Morpho-anatomical characterization and DNA barcoding of Artemesia vulgaris L.

Artemisia vulgaris L. belongs to Asteraceae, is a herbal plant that has various benefits in the medical field, so that its use in the medical field can be explored optimally, the plant must be thoroughly identified. This study aims to identify A. vulgaris both in terms of descriptive morpho-anatomy and DNA barcoding using BLAST and phylogenetic tree reconstruction. The morpho-anatomical character was observed on root, stem, and leaf. DNA barcoding analysis was carried out through amplification and alignment of the rbcL and matK genes. All studies were conducted on three samples from Taman Husada (Medicinal Plant Garden) Graha Famili Surabaya, Indonesia. The anatomical slide was prepared by the paraffin method. Morphological studies revealed that the leaves of A. vulgaris both on the lower-middle part and on the upper part of the stem have differences, especially in the character of the stipules, petioles, and incisions they have. Meanwhile, from the study of anatomy, A. vulgaris has an anomocytic type of stomata and its distribution is mostly on the ventral part of the leaves. Through the BLAST process and phylogenetic tree reconstruction, the plant sequences being studied are closely related to several species of the genus Artemisia as indicated by a percentage identity above 98% and branch proximity between taxa in the reconstructed phylogenetic tree.

Addressing tuberculosis control in fragile states: Urban DOTS experience in Kabul, Afghanistan, 2009-2015.

Tuberculosis (TB) is a major public health problem in Afghanistan, but experience in implementing effective strategies to prevent and control TB in urban areas and conflict zones is limited. This study shares programmatic experience in implementing DOTS in the large city of Kabul. We analyzed data from the 2009-2015 reports of the National TB Program (NTP) for Kabul City and calculated treatment outcomes and progress in case notification using rates, ratios, and confidence interval. Urban DOTS was implemented by the NTP in partnership with United States Agency for International Development (USAID)-funded TB projects, the World Health Organization (WHO), and the private sector. Between 2009 and 2015, the number of DOTS-providing centers in Kabul increased from 22 to 85. In total, 24,619 TB patients were enrolled in TB treatment during this period. The case notification rate for all forms of TB increased from 59 per 100,000 population to 125 per 100,000. The case notification rate per 100,000 population for sputum-smear-positive TB increased from 25 to 33. The treatment success rate for all forms of TB increased from 31% to 67% and from 47% to 77% for sputum-smear-positive TB cases. The treatment success rate for private health facilities increased from 52% in 2010 to 80% in 2015. In 2013, contact screening was introduced, and the TB yield was 723 per 100,000-more than two times higher than the estimated national prevalence of 340 per 100,000. Contact screening contributed to identifying 2,509 child contacts of people with TB, and 76% of those children received isoniazid preventive therapy. The comprehensive urban DOTS program significantly improved service accessibility, TB case finding, and treatment outcomes in Kabul. Public- and private-sector involvement also improved treatment outcomes; however, the treatment success rate remains higher in private health facilities. While the treatment success rate increased significantly, it remains lower than the national average, and more efforts are needed to improve treatment outcomes in Kabul. We recommend that the urban DOTS approach be replicated in other countries and cities in Afghanistan with settings similar to Kabul.

Stable heterologous expression of biologically active terpenoids in green plant cells.

Plants biosynthesize a great diversity of biologically active small molecules of interest for fragrances, flavors, and pharmaceuticals. Among specialized metabolites, terpenoids represent the greatest molecular diversity. Many terpenoids are very complex, and total chemical synthesis often requires many steps and difficult chemical reactions, resulting in a low final yield or incorrect stereochemistry. Several drug candidates with terpene skeletons are difficult to obtain by chemical synthesis due to their large number of chiral centers. Thus, biological production remains the preferred method for industrial production for many of these compounds. However, because these chemicals are often found in low abundance in the native plant, or are produced in plants which are difficult to cultivate, there is great interest in engineering increased production or expression of the biosynthetic pathways in heterologous hosts. Although there are many examples of successful engineering of microbes such as yeast or bacteria to produce these compounds, this often requires extensive changes to the host organism's metabolism. Optimization of plant gene expression, post-translational protein modifications, subcellular localization, and other factors often present challenges. To address the future demand for natural products used as drugs, new platforms are being established that are better suited for heterologous production of plant metabolites. Specifically, direct metabolic engineering of plants can provide effective heterologous expression for production of valuable plant-derived natural products. In this review, our primary focus is on small terpenoids and we discuss the benefits of plant expression platforms and provide several successful examples of stable production of small terpenoids in plants.

Public health surveillance at a mass gathering: urs of Baba Farid, Pakpattan district, Punjab, Pakistan, December 2010.

The objective of this study was to identify health related problems encountered during an annual mass gathering in Pakpattan, Pakistan, the anniversary of the death (urs) of Baba Farid, and to make recommendations for planning and prevention activities. A surveillance system was established to capture health related data for the event. A list of reportable diseases was developed. Data were collected pre-, during, and post-event by health care workers trained for the purpose. A total of 5918 people reported to the 15 health care facilities providing services during the event; 58% of consultations were because of communicable diseases, 21% of which were respiratory tract infections and 26% gastrointestinal illness. Injuries accounted for 31% of cases and noncommunicable diseases for 11%. Prevalence of reportable disease during the event showed sizeable increases. No major disease outbreak was observed.

Enhanced production and characterization of a beta-glucosidase from Bacillus halodurans expressed in Escherichia coli.

A putative beta-glucosidase gene from the genome of Bacillus halodurans C-125 was expressed in E. coli under the regulation of T7lac promoter. On induction with isopropyl-beta-D-1-thiogalactopyranoside, the enzyme expressed at approximately 40% of the cell protein producing 238 mg/liter culture. With increase in culture cell density to A(600) 12 in auto-inducing M9NG medium, beta-glucosidase production increased 3-fold. Approximately 70% of the expressed enzyme was in a soluble form, while the rest was in an insoluble fraction of the cell lysate. The soluble and active form of the expressed enzyme was purified by ammonium sulfate precipitation followed by ion-exchange chromatography to a purity >98%. The mass of the enzyme as determined by MALDI-TOF mass spectrometry was 51,601 Da, which is nearly the same as the calculated value. Phylogenetic analysis of the beta-glucosidase of B. halodurans was found to cluster with members of the genus Bacillus. Temperature and pH optima of the enzyme were found to be 45 degrees C and 8.0, respectively, under the assay conditions. K(m) and k(cat) against p-nitrophenyl-beta-D-glucopyranoside were 4 mM and 0.75 sec(-1), respectively. To our knowledge, this is the first report of high-level expression and characterization of a beta-glucosidase from B. halodurans.

Spin-polarized structural, electronic, and magnetic properties of diluted magnetic semiconductors Cd(1-x)Mn(x)S and Cd(1-x)Mn(x)Se in zinc blende phase.

We studied the structural, spin-polarized electronic band structures, density of states, and magnetic properties of the diluted magnetic semiconductors (DMSs) Cd(1-x)Mn(x)S and Cd(1-x)Mn(x)Se in zinc blende phase (B3) with 25% Mn by using the ab initio method. The calculations were performed by using the full potential linearized augmented plane wave plus local orbitals (FP-L/APW+lo) method within the spin-polarized density functional theory and the local spin density approximation (LSDA). Calculated electronic band structures and the density of states of these DMSs are discussed in terms of the contribution of Mn 3d(5)4s(2), Cd 4d(10)5s(2), S 3s(2)3p(4), and Se 4s(2)4p(4) partial density of states and we also compute the local magnetic moments. We estimated the spin-exchange splitting energies, Delta(x)(d) and Delta(x)(p-d), produced by the Mn 3d states, and we found that the effective potential for the minority spin is more attractive than that for the majority spin. We determine the s-d exchange constant N(0)alpha and p-d exchange constant N(0)beta, which resembles a typical magneto-optical experiment. The calculated total magnetic moment is found to be 5.0020 and 5.00013 mu(B) for Cd(1-x)Mn(x)S and Cd(1-x)Mn(x)Se, respectively. These values indicate that every Mn impurity adds no hole carriers to the perfect CdS and CdSe crystals. Moreover, we found that p-d hybridization reduces the local magnetic moment of Mn from its free space charge value of 5.0micro(B) and produces small local magnetic moments on the nonmagnetic Cd and S sites.

Clinical and laboratory features of systemic lupus erythematosus (SLE) in Pakistani patients.

OBJECTIVES: To study the clinico-pathological features of systemic lupus erythematosus (SLE) in Pakistani patients at the time of presentation in the four teaching hospitals of Rawalpindi-Islamabad area. SETTINGS: Department of Immunology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, Military Hospital (MH), Rawalpindi, Armed Forces Institute of Urology (AFIU), Rawalpindi, Combined Military Hospital (CMH), Rawalpindi, Department of Rheumatology, Pakistan Institute of Medical Sciences (PIMS), Islamabad. SUBJECTS: Patients suffering from SLE, diagnosed on the basis of the ARA criteria. MAIN OUTCOME MEASURES: Clinico-pathological features at the time of presentation. RESULTS: Twelve male patients in age range 5-30 years (mean 21.6 years) and 38 female patients in age range 8-37 years (mean 22.2 years) were included in the study, showing a female to male ratio of 3:1. Fever was the most common presenting feature (100%), followed by arthralgias/arthritis (98%), malar rash (64%) and oral ulcers (58%). Laboratory results showed antinuclear antibody to be positive in all patients followed by anti double stranded DNA antibodies (anti DNA antibodies: 64%), anti extractable nuclear antigen antibodies (anti ENA antibodies: 64%) and evidence of complement consumption in 64% of the patients. Erythrocyte Sedimentation Rate (ESR) was found to be increased with a mean of 64 mm first hour fall in males and a mean of 87 mm first hour fall in the female patients. The C reactive protein was detected to be normal or marginally increased inspite of the increased ESR with mean of 7 mg/l for male patients and 5 mg/1 for the females. CONCLUSION: The subject patient population presented almost universally with fever and arthralgias or arthritis in combination with malar rash or oral ulcers and in some patients a combination of all of the above was observed. A combination of positive anti nuclear antibody test, evidence for consumption of the complement in form of low C4 and C3, increased ESR and low C reactive protein were found to be a sensitive and cost effective set of laboratory findings for the diagnosis of patients suffering from SLE. The above mentioned set of clinical and laboratory features would help in the correct and early diagnosis of patients suffering from SLE, a relatively rare disease, in the busy medical out patient departments in our set up.

Presenting features in Pakistani patients suffering from the antineutrophil cytoplasmic antibody--classical subtype (c-ANCA) associated vasculitis.

OBJECTIVES: To study the clinicopathological features in c-ANCA positive patients suffering from vasculitis with a view to find out the most common mode of presentation. STUDY DESIGN: Retrospective. SETTINGS: Department of Immunology, AFIP, Rawalpindi, MH Rawalpindi, CMH Rawalpindi, Department of Rheumatology, PIMS, Islamabad, RGH Rawalpindi, FFH, Rawalpindi. SUBJECTS: Seventeen patients suffering from vasculitis and found to be positive for c-ANCA. MAIN OUTCOME MEASURES: Clinico-pathological features at presentation. RESULTS: There were 9 males in age range 11-60 years (mean age 32.5 years) and 8 females in age range 26-42 years (mean age 32.3 years). Common presenting features were a combination of cough, blocked nose and post nasal drip 14/17 (82%) followed by nose bleed and haematuria 11/17 (65%). Six patients were demonstrated to be suffering from Wegener's granulomatosis after biopsy. C-ANCA was detected by indirect immunofluorescence in the titre range of 8-640. The auto antibody levels related to disease activity. CONCLUSION: c-ANCA associated vasculitis is a rare (17 patients in five years) but aggressive form of vasculitis. It must be suspected in patients with persistent respiratory tract related symptoms associated with fever and joint pains which fail to respond to adequate treatment for infections. The c-ANCA estimations can be utilised as sensitive and specific diagnostic and prognostic marker in this form of vasculitis.

The pattern of bone marrow infiltration in non-Hodgkin's lymphomas.

The pattern of bone marrow infiltration in 40 consecutive cases of non-Hodgkin's lymphomas (NHLs) having marrow involvement is presented. For histological break-up, working formulation of NHLs was used. Low grade NHLs (50%) were the commonest, followed by intermediate (32.5%) and high grade NHLs (17.5%), respectively. Histologically, small lymphocytic type (32.5%) was the commonest, followed by diffuse small cleaved cell (15%) type; follicle small cleaved cell, diffuse mixed small and large cell as well as lymphoblastic types were equally frequent (12.5% each). Other histological types were less common. The pattern of infiltration of the bone marrow was diffuse in 76.7% of intermediate, 70% of low and 100% of high grade NHLs. It was focal random (non-paratrabecular) in 15% of low grade and 15.4% of intermediate grade NHLs. Focal paratrabecular pattern was evident in 15% of low grade and 7.7% of intermediate grade NHLs.

Visceral leishmaniasis--a study of 38 cases on the basis of geographical distribution.

Geographical distribution of thirty-eight cases of visceral leishmaniasis (VL), who were diagnosed in the department of Pathology, Rawalpindi Medical College and two private laboratories in Rawalpindi and Islamabad is presented. Majority (84.2%) came from Azad Kashmir, especially from areas around Poonch (e.g., Rawlakot-28.9%; Bagh-23.7%; Dirkot-5.3% and Pallandari-2.6%) and areas around Muzaffarabad (e.g., Chakothi-13.2% and Punja Sharif-5.3%). Only two patients (5.3%) belonged to Gilgit agency. Four cases came from villages around Murree, Rawalpindi and Abbottabad. These places are not known as endemic areas of VL. It is proposed that epidemiological studies for VL should be carried out in areas of Azad Kashmir, especially concentrating on the places from where patients keep on presenting for treatment in various hospitals of Rawalpindi and Islamabad.

A morphological pattern of 234 cases of leukemias.

Morphological pattern of 234 consecutive cases of various types of leukemias is presented. Acute leukemias (62.8%) were commoner than chronic (37.2%). Amongst acute leukemias, myeloid leukemias (AML) were more frequent as compared to lymphoblastic (ALL). AML:ALL ratio was 2.57:1 in adults and 1:3 in children. Amongst AML cases, M4 was the commonest, followed by M2, M1, M3 and M4 and M6 and M7 respectively. In ALL patients, L1 was the commonest, followed by L2 and L3 respectively. Amongst chronic leukemias, myelocytic leukemia (CML) was more common than lymphocytic leukemia (CLL) with a CML:CLL ratio of 3.1:1. In a total of 60 CML cases, two had juvenile CML, four were between 10 and 15 years of age and the remaining 54 were adults. Hairy cell leukemia (2 cases) and lymphoma/leukemia syndrome (5 cases) were uncommon.

Severe aplastic anaemia--an aetiological correlation.

Over 4 years, 43 cases of severe aplastic anaemia (SAA) were seen. Etiologically 58.1% had idiopathic, 39.5% secondary and 2.3% congenital aplastic, anaemia. Idiopathic SAA was more common in patients between 0-15 years of age, whereas secondary SAA was more frequent in 16-60 years age group. Males were more commonly affected with a male:female ratio of 3.3:1. Amongst 17 cases of secondary SAA, chloramphenicol was responsible for 4, septran for 3, insecticides for 2 and anti-scabitic drugs, anti-diabetic drugs and "kushta from hakeem" in one patient each. Hepatitis associated SAA was suspected in 3 cases.

Acute myeloid leukemia-FAB classification and its correlation with clinico-haematological features.

The French-American and British (FAB) classification of 62 consecutive cases of acute myeloid leukemia was undertaken. AML-M2 was the commonest FAB type (32.26%), followed by M1 and M4 (22.58% each), M5 (8.6%) and M6 and M7 (1.61% each), respectively. One of the patients was diagnosed as AML-MO (not a FAB type). The mean age of M1, M2, M3 and M5 cases was between 25 and 29 years, whereas in M4 patients it was 45.6 years. AML-M2, M4 and M5 were commoner in males, M1 in females and M3 equal in both sexes. Feeling of weakness, easy fatiguability and pallor were invariably present in all FAB types. All the patients of M1 and M5, 85% of M2, 64% of M4 and 50% of M3 presented with fever. Bleeding manifestations were most frequent in M3 cases followed by M5, M1, M4 and M2, respectively. Hepatomegaly and splenomegaly were relatively less prominent features in M3 as compared to other FAB types. Amongst the haematological parameters, anaemia was more severe in M1, leucocytosis in M2 and thrombocytopenia in M3 cases as compared to other FAB types.