RNA Interference highlights the role of CCaMK in dissemination of endosymbionts in the aeschynomeneae legume arachis

In legume–rhizobia symbiosis, Ca2+/Calmodulin-dependent Protein Kinase (CCaMK) is essential for rhizobial invasion through infection threads in the epidermis and nodule organogenesis in the cortex. Though CCaMK is actively transcribed in the infected zone of nodules, its role in the later stages of nodule development remain elusive because of the epidermal arrest of “loss-of-function” mutants. In Aeschynomeneae legumes such as Arachis hypogea, rhizobia directly access the cortex, where nodule organogenesis as well as endosymbiont dissemination take place by multiplication of infected cortical cells. We characterized CCaMK (GI:195542474) from A. hypogea and downregulated the kinase through RNA interference (RNAi) to understand its role during organogenesis of its characteristic aeschynomenoid nodules. In CCaMK downregulated plants, the inception of nodules was delayed by approximately 4 weeks and nodulation capacity was decreased (>90%). The infected zones of the RNA interference nodules were scattered with uninfected or binucleated cells as opposed to the homogeneous infection zone in empty-vector-transformed nodules. Symbiosomes in RNAi nodules were pleomorphic with diverse geometrical shapes or arrested during division in the final stages of their fission as opposed to uniform-sized, spherical symbiosomes in empty-vector-transformed nodules. Together, our results reveal CCaMK to be essential for development of functional aeschynomenoid nodules, with a critical role in rhizobial dissemination during nodule organogenesis

A Rare Case of Cerebral Hemangioblastoma Mimicking as Anterior Choroid Plexus Cyst

Abstract A choroid plexus cyst may be present throughout the ventricular system of the brain. A hemangioblastoma is generally found in patients with genetic diseases like Von Hippel-Lindau (VHL) syndrome disease, but it can also be found sporadically in the spinal cord, retina and in rare cases in the cerebrum. An eighteen-year-old male subject was referred for autopsy in Kolkata police morgue on 27/02/2017. Two hours before, the patient had been brought to the hospital for severe headache; but before any diagnosis could be made, he collapsed and died. On autopsy, a 4cm fluid filled cyst was found in the anterior portion of third ventricle and sent for histopathological examination. HPE findings were consistent with a hemangioblastoma, which made this case extremely rare. Sudden death due to increased intracranial tension cases should be properly examined for any neuroepithelial tumours like a hemangioblastoma

Shifting trend of tubercular lymphadenitis over a decade – A study from eastern region of India

Background: The incidence of tuberculosis (TB) is changing in India. The commonest extra pulmonary manifestation of TB is tuberculous lymphadenitis. Aims and objectives: To detect changes in occurrence of TB lymphadenitis by comparing current data with that from 10 years back. Methods: Fine Needle Aspiration Cytology (FNAC) findings of patients presenting with lymphadenopathy between January 2014 and December 2015 were included in the study as current data. Cases of TB lymphadenitis were analysed for cyto-morphological patterns and frequency of acid fast bacilli (AFB) positivity. Cytological data of patients with lymphadenopathy between January 2002 and December 2003, were collected from records. Subsequently, comparison was done between previous data and current observation. Results: Total 302 cases of lymphadenopathy were included in 2014 and 2015. Tuberculous lymphadenitis accounted for 96 (31.7%) cases, among which AFB was present in 47 (49%) cases. Data collected from registers of the years 2002 and 2003 included 318 cases where tuberculous lymphadenitis accounted for 161 (50.6%) cases and AFB was detected in 101 (63%) patients. Maximum AFB positivity was found in smears with caseous necrosis only. Conclusion: Presently occurrence of TB lymphadenopathy has reduced compared to the period of 2002–2003. Despite this declining trend, the disease continues to be one of the leading causes of lymphadenopathy in both periods

Etiology of Pancytopenia: An Observation from a Referral Medical Institution of Eastern Region of India

Background: Pancytopenia is a relatively common hematological condition, the etiological factors of which vary widely in different geographic location. Determining the specific etiology is of immense importance for appropriate management. Aims and Objectives: The present study was undertaken to delineate etiological factors leading to pancytopenia in a Tertiary Care Hospital of West Bengal from Eastern Region of India. Aims and Objectives:The present study was undertaken to delineate etiological factors leading to pancytopenia in a Tertiary Care Hospital of West Bengal from Eastern Region of India. Materials and Methods: A prospective study was conducted for a period of 2 years in which 248 patients were included. After obtaining a relevant clinical history, physical examination was done followed by complete blood count including peripheral blood smears examination, relevant biochemical, and radiological investigations. Afterward, bone marrow aspiration and biopsy were performed and microscopically examined. Results: Among 248 patients studied, 156 (62.9%) were males and 92 (37.09%) were females. The mean age of the patients was 33 years. Aplastic anemia was the most common cause of pancytopenia that was observed in 83 cases (33.47%) followed by megaloblastic anemia in 52 cases (20.97%), leishmaniasis in 34 patients (13.71%), hypersplenism also in 34 patients (13.71%), and tuberculosis and other connective tissue disorders in 18 cases (7.26%). The occurrence of aplastic anemia was statistically significant in pediatric (≤15 years) age group. Conclusion: Aplastic anemia was found to be the most common cause of pancytopenia in this study, which is in contrast to studies conducted from other regions of India. Delineation of etiologies of pancytopenia in various regions can help in defining diagnostic and therapeutic strategies, which is expected to contribute toward the better management of such patients

Giant myelolipoma of left adrenal gland simulating a retroperitoneal sarcoma

Adrenal myelolipoma is a rare benign tumor composed of an admixture of adipose and hematopoietic tissue. Most tumors are asymptomatic, small in size, right-sided, and usually detected incidentally. Though they are often smaller than four cm in diameter, they can reach larger sizes. Giant adrenal myelolipomas are extremely rare tumors having only about 10 reported cases in the literature. Presently, a case is being reported in a 59-year-old lady who presented with abdominal distention and dragging sensation in the upper left abdomen. Computed tomography of the abdomen revealed a retroperitoneal tumor which was suspected to be a retroperitoneal sarcoma. Other fat containing retroperitoneal tumors such as a lipoma, myelolipoma and teratoma were also considered as possibilities. All hormonal studies related to adrenal gland were within normal limits. The patient underwent surgical resection of the tumor. The resected mass was 23 cm × 16 cm ×9 cm in size and weighed 3.3 kg. Histopathological examination revealed the tumor to be an adrenal myelolipoma. This present case of adrenal myelolipoma is unusual in view of its left-sided location and its large size which clinically and radiologically simulated a retroperitoneal sarcoma

Transformed Hairy Roots of Arachis hypogea: A Tool for Studying Root Nodule Symbiosis in a Non–Infection Thread Legume of the Aeschynomeneae Tribe

Arachis hypogea is a non–“infection thread” (IT) legume where rhizobial entry or dissemination in the nodules never involves IT. Rhizobia invade through epidermal “cracks” and directly access the cortical cells to develop the characteristic aeschynomenoid nodules. For investigating these nonclassical nodulation features in Arachis spp., we developed an efficient procedure for Agrobacterium rhizogenes R1000-mediated transformation of this plant. In this study, we optimized the induction of hairy roots and nodulation of composite Arachis hypogea plants in the presence of Bradyrhizobium sp. (Arachis) strain NC92. 35S promoterdriven green fluorescent protein and β-glucuronidase expression indicated transformation frequency to be above 80%. The transformed roots had the characteristic rosettetype root hairs and had normal level of expression of symbiosis- related genes SymRK and CCaMK. The transgenic nodules resembled the wild-type nodules with an exception of 2 to 3%, where they structurally deviated from the wildtype nodules to form nodular roots. A 16S rRNA profile of an infected-zone metagenome indicated that identical populations of bradyrhizobia invaded both composite wild-type plants grown in natural soil. Our results demonstrate that Arachis hairy root is an attractive system for undertaking investigations of the nonclassical features associated with its nitrogen-fixing symbiotic interaction

Spectrum of thalassemias and hemoglobinopathies in West Bengal: A study of 90,210 cases by cation exchange high-performance liquid chromatography method over a period of 8 years

Background: Thalassemias and hemoglobinopathies are highly prevalent in India. Identification of these disorders is important for epidemiologic purposes and for prevention of thalassemia major and clinically severe hemoglobinopathies. Objectives: The aim of this study was to determine the prevalence of thalassemias and hemoglobinopathies in patients of a tertiary care hospital of West Bengal. Materials and Methods: A prospective study was undertaken in which 90,210 cases were included over a period of 8 years. Clinical history and family history were obtained from each patient. The venous blood samples were analyzed for complete blood count, liver function tests, serum iron, ferritin, cobalamin and folate levels. High-performance liquid chromatography (HPLC) was performed on the samples with Biorad Variant using beta thalassemia short program. Confirmatory tests were done whenever required. Results: Normal hemoglobin (Hb) pattern was observed in 79,897 (88.57%) cases and abnormalities were detected in 10,313 (11.43%) patients. β (beta) thalassemia trait was the most common abnormality found in 3870 (4.29%) patients. HbE trait was found in 2418 (2.68%) cases, and then Eβ thalassemia in 1406 (1.56%) patients and β thalassemia major/intermedia in 1135 (1.26%) cases. Other variants detected included sickle cell trait, HbE disease, sickle cell disease, sickle β thalassemia, HbD-Punjab trait, double heterozygous state of HbS and HbE, double heterozygous state of HbS and HbD, Hb Lepore, HbJ-Meerut and HbH. Conclusion: Premarital and antenatal screenings are important measures to prevent birth of children with severe Hb disorders. HPLC is a rapid and reliable technique for identification of various Hb fractions

Transformed hairy roots of Aachis hypogea: a tool for studying root nodule symbiosis in a non–infection thread legume of the Aeschynomeneae tribe

Arachis hypogea is a non–“infection thread” (IT) legume where rhizobial entry or dissemination in the nodules never involves IT. Rhizobia invade through epidermal “cracks” and directly access the cortical cells to develop the characteristic aeschynomenoid nodules. For investigating these nonclassical nodulation features in Arachis spp., we developed an efficient procedure for Agrobacterium rhizogenes R1000-mediated transformation of this plant. In this study, we optimized the induction of hairy roots and nodulation of composite Arachis hypogea plants in the presence of Bradyrhizobium sp. (Arachis) strain NC92. 35S promoter-driven green fluorescent protein and β-glucuronidase expression indicated transformation frequency to be above 80%. The transformed roots had the characteristic rosette-type root hairs and had normal level of expression of symbiosis-related genes SymRK and CCaMK. The transgenic nodules resembled the wild-type nodules with an exception of 2 to 3%, where they structurally deviated from the wild-type nodules to form nodular roots. A 16S rRNA profile of an infected-zone metagenome indicated that identical populations of bradyrhizobia invaded both composite wild-type plants grown in natural soil. Our results demonstrate that Arachis hairy root is an attractive system for undertaking investigations of the nonclassical features associated with its nitrogen-fixing symbiotic interactions

Histopathological study of adrenocortical carcinoma with special reference to the Weiss system and TNM staging and the role of immunohistochemistry to differentiate it from renal cell carcinoma

Background: Adrenocortical carcinomas (ACC) are rare tumors with an incidence of 1-2/million/year. They account for 0.05-0.2% of all malignancies. Aims and Objectives: This study was aimed to evaluate the Weiss system to diagnose ACCs and to compare it with TNM staging. The role of immunohistochemistry (IHC) was also evaluated to differentiate ACC from other differential diagnoses especially, renal cell carcinoma (RCC). Materials and Methods: A total of 10 ACCs were included from April, 2000 to March, 2012. All the relevant information like weight, tumor size, gross features was recorded. Multiple sections were taken for histologic examination. Immunomarkers like vimentin, synaptophysin, Melan-A, calretinin, inhibin, EMA, cytokeratin, and Ki-67 were used. Results: Out of 10 cases of ACCs (diagnosed by the Weiss system), the tumors were in TNM stage I (1 case), stage II (2 cases), stage III (5 cases), and stage IV (2 cases). The Weiss score in stages I-IV was 4; 5-7; 6-8; and 5-9 respectively. ACCs were positive for vimentin, inhibin, Melan-A, calretinin and negative for EMA and cytokeratin. Proliferative index (Ki-67/MIB-1) was ≥ 20 % (20-65%). Conclusion: Both the Weiss system and TNM staging are useful in predicting the malignant behavior and prognosis of ACC. Weight and tumor size, though originally not included in the Weiss system, are also important parameters. In lower stages (stages I and II), the Weiss score is low (4-7) while the score is high (6-9) when the tumor is in higher stages (stages III and IV). IHC plays a vital role to confirm the diagnosis and to exclude the possibility of RCC

Clinicopathological study of annexin A5 & apelin in pre-eclamptic placentae with emphasis on foetal outcome

Background & objectives: Pre-eclampsia has remained an elusive disease with serious impacts on both maternal and foetal health. Two novel markers, annexin A5 (ANXA5) and apelin are currently of considerable interest. The present study aimed to determine the placental expression of ANXA5 and apelin in pre-eclamptic placentae and also to elucidate if there is any correlation between the expression of these markers and the clinical features of both, mother and neonate. The comparison between gross and histopathological features of pre-eclamptic placentae and controls was another objective. Methods: A prospective, observational study was undertaken for one year. Placentae of pre-eclamptic patients and matched controls (matched for age, ethnic and socio-economic background) were collected along with the clinical data. Gross and histopathological analyses were done and immunohistochemical study of placental sections with ANXA5 and apelin was also undertaken. Results: 79 pre-eclamptic patients and equal numbers of matched controls were included in the study. The difference in weight and dimensions of placentae between the pre-eclampsia group and matched controls was significant. Histopathological features noted in the pre-eclamptic placentae included decidual vasculopathy, infarction, perivillous fibrin deposition, increased syncytial knots and distal villous hypoplasia. There was a significant reduction in the expression of both ANXA5 and apelin in pre-eclamptic placentae compared to controls. Among pre-eclamptic patients, the intensity of ANXA5 and apelin expression showed a significant association with respect to neonatal resuscitation. Furthermore, the intensity of apelin showed expression a significant correlation with the requirement of sick neonatal care unit treatment. Interpretation & conclusions: The results of the present study suggest that both ANXA5 and apelin levels are reduced in pre-eclamptic placentae. Hence, it is recommended to further explore the impact of these markers on pregnancy outcomes by undertaking randomized controlled trials