Symphytum officinale augments osteogenesis in human bone marrow-derived mesenchymal stem cells in vitro as they differentiate into osteoblasts.

ETHNOPHARMACOLOGICAL RELEVANCE: Mesenchymal stem cells (MSCs) are multipotent stem cells possessing regenerative potential. Symphytum officinale (SO) is a medicinal plant and in homoeopathic literature, believed to accelerate bone healing. AIM OF THE STUDY: This study aimed to determine if homoeopathic doses of SO could augment osteogenesis in MSCs as they differentiate into osteoblasts in vitro. MATERIALS AND METHODS: Bone marrow samples were obtained from patients who underwent bone grafting procedures (n = 15). MSCs were isolated, expanded and characterized by flow cytometry (CD90, CD105). Cytotoxicity of SO was evaluated by MTT assay. Osteogenic differentiation was induced in MSCs with ß-glycerophosphate, ascorbic acid and dexamethasone over 2 weeks. Different homoeopathic doses of SO (MT, 3C, 6C, 12C and 30C) were added to the basic differentiation medium (BDM) and efficiency of MSCs differentiating into osteoblasts were measured by evaluating expression of Osteocalcin using flow cytometry, and alkaline phosphatase activity using ELISA. Gene expression analyses for osteoblast markers (Runx-2, Osteopontin and Osteocalcin) were evaluated in differentiated osteoblasts using qPCR. RESULTS: Flow cytometry (CD90, CD105) detected MSCs isolated from bone marrow (93-98%). MTT assay showed that the selected doses of SO did not induce any cytotoxicity in MSCs (24 hours). The efficiency of osteogenic differentiation (2 weeks) for different doses of Symphytum officinale was determined by flow cytometry (n = 10) for osteoblast marker, Osteocalcin, and most doses of Symphytum officinale enhanced osteogenesis. Interestingly, gene expression analysis for Runx-2 (n = 10), Osteopontin (n = 10), Osteocalcin (n = 10) and alkaline phosphatase activity (n = 8) also showed increased osteogenesis with the addition of Symphytum officinale to BDM, specially mother tincture. CONCLUSIONS: Our findings suggest that homoeopathic dose (specially mother tincture) of Symphytum officinale has the potential to enhance osteogenesis.

Serum and blister-fluid elevation and decreased epidermal content of high-mobility group box 1 protein in drug-induced Stevens-Johnson syndrome/toxic epidermal necrolysis.

BACKGROUND: High-mobility group box 1 (HMGB1) is a damage-associated molecular-pattern protein. Stevens-Johnson syndrome (SJS)/toxic epidermal necrolysis (TEN) are serious, immune-mediated skin-blistering conditions. OBJECTIVES: To determine serum and/or blister-fluid total HMGB1 levels in SJS/TEN cohorts, and HMGB1 expression in formalin-fixed, paraffin-embedded (FFPE) SJS/TEN skin vs. healthy and maculopapular exanthema (MPE) skin. Methods Serum HMGB1 was quantified in Malawian nevirapine-induced hypersensitivity, Taiwanese SJS/TEN and Spanish SJS/TEN cohorts. FFPE skin (healthy skin, MPE, SJS/TEN) was stained and assessed for HMGB1 expression. RESULTS: Serum total HMGB1 was not significantly elevated in patients with nevirapine-induced SJS/TEN (3·98 ± 2·17 ng mL-1 ), MPE (3·92 ± 2·75 ng mL-1 ) or drug reaction with eosinophilia and systemic symptoms (4·73 ± 3·00 ng mL-1 ) vs. tolerant controls (2·97 ± 3·00 ng mL-1 ). HMGB1 was significantly elevated in Taiwanese patients with SJS/TEN, highest during the acute phase (32·6 ± 26·6 ng mL-1 ) vs. the maximal (19·7 ± 23·2 ng mL-1 ; P = 0·007) and recovery (24·6 ± 25·3 ng mL-1 ; P = 0·027) phases. In blister fluid from Spanish patients with SJS/TEN, HMGB1 (486·8 ± 687·9 ng mL-1 ) was significantly higher than in serum (8·8 ± 7·6 ng mL-1 ; P <0·001). Preblistered SJS/TEN skin showed decreased epidermal nuclear HMGB1 expression in upper epidermis vs. healthy or MPE skin but retained basal/suprabasal expression. CONCLUSIONS: Epidermal HMGB1 expression was decreased in SJS/TEN skin. Retained basal/suprabasal epidermal HMGB1 expression may exacerbate localized injury in SJS/TEN.

Hemophagocytosis on bone marrow aspirate cytology: single center experience in north himalayan region of India.

BACKGROUND: The differentiation of hemophagocytic lymphohistiocytosis (HLH) with other causes of hemophagocytosis is essential as HLH is life-threatening condition and requires definite clinical and biochemical criteria for its diagnosis. AIM: The present study was conducted to study hemophagocytosis on bone marrow aspirates and to observe if there is any difference on bone marrow examination between HLH and non HLH cases showing hemophagocytosis. MATERIALS AND METHODS: The study reviewed all the cases of bone marrow aspirate for evidence of hemophagocytosis in which at least three smears and 500 nucleated cells were observed for every case. The cases were provisionally diagnosed as HLH according to the proposed HLH diagnostic criteria, 2009. RESULTS: A total of 80 cases showed hemophagocytosis in the present study with infections followed by HLH being the most common cause. Pancytopenia and erythroid hyperplasis were common hematological presentation. Moderate to severe hemophagocytosis was observed in HLH cases while mild grade in non HLH cases. CONCLUSION: The study concludes that hemophagocytosis even if observed in single cell should always be documented in the bone marrow reports. It may be the only indicator of subtle infection in the marrow. Pancytopenia and higher grade of hemophagocytosis on bone marrow examination may be helpful in establishing an early differentiation of HLH and non HLH cases.

Ultrasound measurement for abdominal aortic aneurysm screening: a direct comparison of the three leading methods.

OBJECTIVES: Ultrasound (US) is non-invasive and cost-effective for screening abdominal aortic aneurysms (AAAs) but there is no universally accepted method to measure the aortic diameter. This study evaluates the accuracy, reproducibility, and repeatability of three methods: inner-to-inner (ITI), leading-to-leading edge (LTL), and outer-to-outer (OTO). The secondary objective of this study was to determine whether aneurysm size or grade of operator had any effect on either intra- or inter-observer variability. METHODS: Fifty static US images were measured by six assessors (2 vascular radiologists, 2 interventional radiology trainees, and 2 sonographers) on two separate occasions 6 weeks apart. Repeatability and reproducibility were calculated and compared with computed tomography (CT) as the gold standard. RESULTS: All three methods have high repeatability and reproducibility when static images are used. The inter-observer reproducibility coefficients between assessors were 0.48 cm, 0.35 cm, and 0.34 cm for ITI, LTL and OTO, respectively. The intra-observer repeatability coefficients between assessors were 0.30 cm, 0.20 cm, and 0.19 cm for ITI, LTL and OTO, respectively. The mean difference between CT and OTO, LTL, and ITI was 1 mm, 3 mm, and 5 mm, respectively (all underestimations) (p < .0001). CONCLUSIONS: US consistently underestimates aortic size when compared with CT, with ITI demonstrating the greatest underestimation (on average 5 mm). In the UK, this underestimation by the NHS Abdominal Aortic Aneurysm screening programme reduces the sensitivity of the screening test and may impact on the way in which vascular specialists interpret the findings of the screening programme.

Term vaginal delivery following fetoscopic laser photocoagulation of type II vasa previa.

Unrecognized vasa previa carries a significant risk of fetal mortality. Advances in ultrasound have improved detection of vasa previa and led to a dramatic reduction in fetal morbidity and mortality. However, current management strategies require prolonged hospitalized surveillance, preterm delivery prior to the onset of labor or rupture of membranes, and a cesarean delivery. Fetoscopic laser ablation of type II vasa previa allows for the possibility of term vaginal delivery. We present a patient who underwent successful laser photocoagulation of a type II vasa previa at 32(5)/7 weeks' gestation. She subsequently delivered vaginally at term without complications. The potential benefits of definitive in utero treatment of non-type I vasa previa, such as vaginal delivery at term, must be weighed against the procedure-related risks of operative fetoscopy.

Penetrating atherosclerotic ulcer of the aorta: a continuing debate.

Aortic penetrating atherosclerotic ulcer (PAU) is a relatively common incidental finding on thoracic computed tomography (CT) examinations. This is likely to relate to the steady increase in the number of CT examinations performed and also due, in part, to the increasing age of the general population. There is as yet no consensus on the management of incidental PAUs in asymptomatic patients. This article aims to review the literature and discuss the natural history, prognosis, and management of incidental PAU.

Primary infrarenal aortic stenting with or without iliac stenting for isolated and aortoiliac stenoses: single-centre experience with long-term follow-up.

PURPOSE: The purpose of this study was to evaluate the technical success, complications, long-term clinical outcome, and patency after primary infrarenal aortic stenting for aortic and aortoiliac stenosis. Between January 1999 and January 2006, 22 consecutive patients underwent endovascular treatment because of infrarenal aortic stenosis with and without common iliac stenosis (10 men; mean age 64 ± 14 years). Eleven (11 of 22) patients had an isolated aortic stenosis, whereas 11 of 22 had aortic stenosis that extended into the common iliac arteries (CIAs). Thirteen patients were Rutherford classification type 3, and 9 patients were type 4. Statistical analysis included paired Student t test and Kaplan-Meier life table analysis; p < 0.05 was considered significant. Technical and initial clinical success was achieved in all patients. There were three (14 %) procedure-related complications, which included two access-point pseudoaneurysms and one non-flow-limiting left external iliac dissection. Patients were followed-up for a mean period of 88 months (range 60-132). Mean preprocedure ankle brachial pressure indexes (ABPI) were 0.60 ± -0.15 (right) and 0.61 ± -0.16 (left). After the procedure they were 0.86 ± -0.07 (right) and 0.90 ± -0.09 (left). The increase in ABPI was significant (p < 0.05), and this continued throughout follow-up. Four (18 %) patients had recurrence of symptoms during follow-up. These occurred at 36, 48, 48, and 50 months after the original procedure. All four patients were successfully treated with repeat angioplasty procedures. There was a significant difference in primary patency between isolated aortic stenosis (100 %) and aortoiliac stenosis (60 %) (p = 0.031). Cumulative follow-up was 1920 months yielding a reintervention rate of 0.025/events/year. CONCLUSION: Primary stenting of infrarenal stenosis is safe and successful with a low reintervention rate. It should be considered as first-line treatment for patients with infrarenal aortic stenotic disease.

A prospective study of wound infections after laparotomy in obstetrics and gynaecology department.

Postoperative wound infection is of great importance to both surgeon and patient. All surgeons know that postoperative wound infection means morbidity, anxiety, longer hospitalization, higher cost; not to forget the embarrassment to the surgeons. However, it is still a common surgical complication despite other advances in modern medicine. This study was conducted to find out the incidence of postoperative wound infection and to survey the risk factors for wound infection in obstetrics and gynaecology department of Nepal medical college teaching hospital (NMCTH). A prospective study of all the cases with infected wound after laparotomy and lower segment cesarean section done in NMCTH in obstetrics and gynaecology department was carried out in this study from Chaitra 2068 to Falgun 2069. The frequency of wound infection was 5.87%. Most of the wound infection after lower segment cesarean section occurred in emergency cases (16 out of 19). Seventy percent of patients had preoperative hospital stay range of 0-2 days whereas the range was 3-18 days for the rest 30%. The range of blood loss was 150-300 ml in nearly 75.7 % of patients. Duration of operation was 1-2 hours in 89.2% cases. Only 48.6% of wound infection was identified within 8th postoperative day. Number of infected cases getting prophylactic antibiotic was 25 (67.6%). BMI was >25 in 62.2% of patients. Type of skin incision was pfannenstiel in 94.6%. Skin was closed subcuticularly with vicryl no. 1 in 81.1%. Staphylococcus aureus was isolated in swab culture in 24.3%. Resuturing was required in 18.9% of cases.

Surgical abortion in second trimester: initial experiences in Nepal.

INTRODUCTION: In spite of legalising abortion and making safe abortion available at affordable price at accessible distance to almost everyone, unsafe abortion especially second trimester abortion is still a big health problem in Nepal. OBJECTIVE: The objective of the study is to fi nd the demographic profile, reasons for seeking abortion and to see the effectiveness of Misoprostol in preparing the cervix. MATERIALS AND METHODS: A prospective study was done in the two second trimester abortion trainings conducted in Maternity hospital, Kathmandu. Total 57 clients had second trimester abortion performed. Information was collected from structured questionnaire and then data was analysed. RESULTS: Commonest reason for seeking abortion was, multiparity (61.4%). Common reasons for second trimester abortion were, completed family size with unwanted pregnancy (61.4%), unwanted pregnancy in married (10.52%) unwanted pregnancy in unmarried (5.26%). CONCLUSIONS: Second trimester abortion is one of the most common procedures performed in reproductive-aged women and when performed by a skilled provider in the appropriate setting, it is one of the safest surgeries, if it is well supported by change in policy of the country and acceptability of the people.

Detection of tubal abnormalities by HSG in Nepalese subfertile women.

Fallopian tube defects are responsible for subfertility in 12.0-33.0% of subfertile couple. Hysterosalpingography (HSG) is a safe and less invasive method of detecting both the tubal and uterine defects. The objective of this study was to find out the incidence of tubal blockage including its site and side diagnosed by HSG in subfertile Nepalese women and to find out the incidence of uterine and other abnormalities detected by this test. This was a prospective study of 1000 cases of subfertility, conducted in Om Hospital, Kathmandu. A short history and HSG report of these cases were obtained from the Radiology department of the hospital. Size and shape of the uterine cavity, evidence of cervical incompetence, tubal visualization, spillage of dye, tubal block with its side and site, evidence of peritoneal adhesion and intravasation of dye in vessels were noted. Quick spillage of the dye in the peritoneal cavity or spillage only after pushing the dye with pressure was also noted. Results were entered in simple tabulations and analyzed. Among 1000 cases, 65.8% had primary and 34.2% had secondary subfertility. 29.0% of the total 1000 cases had abnormal HSG findings. 19.0% of total 1000 cases had tubal blockage. Incidence of tubal blockage in both primary (19.1%) and secondary subfertilty (18.7%) was almost same, in contrary to previous belief. Mullerian defect was present in 3.2% of primary subfertility and 2.0% of secondary subfertility cases. Cervical incompetence was not detected in any case. Evidence of uterine infection was present in 0.7% of primary subfertility and 0.2% of secondary subfertility cases. Abnormal size of uterine cavity was present in 1.2% of primary subfertility and 0.5% of secondary subfertility. Features of phimosis of fimbrial opening, localized spill and intravasation of dye were present respectively in 5.6%, 1.5%, 1.2% in primary subfertility and 4.9%, 1.7% and 1.7% in secondary subfertility.In conclusion; the incidence of tubal blockage detectable by HSG in this study was 19.0%.

Removal of basic dye (methylene blue) from aqueous solution by adsorption using Musa paradisica: a agricultural waste.

Adsorbent prepared from Musa paradisica leaves--a low cost bioadsorbent was successfuly used to remove methylene blue from an aqueous solution in batch study. The effect of contact time, adsorbent dosage, pH of solution, initial dye concentration, particle size of adsorbent and kinetics of adsorption was studied. It has been observed that the optimum pH is 8 and adsorbent dosage required is 4 g/L for 97% removal of methylene blue (100 mg/L). The removal of methylene blue increases with increasing contact time and becomes constant after 30 minutes. The adsorbent dosage was increased from 1 g/L to 24 g/L and percentage removal increases continuously. The adsorption follows Langmuir and Freundlich isotherm models. The comparison was also done with pseudo first order, pseudo second order, and Elovich kinetic models, respectively.

Enhancement of radionuclide induced cytotoxicity by 2-deoxy-D-glucose in human tumor cell lines.

The efficacy of targeted radiotherapy can be enhanced by selective delivery of radionuclide to the tumors and/or by differentially enhancing the manifestation of radiation damage in tumors. Our earlier studies have shown that the 2-deoxy-D-glucose (2-DG), an inhibitor of glucose transport and glycolytic ATP production, selectively enhances the cytotoxicity of external beam radiation in tumor cells. Therefore, it is suggested that 2-DG may also enhance the cytotoxic effects of radionuclides selectively in tumor cells, thereby improving the efficacy of radionuclide therapy. In vitro studies on breast carcinoma (MDA-MB-468) and glioma (U-87) cell lines, has been carried out to verify this proposition. Clonogenicity (macrocolony assay), cell proliferation, cytogenetic damage (micronuclei formation) and apoptosis were investigated as parameters of radiation response. Mean inactivation dose D (dose required to reduce the survival from 1 to 0.37), was 48 MBq/ml and 96 MBq/ml for 99 mTc, treated MDA-MB-468 and U-87, respectively. The dose response of growth inhibition, induction of micronuclei formation and apoptosis observed under these conditions, were correlated well with the changes in cell survival. Presence of 2-DG (5 mM) during radionuclide exposure (24 hrs), reduced the survival by nearly 2 folds in MDA-MB-468 (from 48.5 MBq to 18.5 MBq) and by 1.6 folds in U-87 cells (from 96 MBq to 66 Mbq). These results clearly show that the presence of 2-DG during radionuclide exposure, significantly enhances the cytotoxicity, by increasing mitotic as well as interphase death. Further studies to understand the mechanisms of radio-sensitization by 2-DG and preclinical studies using tumor-bearing animals, are required for optimizing the treatment schedule.

Subfascial endoscopic perforator surgery: a review.

Approximately 1-2% of the UK population suffers from venous ulcers. Incompetent perforator leg veins are thought to be a major contributory factor. Subfascial endoscopic perforator surgery treats incompetent perforators in a minimally invasive fashion with significant improvement in wound healing and reduction in ulcer recurrence rates.

Coronary artery aneurysm with a fistulous connection to the right atrium mimicking a sinus of Valsalva aneurysm.

Coronary artery aneurysms are uncommon and may be complicated by rupture, thromboembolic phenomenon, and more rarely fistulation into one of the cardiac chambers. This case report highlights the difficulty in making a preoperative diagnosis of a coronary artery aneurysm that has fistulated into the right atrium, and lists possible differential diagnoses.

Cadmium chloride-induced changes in hypothalamic 5-hydroxytryptamine, noradrenaline and dopamine levels in the Indian palm squirrel, Funambulus pennanti (Wroughton).

A single intraperitoneal injection of cadmium chloride (CdCl2, 0.32 mg/kg body weight) was administered to 15 adult female palm squirrels (Funambulus pennanti (Wroughton] and the hypothalamic 5-hydroxytryptamine (5-HT), noradrenaline (NA) and dopamine (DA) levels were quantified on the 1st, 7th and 40th day. 5-HT levels significantly decreased on the 1st and 40th day but showed an increase on the 7th day. Similarly, NA levels significantly decreased after 1, 7 and 40 days of treatment, and DA levels decreased on the 1st and 7th day but insignificantly increased on the 40th day. This suggests that CdCl2 interferes in the functioning of the hypothalamo-hypophyseal axis by impairing the hypothalamic 5-HT, NA and DA levels, which may in turn affect the physiology of the peripheral endocrine glands.

Susceptibility of hamsters to caecal amoebiasis by oral infection.

A method is described for successfully establishing caecal amoebiasis in hamsters which were not fed and which were pretreated with 1 ml of magnesium sulphate every 24 hours for three days and then given 12 x 10(5) trophozoites of the HM-1 axenic strain or 18 x 10(5) trophozoites of the HK-9 axenic strain of Entamoeba histolytica by the oral route. All the animals developed diarrhoea within 24 hours of infection. When the animals were killed on the fifth day after infection the caecum was swollen and fused. Large macroscopic ulcers full of pus could be seen in the caecum. None of the control animals showed any of the changes mentioned above.