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Background/Aim. Bacterial vaginosis (BV) is a common cause of vaginal discomfort in women. The aim of this study was to compare Nugent?s scoring system and novel microscopy method, introduced in our laboratory and used in BV diagnosis. Methods. This study included 705 pregnant and asymptomatic women between 24 and 28 weeks of pregnancy. The degree of agreement between methods was determined by the kappa (?) index. The sensitivity, specificity, positive and negative predictive value of the novel microscopy method was compared to Nugent?s score as standard. Results. Based on the scoring system of both methods, Nugent?s and novel microscopy method, BV was diagnosed in 21% and 25% of women, respectively. Despite the disparities among diagnostic criteria, which mainly concerned classification of intermediary samples, the degree of agreement between categories, determined by ? index, was satisfactory: Nugent?s vs. novel microscopy method (? = 0.68; good agreement), and Nugent?s vs. novel microscopy method without intermediary results (? = 0.83; very good agreement). We also demonstrated that compared to Nugent?s method, as the golden standard, the novel microscopy method had high sensitivity and specificity (ranging from 75%?99.3%) and positive and negative predictive values (ranging from 88.8%?99.5%). Our method is based on a relative number of bacterial morphotypes, either rod forms (? 1.5 ?m, lactobacilli) or non-rod forms (< 1.5 ?m, bacterial vaginosis associated bacteria) under 200? magnification, which extends the surface of examined preparation, but without prolongation of observer?s working time. Conclusion. The novel microscopy method in diagnosing BV corresponded well with Nugent?s scoring system which allows it to be an alternative method in diagnosing BV. Further-more, the novel microscopy method is based on a relative number of bacterial morphotypes that appeared to be flexible and can be reorganized in the way to categorize findings into only two groups: normal and BV, which makes it comparable to dichotomous Amsel?s clinical criterion.

Background Age estimation is an important factor in forensic science for human identification. Teeth are considered to play a vital role as they resist decomposition at death unlike other tissues. This resistance and the gradual structural changes that take place throughout the life of an individual have made teeth useful indicators for age estimation. Dental cementum shows continuous apposition throughout the life of an individual. Tooth cemental annulation is a microscopic method for the determination of an individual’s age based on the analysis of incremental lines of cementum. Light microscopy as well as specialized microscopic methods have been employed to enhance the assessment of the cemental annulations. Periodontal disease is the most common dental problem affecting millions of people. Assessing the efficiency of the tooth cemental annulations method in periodontally diseased teeth is an important requisite. This study aims at assessing and evaluating the tooth cemental annulations in normal and periodontally diseased teeth using phase contrast microscopic method for age determination. Results A total of 60 teeth were included in the study and out of which 30 teeth were normal (sound teeth without any associated pathologies) and 30 were periodontally involved teeth respectively. Longitudinal ground sections were prepared and observed under phase contrast microscope. Measurements were made using an image analyzer software. The total width of the cementum was divided by the distance between two incremental lines. The eruption age of the tooth was then added to this to obtain the chronologic age for each individual. The results in the present study showed that tooth cemental annulations are applicable to periodontally sound teeth as well as in periodontally diseased teeth. There was no significant difference of estimated age from the actual age in both periodontally sound and periodontally diseased teeth. Normal teeth showed a reliability value of 92% and periodontally compromised teeth showed 96% respectively. There was no substantial influence of periodontal health on the estimated age. Conclusions The study concludes that the use of phase contrast microscopy in conjunction with image enhancement procedures improves the accuracy of age estimation and may serve as a reliable aid in forensic identification.

Cystoisospora belli (C. belli) is the only pathogenic species of the Cystoisospora genus responsible for severe diarrhea in immunocompromised patients. Most common microscopic method of diagnosis is less sensitive due to intermittent shedding of oocysts. We developed a new single-run polymerase chain reaction (PCR)–based diagnostic assay for C. belli. A new single-run PCR-based diagnostic assay was standardized for the detection of C. belli. Diagnostic reproducibility and repeatability of the PCR assay were evaluated. A cross-sectional analytical study was done on a total of 354 stool samples collected from 331 immunocompromised patients with diarrhea. All the stool samples were tested for the presence of oocysts of C. belli and were also tested by our new PCR assay for C. belli. Three of the representative PCR products were confirmed by sequencing. Fisher’s exact test was used to compare the two proportions. Microscopy detected C. belli in 11/354 (3.1%) of stool samples, and the new PCR-based assay detected C. belli in 16/354 (4.5%). The new single-run PCR-based assay detected C. belli in all the stool samples which were tested positive by microscopy and additionally detected C. belli in five stool samples. The developed PCR assay detected statistically significant proportion of C. belli (p < 0.001) as compared to microscopy. The 795 base pair PCR product from one microscopy positive stool sample and two microscopy negative stool samples were confirmed by sequencing. Our newly developed single-run PCR-based detection assay for C. belli is robust and reproducible. It may be used for molecular diagnosis of cystoisosporiasis especially in transplant, pediatrics, and human immunodeficiency virus (HIV) positive patients.

The success of the anti-helminth mass treatment use Albendazole makes detection of soil transmitted helminth infections even more difficult to do microscopically. It is hoped that the molecular method was able to help increase the detectability of Soil Transmitted Helminth infection. The research aimed is to evaluate effectiveness of Albendazole administration in Bali, to identify the presence of β-tubulin gene as molecular diagnosis of STHs infection among children who treated by Albendazole. This study is a cross-sectional study that recruits elementary school children aged 6-12 years as subjects. Stool examination was carried out using the Kato-Katz technique, then followed by a molecular method using the B-tubulin gene as the target gene. The results showed that only 1 sample out of 140 examined using Kato-Katz was positive for Trichuris trichiura. 30 samples were then extracted from the faeces and performed Polymerase Chain Reaction. A total of 4 positive samples detected the B-tubulin Ascaris lumbricoides gene and 1 positive sample of the B-tubulin Trichuris trichiura gene. In conclusion, albendazole is still effective to treat STH infection, the molecular method has a higher detectability than the microscopic method.

Introduction: Anterior odontoid screw fixation technique for specific types of odontoid fracture has been proven to be an effective, yet challenging procedure because of threatened damage to the structures in the vicinity. There are few articles suggesting the role of percutaneous and endoscopic technique as an alternative approach to the standard microscopic way. This is the first cadaveric study using existing “EASY GO” endoscopic system-assisted odontoid screw placement. Study Design: This was a cadaveric study. Objective: The objective of the study is to use the endoscope as a safer minimally invasive approach than the standard microscopic anterior approach to odontoid. Methodology: This is a pilot study on 10 fresh-frozen formalin-fixed adult human cadavers. The cadaver was positioned in a way to simulate reduced odontoid fracture. Tubular dilators were used to dock at C2–3 disk space after identifying the landmarks through the microscopic method. The EASY GO endoscopic system was then introduced, and a handheld drill was used to mark the entry point and pass the K-wire through the planned trajectory. Results: No vascular or adjacent vital structures' injury was observed in any of the cadavers. The initial difficulty in attaining the alignment was overcome by the appropriate positioning of the scope. Conclusions: Endoscopic-assisted technique for odontoid screw fixation shall provide a minimally invasive, safe, and easy surgery.

Objective: To study thrombocytopenia and elevated ALT levels in malaria patients reporting to Pak Med Level II+ Hospital XII and XIII, UN Mission Liberia, West Africa. Study Design: Cross sectional study. Place and Duration of Study: Departments of Pathology and Medicine, Pak Med Level II+ Hospital XII and XIII, UN Mission Liberia, West Africa, from Feb 2015 to Dec 2016. Methodology: In total of 100 febrile patients of both gender and all ages reported to Pak Med Level II+ Hospital XII and XIII, Liberia with clinical features of malaria and having positive malarial parasite (MP) on any of the methods of immune chromatography (ICT) or microscopic film were included. The febrile patients with typical clinical features of malaria but having negative MP both on ICT and microscopic methods were excluded.Pretreatment whole blood in EDTA was collected for testing MP on ICT and by microscopic method including MP index and platelets, while serum for ALT. Results: Out of total 100 patients, 75 were males, 25 females with mean age of 38 ± 5 years. MP was found positive on MP film in 95% and on ICT in 88% cases. Thrombocytopenia was found in 69% and ALT was elevated in 60% cases. Mean and SD for platelets count was 129 ± 72 x 109/L and of ALT 76 ± 66 U/L. Mean of MP index was 0.49 ranging from 0 to 3.5%. Correlation of MP index with platelets and ALT was found insignificant, although in malaria patients, platelets and ALT was inversely correlated significantly with each other showing low platelets along with elevated ALT levels. Conclusion: In malaria patients, thrombocytopenia and elevated ALT are frequent findings, which may not definitely correlate with MP index. So it is advisable to test for platelet count and ALT in all patients of malaria for early diagnosis and better management.

Plasmodium ovale is a benign tertian malaria parasite that morphologically resembles Plasmodium vivax. P. ovale also shares similar tertian periodicity and can cause relapse in patients without a radical cure, making it easily misidentified as P. vivax in routine diagnosis. Therefore, its prevalence might be underreported worldwide. The present study aimed to quantify the prevalence of P. ovale misidentified as P. vivax malaria using data from studies reporting confirmed P. ovale cases by molecular methods. Studies reporting the misidentification of P. ovale as P. vivax malaria were identified from three databases, MEDLINE, Web of Science, and Scopus, without language restrictions, but the publication date was restricted to 1993 and 2020. The quality of the included studies was assessed using the Quality Assessment of Diagnostic Accuracy Studies (QUADAS). The random-effects model was used to estimate the pooled prevalence of the misidentification of P. ovale as P. vivax malaria by the microscopic method when compared to those with the reference polymerase chain reaction method. Subgroup analysis of participants was also performed to demonstrate the difference between imported and indigenous P. ovale cases. The heterogeneity of the included studies was assessed using Cochran's Q and I2 statistics. Publication bias across the included studies was assessed using the funnel plot and Egger’s test, and if required, contour-enhanced funnel plots were used to identify the source(s) of funnel plot asymmetry. Of 641 articles retrieved from databases, 22 articles met the eligibility criteria and were included in the present study. Of the 8,297 malaria-positive cases identified by the PCR method, 453 P. ovale cases were confirmed. The pooled prevalence of misidentification of P. ovale as P. vivax malaria by the microscopic method was 11% (95% CI: 7–14%, I2: 25.46%). Subgroup analysis of the participants demonstrated a higher prevalence of misidentification in indigenous cases (13%, 95% CI: 6–21%, I2: 27.8%) than in imported cases (10%, 95% CI: 6–14%, I2: 24.1%). The pooled prevalence of misidentification of P. vivax as P. ovale malaria by the microscopic method was 1%, without heterogeneity (95% CI: 0–3%, I2: 16.8%). PCR was more sensitive in identifying P. ovale cases than the microscopic method (p 2: 65%). Subgroup analysis of participants demonstrated the better performance of PCR in detecting P. ovale malaria in indigenous cases (p: 0.0009, OR: 6.92, 95% CI: 2.21–21.7%, I2: 68%) than in imported cases (p: 0.0004, OR: 2.15, 95% CI: 1.41–3.29%, I2: 63%). P. ovale infections misidentified as P. vivax malaria by the microscopic method were frequent and led to underreported P. ovale cases. The molecular identification of P. ovale malaria in endemic areas is needed because a higher rate of P. ovale misidentification was found in endemic or indigenous cases than in imported cases. In addition, updated courses, enhanced training, and refreshers for microscopic examinations, particularly for P. ovale identification, are necessary to improve the microscopic identification of Plasmodium species in rural health centres where PCR is unavailable.

Mycobacterium tuberculosis is a cause of tuberculosis (TB ) disease. In this research Mycobacterium tuberculosis can be detected using the Microscopic and Real-Time PCR. This research aims are to know the difference in the results of the examination Mycobacterium tuberculosis with microscopic method and Real-Time PCR. This research used an experimental research design and was tested using the Chi Square test. This results showed at significant difference (p=0.000) detection of pulmonary TB disease by microscopic and Real-Time PCR methods

When considering methods of detecting Cryptosporidium in patient samples, clinical and public health laboratories have historically relied primarily on microscopy. However, microscopy is time intensive and requires trained personnel to accurately identify pathogens that are present. Even with skilled analysts, the parasitemia level has the potential to fall below the level of detection. In addition, public health laboratories do not always receive specimens in fixatives that are compatible with the desired microscopic method. Antigen-based and molecular methods have proven to be effective at identifying Cryptosporidium at low levels and require less training and hands-on time. Here, we have developed and validated a real-time polymerase chain reaction (RT-PCR) laboratory-developed test (LDT) that identifies Cryptosporidium hominis and Cryptosporidium parvum, and also includes detection at the genus level to identify additional species that occasionally cause disease in humans. Results of the molecular test were compared with those obtained from modified acid-fast microscopy, immunofluorescent microscopy, an antigen-based detection rapid test, and a commercial gastrointestinal panel (GI panel). Of 40 positive samples, microscopy and antigen-based methods were able to detect Cryptosporidium in only 20 and 21 samples, respectively. The GI panel detected 33 of the 40 positive samples, even though not all specimens were received in the recommended preservative. The LDT detected Cryptosporidium in all 40 positive samples. When comparing each method for the detection of Cryptosporidium, our results indicate the LDT is an accurate, reliable, and cost-effective method for a clinical public health reference laboratory.

Mycoplasma suis (M. suis) is an haemotropic Mycoplasma that adheres and invades erythrocytes and is responsible for infectious anaemia of pigs. Infections with M. suis have been reported worldwide. Clinical signs after M. suis infection can be significant particularly for the breeding herd in the period around farrowing but consequences are highly variable with some infected pigs never exhibiting clinical disease. The study aimed to determine the clinical relevance of Giemsa-stained blood smear for the diagnosis of M. suis compared with qPCR results. In our study, the comparison of qPCR results with microscopic investigation of Giemsa-stained blood smears revealed a lower sensitivity of the microscopic method: only 33 out of 102 qPCR positive blood samples were microscopically positive (M. suis visualised). No relationship between mean qPCR loads and microscopic observation was observed. Although more costly, qPCR is probably the best diagnostic tool available today for M. suis diagnosis.

Aim: To compare the effectiveness of endoscopic v/s microscopic excision of a benign pituitary adenoma. Materials and methods: The present prospective comparative was conducted in the Department of General Surgery, Pt. J.N.M. Medical College Raipur, C.G., India among 50 patients diagnosed of noninvasive pituitary Adenoma. Group I (n=28); underwent endoscopic transsphenoidal surgery. Group II (n=22): underwent microscopic transsphenoidal surgery. Results: A total of 50 patients with pituitary noninvasive adenoma were operated transsphenoidally. Endonasal endoscopic transsphenoidal surgery (group I) was carried out among 28 patients and 22 patients were operated by microscopic transsphenoidal surgery (group II). In group I, complete tumor excision was achieved in 16 (57.1%) patients, and in group II, it was achieved in 12 (54.5%) patients. Conclusion: Pituitary noninvasive adenomas can be treated with any approach. In compared to microscopic method, endoscopy is superior for resection and has less post-operative complications.

Blastocystis hominis is the most common intestinal parasite found in humans and many other hosts. Pathogenicity of Blastocystis spp. remains controversial, and it has been suggested that it may be associated with specific subtypes of the organism. This study identified the B. hominis subtypes and their prevalence rates in the northeast of Iran. A total of 1878 samples were collected from the northeast of Iran from January to December 2017. The patients’ demographic details were recorded. Samples were examined by a wet mount, and genomic DNA was extracted from positive samples. Also, PCR was done on the positive samples, and sequencing and phylogenetic analysis were subsequently performed. From 1878 collected stool samples, 152 (8.1%) Blastocystis samples were detected by the microscopic method. Of the 152 samples, Blastocystis spp. were found in 53.6% of the men and 28.9% of the women who showed clinical gastrointestinal symptoms, and a significant relationship was observed between gender and clinical symptoms ( P = 0.002 ). A meaningful relationship was found between the season and infection with this parasite ( P value = 0.003). The results of the sequencing of 22 PCR products showed the dominance of ST3, which was isolated from 10 (45.45%) patients, while ST1, ST2, and ST7 were found in 4 (18.19%), 7 (31.81%), and 1 (4.55%) patients, respectively. In this study, ST7 had a low prevalence in the northeast of Iran, and similar to previous studies, ST3 was the dominant subtype.

Trypanosomes are the causative agents of animal African trypanosomiasis (AAT) and human African trypanosomiasis (HAT), the former affecting domestic animals prevalent in Sub-Saharan Africa. The main species causing AAT in cattle are T. congolense, T. vivax, and T. b. brucei. Northern Uganda has been politically unstable with no form of vector control in place. The return of displaced inhabitants led to the restocking of cattle from AAT endemic areas. It was thus important to estimate the burden of trypanosomiasis in the region. This study was designed to compare the prevalence of animal African trypanosomes in cattle in Lira District using microscopy and polymerase chain reaction amplification (PCR) methods. In this cross-sectional study, a total of 254 cattle from the three villages of Acanakwo A, Barropok, and Acungkena in Lira District, Uganda, were selected by simple random sampling technique and screened for trypanosomiasis using microscopy and PCR methods. The prevalence of trypanosomiasis according to microscopic results was 5/254 (2.0%) as compared to 11/254 (4.3%) trypanosomiasis prevalence according to PCR analysis. The prevalence of trypanosomiasis infection in the animal studied is 11/254 (4.3%). Trypanosoma congolense was the most dominant trypanosome species with a proportion of 9/11 (81.8%), followed by T. vivax 1/11 (9.1%) and mixed infection of T. congolense/T. vivax1/11 (9.1%). Barropok village had the highest prevalence of trypanosomiasis with 6/11 (54.5%). There is a statistically significant relationship ( OR = 6.041 ; 95% CI: 1.634-22.331; p < 0.05 ) between abnormal PCV and trypanosome infection. Polymerase reaction amplification was the most reliable diagnostic method due to its high sensitivity and specificity as compared to the conventional microscopic method. Polymerase reaction amplification appears to have adequate accuracy to substitute the use of a microscope where facilities allow. This study, therefore, underscores the urgent need for local surveillance schemes more especially at the grassroots in Uganda to provide data for reference guideline development needed for the control of trypanosomiasis in Uganda.

Background/aim Conventional viability tests, help to screen the cellular effects of candidate molecules, but the endpoint of these measurements lacks sufficient information regarding the molecular aspects. A non-invasive, easy-to-setup live-cell microscopic method served to in-depth analysis of mechanisms of potential anticancer drugs. Materials and methods The proposed method combining the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) test with time-lapse scanning microscopy (TLS), provided additional data related to the cell-cycle and the dynamic properties of cell morphology. Apoptotic and necrotic events became detectable with these methods. Results Quantification of the results was assisted by image analysis of the acquired image sequences. After demonstrating the potential of the TLS method, a series of experiments compared the in vitro effect of a known and a newly synthesized nucleoside analogue. Conclusion The proposed approach provided a more in-depth insight into the cellular processes that can be affected by known chemotherapeutic agents including nucleoside analogues rather than applying repeated individual treatments.

Background: Among the human parasitic diseases, malaria is the main cause of morbidity and mortality. To prevent the high mortality and tracking malaria elimination efforts, a prompt and sensitive diagnosis is essential. This study aimed to compare High-Resolution Melting (HRM) and microscopic methods to diagnose Plasmodium falciparum and P. vivax. Methods: Eighty-one blood samples were collected from patients with clinical symptoms who were suspect to malaria in Chabahar district, southeastern Iran and also, from those who were referred to Malaria National Laboratory in the Tehran University of Medical Sciences, Tehran, Iran. Microscopic examination and HRM method were used to the diagnosis of Plasmodium parasites simultaneously. Results: Microscopic results revealed 45 positive cases (12 P. falciparum and 33 P. vivax) out of 81 collected samples while according to HRM analysis results 11 and 33 samples were identified as P. falciparum and P. vivax, respectively. HRM analysis also revealed 1 mixed infection of P. falciparum and P. malariae. Conclusion: HRM analysis provides a promising mean for simultaneous detection and discrimination of the Plasmodium spp. especially in mixed infection cases.

Microscopic transsphenoidal surgery is beyond any doubt a widely accepted and highly effective therapy for pituitary adenomas. Currently several centers have converted to an endoscopic transsphenoidal approach; suggesting that this technique provides more complete tumor resection, better visualization and reduce complications. However, there have been few series to document the results of this procedure. This report presents the technical details and compares it to the results of the microscopic method based on the experience of the group and a literature review. The endoscopic transnasal/ transsphenoidal technique is a safe and effective method for removal of pituitary adenomas. The results found in the literature suggest that the endoscope provides more complete tumor removal, and reduces complications. We believe that the advantages of the endoscopic technique will allow this procedure to become the future gold standard surgical therapy for pituitary adenomas.

Background: Dermatophytosis is considered as one of the major public health problems in the world and is the most commonly diagnosed skin disease in Bangladesh. The prevalence and characteristics of dermatophytic infections vary with climatic conditions, age, lifestyle and population migration patterns.Objective: To determine the prevalence of dermatophytic infection and sensitivity of different diagnostic procedures among the patients visiting dermatology outpatient department of Tairunnessa Memorial Medical College & Hospital, Gazipur, Bangladesh (TMMCH).Materials and Methods: This cross-sectional study was done during a period of 12 months from July 2015 to June 2016. Total 80 specimens were collected based on clinical presentations irrespective of age and sex. The diagnosis was confirmed by microscopic examination using 20% potassium hydroxide (KOH) and culture on Sabouraud’s dextrose agor medium.Results: Out of 80 samples, 31(38.75%) were found positive by culture and 21 (26.25%) were found positive by microscopic method which were also found positive by culture. This study found that most (51.62%) of the dermatophyteinfected cases were in the age group of 21−40 years followed by 41−60 years (29.03%) with male and female distribution 58.06% and 41.94% respectively The maximum number of infections was reported from groin followed by hands/legs and feet.Conclusion: The result of this study shows higher prevalence of dermatophytosis in both genders in this area. An accurate diagnosis can help in proper and effective treatment of dermatophytosis.J Enam Med Col 2018; 8(1): 11-15

The article is devoted to the microscopic method of research in modern obstetrical and gynecological practice. Special attention is paid to the value of the method, the diagnostic significance of the results of the examination. There is described the preparation for the study, the purpose and methodology of the exercise. The attached photos show in detail all the stages of sampling a free discharge from the urogenital tract.

The present study conducted on thirty patients with scabies. Blood samples were collected before and after treatment in addition to 30 controls, that used in immunological study to determine the effect of infection with scabies on concentrations of IL-31, β-endorphin, PAR2 (Protease-activated receptor 2) before and after treatment. The sim of this study is to examine the immunological response in patients after becoming infected with scabies. All these cases were collected from Healthy centers of camps Kaparto1and 2, Sharyia and Khanki in Duhok governorate and diagnosed by clinical examination and light microscopic method for examining the Sarcoptes scabiei mite or its eggs, at laboratories of these centers from March 2019 to about 30 January 2020. Immunological parameter including interleukin-31, β-endorphin, PAR2 (Protease-activated receptor 2) were assessed using ELISA. Results of study showed that IL-31, β-endorphin, PAR2 (Protease-activated receptor 2) significantly increasing (P

The present study conducted on thirty male patients who were infected with scabies. Blood samples collected before and after treatment in addition to 30 male controls, were used in immunological study to determine the effect of infection with scabies on concentrations of Tropomyosin, Paramyosin, Melatonin, and Cathepsin-D before and after treatment. All these cases were collected from Healthy centers of camps Kaparto1and 2, Sharyia and Khanki in Duhok governorate and diagnosed by light microscopic method for examining the Sarcoptes scabiei mite or its eggs, at laboratories of these centers from March 2019 to about 30 January 2020. The results of study revealed the levels of Tropomyosin, Paramyosin, Cathepsin-D were significantly increasing (P

Tuberculosis is still an important health problem in Indonesia, being the third place in the world. The diagnosis could be done bydirect microscopic method of slides stained with Ziehl Neelsen. The finding of AFB could be improved by decontaminating the specimensto provide a concentrated sample. To analyze the findings of AFB through microscopy and concentrated smear methods, comparing ofeach to bacteriological culture. Diagnosis was done on 148 patients during May to September 2005, using microscopic method (directmicroscopy and concentrated smear) and culture. Analysis of results was done by using SPSS for Window v. 11.5. Of the 148 specimens,15 (10.1%) and 55 (37.2%) positive AFB were found in direct microscopy and concentrated smears respectively. The sensitivities indirect microscopy and concentrated smears were 22% and 64.8%, while the specificities were 96.8% and 78.7%, positive predictivevalues (PPV) were 80% and 63.6%, negative predictive values (NPV) 68.4% and 79.5%, accuracy values were 69.5% and 73.6%. Thespecificity and PPV were higher in direct microscopic method, while positive value, sensitivity, NPV and accuracy value were higher indecontamination, concentrated sample method. Concentrated sample method will increase the accuracy of the finding of Ziehl NeelsenstainedAFB so that this method could be assumed better for use.

Background: Malaria and hepatitis B virus (HBV) are two diseases of major medical importance in Nigeria. Methods: The study determined the preponderance of malaria, HBV and their co-infection among 600 outpatients of three major hospitals in Port Harcourt, Nigeria, using standard microscopic method and rapid serology assay diagnostic kit. Results: Total preponderance rate of malaria, HBV, and co-infection was observed to be 41.0%, 6.0%, and 4.33%, respectively. The female gender was more infected with malaria (44.67%) than the male counterparts (37.54%), while the males were more infected with HBV (7.12%) and co-infection (5.18%) than the females with 4.8% and 3.44%, respectively. The infection rate between genders was not statistically significant (P > 0.05). The prevalence of malaria was statistically high among the age group of 21 - 30 years (55.66%) (P < 0.05). HBV and co-infection prevalence rate was statistically the highest among the age group of 21 - 30 years with 11.76% and 10.3% rates of infection, respectively. Conclusions: The high rate of malaria prevalence and the presence of HBV and co-infection among the study subjects imply that these infections are threats to the human health and strategies should be adopted to help combat and control these infections.

Background and aims Diagnosis of thrombotic microangiopathy (TMA) relies on microscopic schistocyte determination by an experienced microscopist. In addition, schistocytes can be found in non‐TMA–related disorders such as thalassaemia. We aimed to compare the accuracy of the automated haematology analyser Sysmex XN‐3000 for schistocyte detection, to that of the microscopy approach, in patients suspected of having schistocytosis. Methods Consecutive blood samples were collected between April 2016 and March 2017 at Siriraj Hospital, Mahidol University, Bangkok, Thailand. Specimens were collected from adults with suspected TMA or with thalassaemia trait and/or disease. All blood samples were examined by both microscopy and the analyser. Samples were considered to be positive for schistocytes (ie, schistocytosis) if they had a schistocyte count ≥1% by microscopy. The analyser's ability to determine schistocytosis was assessed by receiver operating characteristic (ROC) curve. Sensitivity, specificity, positive (PPV), and negative predictive value (NPV) of an appropriate cut‐off point were calculated, with manual microscopy as the standard. Quantitative agreement in schistocyte counts between the two approaches was assessed using 95% limits of agreement, Bland‐Altman plots, intraclass correlation coefficient, and concordance correlation coefficient. Results Ninety‐seven blood samples (62 suspected TMA and 35 thalassaemia) were collected. ROC curve analysis of the analyser for determining schistocytosis showed an area under the curve of 0.803 (95% confidence interval, 0.689‐0.917, P < 0.001). A cut‐off point of 0.6% yielded 86.1% sensitivity, 77.8% specificity, 94.4% PPV, and 56.0% NPV. The automated schistocyte count did not quantitatively agree with schistocyte counts by microscopy, neither in all blood specimens (mean of difference: −1.09; 95% limits of agreement, −11.9 to 9.7) nor in the subgroups (TMA, −0.88; 95% limits of agreement, −6.60 to 4.84; thalassaemia, −2.4; 95% limits of agreement, −14.10 to 9.30). The differences in the estimation of fragmented red blood cells between the methods tended to increase at higher schistocyte counts. Conclusion Sysmex XN‐3000 can be used for qualitative measurement of schistocytosis, but should not be used as a quantitative tool for schistocyte counting. Improvements are needed before this analyser's schistocyte detection feature can be recommended for use in clinical practice.

Summary Introduction The CellaVision Advanced Red Blood Cell (RBC) Software Application is a new software for advanced morphological analysis of RBCs on a digital microscopy system. Upon automated precharacterization into 21 categories, the software offers the possibility of reclassification of RBCs by the operator. We aimed to define the optimal cut-off to detect morphological RBC abnormalities and to evaluate the precharacterization performance of this software. Methods Thirty-eight blood samples of healthy donors and sixty-eight samples of hospitalized patients were analyzed. Different methodologies to define a cut-off between negativity and positivity were used. Sensitivity and specificity were calculated according to these different cut-offs using the manual microscopic method as the gold standard. Imprecision was assessed by measuring analytical within-run and between-run variability and by measuring between-observer variability. Results By optimizing the cut-off between negativity and positivity, sensitivities exceeded 80% for ‘critical’ RBC categories (target cells, tear drop cells, spherocytes, sickle cells, and parasites), while specificities exceeded 80% for the other RBC morphological categories. Results of within-run, between-run, and between-observer variabilities were all clinically acceptable. Conclusion The CellaVision Advanced RBC Software Application is an easy-to-use software that helps to detect most RBC morphological abnormalities in a sensitive and specific way without increasing work load, provided the proper cut-offs are chosen. However, evaluation of the images by an experienced observer remains necessary.

Microscopic examination of urine sediment is necessary for evaluation of renal and urinary tract diseases. In this study, we evaluated and compared analytic and diagnostic performances of DIRUI FUS-200 and a new image-based automated urine sediment analyzer Urised 3.A total of 440 urine samples, submitted to our laboratory, were evaluated by two automated urine sediment analyzers and a standardized manual microscopy. Precision, linearity and method comparison studies were performed according to CLSI guidelines.Considering the red blood cell (RBC) and white blood cell (WBC) counts, strong correlations existed between FUS-200 and manual microscopy (r=0.993 vs 0.861), Urised 3 and manual microscopy (r=0.962 vs 0.818), FUS200 and Urised 3 (r=0.961 vs 0.961). Clinical non-concordance ranged from 7% to 14.16% among all methods.The concordance between the analyzers and manual microscopy for WBC was better than that of RBC. The concordance between the two analyzers was better for WBC and RBC, with respect to the manual microscopy. Although the Urised 3, FUS-200 and manual microscopy counts were in agreement; confirmation of the results of automated analyzers with manual microscopy is particularly helpful, for pathological samples with near cut-off values.Böbrek ve idrar yolu hastalıklarının değerlendirilmesinde idrar sedimentinin mikroskobik analizi gereklidir. Bu çalışmada, DIRUI FUS-200’ün ve yeni bir görüntü tabanlı otomatik idrar sediment analizörü olan Urised 3’ün analitik ve diagnostik performanslarını değerlendirdik ve karşılaştırdık.Laboratuvarımıza gönderilen 440 idrar örneği her iki otomatik idrar sediment analiz cihazı ve standart manuel mikroskopi ile değerlendirildi. Kesinlik, linearite ve yöntem karşılaştırma çalışmaları CLSI kılavuzlarına göre yapıldı.Eritrosit (RBC) ve lökosit (WBC) sayımları düşünüldüğünde; FUS-200 ile manuel mikroskopi arasında (sırasıyla r=0,993 ile 0,861); Urised 3 ve manuel mikroskopi arasında (sırasıyla r=0,962 ve 0,818); FUS200 ve Urised 3 arasında (sırasıyla r=0,961 ve 0,961) güçlü korelasyon vardı. Klinik uyumsuzluk tüm yöntemler arasında 7% ile 14,16% arasında değişmekteydi.WBC için analizörler ve manuel mikroskopi arasındaki uyum, RBC’den daha iyiydi. Analizörler arasındaki uyum WBC’de ve RBC’de, manuel mikroskopiye göre daha iyiydi. Her ne kadar Urised 3, FUS-200 ve manuel mikroskopi sonuçları belli bir uyum içinde olsa da, otomatize yöntemlerin sonuçlarının manuel mikroskopi ile teyit edilmesi, özellikle de kesme değerlerine yakın patolojik örnekler için faydalıdır.

Background/aims: Ankaferd blood stopper (ABS) is a mixture of certain ratios of 5 different plant roots (Thymus vulgaris, Glycyrrhiza glabra, Vitis vinifera, Alpinia officinarum, and Urtica dioica). The aim of this study is to evaluate the histopathological effects of ABS on articular cartilage in vitro. Materials and methods: Twenty-one albino Sprague Dawley rats were randomly allocated to 3 groups: 0.1 mL of saline was injected in the first group, 0.1 mL of ABS was injected in the second group, and 0.1 mL of blood and 0.1 mL of ABS were injected in the third group. One month later all rats were sacrificed. Specimens were obtained for histopathological evaluation. Results: Significant results were detected in the groups with respect to International Cartilage Repair Society and synovial proliferation scores (P < 0.05 and P < 0.01). According to inflammatory cell infiltration and fibrin formation scores, there was no significant difference between group 1 and group 2 (P < 0.01), although there was significant difference between group 3 and the other groups (P > 0.05). Conclusion: ABS and hemarthrosis had toxic effects on knee cartilage. The side effects were increased with the combination of hemarthrosis and ABS. As a result, ABS had unexpected effects on experimental hemarthrosis.

Objective: To assess the efficacy of the combined medication Vagiferon® for topical application, containing antimicrobial, antifungal, antiviral and immunomodulatory components in the treatment of non-pregnant women with bacterial vaginosis. Study design: This paper presents the experience of use of the medication Vagiferon® in the treatment of 29 non-pregnant women aged 22-45 with bacterial vaginosis. The medication Vagiferon® was administered vaginally as 1 suppository 1 time per day at bedtime for 10 days. The patients underwent a standard pelvic exam, which included the study of their vaginal microflora content by the microscopic method (Gram staining). The study was undertaken before therapy, in 14 days after its start, and in three months after its completion. Results: According to our observations of patients with bacterial vaginosis, in 93,1 percent of cases complete clinical remission was noted. Microbiological efficacy of the treatment was 86,2 percent. The signs of disease recurrence as well as any side effects or rejection of the use of the medication were not registered. Conclusion: Based on the combination of high efficacy and safety of Vagiferon®, this medicine can be recommended for wide use in the gynaecological practice.

In assessing abstinence from drug or alcohol abuse, hair analysis plays an important role. Cosmetic hair treatment influences the content of deposited drugs which is not always detectable during analysis. Since oxidation of melanin leads to an increase in fluorescence, a microscopic method was developed to distinguish natural from cosmetically treated hair. For validation, natural hair samples were treated with different types of cosmetics and inspected by fluorescence microscopy. Hair samples from 20 volunteers with documented cosmetic treatment and as a proof of concept 100 hair samples from forensic cases were analyzed by this method. Apart from autofluorescence with excitation at 365 nm, no obvious fluorescence was observed in untreated hair samples. Tinting and a natural plant product had no influence on fluorescence, but dyeing procedures including oxidation led to a marked increase in fluorescence. Proof of cosmetic treatment was achieved in hair samples from the 20 volunteers. In 100 forensic cases, 13 samples were characterized as oxidatively treated, which was in accordance with the respective disclosure except for one case where treatment was not admitted. This fluorescence microscopic procedure proved to be fast, easy, and reliable to identify oxidatively treated hair samples, which must be considered especially in evaluating cases of negative drug results. Copyright © 2015 John Wiley & Sons, Ltd.

Background/Aim. The Nugent?s score is still the gold standard in the great majority of studies dealing with the assessment of vaginal flora and the diagnosis of bacterial vaginosis (BV). The aim of this study was to show that the analysis of Gram-stained vaginal samples under microscope at the magnification of ?200 (a novel microscopic method - NMM), as a fast and simple tool, easily applicable in everyday practice, better reflects complexity of vaginal microflora than the Nugent?s methodology (?1000). Methods. Gramstained vaginal smears from 394 asymptomatic pregnant women (24-28 week of pregnancy) were classified according to the Nugent?s microscopic criteria (immersion, magnification ?1000). The smears were then reexamined under immersion but at magnification ?200. All samples were classified into 6 groups according to semiquanititative assessment of numbers (cellularity) and the ratio of rod (length < 1.5 ?m) and small bacterial (< 1.5 ?m) forms: hypercellular (normal full - NF), moderately cellular (normal mid - NM), hypocellular (normal empty - NE), bacterial vaginosis full (BVF), bacterial vaginosis mid (BVM), and bacterial vaginosis empty (BVE). Also yeasts, coccae, bifido and lepto bacterial forms as well polymorphonuclear (PMN) leukocytes were identified. Results. According to the Nugent?s scoring, BV was found in 78, intermediate findings in 63, and yeasts in 48 patients. By our criteria BV was confirmed in 88 patients (37 BVF, 24 BVM, and 27 BVN). Generally, both tools proved to be highly concordant for the diagnosis of BV (Lin?s concordance correlation coefficient = 0.9852). In 40% of the women mixed flora was found: yeasts in 126 (32%), coccae in 145 (37%), bifido forms in 32 (8%) and lepto forms in 20 (5%). Almost a half of BV patients had also yeasts (39/88). Elevated PMN numbers were found in 102 (33%) patients with normal and in 36 (41%) women with BV. Conclusion. The newly described methodology is simpler to apply and much better reflects diversity of vaginal microflora. In this way it may be more valuable to molecular biologists and their attempts based on quantitative polymerase chain reaction (PCR) to define formulas for molecular diagnosis of bacterial vaginosis.

Background Fully automated urine analyzers now play an important role in routine urinalysis in most laboratories. The recently introduced UriSed 3 has a new automated digital imaging urine sediment analyzer with a phase contrast feature. The aim of this study was to compare the performance of the UriSed 3 and UX-2000 automated urine sediment analyzers with each other and with the results of the manual microscopic method. Methods Two hundred seventy-seven (277) samples of leftover fresh urine from our hospital's central laboratory were evaluated by two automated urine sediment analyzers—UriSed 3 and UX-2000. The results of urine sediment analysis were compared between the two automated analyzers and against the results of the manual microscopic method. Results Both devices demonstrated excellent agreement for quantitative measurement of red blood cells and white blood cells. UX-2000 had a lower coefficient correlation and demonstrated slightly lower agreement for squamous epithelial cells. Regarding semiquantitative analysis, both machines demonstrated very good concordance, with all applicable rates within one grade difference of the other machine. UriSed 3 had higher sensitivity for small round cells, while UX-2000 showed greater sensitivity for detecting bacteria and hyaline casts. UriSed 3 demonstrated slightly better specificity, especially in the detection of hyaline and pathological casts. Conclusions Both instruments had nearly similar performance for red blood cells and white blood cells measurement. UriSed 3 was more reliable for measuring squamous epithelial cells and small round cells, while the UX-2000 was more accurate for detecting bacteria and hyaline casts.

Background: Periodontitis is caused by specific microorganisms or group of specific microorganisms. Spirochetes have been implicated to play a role in the pathogenesis of the periodontal disease. Objectives: The aim is to detect the prevalence of total spirochetes in patients with healthy periodontium, gingivitis, and periodontitis using microscopy and polymerase chain reaction (PCR) techniques. And to compare microscopy (silver nitrate staining) technique and PCR technique in the detection of spirochetes. Subjects and Methods: A total of 150 patients were selected and divided into healthy, gingivitis, and periodontitis groups with 50 patients in each group. Subgingival plaque samples were collected using sterile Gracey curettes. The plaque samples were analyzed for the presence of spirochetes using microscopy and PCR techniques. Statistical analysis for the prevalence of spirochetes was performed using the Chi-square test. The sensitivity and specificity of the microscopy and PCR techniques were compared. Results: The prevalence of spirochetes increased from healthy to gingivitis to periodontitis patients. The prevalence of spirochetes was statistically significantly higher in the periodontitis group as compared to the healthy and gingivitis group. PCR was more sensitive in detecting spirochetes than microscopy, but that microscopy showed slightly greater specificity than PCR. Conclusion: There is a strong association between the presence of spirochetes with periodontal disease as well as its severity. The microscopic method by silver nitrate staining can be a very useful, simple, and rapid screening method for the evaluation of oral spirochetes, but PCR technique may serve as more sensitive indicator of subjects at risk for periodontal disease, thus allowing preventive measures.

Introduction: The objective of the study was to comparatively evaluate the porosity distribution in the root canals obturated using the single-cone technique and thermoplasticized technique. Materials and Methods: A total of 42 permanent single-rooted human extracted teeth were divided equally into two groups based on the technique employed: single-cone technique and thermoplasticized technique. Following postobturation restoration, void volume was analyzed by cone-beam computed tomography (CBCT) and void location was evaluated by microscopic method at three levels: coronal, middle, and apical. Independent Student't-test and Chi-square test were used for analyzing the obtained results. Results: On comparing the void volumes as seen in CBCT images between the two groups, the thermoplasticized technique showed greater but insignificant void volume (P = 0.18). When the association of the location of the void with the void area percentage was analyzed between the two techniques, significant associations were seen in the middle (P = 0.016) and the apical levels (P = 0.031). Conclusion: The single-cone technique showed a comparable seal with the thermoplasticized technique. Voids were predominantly seen at the sealer–cone interface and sealer–root canal wall interface for single-cone technique and in between the condensed gutta-percha for thermoplasticized technique.

As parasitological or microscopic method is the gold standard and the best method for diagnosis of malaria, so fixed Geimsa-stained blood slides in the form of thick and thin blood smears are the most important data collections of malaria, especially historical slides. The parasites are dead but their DNA is valuable for many molecular biologic researches. A simple and efficient method for catching and extraction malaria parasites' DNA with a desired yield from dried and stained blood on slides is the first and major step. Introduction of an applicable, efficient and inexpensive DNA catching method and assessment of its performance in following molecular applications was the main objective of present study.

Hemoglobin C (HbC) is one of the most prevalent hemoglobinopathies worldwide along with HbS (sickle cell) and HbE. HbC disease (HbCC) produces mild symptoms but is a life-threatening disorder if inherited with HbS (HbSC). HbS and HbC are most prevalent in sub-Saharan countries underequipped to diagnose its presence through gold standard methods like electrophoresis, HPLC or isoelectric focusing. This study aims to create a simple and inexpensive method to identify the presence of HbC in human blood using limited resources with the potential to determine zygosity. HbC crystals that form when blood is incubated in a hypertonic salt solution become visible microscopically when stained with New Methylene Blue. The method was optimized by modifying the salt type, salt concentration, incubation time, and incubation temperature. The optimized method incubates RBCs in a 5x Dulbecco9s phosphate buffered saline (DPBS) at 37°C for 4 hours. Blood samples with the HbSC genotype yielded between 600-750 HbC crystals for every 1000 RBCs counted while negative control samples (AA) do not produce HbC crystals. Preliminary results indicate that AS samples produce between 80-150 HbC crystals/1,000 RBCs. Future studies will include testing of blood samples of AC, CC, SC, and AA genotypes to determine if the number of crystals that form using this method will differentiate genotype. If so, this inexpensive, simple, and relatively rapid method can be used to identify patients with HbC and determine genotype in underdeveloped countries where HbC is prevalent.

The objective of this study was to develop a simple, inexpensive, and rapid confirmatory test using a sodium metabisulfite microscopic method to distinguish AS from SS genotype in patients testing positive for hemoglobin S. Equal volumes of de-identified EDTA blood and 2% sodium metabisulfite were mixed, placed on a microscope slide with coverslip, and observed for sickling at 30-minute intervals over 3 hours. Sickle cells were enumerated/200 RBCs under 100x oil immersion and placed into 4 Likert categories (1+-4+) based on degree of sickling. In AS samples, 2+ and 3+ sickle cells rose most rapidly while 4+ sickle cells showed the steepest rise in the SS samples. Based on these data the number of 4+ sickle cells were counted every 30-minutes over 3 hours in 5 AS and 28 SS samples at 37 degrees Celsius. Mean number of 4+ sickle cells in AS samples were 4.75/200 at 2 hours and 17.75/200 at 3 hours whereas SS samples yielded 78.29/200 at 2 hours and 115.43/200 at 3 hours. Two-hour incubation showed a statistical difference (P=0.00024) between groups in the shortest time and may be used to distinguish SS from AS genotypes. More testing is needed to determine cutpoints to distinguish genotype.

Objective This study surveyed the prevalence of cryptosporidiosis in patients who were treated in Dumlupinar University Medical Faculty Hospital for various gastrointestinal complaints such as abdominal pain, nausea, vomiting and diarrhea. Water samples were taken from various homes and fountains. Methods Stool samples were investigated for the presence of oocysts with Kinyoun's carbol fuchsin staining method. After miscroscopic analysis, samples were preserved. These frozen samples were thawed promptly for antigenic diagnosis (Ridascreen® , R-Biopharm AG, Darmstadt Germany). Thirty units of ten-liter water samples were filtered by a vacuum pump (45 μm). Filtered materials were stained through Kinyoun's carbol fuchsin staining method and examined. Results Stool samples taken from a total of 146 patients were examined for oocyst presence and 4 individuals (2.7 percent) were found to be positive. Elisa tests of the same samples proved 5 individuals (3.4 percent) as positive for specific antigens. Only one water sample was diagnosed as positive for oocysts. Conclusion Cryptosporidiosis was found to be present in 3.4 percent of this group of people, and the distribution of infection among subjects was understood to be not statistically meaningful by gender, age and identification methods (p > 0.05). Finding of oocysts in only one sample out of 30 (3.3%) led to a need for reconsideration of other causal factors in the epidemiology.

Transfusion reactions in the thrombocyte concentrate (TC) manual is often associated with the storage time due to the release of cytokines by leukocytes during storage. The results showed the majority of transfusion reactions include fever/febrile non-haemolytic transfusion reaction (FNHTR). The quality control of blood components is necessary for the determination a component of blood in the bag already qualified for or not for transfusion. The aims of this study is to determine the residual leukocytes in the TC manual of Blood Donor Unit (UDD) at PMI Semarang with the shelf life of 3−5 days in compliance with the requirements of TC quality criteria of the European Union Council (EU).A Descriptive study is conducted at the Blood Donor Unit (UDD) PMI Semarang. The TC sample consist of 30 bags and manuals from the transfusion bag, which has been stored between 3−5 days. The examination of the residual leukocytes from the TC bag is carried out by manual microscopic method as well as the counting. The results obtained from the O blood group are 13 bags (43%) of TC, the B blood group seven (7) bags (33%), A blood group two bags (7%), and AB five (5) bags (17%). The range of the residual values of white blood cells in the TC manual is from 0 up to 0.0086í—109/unit. According to the EU Council of leukocytes in the TC control value is

In vitro susceptibility tests provide information on the intrinsic response of Plasmodium vivax to antimalarials, free from confounding factors such as host immunity or relapse. This study examined the utility of radioisotope and PicoGreen assays as alternatives to the traditional microscopic examination for assessing response of P. vivax to antimalarial drugs. There was no significant difference in the mean chloroquine IC50 of P. vivax (n = 40) as determined by the microscopic (33.4 ng/ml), isotopic (33.6 ng/ml), and PicoGreen (39.1 ng/ml) assays, respectively (F = 0.239, df = 2, 51, and p = 0.788). However measurement of IC50s by the microscopic method was slightly more successful in producing valid assays (57%), compared to the isotopic (32.5%) and PicoGreen (45.5%) methods. In a paired comparison of 20 fresh and cryopreserved isolates as examined by the microscopic method, there were no significant differences between the mean IC50 responses (T = 1.58, df = 15, and p = 0.34). Detailed methodologies for the short time culture of field and cryopreserved P. vivax are described. Although the microscopic in vitro assay provides a useful method for characterizing the drug susceptibility phenotype of P. vivax isolates, its utility is limited by a laborious methodology and need for highly skilled microscopists. Future efforts should focus on further development of high throughput assays such as the PicoGreen assay as described in this study.

Ex vivo antimalarial sensitivity testing in human malaria parasites has largely depended on microscopic determination of schizont maturation. While this microscopic method is sensitive, it suffers from poor precision and is laborious. The recent development of portable, low-cost cytometers has allowed us to develop and validate a simple, field-optimized protocol using SYBR green and dihydroethidium for the accurate and objective determination of antimalarial drug sensitivity in freshly isolated Plasmodium vivax and Plasmodium falciparum .

Background/aims: Epidermal thickness is frequently measured by light microscopy. The preservation and staining methods may alter the proportions of the specimens and thereby influence the measurements. The aim of the present study was to describe: 1) a standardized light microscopic method to quantify the thicknesses of the stratum corneum and the cellular epidermal layers, 2) the variations according to preparation and staining techniques, and 3) the observer variability. Methods: One hundred and sixty skin biopsies from 67 human volunteers were included. The cellular epidermis and the stratum corneum were estimated in sections preserved by freezing and subsequent preparation with cryostat or formalin-paraffin techniques. The slides were stained with haematoxylin-eosin or er-ythrocin, and thicknesses of the stratum corneum and the cellular epidermis were measured by a calibrated ruler and an ocular grid, respectively. Results: The formalin fixation gave slightly higher values for the cellular epidermis than the cryostat technique. In comparison to erythrocin staining, haematoxylin-eosin gave a significantly thinner stratum corneum. No significant inter- or intra-observer variation was found for the thickness of the stratum corneum assessed twice by two experienced observers. However, the two observers differed slightly from each other on the thickness of the cellular epidermis. Conclusion: It is found that thickness measurements of the stratum corneum and the cellular epidermis are reliably performed on cryostatic cut sections stained with haematoxylin-eosin.

The villous tree of the human placenta is a complex three-dimensional (3D) structure with branches and nodes at the feto-maternal border in the key area of gas and nutrient exchange. Recently we introduced a novel, computer-assisted 3D light microscopic method that enables 3D topological analysis of branching patterns of the human placental villous tree. In the present study we applied this novel method to the 3D architecture of peripheral villous trees of placentas from patients with intrauterine growth retardation (IUGR placentas), a severe obstetric syndrome. We found that the mean branching angle of branches in terminal positions of the villous trees was significantly different statistically between IUGR placentas and clinically normal placentas. Furthermore, the mean tortuosity of branches of villous trees in directly preterminal positions was significantly different statistically between IUGR placentas and clinically normal placentas. We show that these differences can be interpreted as consequences of morphological adaptation of villous trees between IUGR placentas and clinically normal placentas and may have important consequences for the understanding of the morphological correlates of the efficiency of the placental villous tree and their influence on fetal development.

Diffuse gallbladder wall thickening is a nonspecific alteration caused by both intrinsic and extracholecystic diseases. The incidence of gallbladder illness in our country is increasing day by day. Details morphological and histomorphological knowledge is essential for proper diagnosis & management of the gallbladder diseases. This is a cross sectional descriptive type of study which was carried out in the Department of Anatomy, Sir Salimullah Medical College, Dhaka from July 2010 to June 2011. The number of sample was 62 postmortem human gallbladder which were collected from unclaimed dead bodies of the morgue of Dhaka Medical College and Sir Salimullah Medical College. The samples were divided into three different age groups. Morphological study was carried out on all samples by fine dissection method and histomorphological study was carried out on 18 relatively fresh samples by light microscopic method. There was statistically significant difference found when values were compared between different age groups. This study has revealed an age related change in the thickness of the gallbladder wall. DOI: http://dx.doi.org/10.3329/jdnmch.v18i1.12238 J. Dhaka National Med. Coll. Hos. 2012; 18 (01): 34-38

The purpose of this study was to investigate usefulness of the pine needles extract on Atopic Dermatitis(AD)-like skin lesions. To investigate the effect of pine needles extract in vivo, atopic dermatitis (NC/Nga) mice using DNCB (2.4-Dinitrochlorbenzene) was used. NC/Nga mice were challenged with DNCB during 2 weeks to develope AD-like skin lesion. After that, pine needles extract was applied to AD-like skin lesion on the backs of the NC/Nga mice during 3 weeks. The efficacy of pine needles extract in the NC/Nga mice was evaluated by measurement of the skin lesion severity(NC mouse score), the serum IgE level, epidermal thickness changes, and mast cell number. Blood was collected from the retro-orbital area and the level of IgE in the blood was measured. The epidermal thickness and mast cell number were observed by microscopic method after H&E stain. The serum IgE levels were decreased after treatment with pine needles extract. The epidermal thickness and mast cell number were decreased after treatment with pine needles extract. To conclude, the topical application of pine needles extract suppressed the progression of AD-like skin lesion.

Although many variables that skeletal biologists examine have been standardized, the actual techniques used to collect these data from bone thin sections vary. This project compares different methods of obtaining data (relative cortical area values) for histomorphometric research. One visual and three digital methods of histomorphometric data collection are compared: (i) Merz microscopic eyepiece counting reticule, (ii) flatbed scanner, (iii) overlaying multiple images of a thin section, and (iv) digital SLR camera with macro settings. The discussion includes a comparison of usability factors such as cost, time, user-experience, and ease-of-use, which vary for each method. Values from the different methods are compared using ANOVA tests to evaluate inter-method, inter-observer, and intra-observer variability. Intra-observer error was greater for the microscopic method, although the error values are concomitant with experience. We found no statistically significant differences between the four methods examined, but certain caveats must be addressed when these methods are used. Language: en

Tooth cementum annulation, the microscopic method for the determination of an individual’s age, gives results that are highly correlated with the chronological age of an individual. Nevertheless, this method is still rarely used for age estimation in archaeological populations. In this study, using the tooth cementum annulation method, teeth of 21 individuals of the Djerdap anthropological series, dated to a period from the 10th to the 6th millennium BC, were analysed. The obtained data are important for overcoming some methodological issues in anthropology of the Danube Gorges, as well as with the precise age estimation of old individuals and with the assessment of age in cases where the skeletal material has been very poorly preserved. The only obstacle to the full application of the tooth cementum annulation method is the taphonomy changes of tooth cementum which were detected on several teeth in this study. [Projekat Ministarstva nauke Republike Srbije, br. 47 001: Bioarchaeology of ancient Europe - people, animals and plants in Serbian prehistory]

Sputum smear microscopy is an easy, inexpensive, and rapid method for detecting tubercle bacilli when there are more than 10,000 bacilli/ml in the original sputum. Furthermore, because the microscopic method provides not only quantitative, but also qualitative information, such as the shape of bacilli, it has remained significant. We have previously developed and reported panel test slides made from polyacrylamide-based artificial sputum (PBAS) mixed with both cultured THP-1 cells and nonpathogenic mycobacteria. In this paper, we report an improved preparation method for PBAS for panel test slides that provides a simplified method and enhanced availability with high consistency in each grade and in which only negative PBAS is prepared from polyacrylamide and cultured THP-1 cells and mixed with graded formalin-fixed Mycobacterium tuberculosis solution (FFTBS) containing oral flora and Pseudomonas aeruginosa on the slides. In the smears prepared using this improved method, the numbers (average ± standard deviation [SD]) of acid-fast bacilli (AFB) in 300 fields (2- by 3-cm smear) in eight smears of each grade ranged from 5 to 9 (6.4 ± 1.4), from 59 to 88 (74.6 ± 10.0), from 503 to 912 (705.0 ± 145.7), and from 1,819 to 3,256 (2133.3 ± 478.0) in ±, +, ++, and +++ smears, respectively. In addition, this preparation method provided high similarity to the microscopic appearance of bacilli and background seen in the actual patient sputum, with high feasibility. These results revealed that our new PBAS had high authenticity in the appearance and consistency in each grade, which could make it valuable as a reliable artificial sputum for the training of microscopists.