Longitudinal analysis of 25 sequential sample-pairs using a custom multiple myeloma mutation sequencing panel (M(3)P).

Recent advances in genomic sequencing technologies now allow results from deep next-generation sequencing to be obtained within clinically meaningful timeframes, making this an attractive approach to better guide personalized treatment strategies. No multiple myeloma-specific gene panel has been established so far; we therefore designed a 47-gene-targeting gene panel, containing 39 genes known to be mutated in ≥3 % of multiple myeloma cases and eight genes in pathways therapeutically targeted in multiple myeloma (MM). We performed targeted sequencing on tumor/germline DNA of 25 MM patients in which we also had a sequential sample post treatment. Mutation analysis revealed KRAS as the most commonly mutated gene (36 % in each time point), followed by NRAS (20 and 16 %), TP53 (16 and 16 %), DIS3 (16 and 16 %), FAM46C (12 and 16 %), and SP140 (12 and 12 %). We successfully tracked clonal evolution and identified mutation acquisition and/or loss in FAM46C, FAT1, KRAS, NRAS, SPEN, PRDM1, NEB, and TP53 as well as two mutations in XBP1, a gene associated with bortezomib resistance. Thus, we present the first longitudinal analysis of a MM-specific targeted sequencing gene panel that can be used for individual tumor characterization and for tracking clonal evolution over time.

Genomic characterization of high-count MBL cases indicates that early detection of driver mutations and subclonal expansion are predictors of adverse clinical outcome.

High-count monoclonal B-cell lymphocytosis (MBL) is an asymptomatic expansion of clonal B cells in the peripheral blood without other manifestations of chronic lymphocytic leukemia (CLL). Yearly, 1% of MBLs evolve to CLL requiring therapy; thus being critical to understand the biological events that determine which MBLs progress to intermediate/advanced CLL. In this study, we performed targeted deep sequencing on 48 high-count MBLs, 47 of them with 2-4 sequential samples analyzed, exploring the mutation status of 21 driver genes and evaluating clonal evolution. We found somatic non-synonymous mutations in 25 MBLs (52%) at the initial time point analyzed, including 12 (25%) with >1 mutated gene. In cases that subsequently progressed to CLL, mutations were detected 41 months (median) prior to progression. Excepting NOTCH1, TP53 and XPO1, which showed a lower incidence in MBL, genes were mutated with a similar prevalence to CLL, indicating the early origin of most driver mutations in the MBL/CLL continuum. MBLs with mutations at the initial time point analyzed were associated with shorter time-to-treatment (TTT). Furthermore, MBLs showing subclonal expansion of driver mutations on sequential evaluation had shorter progression time to CLL and shorter TTT. These findings support that clonal evolution has prognostic implications already at the pre-malignant MBL stage, anticipating which individuals will progress earlier to CLL.

Chronic ulcerative stomatitis: diagnostic and management challenges--four new cases and review of literature.

Chronic ulcerative stomatitis (CUS) is a recently described mucocutaneous disease characterized by involvement of mucosal surfaces and skin. The disease usually presents in the form of painful oral ulcers and has been seen predominantly among older women. Clinically, CUS patients may exhibit erosive or ulcerative lesions of oral mucosa resembling erosive lichen planus and/or other vesiculobullous lesions. Routine histology may exhibit features of lichenoid mucositis and is often nondiagnostic. Direct immunofluorescence studies of mucosal or skin biopsies usually reveal a unique pattern of IgG bound to nuclei of keratinocytes of the basal and parabasal cell layers. Chronic ulcerative stomatitis exhibits a unique resistance to standard treatments available, including topical and systemic corticosteroids and immunomodulatory medications. However, long-lasting favorable clinical responses may be achieved with hydroxychloroquine pharmacotherapy. We present the clinicoimmunopathologic findings of four cases of CUS and discuss the varied clinical, microscopic spectrum, and treatment for this chronic, debilitating, and often recalcitrant condition. A systematic review of the literature, including 35 previously reported cases, is presented.

Cytochemical differentiation of red and white fibers in the respiratory muscles of a fresh-water mud-eel, Macrognathus aculeatum (Bloch).

1. Functional morphology including the origin, insertion, innervation and action of the respiratory muscles in relation to buccal pressure and opercular suction pumps in a fresh-water mud-eel, Macrognathus aculeatum have been studied. 2. Cytochemical studies were made on the SDH activity of Adductor maxillaris, Adductor mandibularis, Protractor hyoideus, Dilator-operculi, Levator operculi, Adductor operculi, Inferior and Superior Hyohyoideus and the adductor muscles of gill filaments. The intensity of reaction reveals the composite nature of the respiratory muscles. Adductor mandibularis is made up entirely of white muscle fibers whereas others are composed of red as well as white muscle fibers. The Adductor maxillaris consists of superficial white muscle fibers whereas the interior is dominated by red muscle fibers. 3. The muscles innervated by the branches of VIIth facialis is dominated by red fibers whereas those innervated by the Vth trigeminal are dominated by white muscle fibers. 4. The adductor muscle units of the gills are made up entirely of red muscle fibers. This suggests the continued role of adductor muscle during the ventilation of gill lamellae.

Comparative studies of boerhaavia diffusa L. And boerhaavia verticillata poir. (Nyctaginaceae).

Comparative taxonomical and phytochemical studies of B diffusa ands B. verticillata are described in this article.

Evaluation of the diuretic activity of boerhaavia verticillata.

The present investigation was undertaken to evaluate the potential diuretic activity of Boerhaavia verticillata . The ethanolic and aqueous extracts of B. verticillata were administered in doses of 50 and 100 mg/kg, i.p., to normal and adult albino rats, which were monitored over a period of 24 h. Experimental results confirmed that both the ethanol and aqueous extracts have diuretic properties, but the ethanol extract seems to have less diuretic and natriuretic activity than the aqueous extract.

Cytochemical differentiation of muscle fibers by succinic dehydrogenase (SDH) activity in the respiratory muscles of an air-breathing fish, Channa punctatus (Bloch).

1. A cytochemical study on the succinic dehydrogenase activity of the respiratory muscles of Channa punctatus reveals that the Adductor mandibularis is made up entirely of white muscle fibers. 2. The Adductor, Dilator and Levator operculi are composite muscles and composed of white as well as red fiber bundles. However, these composite muscles are dominated by white muscle fibers. 3. The interior of the Protractor hyoideus is dominated by red and medium fibers whereas its superficial part is dominated by white muscle fibres. 4. The muscles innervated by the branches of V trigeminal nerve (Adductor mandibularis, Dilator operculi) are dominated by white fibers, whereas those innervated by the VII facialis (Adductor and Levator operculi and Protractor hyoideus) show dominance of red fibers.

A review of the history of prameha and diabetes mellitus.

Several diseases of polyuric nature were described in Ayurveda. Collectively called Prameha, this group includes an entity called Madhumeha which is the equivalent of diabetes mellitus. The medical history of the two diseases is described in the article.

Oxygen uptake capacity of gills and skin in relation to body weight of the air-breathing siluroid fish, Clarias batrachus (Linn.).

Oxygen consumption through gills and skin in relation to body weight was estimated in the air-breathing catfish, Clarias batrachus, under two experimental conditions, viz., (i) when access to air was allowed and (ii) when air-breathing was prevented. There was a positive correlation between VO2 (ml/hr) and body weight in both experimental conditions. Oxygen consumption (ml/hr) increased by a power of 0.869 when access to air was allowed whereas the power was slightly less (b = 0.841) when air-breathing was prevented. As the values for exponent (b) were less than 1.0, the weight specific VO2 (ml/kg/hr) decreased with increasing body weight. The decrease was more marked (b = - 0.180) in fishes which were not allowed air than in those where access to air was allowed (b = - 0.148). Under normal conditions of water and air-breathing the rate of VO2 (ml/kg/hr) via gills and skin from water ranged from 39.7 +/- 3.21 to 76.7 +/- 9.01 and this increased to 42.17 +/- 6.2 to 105.9 +/- 8.33 when air-breathing was prevented. The increase in the rate of VO2 was perhaps associated with the increase in the volume of water irrigating the gills per unit time.

Succinic dehydrogenase activity of the respiratory muscles of a fresh-water teleost, Bagarius bagarius (Ham.) (Sisoridae, Pisces).

Functional morphology including the origin, insertion, and innervation of the respiratory muscles in relation to buccal pressure pump and opercular suction pumps in a fresh-water bottom dwelling siluroid fish, Bagarius bagarius have been studied. Histochemical studies were made on the succinic dehydrogenase activity of adductor mandibulae, retractor tentaculi, levator operculi, dilatator operculi, adductor operculi, intermandibularis, interhyoideus, hyohyoideus superior and constrictor branchialis. The intensity of reaction reveals the presence of three types of muscle fibres in some of the respiratory muscles. The muscle containing red muscle fibres are mostly innervated by the branches of the VIIth cranial nerve. The retractor tentaculi consists of superficial white muscle fibres and the interior part is dominated by red muscle fibres. The muscles (adductor operculi, levator operculi, dilatator operculi, interhyoideus, hyohyoideus superior) concerned with the opercular suction pumps are of mixed type and consist of white and red muscle fibres, whereas adductor mandibulae and intermandibularis are made up entirely of white muscle fibres. The adductor muscle bundles of the constrictor branchialis, which are responsible for movement of gill filaments, are dominated by the red muscle fibres. The abductor part, however, is made up entirely of white muscle fibres.

Comparative histochemical study of the succinic dehydrogenase (SDH) activity in the respiratory muscles of an air-breathing siluroid fish, Clarias batrachus (Linn.).

1. Funcitonal morphology including origin, insertion, innervation and action of the respiratory muscles in relation to the respiratory pumps and the movement of barbels and gill filaments have been studied in a fresh-water air-breathing siluroid fish, Clarias batrachus (Linn.). 2. Cytochemical studies were made on the succinic dehydrogenase (SDH) activity of:--Abductor hyomandibularis, Adductor arcus palatini, Adductor hyomandibularis, Adductor mandibularis, Intermandibularis anterior and posterior, Interhyoideus, and Rectus cervicis (all concerned with buccal pressure pump), Adductor operculi, Dilator operculi, Levator operculi, Inferior hyohyoideus, Superior hyohyoieus (all concerned with opercular suction pumps), Extensor tentaculi, Protractor tentaculi, Retractor tentaculi (movement of barbels), adductor and abductor muscle sheets of the Constrictor branchialis (movement of gill filaments). 3. The intensity of SDH reaction reveals that except the Adductor mandibularis, Intermandibularis anterior and the Protractor tentaculi all muscles are made up of red as well as white muscle fibers. 4. The muscle innervated by the branches of Vth (trigeminal) nerve is dominated by white fibres, whereas those innervated by the branches of VIIth (facialis) contain more red muscle fibres. 5. The Retractor tentaculi (concerned with the movement of maxillary barbel) consists of superficial white fibers and its interior deeper part is dominanted by red and intermediate muscle fibres. 6. The adductor muscle sheet of the Constrictor branchialis is made up of red fibres, whereas the abductor muscle sheet is entirely made up of white muscle fibres.

Histochemical localization of acid mucopolysaccharide in the respiratory muscles of a fresh-water air-breathing siluroid fish, Clarias batrachus (Linn.).

Respiratory muscles involved in gill ventilation (= irrigation) of an amphibious siluroid fish, Clarias batrachus (Linn.) were studied by phase contrast and light microscopy after the treatment with PAS. Alcian Blue at pH 2.5 and 1.0, dialyzed iron and Toludine Blue. The transverse muscle bands lightly stained with PAS, Alcian Blue at pH 2.5 and 1.0 and Dialyzed Iron suggesting that the mucopolysaccharide occured in relatively low concentrations. Phase contrast microscopy indicated that the transverse bands stained by the above mentioned reagents correspond to the I-bands. Methylation for 4 hours at 60 degrees C prevented I-band staining with Alcian Blue in the muscles studied. Saponification alone left I-band alcianophilia intact. These findings reveal that myofibrillar I-bands of respiratory muscles contain sulphated acid mucosubstances.

Effect of indigenous drug (Pushkarmula) on experimentally induced myocardial infarction in rats.

In the present study, myocardial infarction was induced experimentally in rats by isoprenaline injection. Circulating GOT, LDH, CPK, cAMP, Cortisol, pyruvate, lactate glucose and cardiac cAMP adenyl cyclase levels were gradually increased and serum and cardiac cAMP-PDE levels were gradually decreased from 1 hour to 120 hours after the first injection of isoprenaline. In the rats pretreated with ciplar (beta blocker) or Pushkarmula (indigenous drug) these changes were less when compared to untreated infarcted rats. Similar type of results were also observed in the infarcted rats post treated with Pushkarmula. The pretreatment with Pushkarmula was found to be more effective than post treatment which gives a preventive and curative bearing of the drug in myocardial infarction.

Vascular organization of the head and respiratory organs of the air-breathing catfish, Heteropneustes fossilis.

Light and scanning electron microscopy of vascular replicas from the facultative air-breathing fish Heteropneustes fossilis show modifications in the macrocirculation of the respiratory organs and systemic circulation, whereas, gill microcirculation is similar to that found in typical water-breathing fish. Three and sometimes four ventral aortae arise directly from the bulbus. The most ventral vessel supplies the first pair of arches. Dorsal to this another aorta supplies the second gill arches, and a third, dorsal to, and larger than the other two, supplies the third and fourth arches and the air sacs. Occasionally a small vessel that may be the remnant of a primitive aortic arch arises from the first ventral aorta and proceeds directly to the mandibular region without perfusing gill tissue. The air sac is perfused by a large-diameter extension of the afferent branchial artery of the fourth gill arch and its circulation is in parallel with the gill arches. Blood drains from the air sac into the fourth arch epibranchial artery. A number of arteries also provide direct communication between the efferent air sac artery and the dorsal aorta. All four gill arches are well developed and contain respiratory (lamellar) and nonrespiratory (interlamellar and nutrient) networks common to gills of water-breathing fish. Air sac lamellae are reduced in size. The outer 30% of the air sac lamellar sinusoids are organized into thoroughfare channels; the remaining vasculature, normally embedded in the air sac parenchyma, is discontinuous. A gill-type interlamellar vasculature is lacking in the air sac circulation. Despite the elaborate development of the ventral aortae, there is little other anatomical evidence to suggest that gill and air sac outflow are separated and that dorsal aortic oxygen tensions are maintained when the gills are in a hypoxic environment. Physiological adjustments to hypoxic water conditions probably include temporal regulation of gill and air sac perfusion to be effective, if indeed they are so.

Vasculature of the head and respiratory organs in an obligate air-breathing fish, the swamp eel Monopterus (=Amphipnous) cuchia.

Methyl methacrylate vascular corrosion replicas were used to examine the macrocirculation in the head region and the microcirculation of respiratory vessels in the air-breathing swamp eel Monopterus cuchia. Fixed respiratory tissue was also examined by SEM to verify capillary orientation. The respiratory and systemic circulations are only partially separated, presumably resulting in supply of mixed oxygenated and venous blood to the tissues. A long ventral aorta gives rise directly to the coronary and hypobranchial arteries. Two large shunt vessels connect the ventral aorta to the dorsal aorta, whereas the remaining ventral aortic flow goes to the respiratory islets and gills. Only two pairs of vestigial gill arches remain, equivalent to the second and third arches, yet five pairs of aortic arches were identified. Most aortic arches supply the respiratory islets. Respiratory islet capillaries are tightly coiled spirals with only a fraction of their total length in contact with the respiratory epithelium. Valve-like endothelial cells delimit the capillary spirals and are unlike endothelial cells in other vertebrates. The gills are highly modified in that the lamellae are reduced to a single-channel capillary with a characteristic three-dimensional zig-zag pathway. There are no arterio-arterial lamellar shunts, although the afferent branchial artery supplying the gill arches also supplies respiratory islets distally. A modified interlamellar filamental vasculature is present in gill tissue but absent or greatly reduced in the respiratory islets. The macro- and micro-circulatory systems of M. cuchia have been considerably modified presumably to accommodate aerial respiration. Some of these modifications involve retention of primitive vessel types, whereas others, especially in the microcirculation, incorporate new architectural designs some of whose functions are not readily apparent.