm on mammography were biopsied with 11-gauge or 12-gauge devices whereas lesions >15 mm were targeted with 8-gauge and 9-gauge. Mammotome(®) system 8-gauge device was used in 34 patients, the 11-gauge system in 37 patients. The ATEC(®) system 9-gauge system was used in 37 patients and 12-gauge in 38 patients. Time was taken, focusing on preparing the system, time of collecting the samples, preparing the samples, and cleaning the site. During the biopsies 24 samples were taken. The histologic quality of the tissue samples was judged by a pathologist in a blinded fashion according to a specimen grading classification concerning tissue fragmentation, artefacts, and the adequacy of the tissue for diagnosis. The median overall time for the Mammotome(®) system was 879 s (11-gauge) and 934 s (8-gauge) and for the ATEC(®) system 671 s (12-gauge) and 673 s (9-gauge). The ATEC(®) system displays a significantly shorter overall time for small and large biopsy devices (U-test, P < 0.001). Concerning the mean time difference of the overall time comparing small and large systems the ATEC(®) system was 267.6 s faster using the small and 244.8 s faster using the large system. Comparing the histologic quality of tissue samples the Mammotome(®) system shows significantly higher values for the large and the small system (Chi-square test, P < 0.001). Both biopsy systems meet all requirements for daily practice and confirm the diagnosis of suspicious microcalcifications. The ATEC(®) system was observed to be faster but this difference of about 250 s might not be relevant in daily practice. The Mammotome(®) system provides a better histologic quality of tissue samples.

The role of IL-6 in bone marrow (BM)-derived mesenchymal stem cells (MSCs) proliferation and chondrogenesis

Rheumatoid arthritis (RA) is the most common degenerative arthritic cartilage and represents a disease where the prospect of stem cell therapy offers considerable hope. Currently, bone marrow (BM) represents the major source of mesenchymal stem cells (MSCs) for cell therapy. In the pathology of RA, the pro-inflammatory cytokines, such as interleukin 6 (IL-6) play a pivotal role. To investigate the direct role of IL-6 in the chondrogenic differentiation of murine MSCs (mMSCs), we isolate MSCs from the murine bone marrow, and induce MSCs chondrogenesis with different concentrations of IL-6 in vitro. Through detecting the histological and histochemical qualities of the aggregates, we demonstrate that IL-6 inhibited the differentiation of MSCs into chondrocytes in the dose-dependence manner. These findings suggest that possible strategies for improving the clinical outcome of cartilage repair procedures.

Adenoma Detection Rates in an Urban Hospital with African American Majority

Purpose: The adenoma detection rate (ADR) is an important quality indicator of screening colonoscopy. Grady memorial hospital is an urban hospital that serves a large indigent population of African American majority. The aim of this study is to measure the overall adenoma detection rates, and compare ADR in black and non black patients. Methods: This is a retrospective analysis of all asymptomatic patients who underwent first-time screening colonoscopy between June 1st 2011 and June 1st 2012. Data about age, race, gender, quality of colon preparation, adenoma detection, size and histology were recorded and analyzed. Results: A total of 2,202 screening colonoscopies (1,429 women and 773 men) were performed during the study period. 2,038 patients (92.6%) had an adequate and complete examination, while 146 patients (6.6%) had an inadequate/poor colon preparation, and 18 patients (0.8%) had failed cecal intubation. Among patients with adequate and complete examination, 711 (35%) were men and 1,327 (65%) were women. Mean age was 58 years. The majority of patients were black (n=1,760 patients, 86.4%), followed by white (n=124, 6.1%), Hispanic (n=66, 3.2%), Asian (n=45, 2.6%) and other races (n=43, 2.4%). The overall ADR was 29.2% (595/2,038). The ADR was 35.3% in men and 25.9% in women. The ADR was comparable in black patients (28.6%, 503/1,760) versus non black (33.1%, 92/186); p=0.136. The prevalence of adenomas sized more than 9 mm in diameter was similar between black (107/1,760, 6.1%) and non black patients (20/278, 7.2%); p=0.5. The prevalence of adenomas with advanced histology (high grade dysplasia, villous, tubulovillous or carcinoma) was similar between black (72/1,760, 4.1%) and non black patients (10/278, 3.6%); p=0.87. The ADR increased with increasing age among all racial groups.Table: [1939] Prevalence of adenomas and advanced lesions by race and genderConclusion: Cecal intubation rates and ADR compared favorably to accepted quality standards. Contrary to previous reports, the prevalence of adenomas, adenomas >9 mm in size, and adenomas with advanced histology was similar in black and non black patients.

Non-formalin fixative versus formalin-fixed tissue: A comparison of histology and RNA quality

Preanalytical handling of tissue samples can influence bioanalyte quality and ultimately outcome of analytical results. The aim of this study was to compare RNA quality, performance in real time RT PCR and histology of formalin-fixed tissue to that of tissue fixed and stabilized with a formalin-free fixative, the PAXgene Tissue System (PAXgene), in an animal model under highly controlled preanalytical conditions. Samples of rat liver, kidney, spleen, intestine, lung, heart muscle, brain, and stomach tissue were either fixed in formalin or fixed in PAXgene or fresh frozen in liquid nitrogen. RNA was extracted from all samples, examined for integrity in microcapillary electrophoresis, and used in a series of quantitative RT PCR assays with increasing amplicon length. Histology of paraffin-embedded samples was determined by staining with hematoxylin and eosin.Histology of all formalin-fixed and PAXgene fixed samples was comparable. RNA with acceptable integrity scores could be isolated from all embedded tissues, 4.0 to 7.2 for formalin and 6.4 to 7.7 for PAXgene, as compared to 8.0 to 9.2 for fresh frozen samples. While RNA with acceptable RINs (RNA integrity number) could be isolated from formalin-fixed samples, in microcapillary electrophoresis this RNA separated with a slower migration rate and displayed diffuse, less focused peaks for ribosomal RNA as compared to RNA from frozen or PAXgene fixed samples. Furthermore, RNA from formalin-fixed tissues exhibited inhibition in quantitative RT PCR assays which increased with increasing amplicon length, while RNA from PAXgene fixed samples did not show such inhibition.In conclusion, our results demonstrate that excluding other preanalytical factors, PAXgene Tissue System preserves histology similarly to formalin, but unlike formalin, does not chemically modify RNA. RNA purified from PAXgene fixed tissues is of high integrity and performs as well as RNA from fresh frozen tissue in RT PCR regardless of amplicon length.

Radiation‐guided breast sentinel lymph node biopsies – is a handling delay for radiation protection necessary?

Radiation-guided sentinel lymph node (SLN) biopsy is a well-established procedure in many countries. However, histopathology protocols in different centres specify widely differing sample handling delays of between 0 and 72 h. Introducing a delay reduces the radiation exposure of pathologists, but has a detrimental effect on the quality and validity of histology. This study aims to show that a sample handling delay is not justified by the radiation doses to pathologists handling samples received directly from surgery. Radiation doses to the body and hands of pathologists handling samples delivered directly from theatres were measured using personal dose meters. These measurements were supplemented by dose assessments undertaken using dose-rate measurements at 1 cm and 30 cm from Tc-99m sources to simulate the processing of samples. The study has shown that radiation doses arising from a zero delay in sample handling represent a negligible radiation risk to pathologists and are well within relevant limits specified in the Ionising Radiations Regulations 1999. This study supports adoption of a zero-delay SLN histopathology protocol. Centres must, however, complete a risk assessment that accounts for local practice and adopt simple precautions to keep doses to pathologists as low as reasonably achievable.

Laser-assisted surgery with different wavelengths: a preliminary ex vivo study on thermal increase and histological evaluation

Since the introduction of laser in clinical practice, different wavelengths have been used for oral surgery on the basis of the different characteristics and affinities of each one. The aim of this study was a comparison of different laser wavelengths in relation to both thermal increase and "histological quality" in a model of soft tissue surgery procedures. Thermal evaluation was realized, during laser-assisted surgery excision performed on a bovine tongue, by a thermal camera device to evaluate thermal increase on the surface of the sample and with four thermocouples to evaluate thermal increase on the depth of the specimen; temperature was recorded before starting surgical procedure and at the peak of every excision. The quality of excision, in terms of tissue damage and regularity, was realized by two blind examiners on the basis of established criteria. The highest superficial thermal increase was recorded for Superpulse 5-W CO2 laser, the lowest one for Er:YAG laser. The highest in depth thermal increase was recorded for 5W Diode laser, the lowest one for Er:YAG laser. The best quality of incision was obtained with a 3-W CO2 laser and 3-W diode laser; epithelial, stromal, and vascular damages were evaluated with different degrees for all the used wavelengths with the best result, in terms of "tissue respect," for Er:YAG laser. In all the surgical procedures performed, thermal increase was evaluated until the end of the procedure; at remaining tissue level, thermal decrease was evaluable in the few seconds after surgery. The Er:YAG laser was the device with a lower influence on thermal increase; CO2 and diode lasers revealed a good histological quality. Further studies may be necessary to test the reliability of laser devices for the excision of all the types of specimens needing histological evaluation and diagnosis.

Histomorphometric Analysis and Regional Variations of Full Thickness Skin Grafts in Dogs

To (1) compare thickness of the epidermis and dermis of anatomically different donor sites in dogs, and (2) evaluate hair follicle damage during full thickness skin grafts (FTSG) preparation from anatomically different donor sites. Another objective was to compare the extent of agreement between gross and histologic quality of graft preparation. Ex vivo study. Healthy Beagle dogs (n = 12). Skin samples were harvested from fresh cadavers free of gross dermatopathology and FTSG prepared. Regional, epidermal and dermal thickness, and hair follicle density in intact skin and FTSG specimens from different regions were determined by histomorphometric analysis. Hair follicle density in intact skin and FTSG specimens, and skin thickness measurements were compared among regions. Mean epidermis and dermis thickness of the dorsal lumbar and lateral neck regions were significantly greater when compared with other sites. Total hair follicle (superficial and deep dermis) densities were significantly less in prepared FTSG versus intact skin specimens. The dorsal lumbar region had the highest hair follicle density. Thickness of the epidermis and dermis is dependent on body region. Hair follicle density in the FTSG specimens was decreased compared with intact skin specimens, which may affect hair regrowth in FTSG.

Elimination Diet Effectively Treats Eosinophilic Esophagitis in Adults; Food Reintroduction Identifies Causative Factors

Adults with eosinophilic esophagitis (EoE) typically present with dysphagia and food impaction. A 6-food elimination diet (SFED) is effective in children with EoE. We assessed the effects of the SFED followed by food reintroduction on the histologic response, symptoms, and quality of life in adults with EoE. At the start of the study, 50 adults with EoE underwent esophagogastroduodenoscopies (EGDs), biopsies, and skin-prick tests for food and aeroallergens. After 6 weeks of SFED, patients underwent repeat EGD and biopsies. Histologic responders, defined by ≤ 5 eosinophils/high-power field (eos/hpf) (n = 32), underwent systematic reintroduction of foods followed by EGD and biopsies (n = 20). Symptom and quality of life scores were determined before and after SFED. Common symptoms of EoE included dysphagia (96%), food impaction (74%), and heartburn (94%). The mean peak eosinophil counts in the proximal esophagus were 34 eos/hpf and 8 eos/hpf, before and after the SFED, and 44 eos/hpf and 13 eos/hpf in the distal esophagus, respectively (P < .0001). After the SFED, 64% of patients had peak counts ≤ 5 eos/hpf and 70% had peak counts of ≤ 10 eos/hpf. Symptom scores decreased in 94% (P < .0001). After food reintroduction, esophageal eosinophil counts returned to pretreatment values (P < .0001). Based on reintroduction, the foods most frequently associated with EoE were wheat (60% of cases) and milk (50% of cases). Skin-prick testing predicted only 13% of foods associated with EoE. An elimination diet significantly improves symptoms and reduces endoscopic and histopathologic features of EoE in adults. Food reintroduction re-initiated features of EoE in patients, indicating a role for food allergens in its pathogenesis. Foods that activated EoE were identified by systematic reintroduction analysis but not by skin-prick tests.

Local Delivery of Recombinant Osteoprotegerin Enhances Postorthodontic Tooth Stability

Relapse after orthodontic tooth movement is a significant problem in orthodontics. The purpose of this study was to examine the efficacy of the osteoclast inhibitor osteoprotegerin-Fc (OPG-Fc) for inhibiting postorthodontic relapse. Rat maxillary molars were moved mesially and allowed to relapse for 24days. Low-dose (1mg/kg) or high-dose (5mg/kg) OPG-Fc or saline was injected adjacent to the molars during relapse. Tooth movement, micro-CT, histologic bone quality, and serum OPG and TRAP-5b were measured. OPG-Fc injections significantly diminished postorthodontic relapse from 63% (0.78/1.20mm) of total movement in vehicle control rats to 31% (0.31/1.00mm) in low-dose and 24% (0.28/1.16mm) in high-dose OPG-Fc groups 24days after appliance removal. Normalization of bone and periodontal tissues occurred as early as 8 and 16days in the high- and low-dose OPG-Fc-treated groups, respectively, while the vehicle-treated group showed only partial tissue recovery 24days following tooth movement. After 24days of relapse, there was complete recovery to pre-tooth-movement values for bone volume fraction (BVF) and tissue mineral density (TMD) in both the low- and high-dose OPG-Fc groups, while BVF recovered only partially and TMD did not recover in the vehicle control group. Greatly elevated serum OPG levels and reduced serum TRAP-5b levels in OPG-Fc-treated animals indicated systemic exposure to locally injected drug. The profound decrease in postorthodontic relapse by local OPG-Fc administration indicates that osteoclasts are critical to bone maturation following tooth movement and points to the potential pharmacologic use of OPG-Fc or other RANKL inhibitors for orthodontic retention.

Quantitative and structural changes of testis and semen quality parameters changes caused by peroral administration of diazinon in rats

The aim of this study was to find the quantitative and structural changes in the rat testis and changes of semen quality after a diazinon administration. METHODS: Rats received diazinon (99% purity) in thier drinking water (40 mg.l-1) with free access. Age of rats at the beginning of the experiment was 30 days and experiment lasted for next 90 days. The histological samples were evaluated by histological and morphometric methods in light microscopy and the samples of semen were evaluated with CASA method. RESULTS: Disintergation of cellular associations in the seminiferous epithelium, germ cells evacuation into the tubule lumen and thier necrosis were mostly observed. Rarely vacuolisation and cracks of epithelium and fibrotisation of interstitial tissue were noted. Morphometric methods have shown extension of epithelium (P&lt;0,01), reduction of tubule lumen (P&lt;0,001) and dilatation of blood vessels (P&lt;0,001). In CASA analysis elevation of all parameters were noted, with statistically significant increase of DSL, VSL, ALH. Dilatation of blood vessels will be probably one of the most significant finding in diazinon toxicity because changes in blood flow in the testis are the key factors of accurate physiological function of testis. The epithelium despite the serious disintegration of germ cells associations and release of these necrotised germ cells to the lumen was significantly extended. This fact indicates the self-reparation compensational function. The same tendency (stimulation effects) has been found in all analysed sperm parameters. It supports previous hypothesis. CONCLUSIONS: Diazinon in this design of experiment causes the disintegration of the germinal epithelium cells associations consequently leading to necrosis and release of these cells to the tubule lumen. Dilatation of blood vessels and unknown stimulation effect on sperm quality parameters are two other common effects of diazinon. We concluded that diazinon in our subchronic low dose test causes middle to moderate histological, morphometric and semen quality changes which were partially compensated with some unknown recovery mechanism. Otherwise, subcellular structures and their functions may be damaged which can lead to subfertility. Further investigation of diazinon is needed for verification of our hypothesis.

Assessment of the histological quality of endoscopic biopsies obtained from the canine gastro-esophageal junction

In the dog biopsy samples from the gastro-esophageal junction (GEJ) are rarely obtained during routine gastroscopy. The aim of this pilot study was to assess the histological quality of endoscopic biopsies sampled from the canine esophagus and cardia. It was hypothesised that it is possible to sample adequate specimens from these sites. For this purpose 10 dogs with an indication for gastroscopy were enrolled in a prospective study. Biopsy samples were obtained with standard biopsy forceps for single use exactly from the GEJ thus containing preferably columnar epithelium from the cardia and squamous epithelium from the esophagus, respectively. In every dog the specimens were examined for size, layers and site, respectively. Study endpoint was reached when specimens originated from cardia and esophagus, showing at least epithelium and lamina propria mucosae, and a diameter >2mm on the slide, respectively. 72 biopsy specimens (median 7, range 5-10) obtained from the GEJ were examined in 10 dogs. Specimens from the esophagus containing squamous epithelium with lamina propria mucosae were found in 5 of 10 (50.0%) dogs. Specimens from the cardia containing columnar epithelium with lamina propria mucosae were found in 10 of 10 (100.0%) dogs. Four of 10 (40.0%), and 10 of 10 (100.0%) dogs showed at least one specimen >2mm on the slide originating from the esophagus, and from the cardia, respectively. Histological quality was found to be adequate in 4 of 10 (40.0%) dogs, showing specimens of adequate size, originating from both esophagus and cardia, and containing at least epithelium and lamina propria mucosae. The pilot study provides evidence that during routine gastroscopy it is possible to sample endoscopic biopsies from the cardia and with limitations from the esophagus showing a quality adequate for histological examination of the epithelium and the lamina propria mucosae.

Cervical cytology and histology in the context of a screening programme

In the UK, the national cervical cancer screening programme is probably the most established, robust and rigorous of all screening programmes that has evolved over 25 years. It has seen a multitude of changes based on evidence obtained from a vast amount of research and observations, as well as from advances in technology. This success has led to a definite impact on cervical cancer incidence with reduced mortality and morbidity related to cervical cancer. More recently, the changes introduced to the programme have been exponential and in the UK we are at the brink of a new direction and a new style of provision of the screening programme. Throughout the years, there has been a continued effort to understand morphological aspects of the cytology and histology in cervical neoplasia and many new entities have been recognised or variations of known entities realised. The intricate measures to ensure quality in the screening programme have led to close working relationships between cytologists, histopathologists, colposcopists, and now, molecular biologists. The UK cervical cancer screening programme (CCsP) is likely to change significantly in the next few years, and although reduced in quantity, cytology will still remain an important aspect of it. The importance of histopathology is unlikely to change, but more onus will be placed on the molecular biologists and colposcopists. In order to discuss and understand the importance of cytology and histology in the cervical screening programme and the absolute requirement of correlation between the two, it is important first to appreciate how the UK CCsP has evolved and how it has taken with it the evolution of new technologies in cytology leading to improved correlation between histology and cytology and quality assurance with obvious benefits to the patient.

Virtual microscopy for histology quality assurance of screen-detected polyps

Virtual microscopy for histology quality assurance of screen-detected polyps