Practical tips to using formalin-fixed paraffin-embedded tissue archives for molecular diagnostics in a South African setting

Background: Formalin-fixed paraffin-embedded (FFPE) tissue archives in hospitals, biobanks, and others offer a vast collection of extensive, readily available specimens for molecular testing. Unfortunately, the use of tissue samples for molecular diagnostic applications is challenging; thus, the forensic pathology FFPE tissue archives in Africa have been a largely unexploited genetic resource, with the usability of DNA obtainable from these samples being unknown.Intervention: The study, conducted from January 2015 to August 2016, determined the usefulness of FFPE tissue as a reliable source of genetic material for successful post-mortem molecular applications and diagnostics. Formalin-fixed paraffin-embedded tissue samples were collected and archived from autopsies conducted over 13 years in the forensic medicine department of the University of Pretoria (Pretoria, South Africa). Deoxyribonucleic acid from FFPE tissue samples and control blood samples was amplified by high-resolution melt real-time polymerase chain reaction before sequencing. The procurement parameters and fixation times were compared with the quantity and quality of the extracted DNA and the efficiency of its subsequent molecular applications.Lessons learnt: This study has shown that FFPE samples are still usable in molecular forensics, despite inadequate sample preparation, and offer immense value to forensic molecular diagnostics.Recommendations: FFPE samples fixed in formalin for more than 24 h should still be used in molecular diagnostics or research, as long as the primer design targets amplicons not exceeding 300 base pairs.

Evaluation du comportement tissulaire péri implantaire après temporisation immédiate unitaire antérieure maxillaire / Evaluation of peri implant tissue behaviour after immediate provisionalization single-tooth implant in anterior maxilla

Introduction. La temporisation immédiate unitaire du secteur antérieur maxillaire aide au raccourcissement des délais thérapeutiques, et contribue à la satisfaction esthétique immédiate quand les conditions anatomiques et chirurgicales le permettent. Cependant, un risque d'échec n'est pas exclu. L'état des tissus péri-implantaires et leurs réactions reflètent le succès ou non de cette thérapeutique. Objectif. Cette étude a pour objectif d'évaluer le comportement des tissus durs et mous dans le secteur antérieur maxillaire, après temporisation immédiate unitaire. Matériel et méthode. Une étude prospective et descriptive a été effectuée sur 44 sites unitaires maxillaires antérieurs, répartis entre les incisives centrales et latérales. Une restauration temporaire a été mise en place immédiatement après chirurgie implantaire en sous-occlusion. Le Pink Esthetic Score (PES) et le white Esthetic score (WES) ont été utilisés pour apprécier le comportement des tissus mous. Le taux de succès de l'ostéointégration et le changement du niveau de l'os marginal ont été analysés. Les tissus durs sont analysés au cône beam computed tomography à 6 et 12 mois. Résultats. Les résultats de cette étude sont : un taux de succès d'ostéointégration de 92.68%, un PES de l'ordre de 8.06 ±1.69 à 36 mois et un WES à 7.87±1.23. Cependant, un gain d'os autour de quelques implants a été observé ainsi qu'une résorption. La moyenne de résorption de l'os marginal péri implantaire est de l'ordre de -0.31mm ± 1.22 à 12 mois. Il ressort de cette étude la mise en évidence de la variable (sexe masculin) comme facteur de risque. Conclusion. La temporisation immédiate a un effet positif sur les tissus mous péri implantaires, permettant ainsi de préparer un profil d'émergence en adéquation avec une restauration d'usage de valeur esthétique. La moyenne de résorption de l'os marginal à 1 an et le taux de succès de l'ostéointégration observés, révèlent l'innocuité de cette temporisation immédiate, une sélection fine des patients ainsi qu'une très bonne préparation des sites à implanter sont prises en considération

Introduction. Due to the poursuit of beauty and the demands of social activities, traditional implant protocols, were modified. The immediate restoration protocol was proposed to reduce the edentulous time and avoid a stage-two operation. despite this avantage , this technique is not without a risk of failure. Quality and behaviour of the tissues around implants, can reflects success or not. Aim. This study investigated in changes the surronding hard and soft tissu outcome of immediate non-occlusal loading for single implants in the maxillary. Materials and Methods. This is a prospective study covered forty-for implants immediatly restored and nonocclusally loaded in a single missing maxillary anterior tooth. The outcome measures were : implant success, esthetic outcome (PES/WES), the outcome of marginal bone loss (MBL) : assesses at 6 mouth and one year after implantation. Result. We achieved an osteointegration success rate of 92.68%. An acceptable PES of the order of : 8.06 +/-1.69 à 36 at 36 months and a WES of : 7.87+/-1.23. We found a bone gain a round some implants and resorptions also. The mean marginal bone loss (MBL) was in the order of : -0.31mm +/- 1.22 at one year. This study shows that the male gender Constitue a risk factor. Conclusion. It appears from this study that :immediate non-occlusal loaded single implant in the anterior maxilla, has a positive effect on hard and soft tissues around implants allowing to prepare an emergence profile in accordance with a definitive restoration of esthetic value.

A comparison study of histochemical staining of various tissues after Carnoy's and formalin fixation

Background: The potential problems related to the use of formalin in histopathology; such as health hazards; deterioration of nuclei acids are well-known. The aim of this study was to evaluate the utilization of a Carnoy's solution fixation in comparison with formalin on subsequent tissue sectioning and histochemical staining. Materials and Methods: Corresponding sections of 25 tissue biopsies of rabbit's different organs were fixed in Carnoy's solution and in 10neutral buffered formalin. Samples were processed using the conventional method and then stained applying five histochemical methods. The degree of the quality of the staining was assessed for each method by scoring system (1-10) depending on comparison of the stained tissue sections with illustrated photomicrographs. Results: For the quality of cutting; the best quality was obtained by Formalin (mean = 4.76) then Carnoy's fixative (mean =3.84). The best quality of Haematoxylin and Eosin staining was obtained by formalin (mean =5.28) then Carnoy's (mean = 4.00). For Alcian blue and Perl's Prussian blue; the best staining qualities were obtained by Formalin (mean = 4.76 and 5.64 respectively) followed by Carnoy's (mean = 2.88 and 3.92 respectively). For periodic Acid Schiff's the best staining quality was obtained following Carnoy's fixation (mean = 4.52) then; the formalin (mean = 3.76). Conclusion: Although; Carnoy's fluid is a safe fixative and can rapidly penetrate the tissues; but it can't be a substitute for formalin

Free Flap Surgery at Mengo Hospital; Uganda - A Review of The First 19 Consecutive Microvascular Free Tissue Transfers

Background: Microvascular free tissue transfer is an important method for reconstructing complex surgical and traumatic defects; allowing single stage reconstruction in most instances. This study reviews the first 19 consecutive free tissue transfer (free flap) reconstructions at Mengo hospital; Department of Plastic and Reconstructive surgery. Methods: A retrospective review of the first 19 surgical reconstructions using microvascular free tissue (free flap) transfer techniques from 2005 to date was done. Results: 10 males and 8 females have had surgical reconstructions with free flaps during this period. Average age of patients was 23yrs; ranging from 7 years to 42 years. The majority of reconstructions were in the head and neck region (n=14; 74Background: Microvascular free tissue transfer is an important method for reconstructing complex surgical and traumatic defects; allowing single stage reconstruction in most instances. This study reviews the first 19 consecutive free tissue transfer (free flap) reconstructions at Mengo hospital; Department of Plastic and Reconstructive surgery. Methods: A retrospective review of the first 19 surgical reconstructions using microvascular free tissue (free flap) transfer techniques from 2005 to date was done. Results: 10 males and 8 females have had surgical reconstructions with free flaps during this period. Average age of patients was 23yrs; ranging from 7 years to 42 years. The majority of reconstructions were in the head and neck region (n=14; 74) followed by lower limb (n=3; 16) and upper limb reconstructions (n=2; 10). Indications for reconstructions using free flaps included extensive loss of scalp; extensive loss of soft tissue following open tibial fractures; bomb blast injuries to the face; Noma; Romberg's disease; ameloblastoma. Overall flap survival rate of 79. Head and neck reconstructions flap survival rate of 83.3; lower limb and upper limb free flap survival rates of 33.3and 100respectively

Joint and Soft Tissue Injection

Injection techniques are helpful for diagnosis and therapy in a wide variety of musculoskeletal conditions. Diagnostic indications include the aspiration of fluid for analysis and the assessment of pain relief and increased range of motion as a diagnostic tool. Therapeutic indications include the delivery of local anaesthetics for pain relief and the delivery of corticosteroids for suppression of inflammation. Side effects are few; but may include tendon rupture; infection; steroid flare; hypopigmentation; and soft tissue atrophy. Injection technique requires knowledge of anatomy of the targeted area and a thorough understanding of the agents used. In this overview; the indications; contraindications; potential side effects; timing; proper technique; necessary materials; pharmaceuticals used and their actions; and post-procedure care of patients are presented

Effects of Cassia occidentalis Aqueous Leaf Extract on Biochemical Markers of Tissue Damage in Rats

Purpose: Among the Hausas of Northern Nigeria; it is claimed by local (traditional) healers that the infusion of C. occidentalis leaves is used as a cure for hepatitis. This study was designed to evaluate the possible negative effects of the aqueous extract of this herb on serum total proteins; albumin; bilirubin; alanine amino transferase (ALT); aspartate amino transferase (AST) and alkaline phosphatase (ALP); as part of a larger study undertaken by these authors to ascertain the hepatoprotective potential of the plant extract. Methods: Fresh leaves of C. occidentalis were extracted with water and screened for phytochemical components. The extract was used for the treatment of 18 albino rats. Serum from these animals was used for the determination of the test parameters. Results: The extract showed hypoproteinaemic effect. ALT; AST and ALP levels were significantly elevated (P 0.05). Conclusion: Hypoproteinaemic effects; and increase in ALT; AST and ALP are indications that the crude extract of C. occidentalis leaves may be slightly toxic as a concoction for liver ailments