Tracking Zone-wise perturbation during unfolding of some globular proteins using Eu(III) complex of Tetracycline as a probe exhibiting Stark splitting.

Enhanced 'Antenna effect' of a suitably designed ternary complex of Eu(III), Tetracycline hydrochloride (TC) and globular proteins viz bovine serum albumin (BSA), human serum albumin (HSA) and ß-lactoglobulin A (BLGA) in aqueous medium is employed to characterize the different partially unfolded states along with investigation of the micro- heterogeneous environment of the proteins during their stepwise unfolding. The zone-wise perturbation for the proteins upon denaturation by Urea and Guanidine hydrochloride (Gdn. HCl) is followed by the emission of Eu(III) through 'Antenna Effect' and that of the tryptophan (Trp) residues of the proteins as a function of denaturants both by steady state and time resolved emission study. With Gdn. HCl as denaturant, both BSA and BLGA show quenching of Eu(III) emission compared to pure protein while HSA exhibits an enhancement of antenna effect during unfolding as compared to that in its absence. In the presence of Urea, HSA and BSA show enhancement of antenna effect accompanied by Stark splitting of the 5D0→7F2 transition of Eu(III) although BLGA follows the similar pattern of quenching of Eu(III) emission as observed with Gdn. HCl without any Stark splitting. The proteins exhibit a two state transition with ΔGD values of ~ 2-3 kcal mol-1. Thus the use of Eu(III) emission as an efficient probe is advocating here to rationalize the microenvironment of the proteins during their stepwise unfolding.

Desmoplastic small round cell tumor of the ovary: A rare but poor prognostic disease in a young woman!

Desmoplastic small round cell tumor (DSRCT) is a rare, aggressive neoplasm of uncertain histogenesis that preferentially involves the abdominal and pelvic cavities. DSRCT mainly develops in adolescent and young adults with a strong male predominance; the male to female ratio is 4:1. Ovarian location is exceptional. DSRCT generally develops in the abdomen and have a tendency towards peritoneal spread, with subsequent metastasis to distant lymph nodes, liver and lungs. It is a poorly understood malignancy with a very characteristic morphology, immunophenotype, cytogenetic features, and poor prognosis. This tumor can co-express epithelial, neuronal, and mesenchymal markers. Despite intensive therapy, including surgery, radiotherapy and chemotherapy, and immunotherapy; the 5-year survival is less than 15%.

Cytomorphology of hepatoblastoma with histological correlation and role of SALL4 immunocytochemistry in its diagnosis, subtyping, and prognostication.

BACKGROUND: Hepatoblastoma (HB) is the most common malignant pediatric liver tumor, and cytology material is often the only tissue available for evaluation before definitive therapy. Subcategorization of HB based on cytomorphological features thus carries an important role in its prognostication. Spalt-like transcription factor 4 (SALL4), a marker of embryonic stem cells that is also found in the fetal liver, is reactivated in certain liver tumors. Limited studies have evaluated its role in HB. This study was aimed at evaluating the cytomorphological features of HB and assessing the utility of SALL4 immunocytochemistry (ICC) in its subtyping and prognostication. METHODS: Pretherapy fine-needle aspiration smears from patients diagnosed with HB over a period of 9 years were retrieved. Aspirates were subclassified on the basis of the cytomorphology and were correlated with the histology wherever it was available. ICC for SALL4 was performed in 33 cases, and nuclear staining was considered positive. RESULTS: A total of 53 HB cases were included with 30 available postchemotherapy resection specimens. All the patients were diagnosed as epithelial HB on cytology, and the cases were subclassified as pure fetal (9 of 53), pure embryonal (2 of 53), or combined epithelial HB (42 of 53). There was good concordance between cytology and histology for subtyping. SALL4 immunostaining displayed strong and diffuse nuclear positivity in the embryonal component while focal and weak to negative staining in fetal cells. CONCLUSIONS: Fine-needle aspiration cytology serves as a rapid and effective tool for a correct diagnosis of HB before the implementation of chemotherapy, and SALL4 may serve as a useful diagnostic and prognostic marker.

M2G1G2 white blood cell flag by three-part automated hematology analyzer: A hint to dengue infection in appropriate clinical context.

BACKGROUND: Automated hematology analyzers often generate many flags which can provide important clues to the underlying hematological abnormality. Although pathologists are generally well versed in recognizing the importance of flags indicating potential leukemic blasts, their utility in hinting toward infectious etiology, especially during epidemics, is less well known. We analyzed any abnormal flags generated by a three-part automated hematology analyzer in serologically confirmed cases of dengue. MATERIALS AND METHODS: The study included 28 patients diagnosed with serologically confirmed dengue infection. The venous samples were run on ABX Miros-60 three-part hematology analyzer. The complete blood count data and any abnormal flags were noted and correlated with peripheral blood film findings in all patients. RESULTS: An abnormal white blood cell (WBC) flag was noted in all 28 patients, including two in whom all other hematological parameters were within normal limits. In 26 (93%) patients, M2G1G2 WBC flag was noted while the samples of the remaining two patients generated M2G1 and L1M2G1G2 WBC flags, respectively. CONCLUSION: An abnormal WBC flag, when correlated in appropriate clinical context, especially during a dengue outbreak, can aid in targeting the cohort of patients who will require immediate referral for serological confirmation.

Quantitative histology-based classification system for assessment of the intestinal mucosal histological changes in patients with celiac disease.

BACKGROUND/AIMS: The existing histological classifications for the interpretation of small intestinal biopsies are based on qualitative parameters with high intraobserver and interobserver variations. We have developed and propose a quantitative histological classification system for the assessment of intestinal mucosal biopsies. METHODS: We performed a computer-assisted quantitative histological assessment of digital images of duodenal biopsies from 137 controls and 124 patients with celiac disease (CeD) (derivation cohort). From the receiver-operating curve analysis, followed by multivariate and logistic regression analyses, we identified parameters for differentiating control biopsies from those of the patients with CeD. We repeated the quantitative histological analysis in a validation cohort (105 controls and 120 patients with CeD). On the basis of the results, we propose a quantitative histological classification system. The new classification was compared with the existing histological classifications for interobserver and intraobserver agreements by a group of qualified pathologists. RESULTS: Among the histological parameters, intraepithelial lymphocyte count of ≥25/100 epithelial cells, adjusted villous height fold change of ≤0.7, and crypt depth-to-villous height ratio of ≥0.5 showed good discriminative power between the mucosal biopsies from the patients with CeD and those from the controls, with 90.3% sensitivity, 93.5% specificity, and 96.2% area under the curve. Among the existing histological classifications, our quantitative histological classification showed the highest intraobserver (69.7%-85.03%) and interobserver (24.6%-71.5%) agreements. CONCLUSIONS: Quantitative assessment increases the reliability of the histological assessment of mucosal biopsies in patients with CeD. Such a classification system may be used for clinical trials in patients with CeD. (Intest Res, Published online).

Antenna effect and phosphorescence spectra to find the location of drug tetracycline in bovine ß-lactoglobulin A.

A ternary system comprising of a Eu(III) complex of the drug Tetracycline hydrochloride (Eu3TC) bound to bovine ß-lactoglobulin variant A (BLGA) in aqueous buffer at physiological pH (pH = 7.4) has been investigated to exploit the enhanced "antenna effect" to locate the bound drug and find the microenvironment of the binding site. Steady-state and time-resolved emission studies at room temperature as well as at 77 K have been carried out to evaluate the binding parameters in the binary system consisting of BLGA and tetracycline hydrochloride (TC). Low-temperature phosphorescence studies at 77 K of pure BLGA confirm Trp 19 to be the emitting residue, while Trp 61 is silent. Enhancement of BLGA phosphorescence emission in the ternary system at 77 K indicates that Trp 19 is very close to Eu(III) in the Eu3TC complex. The molecular docking results further confirm that TC binds close to Trp 19 in a hydrophobic domain. The results thus obtained can provide guidelines to design and synthesize target-oriented drugs as well as suitable bio-probes.

Glottic neurogenic tumor: A highly uncommon site for schwannomas.

Schwannomas are benign encapsulated neurogenic tumors, arising from Schwann cells embedded in neurilemal sheath as multinucleated syncytial network. Head and neck schwannomas account for 25%-45% of all schwannomas, majority developing in parapharyngeal space. Laryngeal schwannoma in itself is a rarity, and a glottic origin further complicates its diagnosis. To the best of our knowledge, this is the second case of glottic schwannoma being reported in medical literature. Symptomatology is attributable to mass effect of a slow-growing laryngeal tumor, ranging from sore throat to stridor. The gold standard method for their excision is still debatable. For small tumors, endoscopic or direct laryngoscopic approach is preferred, but for large glottic tumors, external approach is advocated for better exposure and facilitating mucosal grafting. Index case is being presented not only for its rare site of origin, but also to stress on the importance of meticulous histopathological examination to advocate appropriate treatment.

Mixed Phenotypic Acute Leukemia (mixed myeloid/B-cell) with Myeloid Sarcoma of the Thyroid Gland: A Rare Entity with Rarer Asssociation - Detected on FDG PET/CT.

Mixed phenotypic acute leukemia (MPAL) is a rare clinical entity. MPAL associated with myeloidsarcoma (MS) is still rarer with only three cases mentioned in English literature. MS has been described in myriads of location, most commonly in skin, gums and lymph nodes. Although theoritically possible, it is very rare to find MS involving the thyroid gland. The diagnosis of MS can be elusive, very often masquerades and mislabeled as lymphoma. A high index of clinical suspicion coupled with PET/CT findings along with morphological clues and thorough peripheral blood, and bone marrow evaluation is mandatory for arriving at the definitive diagnosis. We report the case of a 58-year-old female presenting with thyroid swelling that was subsequently diagnosed to be MS of the thyroid with underlying MPAL (mixed myeloid/B-cell) only after 18F-FDG PET/CT, which revealed an unusual abnormal pattern of multifocal intense FDG uptake in the thyroid gland.

Androgen Receptor Expression in Endometrial Stromal Sarcoma: Correlation With Clinicopathologic Features.

Endometrial stromal sarcoma (ESS) is a rare neoplasm comprising only 0.2% to 1% of all uterine malignancies and occurs in women between 42 and 59 yr of age. ESSs frequently express estrogen receptor (ER) and progesterone receptor (PR). However, the published literature contains scant data on the expression and therapeutic/prognostic role of androgen receptor (AR) in ESSs. We undertook this study to characterize the expression of AR along with ER and PR in ESSs and correlate it with clinicopathologic features. The clinical details, slides, and blocks of 25 tumors from 24 patients (September 2010 to February 2016) were retrieved. The diagnosis and grade of ESS were reviewed and immunohistochemistry performed with anti-ER, PR, and AR antibodies. Ages ranged from 18 to 50 yr, with a mean age of 36 yr. Low-grade ESS (LGESS) and high-grade ESS (HGESS) were diagnosed in 15 and 9 patients, respectively. An 18-yr-old woman who initially had LGESS suffered a pelvic recurrence; that exhibited high-grade morphology. Our patients, especially those with HGESS, were much younger compared with published worldwide data. ER, PR, and AR immunoreactivity was observed in 14 (93.3%), 12 (80%), and 11 (73.3%) LGESSs, respectively. This is in contrast to HGESSs, in which 5 (50%) tumors had a triple-negative hormonal profile. AR, like ER and PR, was more frequently expressed in LGESS as compared with HGESS. Whether AR, in addition to ER and PR receptor status, may help guide adjuvant hormonal therapy needs further elucidation.

Umbilical cord ulceration: An underdiagnosed entity.

Umbilical cord ulceration is a rare condition presenting with sudden fetal bradycardia due to fetal hemorrhage and in most cases leading to intrauterine death. A strong association with intestinal atresia has been reported. Most cases present after 30 weeks of gestation, with preterm labor or rupture of membranes followed by sudden fetal bradycardia. We report two such cases of umbilical cord ulceration and review the available literature. One of the cases interestingly presented at 26 weeks, much earlier than what is reported in the world literature. In view of high perinatal mortality and morbidity, awareness of this condition is mandatory for timely and appropriate management to improve the fetal outcome.

Extragonadal yolk sac tumor of the head and neck region: A report of two cases.

Extragonadal germ cell tumors (EGCTs) are rare and head and neck is a rarer primary site with most tumors being benign exhibiting teratoma as the leading histologic type. Yolk sac tumor (YST) is relatively uncommon in this location; most commonly described in association with a teratoma and rarely solo. We report two male children, aged 1 year 7 months and 3 years 5 months, with YST involving the head and neck region.

Severe Aplastic Anemia Manifesting After Complete Remission of Acute Promyelocytic Leukemia: Is it a Fortuitous Association?

Acute leukemia, secondary myelodysplasia and paroxysmal nocturnal hemoglobinuria evolving from severe aplastic anemia (AA) following immunosuppressive therapy are well recognized. However, severe AA occurring after complete remission of acute promyelocytic leukemia (APL) has been documented only once in 2009. We report a case of 30-year-old male diagnosed with APL who achieved complete cytogenetic remission with all-trans retinoic acid based induction regimen and developed severe AA few months later during maintenance therapy.

Primary cutaneous leiomyosarcoma: A rare malignant neoplasm.

Primary cutaneous leiomyosarcoma (PCL) is an exceedingly uncommon malignant superficial soft tissue sarcoma with a predilection for middle-aged to elderly male. Morphologic differential diagnosis includes a host of other malignant spindle cell neoplasms, thereby necessitating the use of a panel of immunohistochemical markers to arrive at a definitive diagnosis. We report a case of PCL arising in the right leg of a 70-year-old male.

Intensive method of assessment and classification of the bone marrow iron status: a study of 80 patients.

BACKGROUND: The increasing prevalence of multiple co-morbidities among anemic patients with chronic diseases have made the use of serum ferritin (which is also an acute phase reactant) and transferrin saturation more challenging in diagnosing iron deficiency. Microscopic examination of bone marrow aspirate is the gold standard" for assessing marrow iron store. However, conventional Gale's method assesses iron in marrow fragments alone which provides little valuable information about functional iron deficiency seen in many chronic diseases. AIM: To perform an intensive bone marrow iron grading by assessing iron in fragments, in macrophages around fragments and in erythroblasts and to correlate the marrow iron store results with serum ferritin. MATERIALS AND METHODS: A descriptive study of Perl's Prussian blue stained bone marrow aspirate smears of 80 adult patients with moderate to severe anemia. Bone marrow iron was assessed by both the Gale's method and the intensive method and correlated with serum ferritin. RESULTS: The intensive grading system revealed normal iron stores in 37.5% cases, depleted iron stores in 16.25% patients while 23.75% and 22.5% patients had functional iron deficiency and combined deficiency, respectively. Mean log ferritin concentration was significantly lower in patients with depleted iron stores (0.91 µg/l) in comparison to those with normal iron stores (2.13 µg/l; P = 0.001), functional iron deficiency (2.65 µg/l; P = 0.000), or combined deficiency (2.04 µg/l; P = 0.002). CONCLUSION: Intense marrow iron examination provides a useful iron status classification which is of particular importance in cases of chronic diseases and inflammation.

Glomerulogenesis: can it predict the gestational age? A study of 176 fetuses.

BACKGROUND: Accurate assessment of gestational age of fetuses is essential from both clinical and medico-legal point of view. Crown-rump length, crown-heel length, foot length, and the weight of the fetus are the commonly used parameters for fetal age assessment. However, this estimate often lacks accuracy and sometimes is necessary to combine other data. An analysis of the embryological development of nephrons in the kidney can assist in this determination. OBJECTIVE: To correlate the gestational age with the histological study of sequential development of nephrons in fetal kidney. MATERIALS AND METHODS: This study included 176 fetuses delivered between June 2009 and June 2011 and aged from 12 to 40 weeks. The number of glomerular generations counted in hematoxylin and eosin-stained microscopic sections of the kidneys were correlated with the reported period of gestation based on obstetrical methods. Regression analysis was used to determine the statistical significance of the correlation. RESULTS: A high degree of statistically significant correlation was observed between the period of gestation and the number of glomerular generations (P value < 0.0001). CONCLUSION: The histological assessment of the number of glomerular generations in kidney can be used as a reliable method of estimating fetal age.