Assessment of severity and feto-maternal outcomes among COVID-19 infected pregnant females hospitalized at tertiary care center in western Nepal.

Objective: There is a paucity of data assessing the severity of coronavirus disease 2019 (COVID-19) infection and feto-maternal outcomes in pregnant women in Nepal, especially during the third wave of the pandemic with the Delta variant. This study aimed to severity as well as maternal-fetal outcomes among pregnant women with COVID-19 infection. Methods: In this cross-sectional study carried out amongst COVID-19 positive pregnant women visiting our institution for 6 months from January through June 2022, data surrounding COVID-19 severity and fetal outcomes were collected in a structured questionnaire. Disease severity was categorized based on the NIH (National Institutes of Health) severity classification. Descriptive statistics were used to describe the outcomes. Results: A total of 105 pregnant females infected with COVID-19 during the third wave were included in our study. The participants ranged from 16 through 40 years, mean gestational age being 36.6 weeks. Most of the women (40.55%) belonged to Brahmin and Chhetri ethnic backgrounds. About half (48.55%) of females were asymptomatic and four patients had severe disease requiring intensive care management. Only 13 pregnant females underwent vaccination. Of the 91 females that underwent delivery at our center, 77 (84.62%) were term. The rate of cesarean delivery was slightly higher (51.65%) than normal delivery. None of the babies born to COVID-positive pregnant females tested positive for COVID-19 and there were no stillbirths or any congenital anomalies. The majority of babies (68.14%) had normal fetal weight. Conclusion: Severe COVID-19 infection and adverse maternal-fetal outcomes during the third wave of COVID-19 infection with the Delta variant in Nepal were less than in other regions of the world despite low rates of vaccination.

Improving maternal and neonatal departments in high and low resource settings: the opinion of local health providers.

AIM: We compared local health caregivers' opinions regarding the priority areas for improving the maternal and neonatal departments in low and high resource countries. METHODS: Personnel involved in maternal and neonatal care operating in level III, teaching hospitals in four countries (Sri Lanka, Mongolia, USA, and Italy) were asked to fill out an anonymous, written questionnaire. RESULTS: The questionnaire was completed by 1112 out of 1265 (87.9%) participants. "Personnel's education" was classified as the first most important intervention by health providers working in high (49.0%) as well as in low (29.9%) resource countries, respectively. Improvement in salary, equipment, internet access, and organizational protocols were considered as the most important interventions by a significantly larger percentage of personnel from low resource countries in comparison with those from high resource countries. Health providers from high resource countries considered organizational aspects (to define specific roles and responsibilities) as a priority more frequently than their colleagues from low resource countries. CONCLUSIONS: Although education of personnel was valued as the highest priority for improving maternal and neonatal departments there are substantial differences in priorities associated with the working setting. Local caregivers' opinion may contribute to better design interventions in settings with high or limited resources.

Quinone reduction via secondary B-branch electron transfer in mutant bacterial reaction centers.

Symmetry-related branches of electron-transfer cofactors-initiating with a primary electron donor (P) and terminating in quinone acceptors (Q)-are common features of photosynthetic reaction centers (RC). Experimental observations show activity of only one of them-the A branch-in wild-type bacterial RCs. In a mutant RC, we now demonstrate that electron transfer can occur along the entire, normally inactive B-branch pathway to reduce the terminal acceptor Q(B) on the time scale of nanoseconds. The transmembrane charge-separated state P(+)Q(B)(-) is created in this manner in a Rhodobacter capsulatus RC containing the F(L181)Y-Y(M208)F-L(M212)H-W(M250)V mutations (YFHV). The W(M250)V mutation quantitatively blocks binding of Q(A), thereby eliminating Q(B) reduction via the normal A-branch pathway. Full occupancy of the Q(B) site by the native UQ(10) is ensured (without the necessity of reconstitution by exogenous quinone) by purification of RCs with the mild detergent, Deriphat 160-C. The lifetime of P(+)Q(B)(-) in the YFHV mutant RC is >6 s (at pH 8.0, 298 K). This charge-separated state is not formed upon addition of competitive inhibitors of Q(B) binding (terbutryn or stigmatellin). Furthermore, this lifetime is much longer than the value of approximately 1-1.5 s found when P(+)Q(B)(-) is produced in the wild-type RC by A-side activity alone. Collectively, these results demonstrate that P(+)Q(B)(-) is formed solely by activity of the B-branch carriers in the YFHV RC. In comparison, P(+)Q(B)(-) can form by either the A or B branches in the YFH RC, as indicated by the biexponential lifetimes of approximately 1 and approximately 6-10 s. These findings suggest that P(+)Q(B)(-) states formed via the two branches are distinct and that P(+)Q(B)(-) formed by the B side does not decay via the normal (indirect) pathway that utilizes the A-side cofactors when present. These differences may report on structural and energetic factors that further distinguish the functional asymmetry of the two cofactor branches.

Calcium binding properties of gamma-crystallin: calcium ion binds at the Greek key beta gamma-crystallin fold.

The beta- and gamma-crystallins are closely related lens proteins that are members of the betagamma-crystallin superfamily, which also include many non-lens members. Although beta-crystallin is known to be a calcium-binding protein, this property has not been reported in gamma-crystallin. We have studied the calcium binding properties of gamma-crystallin, and we show that it binds 4 mol eq of calcium with a dissociation constant of 90 microm. It also binds the calcium-mimic spectral probes, terbium and Stains-all. Calcium binding does not significantly influence protein secondary and tertiary structures. We present evidence that the Greek key crystallin fold is the site for calcium ion binding in gamma-crystallin. Peptides corresponding to Greek key motif of gamma-crystallin (42 residues) and their mutants were synthesized and studied for calcium binding. These peptides adopt beta-sheet conformation and form aggregates producing beta-sandwich. Our results with peptides show that, in Greek key motif, the amino acid adjacent to the conserved aromatic corner in the "a" strand and three amino acids of the "d" strand participate in calcium binding. We suggest that the betagamma superfamily represents a novel class of calcium-binding proteins with the Greek key betagamma-crystallin fold as potential calcium-binding sites. These results are of significance in understanding the mechanism of calcium homeostasis in the lens.

Antioxidant properties of green and black tea, and their potential ability to retard the progression of eye lens cataract.

Aqueous extracts of green and black tea are shown to quench reactive oxygen species such as singlet oxygen, superoxide and hydroxyl radicals, prevent the oxidative cross-linking of test proteins and inhibit single strand breakage of DNA in whole cells. They are also seen to be able to counteract the oxidative insult mounted by cigarette smoke. In rats in which cataract was induced by subcutaneous injection of selenite, administration of green or black tea extracts led to a retardation of the progression of lens opacity, suggesting the potential cataracto-static ability of tea.

beta-carbolines that accumulate in human tissues may serve a protective role against oxidative stress.

beta-Carbolines are tricyclic nitrogen heterocycles formed in plants and animals as Maillard reaction products between amino acids and reducing sugars or aldehydes. They are being detected increasingly in human tissues, and their physiological roles need to be understood. Two beta-carboline carboxylates have been reported to accumulate in the human eye lens. We report here on the identification of another beta-carboline, namely 1-methyl-1-vinyl -2, 3,4-trihydro-beta-carboline-3-carboxylic acid, in the lenses of some cataract patients from India. Analysis of these three lenticular beta-carbolines using photodynamic and antioxidant assays shows all of them to be inert as sensitizers and effective as antioxidants; they quench singlet oxygen, superoxide and hydroxyl radicals and inhibit the oxidative formation of higher molecular weight aggregates of the test protein, eye lens gamma-crystallin. Such antioxidative ability of beta-carbolines is of particular relevance to the lens, which faces continual photic and oxidative stress. The beta-carboline diacid IV is also seen to display an unexpected ability of inhibiting the thermal coagulation of gamma-crystallin and the dithiothreitol-induced precipitation of insulin. These results offer experimental support to earlier suggestions that one of the roles that the beta-carbolines have is to offer protection against oxidative stress to the human tissues where they accumulate.

Depth profiling of mammalian cells by photoacoustic spectroscopy: localization of ligands.

Phase-resolved monitoring of photoacoustic signals can provide information about the depth profile of a sample. We describe an application of this principle to determine the depth profiles of ligands and antitumor agents in mammalian cells. Measurements of the in-phase and quadrature components of the photoacoustic spectra (which yield information from the surface and the interior, respectively) of a tumor cell line, AK-5, treated with the antitumor agent coralyne chloride have been made. They clearly show that the drug accumulates in the cell interior and is not seen on the cell surface, providing in situ evidence for the localization of this drug. Histochemical dyes which stain cells uniformly give identical in-phase and quadrature spectra; spectra of cells incubated with nuclear stains demonstrate a differential staining of the nucleus and the cytoplasm. These results demonstrate the usefulness of phase-resolved photoacoustic spectroscopy in monitoring differential interactions of drugs and other ligands with cells.

Modified helix-loop-helix motifs of calmodulin--The influence of the exchange of helical regions on calcium-binding affinity.

The four calcium-binding sites, called the helix-loop-helix, or the EF-hand motifs, of calmodulin differ in their ion-binding affinities; this has been thought to arise due to the variations in the sequences of the loop regions where the ion binds. We focus attention here on the role of the flanking helical regions on the calcium-binding affinities. Peptides were synthesized in a manner that simulates the E and F helical flanks of site 4 (the strongest calcium-binding site of the calmodulin) to sandwich the loop sequences of sites 1, 2, 3 and 4 so as to produce peptides named 414, 424, 434 and 444, as well as using the helical flanks of site 1 (the weakest site) to produce peptides 111, 121, 131 and 141. Calcium binding was monitored using the calcium-mimic dye Stains-all (4,4,4',5'-dibenzo-3,3'-diethyl-9-methyl-thiacarbocyanine bromide). Binding abilities were seen to increase several-fold when the E and F helices of site 1 were replaced by those of site 4 (i.e., 111-414). In contrast, the intensity of circular dichroism induced in the absorption bands of the bound achiral dye decreased significantly when the helical flanks of site 4 were replaced with those of site 1 (i.e., 444-141). The helical flanks of site 4 impart greater binding ability to a given loop region, while the helical flanks of site 1 tend to weaken it.

Seminal plasmin, a bovine seminal plasma protein, lyses dividing but not resting mammalian cells.

Seminal plasmin, an antimicrobial and transcription-inhibitory protein of bovine seminal plasma, is shown to lyse dividing mammalian cells in vitro. It lyses cells in culture such as CHO, Vero, HeLa and L929. It also lyses regenerating rat liver parenchymal cells and cells of two ascitic tumours of rat--the Zajdela ascitic hepatoma and the AK-5. However, it does not lyse resting cells such as adult liver parenchymal cells, erythrocytes, or resting lymphocytes, though it binds to their cell surface. It can be used, therefore, to distinguish cells that are in the division cycle from cells that are in the resting phase. The cell-lytic activity of seminal plasmin is inhibited by Ca2+.

Regulation of cell division and malignant transformation.

The problem of regulation of cell division is essentially a problem of understanding regulation of transition from the resting state of a cell to the dividing state and vice versa. In malignancy the ability to revert back to a normal resting state is impaired. A model is presented which attempts to explain the control of the above transitions through control of uptake of essential nutrients by a transport-inhibitory protein. Experimental evidence in favour of the model is given.