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.

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.

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.

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.

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.

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.

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.

STUDY OF KUSTHA (SAUSSUREA LAPPA, Clarke) IN ISCHAEMIC HEART DISEASE.

This article reports the results obtained from a study of the usefulness of Saussurea Lappa on the treatment of Ichaemic Heart Diseases.

Cytochemical differentiation of tonic (slow) and phasic (fast) muscle units in the ocular muscles of a fresh-water carp, Catla catla (Ham.).

1. Functional morphology of the ocular muscles with special reference to their topography, innervation and action have been studied in a fresh-water carp Catla catla. An attempt has also been made to differentiate tonic (slow) and phasic (fast) muscle units in the various ocular muscles by their SDH activities and their lipid contents. 2. Catla catla has large eyes (2 cm in diameter) in a fish measuring 30 cms. There are four recti (inferior, superior, anterior and posterior) and two obliquii (superior and inferior) muscles for the synchronized rotation of the eye ball within the orbit. Anterior and posterior myodomes are well developed. The former accommodates two obliquii muscles, while the latter gives space for the posterior and anterior recti. 3. In this fish, all the ocular muscles are composite in nature being made up of tonic (slow) and phasic (fast) muscle units. The former is made up of red fibers whereas the latter have only white fibers. In the tonic muscle units lipid is more concentrated. 4. The superior and the inferior obliquii muscles rotate the eye ball on its antero-posterior axis. The upward and downward movements of the eye balls are achieved by the superior and the inferior recti. The anterior and posterior recti move the eye ball in the antero-lateral and postero-lateral directions. 5. The superior obliquii contains about 41% of the tonic and 59% of phasic muscle units. The inferior oblique is made up of 25% of tonic and 75% of the phasic muscle units. 6. The Anterior rectus contains greater percentage of tonic muscle fibers (48%) than the superior rectus (40%), the Interior rectus (34%) and the posterior rectus (35%).

Gill microcirculation of the air-breathing climbing perch, Anabas testudineus (Bloch):relationships with the accessory respiratory organs and systemic circulation.

The general macrocirculation and branchial microcirculation of the air-breathing climbing perch, Anabas testudineus, was examined by light and scanning electron microscopy of vascular corrosion replicas. The ventral aorta arises from the heart as a short vessel that immediately bifurcates into a dorsal and a ventral branch. The ventral branch distributes blood to gill arches 1 and 2, the dorsal branch to arches 3 and 4. The vascular organization of arches 1 and 2 is similar to that described for aquatic breathing teleosts. The respiratory lamellae are well developed but lack a continuous inner marginal channel. The filaments contain an extensive nutritive and interlamellar network; the latter traverses the filament between, but in register with, the inner lamellar margins. Numerous small, tortuous vessels arise from the efferent filamental and branchial arteries and anastomose with each other to form the nutrient supply for the filament, adductor muscles, and arch supportive tissues. The efferent branchial arteries of arches 1 and 2 supply the accessory air-breathing organs. Arches 3 and 4 are modified to serve primarily as large-bore shunts between the dorsal branch of the ventral aorta and the dorsal aorta. In many filaments from arches 3 and 4, the respiratory lamellae are condensed and have only 1-3 large channels. In some instances in arch 4, shunt vessels arise from the afferent branchial artery and connect directly with the efferent filamental artery. The filamental nutrient and interlamellar systems are poorly developed or absent. The respiratory and systemic pathways in Anabas are arranged in parallel. Blood flows from the ventral branch of the ventral aorta, through gill arches 1 and 2, into the accessory respiratory organs, and then returns to the heart. Blood, after entering the dorsal branch of the ventral aorta, passes through gill arches 3 and 4 and proceeds to the systemic circulation. This arrangement optimizes oxygen delivery to the tissues and minimizes intravascular pressure in the branchial and air-breathing organs. The efficiency of this system is limited by the mixing of respiratory and systemic venous blood at the heart.

Morphology and vascular anatomy of the accessory respiratory organs of the air-breathing climbing perch, Anabas testudineus (Bloch).

The vascular organization and endothelial cell specialization of the air-breathing organs of Anabas testudineus were examined by light and scanning electron microscopy of fixed tissue and vascular corrosion replicas. The vessels supplying blood to the lining of paired suprabranchial chambers and the plicated labyrinthine organs within the chambers are tripartite, having a median artery and paired, lateral veins. Hundreds of respiratory islets, the functional units of gas exchange, cover the surfaces of both the chamber and labyrinthine organ. A median islet artery supplies the central aspect of each islet and gives rise to numerous short arterioles from which the transverse channels are formed. Transverse channels are parallel capillary-sized vessels that extend in two rows away from the medial arterioles and drain laterally into one of two lateral islet veins. Basally situated single rows of endothelial cells lining the transverse channels form thick, evaginated, tongue-like cytoplasmic processes that project freely into the lumen from the tissue side of the channel. Other thin, septate, cytoplasmic extensions of the same cells form valve-like septa that extend across the channel. Both the septa and tongue-like processes appear to direct the red blood cells to the epithelial side of the channel and thus decrease the diffusion distance between the air and red cell. A large sinusoidal space lies under the transverse channels and may support the channels and even elevate them during increased oxygen demand. The epithelium covering the transverse channels is smooth, which enhances air convection and minimizes unstirred layer effects. The epithelium between the channels contains microvilli that may serve to trap bacteria or particulates and to humidify the air chambers.

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.