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Biomedical subjects

S Kumar

Publications and source records attributed to S Kumar.

At least 1,315 records · Page 73Linked to original sources

Irradiation of bovine aortic endothelial cells enhances the synthesis and secretion of sulphated glycosaminoglycans.

The effect of X-irradiation on the synthesis of heparan sulphate (HS) and chondroitin/dermatan sulphate (CS/DS) by bovine aortic endothelial cells (BAEC), was studied by measuring the incorporation of [35S]sulphate and [3H]glucosamine into newly synthesized glycosaminoglycan (GAG) chains. Medium extracts from irradiated cultures (5Gy) were found to contain approx. 130% more HS and 200% more CS/DS than unirradiated controls. Smaller increases were observed in cellular extracts, irradiated cultures (5Gy) containing approx. 60% more HS and 100% more CS/DS than unirradiated controls. Structural studies showed no significant changes occurred upon irradiation in either the amounts or distribution of N- and O-sulphate groups in the HS molecule. Values for N-sulphation of 41.1% control and 41.5% irradiated (5Gy) were obtained, the corresponding values for O-sulphation being 19.9% control and 20.2% irradiated. Isotope incorporation data indicated that sulphation of CS/DS may decrease after irradiation, however, analysis of chondroitin ABC lyase derived disaccharides showed no changes in the proportion of non-sulphated and O-sulphated disaccharides. The present study indicates that X-irradiation stimulates the synthesis and secretion of HS and CS/DS proteoglycans (PGs) by BAEC. This could be relevant to many features which are found to be indicative of radiation-induced damage.

Animals↗

HhaI methyltransferase flips its target base out of the DNA helix.

The crystal structure has been determined at 2.8 A resolution for a chemically-trapped covalent reaction intermediate between the HhaI DNA cytosine-5-methyltransferase, S-adenosyl-L-homocysteine, and a duplex 13-mer DNA oligonucleotide containing methylated 5-fluorocytosine at its target. The DNA is located in a cleft between the two domains of the protein and has the characteristic conformation of B-form DNA, except for a disrupted G-C base pair that contains the target cytosine. The cytosine residue has swung completely out of the DNA helix and is positioned in the active site, which itself has undergone a large conformational change. The DNA is contacted from both the major and the minor grooves, but almost all base-specific interactions between the enzyme and the recognition bases occur in the major groove, through two glycine-rich loops from the small domain. The structure suggests how the active nucleophile reaches its target, directly supports the proposed mechanism for cytosine-5 DNA methylation, and illustrates a novel mode of sequence-specific DNA recognition.

5-Methylcytosine↗

The DNA (cytosine-5) methyltransferases.

The m5C-MTases form a closely-knit family of enzymes in which common amino acid sequence motifs almost certainly translate into common structural and functional elements. These common elements are located predominantly in a single structural domain that performs the chemistry of the reaction. Sequence-specific DNA recognition is accomplished by a separate domain that contains recognition elements not seen in other structures. This, combined with the novel and unexpected mechanistic feature of trapping a base out of the DNA helix, makes the m5C-MTases an intriguing class of enzymes for further study. The reaction pathway has suddenly become more complicated because of the base-flipping and much remains to be learned about the DNA recognition elements in the family members for which structural information is not yet available.

Amino Acid Sequence↗

A gradient relationship between low birth weight and IQ at age 6 years.

OBJECTIVE: To test for a suspected adverse effect of low birth weight (< or = 2500 g) on IQ at age 6 years in two socioeconomically disparate populations. DESIGN: Nonconcurrent prospective study. SETTING: An urban, predominantly disadvantaged population and a suburban middle-class population in southeastern Michigan. PARTICIPANTS: Low-birth-weight (N = 473) and normal-birth-weight (N = 350) subjects, randomly selected from the 1983 through the 1985 newborn lists of two major hospitals, one serving an urban and the other a suburban population. MAIN OUTCOME MEASURES: The Wechsler Intelligence Scale for Children-Revised to measure IQ. RESULTS: The mean full-scale IQ of low-birth-weight children was 4.9 points lower than that of normal-birth-weight children, controlling for population site, maternal IQ, maternal education, and race (95% confidence interval [CI], 3.0 to 6.8). There was no evidence of low-birth-weight interaction with population site, and the same IQ difference was detected in both populations. In the urban population, low birth weight was associated with an increased risk for IQ below 85 (odds ratio, 2.2; 95% CI, 1.3 to 3.7). In the suburban population, low birth weight was associated with an increased risk for IQ below 100 (odds ratio, 2.0; 95% CI, 1.2 to 3.4). A gradient relationship with full-scale IQ was observed, with the largest deficit in those born weighing 1500 g or less, an intermediate deficit in those born weighing 1501 through 2000 g, and the least pronounced deficit in those born weighing 2001 through 2500 g. CONCLUSIONS: The overall effect of low birth weight represents an increase of approximately 10% of low-birth-weight children with an IQ of more than 1 SD below the population's mean. Children born weighing 2000 g or less bore the major burden of the IQ deficits.

Child↗

Sporotrichosis as a presenting manifestation of hairy cell leukemia.

An infectious episode is the presenting manifestation of hairy cell leukemia (HCL) in approximately 30% of cases. Most often this is bacterial and only rare cases of opportunistic fungal infection are described. We report a patient who presented with sporotrichal involvement of multiple cutaneous sites and lymph nodes. The lesions resolved following antifungal therapy, but persisting pancytopenia and splenomegaly necessitated further hematological evaluation. A diagnosis of HCL was suspected based on morphologically characteristic hairy cells in the peripheral blood that contained tartrate resistant acid phosphatase. A bone marrow biopsy specimen had a normocellular marrow with an increase in interstitial lymphoid cells that stained with L26, MB2, and LN2 antibodies. On flow cytometry these cells were positive for the leukocyte common antigen, B cell markers, and the CD11c antigen confirming the diagnosis of HCL. We believe that this is the first report of sporotrichosis infection as a presenting manifestation of HCL.

Humans↗

Cutaneous T-cell lymphoma diagnosed by fine-needle aspiration: a case report with clinical, cytological, and immunophenotypic features.

A case study of cutaneous T-cell lymphoma in a 40-yr-old Indian male is presented. Prominent clinical features consisted of large skin tumours, lymphadenopathy, and a rapidly progressive course of development. Aspiration smears from tumours, lymph nodes, and bone marrow showed pleomorphic Sezary cells and lymphoid blasts. The neoplastic cells stained positively with anti-HLA DR, CD1, and CD2 antibodies and showed lack of T-cell subset antigens CD4 and CD8. Numerous proliferating cells were identified with 56% of bone marrow cells and 57% of cells in the dermis staining positively with Ki-67 and PC10 antibodies, respectively. This case is reported in view of its uncommon features.

Adult↗

Hepatic uptake and metabolism of 2,3,7,8-tetrachlorodibenzo-p-dioxin and 2,3,7,8-tetrachlorodibenzofuran.

The pharmacokinetics of TCDD and related compounds is congener, dose, and species specific, with urinary and biliary excretion being dependent on the metabolism of these compounds. Isolated hepatocytes and liver slices in suspension culture and hepatic microsomes were used as in vitro models to assess the hepatic uptake and metabolism of [3H]- and [14C]TCDD and [3H]TCDF (0.01-1.0 microM) in control and induced (5 micrograms TCDD/kg, 3 days earlier) male Sprague-Dawley rats. TCDD pretreatment, with an increase in cytochromes P450 1A1 and 1A2 (CYP1A1, CYP1A2), produced an increase in the hepatic uptake of TCDD, while no increase in the hepatic uptake of TCDF was observed. The results are consistent with CYP1A2 serving as a hepatic binding protein for TCDD but not for TCDF. The rates of metabolism of TCDD and TCDF were directly proportional to their concentrations, indicating that the reaction follows first order kinetics at concentrations from 0.01 to 1.0 microM. Very limited metabolism of TCDD and TCDF was observed in control rat liver (0.45 and 3.2 pmol/hr/g hepatocyte wet wt at 0.1 microM, respectively). TCDD induced its own rate of metabolism about two- to fivefold at 1.0 microM but no induction was observed at 0.01 and 0.1 microM. In contrast, TCDD markedly induced the rate of TCDF metabolism at all substrate concentrations. While the results support the role of rat CYP1A1 in TCDF metabolism, the data suggest that CYP1A1 or CYP1A2 may not metabolize TCDD. These results also support the hypothesis that the more rapid metabolism and excretion of TCDF accounts for the relative resistance of the rat to the acute toxicity of TCDF. Comparative studies in rat and human liver microsomes found that TCDF metabolism exhibited first order kinetics in both species. Furthermore, the rate of TCDF metabolism in human liver microsomes was similar to that of control rat liver microsomes. Together the results suggest that TCDF will be far more persistent in rats, and possibly humans, following exposure at low doses which do not significantly induce cytochrome P450 1A1 and/or 1A2.

Animals↗

Chemotherapy followed by radiotherapy versus radiotherapy alone in locally advanced cervical cancer: a randomized study.

Between August 1990 and January 1992, 184 patients with squamous cell carcinoma of the cervix, FIGO stage IIB-IVA, were randomized to receive either two cycles of bleomycin, ifosfamide-mesna, and cis-platinum (BIP) chemotherapy (CT) followed by radiotherapy (RT) ("CT-RT group," n = 94) or RT alone (RT group, n = 90). In the CT-RT group, of 89 evaluable patients, 64 responded: complete response (CR) 4 (4.5%) and partial response 60 (67.5%). Of the remaining 25 patients, 23 had stable disease and 2 progressed. Eighty of 89 patients completed RT as planned. Following RT 56 (70%) achieved CR, 19 (23.7%) had residual disease, and 5 (6.3%) had progressed. CT responders had a better response to RT: 83% (49/59) vs 33.3% (7/21), P < 0.01). The stage of disease, histologic grade, duration of symptoms, and history of smoking had no influence on the response to CT. Patients aged > 45 years and those with Hb > 10 g/dl had significantly better response. Nausea/vomiting, alopecia, grade I-II myelosuppression, diarrhea, and mucositis were the major side effects of CT. Two patients died of CT toxicity. In the RT group, 88 patients were evaluable: 61 (69.3%) patients achieved CR, 25 had residual disease, and 2 progressed. The side effects of RT were cystitis, proctitis, and local skin reaction. These were equally distributed between the two groups. There was no significant difference in overall and disease-free survival in the two groups.

Adult↗

MRI in intraspinal tuberculosis.

We studied 20 patients with intraspinal tuberculosis (TB), to characterise the MRI features of tuberculous meningitis and myelitis. MRI leptomeningitis and intramedullary involvement in 11 patients, intramedullary lesions alone in 5, leptomeningitis alone in 2, and isolated extradural disease in 2. TB leptomeningitis was characterised by loculation of the cerebrospinal fluid (CSF), nerve root thickening and clumping (seen only in the lumbar region) or complete obliteration of the subarachnoid space on unenhanced images. Gd-DTPA-enhanced images proved useful in 6 cases, revealing linear enhancement of the surface of the spinal cord and nerve roots or plaque-like enhancement of the dura-arachnoid mater complex. Intramedullary lesions included tuberculomas (8), cord oedema (5) and cavitation (3). In seven cases of intramedullary tuberculoma multiple lesions with skip areas were seen, without significant cord swelling. One patient had an isolated lesion in the conus medullaris. The lesions were iso- or hypointense on T1-weighted images, iso-, hypo- or hyperintense on T2-weighted images and showed rim or nodular enhancement with contrast medium.

Adolescent↗

Modulation of proteases and their inhibitors in immortal human osteoblast-like cells by tumor necrosis factor-alpha in vitro.

Inflammatory cytokines like interleukin-1 (IL-1) and tumor necrosis factor-alpha (TNF-alpha) are linked to abnormal cartilage and bone loss in a variety of pathological conditions. We have investigated the effect of TNF-alpha on the synthesis and/or steady-state mRNA levels of collagen, alkaline phosphatase (ALP), plasminogen activators (PAs) and their inhibitor PAI-1, and collagenases (MMPs) and their inhibitor TIMP-1 by human osteoblastic, HOS TE85, cells in monolayer cultures. HOS TE85 cells possess approximately 2000 TNF-alpha receptors per cell with a Kd value of 0.67 nM and receptor of approximately 60 kDa. TNF-alpha enhances urokinase-plasminogen activator (u-PA) activity and steady-state mRNA levels twofold without affecting tissue-plasminogen activator (t-PA) or PAI-1. The increase in u-PA mRNA is due to enhanced transcription of this gene. mRNA levels or activities of collagenase 1 (MMP-1), 72- and 92-kDa gelatinases (MMP-2 and MMP-9) are also nearly doubled with little change in the level of expression of TIMP-1. TNF-alpha does not significantly affect the activity or mRNA levels of ALP. TNF-alpha decreases collagen as well as general protein synthesis. However, the steady-state mRNA for the alpha 2 chain of collagen type I is increased three- to fourfold. These results show that TNF-alpha may increase pathological bone turnover by enhancing the rate of transcription of proteases capable of degrading the nonmineralized osteoid layer and decelerating the maturation of the extracellular matrix formed by osteoblasts.

Cell Line↗

Formation and mineralization of extracellular matrix secreted by an immortal human osteoblastic cell line: modulation by tumor necrosis factor-alpha.

Tumor necrosis factor-alpha (TNF-alpha), a 17-kDa cytokine produced by stimulated macrophages/monocytes, modulates the functions of a variety of cells and has been shown to induce bone resorption in vitro. However, the effects that TNF-alpha may have on the process of bone formation are not completely understood. In order to study the effects of TNF-alpha on matrix development and mineralization, we utilized a human osteoblastic cell line, HOS TE85. Our results show that HOS TE85, which has been shown to be responsive to hormones active on normal osteoblasts, forms an extensive extracellular matrix (ECM) that mineralizes during extended culture. Treatment during the development of the matrix with TNF-alpha has little effect on cell number and DNA synthesis, showing thereby that TNF-alpha is not cytotoxic to the cells. However, TNF-alpha inhibits the formation of alkaline phosphatase (AP)-positive foci in a dose-dependent manner at concentrations of 0.1-10 ng/ml. TNF-alpha treatment caused a significant decrease in the incorporation of collagen into the developing matrix. In addition, TNF-alpha treatment resulted in a significant decrease in the synthesis of AP by HOS TE85 cells during the process of ECM formation and resulted in a pronounced lack of mineralization of the ECM. These results indicate that TNF-alpha may be acting as an uncoupler by decreasing the synthesis and incorporation of proteins required for bone formation, and inhibiting matrix formation and mineralization in vitro.

Alkaline Phosphatase↗

Establishment of human parotid pleomorphic adenoma cells in culture: morphological and biochemical characterization.

This study reports the establishment of alpha-amylase-producing human parotid pleomorphic adenoma cell lines (2HP and 2HP1) which have been maintained in culture for over 1 yr. The procedures required preparation of cellular clumps from tumor tissue and plating them on plasma clot or precoated dishes. During the initial phase of growth they required modified MCDB-153 medium without serum. When cells showed signs of degeneration they were changed to MCDB-153 medium containing first 2% and then 10% heat inactivated fetal bovine serum. Although cells grew well in MCDB-153 containing 10% serum, the epithelial cell morphology was not distinct. Therefore, the growth and morphology of cells grown in MCDB-10% serum were compared with those in RPMI growth medium containing 10% fetal bovine serum and F12 containing 10% agammaglobulin newborn bovine serum. Although the growth of cells was a little slower in F12 medium than those in MCDB and RPMI, the epithelial cell morphology was maintained better than in other growth media. The cells of 2HP and 2HP1 produce low levels of alpha-amylase and relatively high levels of alpha-amylase mRNAs of 1176 and 702 bp and contain neurofilament-160, a neuronal-specific marker. The cells of 2HP1 are tumorigenic when tested in athymic mice, but the cells of 2HP are not. The establishment of amylase-producing human parotid adenoma cell lines of different characteristics in culture provides a new opportunity to study the mechanisms of differentiation and transformation, and regulation of alpha-amylase in these cells.

Adenoma, Pleomorphic↗

Establishment and characterization of immortalized cell lines from rat parotid glands.

This study reports for the first time the establishment of immortalized cell lines from normal adult rat parotid glands. The freshly prepared cellular clumps obtained from parotid glands of isoproterenol-treated rats were incubated in 0.2% trypsin solution without EDTA. These clumps were transfected with plasmid vectors pSV3neo and pSV5neo by electroporation and calcium phosphate-Co-DNA-precipitation techniques. The untransfected and transfected cellular clumps were plated in precoated dishes containing modified MCDB-153 medium. Epithelial cells grew from the clumps that were attached. All epithelial cells from untransfected culture died within 6 to 8 wk. Two cell lines which were isolated from transfected cultures subsequently grew on regular tissue culture dishes. One of them, which was isolated from pSV5neo transfected cultures, exhibited non-epithelial cell morphology, but at confluency, many cells mature to acinar-like cells containing numerous granules. The other cell line (2RS), which was isolated from pSV3neo transfected culture, contained cells of non-epithelial and epithelial morphology. During the initial phase of the growth, MCDB-153 medium was essential; however, at a later time, RPMI medium was better than MCDB-153 or F12 medium for maintaining morphology and growth of these cells. The immortalized cells grew in RPMI with a doubling time of about 25 h, synthesize T-antigen, alpha-amylase mRNAs of 1176 and 702 bp, and alpha-amylase and were non-tumorigenic. These amylase-producing cells can be a useful model to study the mechanisms of regulation of growth and differentiation in these cells.

Animals↗

Effect of isoniazid on glutathione biosynthesis and degradation in Mycobacterium smegmatis.

The mechanism of glutathione (GSH) depletion by isoniazid (INH) was studied in M. smegmatis. INH increased the activity of gamma-glutamyl transferase (GGT) whether added before medium inoculation or to actively growing cells. The activity of GGT in cells grown from the beginning in INH-containing medium increased significantly on growth days 2-6. Three-day old M. smegmatis cells treated with INH exhibited a 30-65% increase in the activity of GGT. The activities of gamma-glutamyl-cysteine synthase (GGCS) and GSH synthase (GS) were lowered by 50 and 56% respectively on the second day of growth when M. smegmatis was grown in a medium supplemented with 1.5 mg INH per L. In 3-day old M. smegmatis, INH significantly inhibited the activities of GSH biosynthetic enzymes. The results demonstrate that the increased activity of GGT and decreased activities of GSH biosynthetic enzymes are responsible for GSH depletion by INH in M. smegmatis.

Glutamate-Cysteine Ligase↗

Apparent feeding behaviour of ensheathed third-stage infective larvae of human hookworms.

The third-stage ensheathed infective larvae (L3) of hookworms are reputed to be non-feeding stages, and the L2-derived sheath is considered to be a barrier to macromolecular uptake and secretion. During a study of the lectin-binding characteristics of en- and exsheathed L3 larvae of Ancylostoma duodenale, fluorescein isothiocyanate conjugated lectins (con A and Ulex europaeus agglutinin) were repeatedly seen in the oesophageal lumen, as well as on the surface of L3 larvae encased in the L2 sheaths. Repetition of these experiments using Necator americanus L3 larvae yielded similar results. These observations suggest that ensheathed hookworm larvae can obtain/utilise and secrete macromolecules, despite the presence of a sheath.

Ancylostoma↗

Insudative lesions--their pathogenesis and association with glomerular obsolescence in diabetes: a dynamic hypothesis based on single views of advancing human diabetic nephropathy.

Kidneys from 74 consecutive, primarily non-insulin-dependent diabetics at autopsy and 59 age-, sex, and ethnic group-matched controls were examined qualitatively and semiquantitatively to determine the prevalence and severity of insudative lesions (ILs) and obsolescent glomeruli with (OGcFC) and without (OGsFC) insudative (fibrin cap) lesions. A subset of 25 cases with advanced diabetic changes was examined using serial sections, immunohistochemical stains, and electron microscopy to determine the pathogenesis of ILs and OGcFCs. Insudative lesions consisted of intramural accumulations (hereafter called deposits) of presumably imbibed plasma proteins and lipids within renal arterioles, glomerular capillaries, Bowman's capsule, and proximal convoluted tubules. Insudative lesions in Bowman's capsule are called capsular drop lesions (CDs), in glomerular capillaries they are called fibrin cap lesions (FC), and in afferent and efferent arterioles they are called hyalinized afferent (HA) and hyalinized efferent (HE) arterioles, respectively. All ILs were much more numerous and/or larger in diabetics than in controls. Contrary to previous opinion, CDs and HE arterioles were not specific for diabetes, being present in small numbers in nine (15%) controls. Controls with CD/HE arterioles had far more HA arterioles and focal mesangiolyses (FMs) than those without. Insudative lesions consisted of the well known homogenous eosinophilic deposits (homogenous eosinophilic ILs) and the less familiar foamy, reticulated, and vacuolar deposits (heterogenous lucent ILs). Homogenous eosinophilic ILs were predominant in afferent arterioles and more so in efferent arterioles, and were segregated into globules of varying density with the denser deposits located peripherally. Two types of CDs, which differed sharply in location and composition, were found. The first was mostly homogenous eosinophilic, usually without capsular adhesions and located near the vascular pole close to preglomerular arterioles. The second was mostly heterogenous lucent, located away from the vascular pole, and consistently connected by adhesions to the capillary tuft usually near FMs and/or Kimmelstiel-Wilson (KW) nodules. The latter ILs sometimes extended in continuity along the internal surface of the basement membrane from Bowman's capsule into the proximal convoluted tubule. It was hypothesized that ILs traveled centrifugally through the walls of preglomerular arterioles to form the first type of CD and longitudinally within the walls of afferent arterioles and glomerular capillaries and through adhesions to form the second. Contrary to previous opinion, FCs were consistently intramural. When numerous, FCs were associated with a form of glomerular obsolescence called OGcFC.(ABSTRACT TRUNCATED AT 400 WORDS)

Age Factors↗

Conformation of a T-cell stimulating peptide of interleukin-1 beta protein: circular dichroism studies.

A T-cell stimulating peptide Val-Gln-Gly-Glu-Glu-Ser-Asn-Asp-Lys-OH, the 163-171 fragment epitope of interleukin-1 beta (IL-1 beta), has been synthesized in solution phase and purified by reverse-phase high-performance liquid chromatography (RP-HPLC). The backbone conformation of the synthetic fragment, investigated in aqueous solution by circular dichroism (CD) spectroscopy, is qualitatively a mixture of beta-turns and random coil. Quantification of the CD spectra revealed the presence of a 9% beta-turn fraction in water at pH 7.0, suggesting the occurrence of the conformation for the epitope fragment in aqueous solution necessary for T-cell stimulation and antigenicity. Concomitant changes in CD spectra were observed with increases in the trifluoroethanol (TFE) concentration in water, and the beta-turn fraction in peptide increased to 28% at a concentration of 90% TFE. This helicogenic solvent, as well as other solvents such as methanol, acetonitrile and dioxane (all favouring an ordered structure in peptides), failed to induce any alpha-helical conformation in the IL-1 beta (163-171) fragment, and CD spectra were attributed to only beta-turn ordered structure. This beta-turn structure has also been found to be a theoretically preferred conformation using Chou-Fasman proclivity data and is in accordance with the presence of an all-beta-globular conformation for its parent molecule IL-1 beta. Thus, the beta-turn conformation is probably involved in retention of T-cell stimulation activity in this synthetic epitope.

Amino Acid Sequence↗

Human chorionic gonadotropin (hCG) increases cytosolic free calcium in adult rat Leydig cells.

Regulation of steroidogenesis by luteinizing hormone is mediated by cAMP and calcium. We have investigated changes in cytosolic free calcium ([Ca2+]i) in Leydig cells by using Fura-2 as the fluorescent calcium indicator. Purified Leydig cells were plated on polylysine coated glass coverslips, cultured for 24 h in DMEM/F12 and loaded with Fura-2 at 37 degrees C. [Ca2+]i measurements were made fluorometrically by placing coverslips into 3 ml cuvettes with PBS+calcium. Addition of hCG increased [Ca2+]i gradually after a lag of about 2-3 min and plateaued by 5-6 min. The plateau level was sustained for at least 15 min. Absence of external Ca2+ in the medium or presence of diltiazem or nicardipine or cobalt chloride abolished the rise. Addition of BAY K 8644 or KCl caused a small but significant increase of [Ca2+]i. 8-Br-cAMP, forskolin or cholera toxin produced a gradual rise in [Ca2+]i that plateaued after 5-6 min similar to that observed with hCG. The action of hCG was inhibited by protein kinase A inhibitor (20-residues peptide) but not by protein kinase C inhibitor (staurosporine). We conclude that binding of hCG to its receptors would transmit the signal through G proteins to adenylyl cyclase to increase cAMP which would increase Ca2+ influx into cytosol across plasma membrane Ca2+ channels. Therefore, it appears that the primary action of hCG is to increase cytosolic cAMP which would then regulate [Ca2+]i as well as steroidogenesis.

Animals↗